Identificação dos microorganismos presentes na saliva, na coroa dental e no canal radicular de dentes indicados ao retratamento endodontico e analise da suscetibilidade antimicrobiana, dos fatores de virulencia e da diversidade genetica dos Enterococcus faecalis isolados / Identification of microorganism present in saliva, crown and root canal of endodontically treated teeth and analysis of antimicrobial susceptibility, virulence factors and diversity of clonal types of Enterococcus faecalis isolated

AUTOR(ES)
DATA DE PUBLICAÇÃO

2009

RESUMO

The aims of this study were: (a) to identify Enterococcus spp., Staphylococcus spp., Candida spp. and enterobacteria in saliva of patients with failure of the endodonticallytreated teeth, (b) to investigate the microbiota in the restorative material and within root canals of the same subjects, (c) to determine the antimicrobial susceptibility of E. faecalis strains isolated from the root canals, (d) to study the E. faecalis virulence factors, and (e) to determine the similarity of the E. faecalis genotypes from different oral sites samplings. Forty-two cases of endodontic failure were selected. Samples were collected during the endodontic procedures. Anaerobic techniques and commercial kits were used to culture and identify the microorganisms. E-test System was conducted to test the antimicrobial susceptibility of the E. faecalis isolates (n=14). PCR (16S rRNA) was used to confirm the presence of E. faecalis isolated from the saliva, restorative materials and root canals. The genotypic studies involved DNA extractions of 74 E. faecalis strains using Qiamp DNA Mini Kit. Afterwards, the virulence factors were also verified using specific primers to detect: aggregation substances (asa e asa373), adherence factors (ace e esp), cytolysin activator (cylA) and gelatinase (gelE). Phenotypic tests were also used for production of gelatinase and cytolysin. Afterwards, the REP-PCR and AP-PCR were processed to determine the genome impression. Enterococcus spp. (62%), Staphylococcus spp. (45%), Candida spp. (21%) and enterobacteria (38%) were detected in saliva. The most prevalent genera identified in crowns and root canals were: Staphylococcus (31%, 43%), Streptococcus (50%, 33%), Enterococcus (28.6%, 33.3%), Actinomyces (23.8%, 31%) and Gemella (23.8%, 21.4%), respectively. E. faecalis were 100% susceptible to amoxicillin + clavulanate. The E. faecalis strains tested (n=74) expressed most of the potential virulence factors investigated. Homogeneous E. faecalis genotypes were observed in different subjects and particularly in multiple colonies isolated from each sampling site. Seven different genotypes (G) were identified using REP-PCR and 7 among the 74 E. faecalis strains using AP-PCR. In conclusion, Enterococcus spp., Staphylococcus spp., Candida spp. and enterobacteria could also be detected in the saliva and in the root canals. The effective antibiotic for E. faecalis was amoxicillin combined with clavulanate. Adherence factors (ace) and gelatinase (gelE) were expressed in 100% of the E. faecalis. Furthermore, multiple E. faecalis showed similarity between genotypes of the same sites and subjects.

ASSUNTO(S)

antibioticos endodontia microbiology antibiotics endodontics microbiologia

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