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PERIODONTOPATHIC BACTERIA AND ANTIBIOTIC RESISTANCE GENES OF ORAL BIOFILMS IN CHILDREN

어린이 치면세균막에서 치주질환원인균과 항생제 내성유전자의 출현율

  • Kim, Seon-Mi (Department of Pediatric Dentistry and Dental Research Institute, School of Dentistry, Chonnam National University) ;
  • Choi, Nam-Ki (Department of Pediatric Dentistry and Dental Research Institute, School of Dentistry, Chonnam National University) ;
  • Cho, Seong-Hoon (Department of Pediatric Dentistry and Dental Research Institute, School of Dentistry, Chonnam National University) ;
  • Lee, Seok-Woo (Department of Periodontics and Dental education, School of Dentistry, Chonnam National University) ;
  • Lim, Hoi-Jeong (Department of Orthodontics, School of Dentistry, Chonnam National University) ;
  • Lim, Hoi-Soon (Department of Dental education, School of Dentistry, Chonnam National University) ;
  • Kang, Mi-Sun (Department of Microbiology, College of Medicine, Chonnam National University) ;
  • Oh, Jong-Suk (Department of Microbiology, College of Medicine, Chonnam National University)
  • 김선미 (전남대학교 치의학전문대학원 소아치과학교실 및 치의학연구소) ;
  • 최남기 (전남대학교 치의학전문대학원 소아치과학교실 및 치의학연구소) ;
  • 조성훈 (전남대학교 치의학전문대학원 소아치과학교실 및 치의학연구소) ;
  • 이석우 (전남대학교 치의학전문대학원 치의학교육학교실 및 치주학교실) ;
  • 임회정 (전남대학교 치의학전문대학원 교정학교실) ;
  • 임회순 (전남대학교 치의학전문대학원 치의학교육학교실) ;
  • 강미선 (전남대학교 의과대학 미생물학교실) ;
  • 오종석 (전남대학교 의과대학 미생물학교실)
  • Received : 2011.04.13
  • Accepted : 2011.05.10
  • Published : 2011.05.31

Abstract

The purpose of this study was to assess the prevalence of periodontopathic bacteria and resistance determinants from oral biofilm of children. Subgingival dental plaque was isolated from 87 healthy children, and PCR was performed to determine the presence of 5 periodontal pathogens including P. gingivalis, T. forsythia, T. denticola, F. nucleatum, A. actinomycetemcomitans, and nine resistance genes including tet(Q), tet(M), ermF, aacA-aphD, cfxA, $bla_{SHV}$, $bla_{TEM}$, vanA, mecA. 1. The prevalence of F. nucleatum, T. forsythia. and P. gingivalis was 95.4%, 55.2%, and 40.2%, respectively. In addition. the prevalence of A. actinomycetemc omitans was 5.7%, while T. denticola was 3.4%. 2. In analysis of antibiotic resistance determinants. cfxA, $bla_{TEM}$ and tet(M) were detected in all the samples tested. It was also found that the prevalence of tet(Q) showing tetracycline resistance. $bla_{SHV}$ associated with resistance to ${\beta}$-lactams, ermF exhibiting erythromycin resistance, and, vanA resulting vancomycin resistance was 88.5%, 29.9% 87.4%, and 48.5%, respectively. The aacA-aphD gene showing resistance to aminoglycosides and mecA gene harboring methicillin resistance exhibited the lowest prevalence with 9.2%. 3. In a correlation analysis between periodontopathic pathogens and antibiotic resistance determinants, it was found that there was a significant correlation between T. forsythia and $bla_{SHV}$. Also, P. gingivalis and vanA showed a correlation. Finally, tet(Q) and ermF showed a significant correlation (phi: 0.514) while mecA and vanA also showed a correlation(phi: 0.25).

Keywords

Periodontopathic bacteria;Antibiotic resistance;Antibiotic resistance determinants;Oral biofilm;Polymerase chain reaction

Acknowledgement

Supported by : 전남대학교

References

  1. Kolenbrander PE : Oral microbial communities: biofilms, interactions and genetic systems. Annu Rev Microbiol, 54: 413-37, 2000. https://doi.org/10.1146/annurev.micro.54.1.413
  2. Mandell RL : A longitudinal microbiological investigation of Actinobacillus actinomycetemcomitans and Eikenella corrodens in juvenile periodontitis. Infect Immun, 45:778-780, 1984.
  3. Haffajee AD and Socransky SS : Microbial etiological agents of destructive periodontal diseases. Periodontol, 5:78-111, 1994. https://doi.org/10.1111/j.1600-0757.1994.tb00020.x
  4. van Winkelhoff AJ, Rams TE, Slots J : Systemic antibiotic therapy in periodontics. Periodontol, 10:45-78, 1996. https://doi.org/10.1111/j.1600-0757.1996.tb00068.x
  5. Clark WB and Loe H : Mechanisms of initiation and progression of periodontal disease. Periodontol, 2:72-82, 1993. https://doi.org/10.1111/j.1600-0757.1993.tb00221.x
  6. Christersson CE, Dunford RG, Glantz PO, et al. : Effect of critical surface tension on retention of oral microorganisms. Scand J Dent Res, 97(3):247-56, 1989.
  7. Aitken S, Birek P, Kulkarni GV, et al. : Serial doxycycline and metronidazole in prevention of recurrent periodontitis in high-risk patients. J Periodontol, 63(2): 87-92, 1992. https://doi.org/10.1902/jop.1992.63.2.87
  8. Haffajee AD, Dibart S, Kent RL Jr, et al. : Clinical and microbiological changes associated with the use of 4 adjunctive systemically administered agents in the treatment of periodontal infections. J Clin Periodontol, 22(8):618-27, 1995. https://doi.org/10.1111/j.1600-051X.1995.tb00815.x
  9. Medeiros AA : Evolution and dissemination of betalactamases accelerated by generations of beta-lactam antibiotics. Clin Infect Dis, Suppl 1:19-45, 1997.
  10. Chopra I and Roberts M : Tetracycline antibiotics: mode of action, applications, molecular biology, and epidemiology of bacterial resistance. Microbiol Mol Biol Rev, 65(2):232-60, 2001. https://doi.org/10.1128/MMBR.65.2.232-260.2001
  11. Roberts MC : Epidemiology of tetracycline-resistance determinants. Trends Microbiol, 2(10):353-7, 1994. https://doi.org/10.1016/0966-842X(94)90610-6
  12. Kononen E : Development of oral bacterial flora in young children. Ann Med, 32(2):107-12, 2000. https://doi.org/10.3109/07853890009011759
  13. 이해송, 김선미, 최남기 등. : 정상인과 정신지체인, 다운증후군 환자에서 치주질환 원인균의 출현율. 대한소아치과학회지, 33(3):457-468, 2006.
  14. 김선미, 양규호, 최남기 등. : Real-time PCR을 이용한 치주질환 원인균의 정량적 분석. 대한소아치과학회지, 35(3):494-503, 2008.
  15. Sweeney EA, Alcoforado GA, Nyman S, et al. : Prevalence and microbiology of localized prepubertal periodontitis. Oral Microbiol Immunol, 2(2):65-70, 1987. https://doi.org/10.1111/j.1399-302X.1987.tb00292.x
  16. Sjodin B, Crossner CG, Unell L, et al. : A retrospective radiographic study of alveolar bone loss in the primary dentition in patients with localized juvenile periodontitis. J Clin Periodontol, 16(2):124-7, 1989. https://doi.org/10.1111/j.1600-051X.1989.tb01625.x
  17. Golub LM, Wolff M, Lee HM, et al. : Further evidence that tetracyclines inhibit collagenase activity in human crevicular fluid and from other mammalian sources. J Periodontal Res, 20(1):12-23, 1985. https://doi.org/10.1111/j.1600-0765.1985.tb00405.x
  18. Stabholz A, Kettering J, Aprecio R, et al. : Antimicrobial properties of human dentin impregnated with tetracycline HCl or chlorhexidine. An in vitro study. J Clin Periodontol, 20(8):557-62, 1993. https://doi.org/10.1111/j.1600-051X.1993.tb00771.x
  19. Rams TE and Keyes PH : A rationale for the management of periodontal diseases: effects of tetracycline on subgingival bacteria. J Am Dent Assoc, 107 (1): 37-41, 1983. https://doi.org/10.14219/jada.archive.1983.0177
  20. McCulloch CA, Birek P, Aitken S, et al. : Efficacy of doxycycline in prevention of recurrent periodontitis. J Dent Res, 68:916, 1989.
  21. McCulloch CA, Birek P, Overall C, et al. : Randomized controlled trial of doxycycline in prevention of recurrent periodontitis in high-risk patients: antimicrobial activity and collagenase inhibition. J Clin Periodontol, 17(9):616-22, 1990. https://doi.org/10.1111/j.1600-051X.1990.tb01682.x
  22. Soder PO, Frithiof L, Wikner S, et al. : The effects of metronidazole in treatment of young adults with severe periodontitis. J Dent Res, 68:710, 1989.
  23. Soder PO, Frithiof L, Wikner S, et al. : The effect of systemic metronidazole after non-surgical treatment in moderate and advanced periodontitis in young adults. J Periodontol, 61(5):281-88, 1990. https://doi.org/10.1902/jop.1990.61.5.281
  24. Ohta Y, Okuda K, Takazoe I : Microbiological and clinical effects of systemic antibiotic administration in advanced periodontitis. Bulletin of the Tokyo Dental College 27:139-148, 1986.
  25. Kornman KS, Newman MG, Flemming T, et al. : Treatment of refractory periodontitis with metronidazole plus amoxicillin or Augmentin. J Dent Res, 68:917, 1989.
  26. Pavicic MJ, van Winkelhoff AJ, Douque NH, et al. : Microbiological and clinical effects of metronidazole and amoxicillin in Actinobacillus actinomycetemcomitans associated periodontitis. A 2-year evaluation. J Clin Periodontol, 21(2):107-12, 1994. https://doi.org/10.1111/j.1600-051X.1994.tb00287.x
  27. Anwar H, Dasgupta MK, Costerton JW : Testing the susceptibility of bacteria in biofilms to antibacterial agents. Antimicrob Agents Chemother, 34(11): 2043-6, 1990. https://doi.org/10.1128/AAC.34.11.2043
  28. Anwar H, Strap JL, Costerton JW : Establishment of aging biofilms: possible mechanism of bacterial resistance to antimicrobial therapy. Antimicrob Agents Chemother, 36(7):1347-51, 1992. https://doi.org/10.1128/AAC.36.7.1347
  29. Hedberg M, Lindqvist L, Bergman T, et al. : Purification and characterization of a new beta-lactamase from Bacteroides uniformis. Antimicrob Agents Chemother, 39(7):1458-61, 1995. https://doi.org/10.1128/AAC.39.7.1458
  30. Parker AC and Smith CJ : Genetic and biochemical analysis of a novel Ambler class A beta-lactamase responsible for cefoxitin resistance in Bacteroides species. Antimicrob Agents Chemother, 37(5):1028-36, 1993. https://doi.org/10.1128/AAC.37.5.1028
  31. Fosse T, Madinier I, Hannoun L, et al. : High prevalence of cfxA beta-lactamase in aminopenicillin- resistant Prevotella strains isolated from periodontal pockets. Oral Microbiol Immunol, 17(2):85-8, 2002. https://doi.org/10.1046/j.0902-0055.2001.00096.x
  32. Lacroix JM and Walker CB : Detection and prevalence of the tetracycline resistance determinant Tet Q in the microbiota associated with adult periodontitis. Oral Microbiol Immunol, 11(4):282-8, 1996. https://doi.org/10.1111/j.1399-302X.1996.tb00182.x
  33. Hawley RJ, Lee LN, LeBlanc DJ : Effects of tetracycline on the streptococcal flora of periodontal pockets. Antimicrob Agents Chemother, 17(3):372-8, 1980. https://doi.org/10.1128/AAC.17.3.372
  34. Bragd L, Wikstrom M, Slots J : Clinical and microbiological study of "refractory"adult periodontitis. J Dent Res, 64:234, 1985.
  35. Speer BS, Shoemaker NB, Salyers AA : Bacterial resistance to tetracycline: mechanisms, transfer, and clinical significance. Clin Microbiol Rev, 5(4): 387-99, 1992.
  36. Roberts MC : Tetracycline resistance determinants: mechanisms of action, regulation of expression, genetic mobility, and distribution. FEMS Microbiol Rev, 19(1):1-24, 1996.
  37. Williams BL, Osterberg SK, Jorgensen J : Subgingival microflora of periodontal patients on tetracycline therapy. J Clin Periodontol, 6(4):210-221, 1979. https://doi.org/10.1111/j.1600-051X.1979.tb01923.x
  38. Jost BH, Field AC, Trinh HT, et al. : Tylosin resistance in Arcanobacterium pyogenes is encoded by an Erm X determinant. Antimicrob Agents Chemother, 47(11):3519-24, 2003. https://doi.org/10.1128/AAC.47.11.3519-3524.2003
  39. Roberts MC : Acquired tetracycline and/or macrolide-lincosamides -streptogramin resistance in anaerobes. Anaerobe, 9(2):63-9, 2003. https://doi.org/10.1016/S1075-9964(03)00058-1
  40. Roberts MC, Sutcliffe J, Courvalin P, et al. : Nomenclature for macrolide and macrolide-lincosamide- streptogramin B resistance determinants. Antimicrob Agents Chemother, 43(12):2823-30, 1999.
  41. Byrne ME, Rouch DA, Skurray RA : Nucleotide sequence analysis of IS256 from the Staphylococcus aureus gentamicin-tobramycin-kanamycin-resistance transposon Tn4001. Gene, 81(2):361-367, 1989. https://doi.org/10.1016/0378-1119(89)90197-2
  42. Rouch DA, Byrne ME, Kong YC, et al. : The aacAaphD gentamicin and kanamycin resistance determinant of Tn4001 from Staphylococcus aureus: expression and nucleotide sequence analysis. J Gen Microbiol, 133(11):3039-52, 1987.
  43. Utsui Y and Yokota T : Role of an altered penicillinbinding protein in methicillin-and cephem-resistant Staphylococcus aureus. Antimicrob Agents Chemother, 28(3):397-403, 1985. https://doi.org/10.1128/AAC.28.3.397
  44. Gaetti-Jardim EC, Marqueti AC, Faverani LP, et al. : Antimicrobial resistance of aerobes and facultative anaerobes isolated from the oral cavity. J Appl Oral Sci, 18(6):551-559, 2010. https://doi.org/10.1590/S1678-77572010000600004

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