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Characterization of A. actinomycetemcomitans strains in subgingival samples from periodontitis subjects in Morocco

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Abstract

Objective

Aggregatibacter actinomycetemcomitans, specially its highly leucotoxic strain (JP2 clone), represents an etiological factor for the onset and progression of aggressive types of periodontitis. The aims of this investigation were to investigate the most relevant periodontal pathogens in the subgingival microbiota of periodontitis patients from Morocco and to describe the clinical and microbiological characteristics of subjects positive for A. actinomycetemcomitans, including serotype, leukotoxin gene, and operon of the cytolethal distending toxin (cdt) distribution.

Material and methods

In consecutive Moroccan subjects diagnosed of periodontitis, subgingival samples were taken and processed by culture. From the positive samples for A. actinomycetemcomitans, one to three isolates were subcultured and characterized by means of polymerase chain reaction (PCR), assessing their specific serotype distribution, the variation in the sequences of the leukotoxin gene, and the operon of the cdt.

Results

Twenty-one (35.6 %) out of 59 periodontitis patients harbored A. actinomycetemcomitans. These patients demonstrated statistically significant deeper pockets (p = 0.035) and higher proportions of P. micra (p = 0.045) than did the negative group. The 39 studied isolates were serotype “b”; in 16 out of 17 patients, there was mono-colonization with this serotype. Five isolates, from two patients, presented the 530-bp deletion in the leukotoxin’s promoter region. Thirty-two isolates (78 % of the strains) were cdt-positive.

Conclusion

A. actinomycetemcomitans was frequently found (35.6 %) in our sample. All strains were serotype “b,” and most (78 %) were also cdt-positive. The JP2 strain type was only detected in 12.2 % of the strains.

Clinical relevance

A. actinomycetemcomitans can be frequently found in Morocco. This fact can influence the therapeutic approach of this type of patients.

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References

  1. Listgarten MA (1976) Structure of the microbial flora associated with periodontal health and disease in man. A light and electron microscopic study. J Periodontol 47:1–18

    Article  PubMed  Google Scholar 

  2. Consensus report (1996) Periodontal diseases: pathogenesis and microbial factors. Ann Periodontol 1:926–932

  3. Asikainen S, Chen C, Slots J (1995) Actinobacillus actinomycetemcomitans genotypes in relation to serotypes and periodontal status. Oral Microbiol Immunol 10:65–68

    Article  PubMed  Google Scholar 

  4. Poulsen K, Theilade E, Lally ET, Demuth DR, Kilian M (1994) Population structure of Actinobacillus actinomycetemcomitans: a framework for studies of disease-associated properties. Microbiology 140:2049–2060

    Article  PubMed  Google Scholar 

  5. Paju S, Carlson P, Jousimies-Somer H, Asikainen S (2000) Heterogeneity of Actinobacillus actinomycetemcomitans strains in various human infections and relationships between serotype, genotype, and antimicrobial susceptibility. J Clin Microbiol 38:79–84

    PubMed  PubMed Central  Google Scholar 

  6. Kanasi E, Dogan B, Karched M, Thay B, Oscarsson J, Asikainen S (2010) Lack of serotype antigen in A. actinomycetemcomitans. J Dent Res 89:292–296

    Article  PubMed  Google Scholar 

  7. Cortelli JR, Aquino DR, Cortelli SC, Roman-Torres CV, Franco GC, Gomez RS et al (2012) Aggregatibacter actinomycetemcomitans serotypes infections and periodontal conditions: a two-way assessment. Eur J Clin Microbiol Infect Dis 31:1311–1318

    Article  PubMed  Google Scholar 

  8. Holtta P, Alaluusua S, Saarela M, Asikainen S (1994) Isolation frequency and serotype distribution of mutans streptococci and Actinobacillus actinomycetemcomitans, and clinical periodontal status in Finnish and Vietnamese children. Scand J Dent Res 102:113–119

    PubMed  Google Scholar 

  9. Celenligil H, Ebersole JL (1998) Analysis of serum antibody responses to periodontopathogens in early-onset periodontitis patients from different geographical locations. J Clin Periodontol 25:994–1002

    Article  PubMed  Google Scholar 

  10. Baehni PC, Tsai CC, McArthur WP, Hammond BF, Shenker BJ, Taichman NS (1981) Leukotoxic activity in different strains of the bacterium Actinobacillus actinomycetemcomitans isolated from juvenile periodontitis in man. Arch Oral Biol 26:671–676

    Article  PubMed  Google Scholar 

  11. Zambon JJ, Christersson LA, Slots J (1983) Actinobacillus actinomycetemcomitans in human periodontal disease. Prevalence in patient groups and distribution of biotypes and serotypes within families. J Periodontol 54:707–711

    Article  PubMed  Google Scholar 

  12. Haubek D, Ennibi OK, Poulsen K, Vaeth M, Poulsen S, Kilian M (2008) Risk of aggressive periodontitis in adolescent carriers of the JP2 clone of Aggregatibacter (Actinobacillus) actinomycetemcomitans in Morocco: a prospective longitudinal cohort study. Lancet 371:237–242

    Article  PubMed  Google Scholar 

  13. Yamano R, Ohara M, Nishikubo S, Fujiwara T, Kawamoto T, Ueno Y et al (2003) Prevalence of cytolethal distending toxin production in periodontopathogenic bacteria. J Clin Microbiol 41:1391–1398

    Article  PubMed  PubMed Central  Google Scholar 

  14. Benrachadi L, Bouziane A, Azziman Z, Bouziane-Ouartini F, Ennibi O (2012) Screening for periodontopathogenic bacteria in severe chronic periodontitis in a Moroccan population. Med Mal Infect 42:599–602

    Article  PubMed  Google Scholar 

  15. Armitage GC (1999) Development of a classification system for periodontal diseases and conditions. Ann Periodontol 4:1–6

    Article  PubMed  Google Scholar 

  16. Mombelli A, McNabb H, Lang NP (1991) Black-pigmenting gram-negative bacteria in periodontal disease. II Screening strategies for detection of P gingivalis. J Periodontal Res 2:308–313

    Article  Google Scholar 

  17. Syed SA, Loesche WJ (1972) Survival of human dental plaque flora in various transport media. Appl Microbiol 24:638–644

    PubMed  PubMed Central  Google Scholar 

  18. Alsina M, Olle E, Frias J (2001) Improved, low-cost selective culture medium for Actinobacillus actinomycetemcomitans. J Clin Microbiol 39:509–513

    Article  PubMed  PubMed Central  Google Scholar 

  19. Yorgancioglu A, Turktas H, Kalayci O, Yardim N, Buzgan T, Kocabas A et al (2009) The WHO global alliance against chronic respiratory diseases in Turkey (GARD Turkey). Tuberk Toraks 57:439–452

    PubMed  Google Scholar 

  20. Kaplan JB, Schreiner HC, Furgang D, Fine DH (2002) Population structure and genetic diversity of Actinobacillus actinomycetemcomitans strains isolated from localized juvenile periodontitis patients. J Clin Microbiol 40:1181–1187

    Article  PubMed  PubMed Central  Google Scholar 

  21. Haubek D, Dirienzo JM, Tinoco EM, Westergaard J, Lopez NJ, Chung CP et al (1997) Racial tropism of a highly toxic clone of Actinobacillus actinomycetemcomitans associated with juvenile periodontitis. J Clin Microbiol 35:3037–3042

    PubMed  PubMed Central  Google Scholar 

  22. Haubek D, Ennibi OK, Poulsen K, Poulsen S, Benzarti N, Kilian M (2001) Early-onset periodontitis in Morocco is associated with the highly leukotoxic clone of Actinobacillus actinomycetemcomitans. J Dent Res 80:1580–1583

    Article  PubMed  Google Scholar 

  23. Van der Velden U, Abbas F, Armand S, Loos BG, Timmerman MF, Van der Weijden GA et al (2006) Java project on periodontal diseases. The natural development of periodontitis: risk factors, risk predictors and risk determinants. J Clin Periodontol 33:540–548

    Article  PubMed  Google Scholar 

  24. Stingu CS, Jentsch H, Eick S, Schaumann R, Knofler G, Rodloff A (2012) Microbial profile of patients with periodontitis compared with healthy subjects. Quintessence Int 43:23–31

    Google Scholar 

  25. Minguez M, Pousa X, Herrera D, Blasi A, Sanchez MC, Leon R et al (2014) Characterization and serotype distribution of Aggregatibacter actinomycetemcomitans isolated from a population of periodontitis patients in Spain. Arch Oral Biol 59:1359–1367

    Article  PubMed  Google Scholar 

  26. Aberg CH, Kwamin F, Claesson R, Johansson A, Haubek D (2012) Presence of JP2 and Non-JP2 Genotypes of Aggregatibacter actinomycetemcomitans and attachment loss in adolescents in Ghana. J Periodontol 83:1520–1528

    Article  PubMed  Google Scholar 

  27. Elamin A, Albandar JM, Poulsen K, Ali RW, Bakken V (2011) Prevalence of Aggregatibacter actinomycetemcomitans in Sudanese patients with aggressive periodontitis: a case–control study. J Periodontal Res 46:285–291

    Article  PubMed  Google Scholar 

  28. Aberg CH, Sjodin B, Lakio L, Pussinen PJ, Johansson A, Claesson R (2009) Presence of Aggregatibacter actinomycetemcomitans in young individuals: a 16-year clinical and microbiological follow-up study. J Clin Periodontol 36:815–822

    Article  PubMed  Google Scholar 

  29. Yoshida Y, Suzuki N, Nakano Y, Shibuya K, Ogawa Y, Koga T (2003) Distribution of Actinobacillus actinomycetemcomitans serotypes and Porphyromonas gingivalis in Japanese adults. Oral Microbiol Immunol 18:135–139

    Article  PubMed  Google Scholar 

  30. Chen C, Wang T, Chen W (2010) Occurrence of Aggregatibacter actinomycetemcomitans serotypes in subgingival plaque from United States subjects. Mol Oral Microbiol 25:207–214

    Article  PubMed  Google Scholar 

  31. Zambon JJ, Slots J, Genco RJ (1983) Serology of oral Actinobacillus actinomycetemcomitans and serotype distribution in human periodontal disease. Infect Immun 41:19–27

    PubMed  PubMed Central  Google Scholar 

  32. Chung HJ, Chung CP, Son SH, Nisengard RJ (1989) Actinobacillus actinomycetemcomitans serotypes and leukotoxicity in Korean localized juvenile periodontitis. J Periodontol 60:506–511

    Article  PubMed  Google Scholar 

  33. Mombelli A, Gmur R, Lang NP, Corbert E, Frey J (1999) Actinobacillus actinomycetemcomitans in Chinese adults. Serotype distribution and analysis of the leukotoxin gene promoter locus. J Clin Periodontol 26:505–510

    Article  PubMed  Google Scholar 

  34. Hoglund Aberg C, Antonoglou G, Haubek D, Kwamin F, Claesson R, Johansson A (2013) Cytolethal distending toxin in isolates of Aggregatibacter actinomycetemcomitans from Ghanaian adolescents and association with serotype and disease progression. PLoS One 8:65781

    Article  Google Scholar 

  35. Ennibi OK, Benrachadi L, Bouziane A, Haubek D, Poulsen K (2012) The highly leukotoxic JP2 clone of Aggregatibacter actinomycetemcomitans in localized and generalized forms of aggressive periodontitis. Acta Odontol Scand 70:318–322

    Article  PubMed  Google Scholar 

  36. Poulsen K, Ennibi OK, Haubek D (2003) Improved PCR for detection of the highly leukotoxic JP2 clone of Actinobacillus actinomycetemcomitans in subgingival plaque samples. J Clin Microbiol 41:4829–4832

    Article  PubMed  PubMed Central  Google Scholar 

  37. Haubek D, Poulsen K, Asikainen S, Kilian M (1995) Evidence for absence in northern Europe of especially virulent clonal types of Actinobacillus actinomycetemcomitans. J Clin Microbiol 33:395–401

    PubMed  PubMed Central  Google Scholar 

  38. Sakellari D, Katsikari A, Slini T, Ioannidis I, Konstantinidis A, Arsenakis M (2011) Prevalence and distribution of Aggregatibacter actinomycetemcomitans serotypes and the JP2 clone in a Greek population. J Clin Periodontol 38:108–114

    Article  PubMed  Google Scholar 

  39. Ahmed HJ, Svensson LA, Cope LD, Latimer JL, Hansen EJ, Ahlman K et al (2001) Prevalence of cdtABC genes encoding cytolethal distending toxin among Haemophilus ducreyi and Actinobacillus actinomycetemcomitans strains. J Med Microbiol 50:860–864

    Article  PubMed  Google Scholar 

  40. Tan KS, Woo CH, Ong G, Song KP (2001) Prevalence of Actinobacillus actinomycetemcomitans in an ethnic adult Chinese population. J Clin Periodontol 28:886–890

    Article  PubMed  Google Scholar 

  41. Fabris AS, DiRienzo JM, Wikstrom M, Mayer MP (2002) Detection of cytolethal distending toxin activity and cdt genes in Actinobacillus actinomycetemcomitans isolates from geographically diverse populations. Oral Microbiol Immunol 17:231–238

    Article  PubMed  PubMed Central  Google Scholar 

  42. van der Reijden WA, Brunner J, Bosch-Tijhof CJ, van Trappen S, Rijnsburger MC, de Graaff MP et al (2010) Phylogenetic variation of Aggregatibacter actinomycetemcomitans serotype e reveals an aberrant distinct evolutionary stable lineage. Infect Genet Evol 10:1124–1131

    Article  PubMed  Google Scholar 

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Acknowledgments

We thank Arancha Llama-Palacios, María José Marín, Itziar González and Ana O’Connor from the Laboratory of Dental Research, University Complutense, Madrid, for their technical assistance.

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Authors and Affiliations

  1. Section of Periodontology, Faculty of Odontology, University Complutense, Plaza Ramón y Cajal S/N, 28040, Madrid, Spain

    M. Mínguez, X. Pousa, M. Sanz & D. Herrera

  2. Clinical Department of Periodontology, Dental School, Mohammed V Souissi University, Rabat, Morocco

    O. K. Ennibi, L. Lakhdar & L. Abdellaoui

  3. Laboratory of Research, Faculty of Odontology, University Complutense, Madrid, Spain

    M. Sánchez

  4. ETEP (Etiology and Therapy of Periodontal Diseases) Research Group, University Complutense, Madrid, Spain

    M. Sanz & D. Herrera

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  1. M. Mínguez
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  2. O. K. Ennibi
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  3. X. Pousa
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  4. L. Lakhdar
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Correspondence to D. Herrera.

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Mínguez, M., Ennibi, O.K., Pousa, X. et al. Characterization of A. actinomycetemcomitans strains in subgingival samples from periodontitis subjects in Morocco. Clin Oral Invest 20, 1809–1818 (2016). https://doi.org/10.1007/s00784-015-1653-7

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  • DOI: https://doi.org/10.1007/s00784-015-1653-7

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