DETERMINATION OF BIOFILM FORMATION AND ASSOCIATED GENE DETECTION IN STAPHYLOCOCCUS GENUS ISOLATED FROM THE ORAL CAVITY UNDER INFLAMMATORY PERIODONTAL DISEASES
DOI: http://dx.doi.org/10.30970/sbi.1403.627
Abstract
Background. Inflammatory periodontal diseases, including chronic periodontitis, are accompanied by a chronic persisting inflammatory process. The latter process and biofilm-forming potential are the factors that contribute to the formation of antibiotic-resistant microorganism strains. The purpose of our work was to study the biofilm-forming ability of antibiotic-resistant biofilm-forming Staphylococcus genus bacteria isolated from the oral cavities of patients suffering from periodontitis, and check the presence of the genes associated with biofilm formation in these bacteria.
Material and methods. Isolates were obtained from patients suffering from chronic periodontitis using differentially diagnostic nutrient media. Chemotaxonomic identification of the antibiotic-resistant isolates was performed on the Microflex LT device. Biofilm-forming potential was studied in plastic Petri dishes by spectrophotometric method. The antibiotic sensitivity of the bacteria and microscopic fungi was identified by the disc diffusion method. The presence of icaA, icaD, icaC, icaB, agrA, srtA genes was detected in two multiplex polymerase chain reactions (mPCR). 16S rRNA gene and icaR genes were amplified in a separate PCR.
Results. Opportunistic microbial associations dominate in the microbiom of the oral cavity under conditions of periodontitis; besides, in the majority of cases, Staphylococcus genus bacteria were present in associations. In 73 % (131/180) of cases, bacteria from Staphylococcus genus represented by six species – S. aureus, S. haemolyticus, S. saprophyticus, S. epidermidis, S. xelosus, and S. hominis were isolated from the nidus during the inflammatory process. From the oral cavities of the patients with inflammatory periodontitis, 51 biofilm-forming strains of S. aureus, 29 strains of S. haemolyticus, 12 strains of S. saprophyticus, and 12 strains of S. epidermidis were isolated. It was established that antibiotic-resistant isolates of S. aureus possess the genetic determinants which are now believed to be connected with icaA, icaC, icaB icaA, icaR, agrA, and srtA biofilm-forming potential. Two non-biofilm forming strains of S. aureus were proven to lack the genes responsible for the biofilm-forming ability.
Conclusion: It has been established that microbial associations with a prevailing share of Staphylococcus genus bacteria, most of which have a biofilm-forming potential, dominate in the microbiom of the oral cavity under conditions of periodontitis.
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1. Al Jehani Y.A. Risk factors of periodontal disease: review of the literature. Int J Dent, 2014; 182513. Crossref • PubMed • Google Scholar | ||||
| ||||
2. Anwar H., Starp J.L., Costerton W.J. Establishment of aging biofilms: possible mechanism of bacterial resistance to antimicrobial therapy. Antimicrobial Agents and Chemotherapy, 1992; 36(7): 1347-1351. Crossref • PubMed • Google Scholar | ||||
| ||||
3. Arciola C.R., Campoccia D., Ravaioli S., Montanaro L. Polysaccharide intercellular adhesin in biofilm: structural and regulatory aspects. Front Cell Infect Microbiol, 2015; 5: 7. Crossref • PubMed • Google Scholar | ||||
| ||||
4. Bascones Martínez A., Figuero Ruiz E. Periodontal diseases as bacterial infection. Av Periodon Implantol, 2005; 17(3): 111-118. Crossref • PubMed • Google Scholar | ||||
| ||||
5. Bereket W., Hemalatha K., Getenet B., Wondwossen T., Solomon A., Zeynudin A., Kannan S. Update on bacterial nosocomial infections. Eur Rev Med Pharmacol Sci, 2012; 16(8): 1039-1044. PubMed • Google Scholar | ||||
| ||||
6. Conlon K.M., Humphreys H., O'Gara J.P. Regulation of icaR gene expression in Staphylococcus epidermidis. FEMS Microbiology Letters, 2002; 216(2): 171-177. Crossref • PubMed • Google Scholar | ||||
| ||||
7. Dancer S.J. Staphylococcus aureus antibiotic resistance. Clinical insights: Staphylococcus aureus antibiotic resistance. Future Medicine Ltd, 2014 Jul; 2-5. Crossref • Google Scholar | ||||
| ||||
8. Fowler V.G. Jr, Fey P.D., Reller L.B., Chamis A.L., Corey G.R., Rupp M.E. The intercellular adhesin locus ica is present in clinical isolates of Staphylococcus aureus from bacteremic patients with infected and uninfected prosthetic joints. Med Microbiol Immunol, 2001; 189(3): 127-31. Crossref • PubMed • Google Scholar | ||||
| ||||
9. Frank K.L., Patel R. Poly-N-acetylglucosamine is not a major component of the extracellular matrix in biofilms formed by icaADBC-positive Staphylococcus lugdunensis isolates. Infection and Immunity, 2007; 75(10): 4728-4742. Crossref • PubMed • Google Scholar | ||||
| ||||
10. Global Action Plan on Antimicrobial Resistance. Microbe Magazine, Anon 2015. 10(9), 354-355. Crossref | ||||
| ||||
11. Grigoryan A.S., Grudyanov A.Y., Rabukhina N.A., Frolova O.A. Periodontal disease. Pathogenesis, diagnosis, treatment. M., 2004. 320 p. (In Russian) | ||||
| ||||
12. Heller D., Silva-Boghossian C.M., do Souto R.M., Colombo A.P. Subgingival microbial profiles of generalized aggressive and chronic periodontal diseases. Arch Oral Biol, 2012; 57(7): 973-80. Crossref • PubMed • Google Scholar | ||||
| ||||
13. Joo H.-S., Otto M. Molecular basis of in vivo biofilm formation by bacterial pathogens. Chem Biol, 2012;19(12): 1503-1513. Crossref • PubMed • Google Scholar | ||||
| ||||
14. Kryvtsova M.V., Kostenko Y.Y. Dominant microbial associations of oral cavat periodontitis and features of their sensitivity to antibacterial drugs. Studia Biologica, 2020; 14(1): 51-62. Crossref • Google Scholar | ||||
| ||||
15. Kukhtyn M., Berhilevych O., Kravcheniuk K., Shynkaruk O., Horiuk Y., Semaniuk N. The influence of disinfectans on microbial biofilms of dairy equipment. Eastern-European Journal of Enterprise Technologies, 2017; 5/11(89): 26-33. Crossref • Google Scholar | ||||
| ||||
16. Kumar S.,Tripathi M. Antibiotic resistant bacteria: A global menace. Virol Immunol J, 2017; 1(3): 000118. Crossref • Google Scholar | ||||
| ||||
17. Li Li. Analysis of biofilm formation and associated gene detection in Staphylococcus isolates from bovine mastitis. African Journal of Boitechnology, 2012; 11(8). Crossref • Google Scholar | ||||
| ||||
18. O'Toole G., Kaplan H.B., Kolter R. Biofilm formation as microbial development. Annu Rev Microbiol, 2000; 54: 49-79. Crossref • PubMed • Google Scholar | ||||
| ||||
19. Otto M. Staphylococcal Biofilms. Bacterial Biofilms. Springer Berlin Heidelber, 2008; 207-228. Crossref • PubMed • Google Scholar | ||||
| ||||
20. Rampelotto R.F., Lorenzoni V.V., Silva D. da C., Coelho S.S., Wust V., Garzon L.R., Nunes M.S., Meneghetti B., Brites P. Ch., Hörner M., Hörner R. Assessment of different methods for the detection of biofilm production in coagulase-negative staphylococci isolated from blood cultures of newborns. Revista da Sociedade Brasileira de Medicina Tropical, 2018; 51(6): 761-767. Crossref • PubMed • Google Scholar | ||||
| ||||
21. Samojlenko A.V. Pathogenic importance of various periodontal microorganisms in development of immunologic and clinical disorders of patients suffering from generalized periodontitis. Ukrayinskyi stomatolohichnyi almanakh, 2001; 6: 44-47. (In Russian) | ||||
| ||||
22. Satorres S.E., Alcaráz L.E. Prevalence of icaA and icaD genes in Staphylococcus aureus and Staphylococcus epidermidis strains isolated from patients and hospital staff. Cent Eur J Public Health, 2007; 15(2): 87-90. PubMed • Google Scholar | ||||
| ||||
23. Stepanović S., Vuković D., Dakić I., Savić B., Švabić-Vlahović M. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. Journal of Microbiological Methods, 2000; 40(2): 175-179. Crossref • PubMed • Google Scholar | ||||
| ||||
24. Stubbings W., Labischinski H. New antibiotics for antibiotic-resistant bacteria. F1000 Biology Reports, 2009; 29: 1. Crossref • PubMed • Google Scholar | ||||
| ||||
25. Vuong C., Saenz H.L., Götz F., Otto M. Impact of agr quorumsensing system on adherence to polystyrene in Staphylococcus aureus. J Infect Dis, 2000; 182: 1688-1693. Crossref • PubMed • Google Scholar | ||||
| ||||
26. Yazdani R., Oshaghi M., Havayi A. Detection of icaAD gene and biofilm formation in Staphylococcus aureus isolates from wound infections. Iran J Public Health, 2006; 35(2): 25-28. Google Scholar | ||||
| ||||
27. Zhang Yu-Zhi, Singh S. Antibiotic stewardship programmes in intensive care units: Why, how, and where are they leading us. World J Crit Care Med, 2015; 4(1): 13-28. Crossref • PubMed • Google Scholar |
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