The profiles of oral streptococci sensitivity to antibacterial drugs may reflect information about the presence of macroorganism resistance determinants. The aim of the work was to isolate the spectrum of oral streptococci from the microbiota of the oral cavity of patients and to determine their sensitivity to a wide range of antibiotics. A total of 342 microbial streptococcal isolates were isolated from saliva samples and a periodontal pocket and tested for antibiotic sensitivity. Species identification of streptococci was carried out using biochemical API test systems. Evaluation of antibiotic resistance was performed using E-tests. Real-time PCR was used to identify the presence of tetracycline and macrolide resistance genes. The study identified six types of oral streptococci: S. oralis, S. salivarius, S. mitis, S. sanguinis, S. anginosus and S. mutans. All streptococci were sensitive to linezolid and meropenem. The proportion of penicillin-resistant streptococci in the subgroup S. oralis / mitis / mutans was 47,8% versus 23,5% in the subgroup S. salivarius / sanguinis / anginosus (p = 0.020). Significant levels of resistance were revealed to macrolides (erythromycin) - 47,9%, tetracyclines (tetracycline) - 44,4% and quinolones (ofloxacin) - 41%. Multiple drug resistance (MDR) was detected in 31,9% of oral streptococcal isolates, a combination of erythromycin, tetracycline and ofloxacin resistance was prevalent in 79 isolates (23,1%). The most common genotypes of macrolides and tetracycline resistant oral streptococci (in 127 streptococcal isolates with combined resistance) were ermB-mefE + and tetM + tetQ-, respectively. Thus, S. oralis / mitis / mutans group streptococci predominated in the structure of antibiotic-resistant oral streptococci, including MDR. So, being in one of the most densely populated biotopes of a macroorganism, oral streptococci can mediate the transfer of resistance determinants to more pathogenic and clinically significant microorganisms, which requires careful monitoring of their level of susceptibility to antimicrobial agents.
The system of antimicrobial peptides (AMP) is one of the most ancient mechanisms of the macroorganism resistance to infectious pathogens invasion. The aim of the study was to determine the role of the antimicrobial peptides system and periodontal pathogenic markers in the development and progression of inflammatory periodontal diseases. Gingival pocket washes (91 samples in total) for the research were received from patients with inflammatory periodontal diseases (chronic periodontitis and gingivitis) and intact periodontium. Using ELISA, the content of antimicrobial peptides was determined: human alpha-defensin (HNP 1-3), beta-defensin (HBD 1-3) and cathelicidin (LL-37). Periodontal pathogenic markers were isolated during RT-PCR. The study revealed differences in AMP concentrations by groups: level of HBD 2 in patients with chronic periodontitis was 1,36 times higher than those in the group of patients with chronic gingivitis (p=0,023) and 2,39 times higher than those in the control group (p<0,001), the content of HNP 1-3 in the group of patients with chronic periodontitis was reduced by 1,23 times compared with the indicators of the group of patients with gingivitis (p=0,045) and by 1,97 times compared with the indicators of the control group (p<0,001). The frequency of detection of periodontal pathogenic bacteria genes was 88,0% in patients with periodontitis, 76,92% in patients with gingivitis and 33,3% in the group with intact periodontium. HBD 2 content moderately correlated with the definition of P. gingivalis (r=0,612; p=0,022), T. forsythensis (r= 0,434; p=0,015), A. actinomycetemcomitans (r=0,483; p=0,006), a moderate negative correlation was detected between the content of HNP 1-3 and the release of periodontal pathogens in associations (P. gingivalis with T. forsythensis and T. denticola) (r=-0,388; p=0,031) in the group of patients with chronic periodontitis. Thus, the revealed relationships and correlations indicate shifts in the processes of reparative regeneration of the oral cavity and the regulation of local immunity in response to microbial invasion.
Relevance. Considering the role of personalized medicine in dental practice, the accumulation of knowledge about the genetic determinants of bacterial resistance and the use of antibacterial drugs, the learning of periodontal microbiota, sensitivity to antimicrobial drugs, the identification of key periodontal pathogens will allow predicting the development mechanisms of inflammatory periodontal diseases and monitoring and prescribing effective antibacterial therapy.Aim. The study aimed to determine the prevalence of pathogens and the occurrence of antibiotic resistance genes in individuals with chronic generalized periodontitis (CGP).Materials and methods. Clinical and laboratory examinations studied 163 subjects aged 18 to 45 years, of which there were 100 patients with inflammatory periodontal diseases and 63 subjects with intact periodontium. The study obtained periodontal pocket and gingival crevice swabs. The real-time PCR isolated marker bacteria and resistance genes to glycopeptide and β-lactam antibiotics.Results. The patients with chronic periodontitis demonstrated a periodontal pathogen detection rate of 96.4 %. Among the isolated periodontal pathogens, bacteria of the red complex were the most common: T. forsythia (81%; p < 0.001), T. denticola (63%; p = 0.054) and P. gingivalis (69%; p < 0.001). In healthy individuals with intact periodontium, P. gingivalis (12.7%), T. denticola (47.62%), and T. forsythia (36.51%) prevailed in the studied material, i.e., gingival fluid. The mean ratio of bone loss in relation to the root length (the Fuchs Index) was 0.83 ± 0.03 in patients with mild CGP and 0.71 ± 0.05 with moderate CGP. The β-lactam antibiotic resistance genes appeared to occur frequently (%). So, TEM and SHV genes were in 72% and 26%. The control group demonstrated the TEM gene in 41.27% (p < 0.001) and the SHV gene in 4.76% (p < 0.001).The group with CGP appeared to have resistance genetic markers: MecA to cephalosporins (15%) and OXA-51 to carbapenems (9%). The control group detected MecA (6.35%; p = 0.0948), while OXA-51 markers were absent (p = 0.014). Both groups did not show resistance genes to other antibiotic groups (AB).Conclusion. The "red complex" microorganisms T. forsythia, T. denticola and P. gingivalis, which were the most frequent during the progression of bone destruction, appeared to play the leading role in the onset and progression of inflammatory periodontal diseases.The oral microbiome can serve as a reservoir for the transfer of resistance genes: the study results indicate a high incidence rate (%) of β-lactam antibiotic resistance genes in the group of patients with chronic periodontitis. The group with chronic periodontitis revealed TEM and SHV genes in 72% and 26%, respectively. The group with intact periodontium also demonstrated a high occurrence rate of the TEM gene in 58.3 % of cases.
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