Investigação de bacterias associadas ao insucesso do tratamento endodontico

Orientador: Brenda Paula Figueiredo de Almeida GomesDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: Vários fatores estão envolvidos no insucesso do tratamento endodôntico, entretanto as bactérias são os principais agentes etiológicos. Tais microrganismos podem ter sobrevivido ao tratamento endodôntico anterior ou reinfectado o canal através das microinfiltrações. Estudos revelam que a microbiota do canal radicular do dente tratado endodonticamente associado à lesão periapical persistente difere substancialmente da microbiota de dentes com polpas necrosadas e não tratados. A microbiota dos canais com insucesso endodôntico é composta por um número bastante limitado de espécies bacterianas, que se apresentam mais resistentes aos métodos de combate à infecção utilizados na Endodontia. O objetivo deste trabalho foi estudar a microbiota de 30 dentes com insucesso do tratamento endodôntico e realizar testes de sensibilidade antimicrobiana das bactérias isoladas. Foram utilizados meios de transporte, cultura e incubação que propiciam o crescimento de bactérias anaeróbias estritas. Microrganismos viáveis estavam presentes em 80% dos casos, sendo que a maioria dos canais apresentava somente 1 ou 2 espécies bacterianas. Do total de 55 espécies bacterianas isoladas, 58% eram bactérias anaeróbias facultativas, 42% anaeróbias estritas, 80% Grani-positivas e 20% Gram-negativas. Os gêneros bacterianos mais freqüentemente isolados dos canais radiculares foram: Enterococcus (36,7%), Streptococcus (33,3%), Peptostreptococcus (23,3%), Actinomyces (13,3%), Prevotella (10%), Staphylococcus (10%), Gemella (10%), Fusobacterium (6,7%), Veil/onella (6,7%), Lactobacil/us (6,7%), Propionibacterium (3,3%) e Haemophilus (3,3%). Espécies dos gêneros Enterococcus e Peptostreptococcus foram testadas quanto à suscetibilidade antimicrobiana através do método do E-test, utilizando as seguintes substâncias: benzilpenicilina, amoxicilina, amoxicilina + ácido clavulânico, eritromicina, azitromicina e clindamicina. Enterococcus faecalis e Peptostreptococcus spp. Foram sensíveis a benzilpenicilina, amoxicilina e amoxicilina + ácido clavulânico. Entretanto, 20% dos Enterococcus faecalis foram resistentes a eritromicina e 60% a azitromicina. Concluímos que a microbiota dos canais de dentes com insucesso do tratamento endodôntico é composta em sua maioria por bactérias anaeróbias facultativas, predominantemente Gram-positivas. Os testes de suscetibilidade antimicrobiana revelaram a presença de resistência bacteriana entre espécies de Enterococcus faecalis spp. aos antibióticos eritromicina e azitromicinaAbstract: Although many failure cases of endodontic therapy are caused by technical problems during treatment, the cause is generally believed to be intracanal infection resisting treatment or microorganisms invading the canal via coronal leakage of the root filling. Studies of the microbiota from the canaIs ofteeth with failure of endodontic therapy have revealed that the flora differ markedly ftom that of untreated necrotic dental pulps. It appears to be a very limited assortment of microorganisms, and studies have indicated difficulties in the elimination of this microbiota during endodontic retreatments. This study aimed to evaluate the microbiota of 30 teeth with failed endodontic treatment and to test the sensivity of this microbiota to antibiotics. Microbial samples, isolation and speciation were done using advanced microbiologic techniques for anaerobic species. Microorganisms were recovered from 80% of the examined teeth. In most of the cases, one or two strains per canal were found. From the 55 microbial species isolated, 58 % were facultative anaerobe species, 42 % strict anaerobes, 80% Gram-positive organisms and 20 % Gram-negatives. The most ftequently bacterial genera recovered ftom the root canaIs were Enterococcus (36,7%), Streptococcus (33,3%), Peptostreptococcus (23,3%), Actinornyces (13,3%), Prevotella (10%), Staphylococcus (10%), Gernella (10%), Fusobacteriurn (6,7%), Veillonella (6,7%), Lactobacillus (6,7%), Propionibacteriurn (3,3%) and Haernophilus (3,3%). Antibiotic sensivity of Enterococcus and Peptostreptococcus species was accomplished with the E-test System. These bacterial isolates were tested for their susceptibility/resistance to benzylpenicilin, amoxicilin, amoxicilin combined with clavulanate, erythromycin, azithromycin, and clindamycin. Enterococcus spp. and Peptostreptococcus spp. Were susceptible to benzylpenicilin, amoxicilin, amoxicilin combined with clavulanate. However, 20% of the Enterococcus spp. isolates were resistant to erytromycin and 60% of these isolates were resistant to azitromycin. It was cocluded that microbial flora in canaIs after failure of the endodontic therapy comprised predominantly facultative anaerobic species and Gram-positive organisms. Antibiotic susceptibility results showed erythromicin and azithromycin resistance among Enterococcus spMestradoEndodontiaMestre em Clínica Odontológic

Comparison of rRNA-based reverse transcription PCR and rDNA-based PCR for the detection of streptococci in root canal infections

Objective: The rDNA-based method is unable to distinguish between alive and dead cells. Alternatively, bacterial viability can be assessed by molecular methods based on ribosomal RNA (rRNA). Therefore, this study aimed to detect viable streptococci in root canal samples using rRNA-based reverse transcription polymerase chain reaction (RT-PCR), compared to an rDNA-based PCR assay. Methodology: Microbiological root canal samples were obtained from 32 teeth with primary endodontic infections before (S1) and after chemomechanical preparation (S2), and after removal of intracanal medication (S3). RNA and DNA were extracted, and complementary DNA (cDNA) was synthesized from RNA using RT reaction. cDNA and genomic DNA were subjected to PCR with primers complementary to the 16S rRNA sequences of Streptococcus spp. McNemar’s test was used to compare the detection rate of both assays (P<0.05). Results: Streptococci were detected in 28.12% (9/32) and 37.5% (12/32) of S1 samples using rRNA- and rDNA-based PCR assays, respectively. In contrast, they were detected in only 6.25% (2/32) of S2 samples using rRNA-based RT-PCR, compared to 15.62% (5/32) using rDNA-based PCR. Finally, in S3 samples, streptococci were not detected by rRNA, whereas rDNA-based PCR still detected the bacteria in 12.5% (4/32) of the samples. The total number of PCR-positive reactions in the rDNA-based PCR was higher than in the rRNA-based assay (P<0.05). Conclusions: The rRNA-based RT-PCR showed a lower detection rate of streptococci when compared to the rDNA-based PCR, suggesting that the latter may have detected dead cells of streptococci in root canal samples

Evaluation Of Pain In Newborn In Intensive Therapy In The Vision Of Health Professionals

Objective: Evaluate how health professionals perceive the pain in the NBs hospitalized in the Neonatal Intensive Care Unit. Method: This is a descriptive, exploratory research with a qualitative approach, developed in the maternity hospital Instituto de Saúde Elpídio de Almeida (ISEA), in the period from February to March 2016, after approval by the Research Ethics Committee (CESED), where the technique of data collection was through an interview with a prior script divided into two parts: the first part deals with socio-demographic issues, while the second part deals with issues related to pain in the NB, where they were recorded and transcribed in their entirety, using the content treatment through content analysis of the thematic type proposed by Laurence Bardin. Results: The results show that all health professionals perceive the pain in the neonate, where three categories emerged: "relates the evaluation of pain with physiological and behavioral measurements"; "know, but do not evaluate with pain score", "do not evaluate with pain score". Regarding the use of some method for pain relief, two categories emerged: "use of non-pharmacological methods for pain relief"; "Use of simultaneous pharmacological and non-pharmacological methods". Conclusion: All the professionals contributed to the growth of this research, being necessary more studies about the pain, mainly with regard to the use of multidimensional scales, since there is a lack of knowledge on the part of the professionals and teams that work in the NICU

Nursing Professionals And Occupational Accidents

The current study has the objective to identify main work accidents that occurs with nursing workers. This is an integrative literature review that enabled to include scientific articles indexed in the databases of Virtual Library in Health (BVS). Searches were made from october to november 2016, using the descriptors: “work accidents”, “nursing professionals”, separated between themselves by the boolean operator AND. The following criteria were adopted: fully available articles, in portuguese language, published in the last five years. Articles that presented some duplicity and didn’t meet the proposed study objectives were excluded. After searches, ten scientific productions about the subject were selected. Results indicate the occurrence of accidents between nursing professionals and the main factors were related with sharp object handling, non-utilization of IPEs, excessive workload and needle resurfacing. It is concluded that strategies are required to minimize these occurrences, as the adoption of standard precautionary measures, the adequacy of staff numbers and better work conditions to this professional category. Besides that, the need of new researches about this subject are emphasized

Microorganisms from canals of root-filled teeth with periapical lesions and the antimicrobial susceptibility to different antibiotics of enterococcus faecalis isolates

Orientador: Brenda Paula Figueiredo de Almeida GomesTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: O presente estudo teve por objetivos: identificar a microbiota de canais radiculares de dentes com insucesso do tratamento endodôntico, e testar, in vitro, a suscetibilidade de cepas de Enterococcus faecalis a diferentes antibióticos. Sessenta dentes tratados endodonticamente com lesões periapicais persistentes foram selecionados para esse estudo. Durante o retratamento endodôntico, o material obturador foi removido e foi realizada a coleta microbiológica dos canais radiculares. Foram utilizados meios de transporte, cultura e incubação que propiciam o crescimento de bactérias anaeróbias estritas. Vinte e uma cepas de Enterococcus faecalis foram testadas quanto à suscetibilidade aos seguintes antimicrobianos: benzilpenicilina, amoxicilina, amoxicilina + ácido clavulânico, clindamicina, eritromicina, azitromicina, vancomicina, cloranfenicol, tetraciclina, doxiciclina, ciprofloxacino e moxifloxacino. As concentrações inibitórias mínimas (CIMs) dos agentes antimicrobianos foram determinadas pelo método do E-test. As cepas também foram testadas quanto à produção de beta-lactamase utilizando nitrocefin. Microrganismos foram isolados em 51 casos. A maioria dos canais apresentava somente 1 ou 2 espécies bacterianas. Do total de espécies bacterianas isoladas, 57,4% eram bactérias anaeróbias facultativas e 83,3% Gram-positivas. Enterococcus faecalis foi a espécie bacteriana mais freqüentemente isolada. Bactérias anaeróbias estritas representavam 42,6% das espécies e, entre eles, Peptostreptococcus spp. foram os mais isolados. Associações significantes foram encontradas entre dor ou história de dor e infecções anaeróbias e polimicrobianas (p256 µg/ml, com apenas 28,5% das cepas sensíveis (CIM =0.5 µg/ml). Suscetibilidade limitada também foi verificada com azitromicina, que foi ativa contra somente 14,2% dos isolados. Nenhuma cepa produziu beta-lactamase. Concluímos que a microbiota dos canais de dentes tratados endodonticamente é composta por um número limitado de espécies microbianas, predominantemente Gram-positivas. Anaeróbios facultativos, especialmente Enterococcus faecalis, são os microrganismos mais comumente isolados, entretanto, infecções polimicrobianas e anaeróbios estritos são freqüentemente encontrados em canais de dentes tratados endodonticamente com sintomatologia clínica. As cepas de Enterococcus faecalis foram sensíveis, in vitro, a amoxicilina, amoxicilina associada ao ácido clavulânico, vancomicina e moxifloxacino. A maioria dos isolados foi sensível aos antibióticos: cloranfenicol, tetraciclina, doxiciclina, ou ciprofloxacino. Eritromicina e azitromicina foram menos eficazesAbstract: The objectives of the present study were to identify the microbial flora within root canals of teeth with failed root canal treatment, and to test, in vitro, the susceptibility of Enterococcus faecalis isolates. Sixty root-filled teeth with persisting periapical lesions were selected for this study. During nonsurgical endodontic re-treatment, the root-filling material was removed and the canals were sampled. Microbial sampling, isolation and species determination were performed using advanced microbiologic techniques for anaerobic species. Additionally, 21 Enterococcus faecalis isolates were tested for their antibiotic susceptibilities to benzylpenicillin, amoxicillin, amoxicillin-clavulanic acid, erythromycin, azithromycin, vancomycin, chloramphenicol, tetracycline, doxycycline, ciprofloxacin and moxifloxacin. Minimal inhibitory concentrations (MICs) for the antimicrobial agents were determined using the E-test System. The strains were also tested for ß-lactamase production with nitrocefin. Microorganisms were recovered from 51 teeth. In most cases, one or two strains per canal were found. Of the microbial species isolated, 57.4% were facultative anaerobic species and 83.3% Gram-positive microorganisms. Enterococcus faecalis was the bacterial species most frequently recovered. Obligate anaerobes accounted for 42.6% of the species and the most frequently isolated genera was Peptostreptococcus. Significant associations were observed between pain or history of pain and polymicrobial infections or anaerobes (p<0.05). All Enterococcus faecalis strains were susceptible to penicillins in vitro, however, the MICs of amoxicillin and amoxicillinclavulanic acid (MIC90= 0.75 µg/ml) were lower than for benzylpenicillin (MIC90= 3.0 µg/ml). All strains studied were also susceptible to vancomycin and moxifloxacin, while 95.2% were susceptible to chloramphenicol. Among the isolates, 85.7% were susceptible to tetracycline and doxycycline, and 80.9% to ciprofloxacin. The MIC of erythromycin ranged from 0.38 to >256 µg/ml; only 28.5% of the strains were susceptible (MIC =0.5 µg/ml). Limited susceptibility was also observed with azithromycin which was active against only 14.2% of isolates. No strains produced ß-lactamase. Therefore, it was concluded that the microbial flora in canals after failure of root canal treatment were limited to a small number of predominantly Gram-positive microbial species. Facultative anaerobes, especially Enterococcus faecalis, were the most commonly isolated microorganisms, however, polymicrobial infections and obligate anaerobes were frequently found in canals of symptomatic root-filled teeth. Enterococcus faecalis isolates were completely susceptible, in vitro, to amoxicillin, amoxicillin-clavulanic acid, vancomycin, and moxifloxacin. Most isolates were susceptible to chloramphenicol, tetracycline, doxycycline, or ciprofloxacin. Erythromycin and azithromycin were least effectiveDoutoradoEndodontiaDoutor em Clínica Odontológic

Capsule Locus Polymorphism among Distinct Lineages of Enterococcus faecalis Isolated from Canals of Root-filled Teeth with Periapical Lesions

Introduction: Although Enterococcus faecalis is a member of the normal microbiota, it is also a major cause of nosocomial infections. Some strains of E. faecalis produce capsule, which contributes to pathogenesis through evasion of host defenses, and its production is dependent on the capsule (cps) operon polymorphism. This study investigated cps locus polymorphism in distinct lineages of E. faecalis isolated from canals of root-filled teeth with periapical lesions. Methods: Twenty-two E. faecalis isolates were evaluated regarding the cps operon polymorphism and genetic diversity. The 3 known CPS types were determined by polymerase chain reaction. This information was correlated with multilocus sequence typing data, which were used to define genetic lineages. Results: cpsA and cpsB were the only detected genes within the cps operon in 62.5% of E. faecalis strains (14/22), indicative of genotype CPS 1, which lacks capsule expression. The essential genes in the cps operon for capsule production were detected in the remaining strains, whereas 3 belonged to genotype CPS 5 and 5 strains to genotype CPS 2. A total of 14 sequence types (STs) were resolved in 22 E. faecalis isolates. Comparison with the E. faecalis international multilocus sequence typing database revealed that 9 STs were previously found, and that the 5 STs were novel. Conclusions: Certain E. faecalis genotypes from canals of root-filled teeth with periapical lesions belong to lineages associated with capsule expression and production of multiple virulence factors, which might account for their increased pathogenic potential. (J Endod 2012;38:58-61)FAPESPFAPESP [09/52661-5

Gemella Morbillorum In Primary And Secondary/persistent Endodontic Infections.

To investigate the presence of Gemella morbillorum by culture or nested PCR in primary and secondary/persistent endodontic infections. Microbial samples were taken from 50 cases with primary and 50 cases with secondary/persistent endodontic infections. Microbiologic techniques were used for culture and identification. The DNA extracted from the samples was analyzed for the presence of the target species using species-specific primers. Culture and polymerase chain reaction (PCR) identified the species in 23 and 77, respectively, of 100 root canals. Culture yielded the test organism in 19 of 50 (38%) of root canal samples from primary and in 4 of 50 (8%) from secondary/persistent infections. PCR yielded the test organisms in 41 of 50 (82%) and 36 of 50 (72%) of the, respectively, primary and secondary/persistent root canal infections studied. Gemella morbillorum was identified more frequently in primary endodontic infections than in secondary/persistent ones. A higher frequency of the target species was detected by PCR than by culture.105519-2

Analysis of genetic lineages and their correlation with virulence genes in enterococcus faecalis clinical isolates from root canal and systemic infections

Introduction: Enterococcus faecalis is a member of the mammalian gastrointestinal microbiota but has been considered a leading cause of hospital-acquired infections. In the oral cavity, it is commonly detected from root canals of teeth with failed endodontic treatment. However, little is known about the virulence and genetic relatedness among E. faecalis isolates from different clinical sources. This study compared the presence of enterococcal virulence factors among root canal strains and clinical isolates from hospitalized patients to identify virulent clusters of E. faecalis. Methods: Multilocus sequence typing analysis was used to determine genetic lineages of 40 E. faecalis\ud clinical isolates from different sources. Virulence clusters were determined by evaluating capsule (cps) locus polymorphisms, pathogenicity island gene content, and antibiotic\ud resistance genes by polymerase chain reaction. Results: The clinical isolates from hospitalized patients formed a phylogenetically separate group and were mostly grouped in the clonal complex 2, which is a known virulent cluster of E. faecalis that has caused\ud infection outbreaks globally. The clonal complex 2 group comprised capsule-producing strains harboring multiple antibiotic resistance and pathogenicity island genes. On\ud the other hand, the endodontic isolates were more diverse and harbored few virulence and antibiotic resistance genes. In particular, although more closely related\ud to isolates from hospitalized patients, capsuleproducing E. faecalis strains from root canals did not carry more virulence/antibiotic genes than other endodontic isolates. Conclusions: E. faecalis isolates from endodontic infections have a genetic and virulence profile different from pathogenic clusters of hospitalized patients’ isolates, which is most likely due to niche\ud specialization conferred mainly by variable regions in the genome.FAPESP, 09/52661-

Bacteriological Study Of Root Canals Associated With Periapical Abscesses.

The aim of this study was to identify microorganisms from root canals with periapical abscesses and to ascertain the susceptibility of Peptostreptococcus prevotii and Fusobacterium necrophorum to antimicrobials. Study design Thirty root canals were microbiologically sampled by using sterile paper points. The concomitant microorganisms were identified through the use of established methods. The susceptibility of P prevotii and F necrophorum to antimicrobials was evaluated by using the E test method. A total of 117 different bacterial strains were recovered, including 75 strict anaerobes or microphilic species. The most frequently isolated strict anaerobes were P prevotii, Peptostreptococcus micros, and F necrophorum. Facultative bacteria such as Gemella morbillorum and Streptococcus mitis were also found, albeit less frequently. The data revealed that P prevotii and F necrophorum were susceptible to the tested antibiotics. Gram-positive anaerobic bacteria predominate in the mixed microbiota of root canals with periapical abscesses. Moreover, P prevotii and F necrophorum are susceptible to the tested antibiotics.96332-