RESUMO
This study assessed the subgingival occurrence of the flagellated, Gram-negative, anaerobic rod Centipeda periodontii in chronic periodontitis and periodontal health/gingivitis with species-specific nucleic acid probes, and evaluated the in vitro resistance of subgingival isolates to therapeutic levels of amoxicillin, metronidazole, and doxycycline. Subgingival plaque biofilm specimens from 307 adults with chronic periodontitis, and 48 adults with periodontal health/localized gingivitis, were evaluated with digoxigenin-labeled, whole-chromosomal, DNA probes to C. periodontii ATCC 35019 possessing a 10(4) cell detection threshold. Fifty-two C. periodontii subgingival culture isolates were assessed on antibiotic-supplemented enriched Brucella blood agar for in vitro resistance to either amoxicillin at 2 µg/ml, metronidazole at 4 µg/ml, or doxycycline at 2 µg/ml. A significantly greater subgingival occurrence of C. periodontii was found in chronic periodontitis subjects as compared to individuals with periodontal health/gingivitis (13.4 vs. 0 %, P < 0.003), although high subgingival counts of the organism (≥ 10(6) cells) were rarely detected (1.3 % of chronic periodontitis subjects). In vitro resistance was not found to amoxicillin or metronidazole, and to doxycycline in only 2 (3.9 %) of the 52 C. periodontii clinical isolates studied. These findings indicate that C. periodontii is not a major constituent of the subgingival microbiome in chronic periodontitis or periodontal health/gingivitis. The potential contribution of C. periodontii to periodontal breakdown in the few chronic periodontitis subjects who yielded high subgingival levels of the organism remains to be delineated. C. periodontii clinical isolates were susceptible in vitro to therapeutic concentrations of three antibiotics frequently used in treatment of human periodontitis.
Assuntos
Periodontite Crônica/microbiologia , Gengivite/microbiologia , Bactérias Anaeróbias Gram-Negativas/patogenicidade , Adulto , Amoxicilina/farmacologia , Antibacterianos/farmacologia , Biofilmes , DNA Bacteriano/análise , Doxiciclina/farmacologia , Feminino , Bactérias Anaeróbias Gram-Negativas/efeitos dos fármacos , Bactérias Anaeróbias Gram-Negativas/isolamento & purificação , Humanos , Masculino , Metronidazol/farmacologia , Testes de Sensibilidade MicrobianaRESUMO
Flexible endoscopy is a widely used diagnostic and therapeutic procedure. Contaminated endoscopes are the medical devices frequently associated with outbreaks of health care-associated infections. Accurate reprocessing of flexible endoscopes involves cleaning and high-level disinfection followed by rinsing and drying before storage. Most contemporary flexible endoscopes cannot be heat sterilized and are designed with multiple channels, which are difficult to clean and disinfect. The ability of bacteria to form biofilms on the inner channel surfaces can contribute to failure of the decontamination process. Implementation of microbiological surveillance of endoscope reprocessing is appropriate to detect early colonization and biofilm formation in the endoscope and to prevent contamination and infection in patients after endoscopic procedures. This review presents an overview of the infections and cross-contaminations related to flexible gastrointestinal endoscopy and bronchoscopy and illustrates the impact of biofilm on endoscope reprocessing and postendoscopic infection.
Assuntos
Infecções Bacterianas/transmissão , Broncoscopia/efeitos adversos , Síndrome de Creutzfeldt-Jakob/transmissão , Endoscopia Gastrointestinal/efeitos adversos , Viroses/transmissão , Infecções Bacterianas/prevenção & controle , Síndrome de Creutzfeldt-Jakob/prevenção & controle , Desinfecção/métodos , Humanos , Doença Iatrogênica/prevenção & controle , Viroses/prevenção & controleRESUMO
Methylobacterium species are a cause of health care-associated infection, including infections in immunocompromised hosts. The ability of Methylobacterium species to form biofilms and to develop resistance to high temperatures, drying, and disinfecting agents may explain the colonization of Methylobacterium in the hospital environment in, e.g., endoscopes. Due to its slow growth, it can be easily missed during microbiological surveillance of endoscope reprocessing. The purpose of this minireview is to present an overview of documented infections and cross-contaminations with Methylobacterium related to endoscopic procedures and to illustrate the health care-associated relevance of this slow-growing bacterium.
Assuntos
Infecção Hospitalar/microbiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Methylobacterium/isolamento & purificação , Atenção à Saúde/métodos , Endoscopia/métodos , Contaminação de EquipamentosRESUMO
OBJECTIVES: Because antimicrobial therapy is often employed in the treatment of infectious dental implant complications, this study determined the occurrence of in vitro antibiotic resistance among putative peri-implantitis bacterial pathogens. METHODS: Submucosal biofilm specimens were cultured from 160 dental implants with peri-implantitis in 120 adults, with isolated putative pathogens identified to species level, and tested in vitro for susceptibility to 4 mg/l of doxycycline, 8 mg/l of amoxicillin, 16 mg/l of metronidazole, and 4 mg/l of clindamycin. Findings for amoxicillin and metronidazole were combined post-hoc to identify peri-implantitis species resistant to both antibiotics. Gram-negative enteric rods/pseudomonads were subjected to ciprofloxacin disk diffusion testing. RESULTS: One or more cultivable submucosal bacterial pathogens, most often Prevotella intermedia/nigrescens or Streptococcus constellatus, were resistant in vitro to clindamycin, amoxicillin, doxycycline, or metronidazole in 46.7%, 39.2%, 25%, and 21.7% of the peri-implantitis subjects, respectively. Only 6.7% subjects revealed submucosal test species resistant in vitro to both amoxicillin and metronidazole, which were either S. constellatus (one subject) or ciprofloxacin-susceptible strains of gram-negative enteric rods/pseudomonads (seven subjects). Overall, 71.7% of the 120 peri-implantitis subjects exhibited submucosal bacterial pathogens resistant in vitro to one or more of the tested antibiotics. CONCLUSIONS: Peri-implantitis patients frequently yielded submucosal bacterial pathogens resistant in vitro to individual therapeutic concentrations of clindamycin, amoxicillin, doxycycline, or metronidazole, but only rarely to both amoxicillin and metronidazole. Due to the wide variation in observed drug resistance patterns, antibiotic susceptibility testing of cultivable submucosal bacterial pathogens may aid in the selection of antimicrobial therapy for peri-implantitis patients.
Assuntos
Resistência Microbiana a Medicamentos , Peri-Implantite/tratamento farmacológico , Peri-Implantite/microbiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Amoxicilina/farmacologia , Biofilmes/efeitos dos fármacos , Ciprofloxacina/farmacologia , Clindamicina/farmacologia , Doxiciclina/farmacologia , Feminino , Humanos , Masculino , Metronidazol/farmacologia , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
Thermal stability is essential for the survival and well-being of preterm neonates. This is achieved in neonatal incubators by raising the ambient temperature and humidity to sufficiently high levels. However, potentially pathogenic microorganisms also can thrive in such warm and humid environments. We therefore investigated whether the level of microbial contamination (i.e., the bacterial load) inside neonatal incubators can be predicted on the basis of their average temperature and relative humidity settings, paying special attention to local temperature differences. Swab samples were taken from the warmest and coldest spots found within Caleo incubators, and these were plated to determine the number of microbial CFU per location. In incubators with high average temperature (≥ 34°C) and relative humidity (≥ 60%) values, the level of microbial contamination was significantly higher at cold spots than at hot spots. This relates to the fact that the local equilibrium relative humidity at cold spots is sufficiently high to sustain microbial growth. The abundance of staphylococci, which are the main causative agents of late-onset sepsis in preterm neonates, was found to be elevated significantly in cold areas. These findings can be used to improve basic incubator hygiene.
Assuntos
Carga Bacteriana , Incubadoras para Lactentes/microbiologia , Staphylococcus/isolamento & purificação , Humanos , Umidade , TemperaturaRESUMO
PURPOSE: The occurrence of in vitro resistance to therapeutic concentrations of spiramycin, amoxicillin, and metronidazole was determined for putative periodontal pathogens isolated in the United States. MATERIALS AND METHODS: Subgingival plaque specimens from 37 consecutive adults with untreated severe periodontitis were anaerobically cultured, and isolated putative periodontal pathogens were identified to a species level. In vitro resistance to spiramycin at 4 µg/ml, amoxicillin at 8 µg/ml, and/or metronidazole at 16 µg/ml was noted when putative periodontal pathogen growth was noted on the respective antibiotic-supplemented primary culture plates. RESULTS: A total of 18 (48.7%) subjects yielded antibiotic-resistant putative periodontal pathogens with spiramycin at 4 µg/ml in drug-supplemented primary isolation plates, as compared to 23 (62.2%) subjects with amoxicillin at 8 µg/ml, and 10 (27.0%) subjects with metronidazole at 16 µg/ml. Spiramycin in vitro resistance occurred among species of Fusobacterium nucleatum (44.4% of organism-positive subjects), Prevotella intermedia/nigrescens (11.1%), Parvimonas micra (10.8%), Streptococcus constellatus (10%), Streptococcus intermedius (10%), Porphyromonas gingivalis (6.7%), and Tannerella forsythia (5.3%). Amoxicillin in vitro resistance was found in P. intermedia/nigrescens (55.5%), T. forsythia (15.8%), S. constellatus (10%), F. nucleatum (5.6%), and P. micra (2.7%). Only S. constellatus (70%) and S. intermedius (40%) exhibited in vitro resistance to metronidazole. When subject-based resistance data for spiramycin and metronidazole were jointly considered, all isolated putative periodontal pathogens were inhibited in vitro by one or the other of the antibiotic concentrations, except for one strain each of S. constellatus and S. intermedius from one study subject. Similarly, either amoxicillin or metronidazole at the drug concentrations tested inhibited in vitro all recovered putative periodontal pathogens, except S. constellatus in one subject. CONCLUSIONS: In vitro spiramycin resistance among putative periodontal pathogens of United States origin occurred in approximately one-half of severe periodontitis patients evaluated, particularly among subgingival F. nucleatum species. In vitro resistance patterns also suggest that therapeutic concentrations of spiramycin plus metronidazole may have potential antimicrobial efficacy in non-Aggregatibacter actinomycetemcomitans-associated periodontitis similar to amoxicillin plus metronidazole, which may be beneficial, where spiramycin is clinically available, for patients hypersensitive to amoxicillin or other beta-lactam antibiotics.
Assuntos
Antibacterianos/farmacologia , Metagenoma/efeitos dos fármacos , Periodontite/microbiologia , Espiramicina/farmacologia , Farmacorresistência Bacteriana , Feminino , Humanos , Masculino , Testes de Sensibilidade Microbiana/métodos , Pessoa de Meia-Idade , Periodontite/tratamento farmacológicoRESUMO
AIM: To test recolonization of periodontal lesions after full-mouth scaling and root planing (FM-SRP) or multiple session-SRP (MS-SRP) in a randomized clinical trial and whether FM-SRP and MS-SRP result in different clinical outcomes. MATERIALS AND METHODS: Thirty-nine subjects were randomly assigned to FM-SRP or MS-SRP groups. At baseline and after 3 months, probing pocket depth (PPD), plaque index (PlI) and bleeding on probing (BoP) were recorded. At baseline, immediately after treatment, after 1, 2, 7, 14 and 90 days, paper point samples from a single site from the maxillary right quadrant were collected for microbiological analysis of five putative pathogens by polymerase chain reaction. RESULTS: FM-SRP and MS-SRP resulted in significant reductions in PPD, BoP and PlI and the overall detection frequencies of the five species after 3 months without significant differences between treatments. Compared with MS-SRP, FM-SRP resulted in less recolonization of the five species, significantly for Treponema denticola, in the tested sites. CONCLUSION: FM-SRP and MS-SRP result in overall clinically and microbiologically comparable outcomes where recolonization of periodontal lesions may be better prevented by FM-SRP.
Assuntos
Bactérias/crescimento & desenvolvimento , Periodontite Crônica/microbiologia , Raspagem Dentária/métodos , Aplainamento Radicular/métodos , Adulto , Idoso , Aggregatibacter actinomycetemcomitans/crescimento & desenvolvimento , Bactérias/classificação , Bacteroides/crescimento & desenvolvimento , Periodontite Crônica/terapia , Protocolos Clínicos , Contagem de Colônia Microbiana , Índice de Placa Dentária , Feminino , Seguimentos , Defeitos da Furca/microbiologia , Defeitos da Furca/terapia , Fusobacterium nucleatum/crescimento & desenvolvimento , Hemorragia Gengival/microbiologia , Hemorragia Gengival/terapia , Humanos , Masculino , Pessoa de Meia-Idade , Índice Periodontal , Bolsa Periodontal/microbiologia , Bolsa Periodontal/terapia , Porphyromonas gingivalis/crescimento & desenvolvimento , Curetagem Subgengival/métodos , Resultado do Tratamento , Treponema denticola/crescimento & desenvolvimentoRESUMO
BACKGROUND: Self-medication with antibiotics occurs among the population in Europe, particularly in southern and eastern countries. We studied the impact of predisposing factors (e.g. attitudes and knowledge concerning antibiotic use and self-medication) and enabling factors (country wealth and healthcare system factors) on self-medication with antibiotics in Europe. METHODS: In this follow-up of a previous European survey, we interviewed a subsample of 1101 respondents. A multilevel analysis with two levels (respondent and country) was performed. Variables that were statistically significantly different between users and non-users of self-medication were considered for inclusion into the multilevel regression analyses. RESULTS: Predisposing factors included individual-level characteristics. High perceived appropriateness of self-medication with antibiotics for bronchitis and an attitude favouring antibiotic use for minor ailments were related to a higher likelihood of self-medication. Enabling factors included individual and country data. At the individual level, perceived availability of antibiotics without a prescription was related to increased probability of self-medication. At the country level, higher gross domestic product (wealth) and exact dispensation of prescribed tablet quantities by pharmacies were independently associated with lower likelihood of self-medication. CONCLUSIONS: Interventions aimed at preventing self-medication should include public education, enforcing regulations regarding the sale of antibiotics, and implementing laws for dispensing exact prescribed tablet quantities in pharmacies. With the included determinants, we explained almost all the variance at the country level, but not at the individual level. Future studies to increase our understanding of determinants of self-medication with antibiotics should focus on individual-level factors such as doctor-patient relationships and patient satisfaction.
Assuntos
Antibacterianos/uso terapêutico , Comparação Transcultural , Atenção à Saúde/organização & administração , Conhecimentos, Atitudes e Prática em Saúde , Adulto , Idoso , Economia , Europa (Continente) , Feminino , Pesquisas sobre Atenção à Saúde , Humanos , Masculino , Pessoa de Meia-Idade , AutomedicaçãoRESUMO
We report the investigation of an outbreak situation of methicillin-resistant Staphylococcus aureus (MRSA) that occurred at the Academic Hospital Paramaribo (AZP) in the Republic of Suriname from April to May 2013. We performed whole genome sequencing with complete gap closure for chromosomes and plasmids on all isolates. The outbreak involved 12 patients and 1 healthcare worker/nurse at the AZP. In total 24 isolates were investigated. spa typing, genome-wide single nucleotide polymorphism (SNP) analysis, ad hoc whole genome multilocus sequence typing (wgMLST), stable core genome MLST (cgMLST) and in silico PFGE were used to determine phylogenetic relatedness and to identify transmission. Whole-genome sequencing (WGS) showed that all isolates were members of genomic variants of the North American USA300 clone. However, WGS revealed a heterogeneous population structure of USA300 circulating at the AZP. We observed up to 8 SNPs or up to 5 alleles of difference by wgMLST when the isolates were recovered from different body sites of the same patient or if direct transmission between patients was most likely. This work describes the usefulness of complete genome sequencing of bacterial chromosomes and plasmids providing an unprecedented level of detail during outbreak investigations not being visible by using conventional typing methods.
Assuntos
Surtos de Doenças , Staphylococcus aureus Resistente à Meticilina/genética , Infecções Estafilocócicas/genética , Adulto , Idoso , Genoma Bacteriano , Humanos , Lactente , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Pessoa de Meia-Idade , Filogenia , Polimorfismo de Nucleotídeo Único , Suriname , Sequenciamento Completo do GenomaRESUMO
OBJECTIVE: To reveal the source of a nosocomial outbreak of colonization and infection with a strain of Serratia marcescens positive for Guiana extended-spectrum beta-lactamase 1 (GES-1) that occurred among patients in a neurosurgical intensive care unit (ICU) in a Dutch university medical center from May 2002 through March 2003. METHODS: Samples from the environment and from the hands of healthcare workers (HCWs) were cultured. A retrospective case-control study was carried out. RESULTS: Fifteen neurosurgical ICU patients who had 1 or more cultures that yielded the epidemic strain of S. marcescens from May 2002 through March 2003 were defined as case patients and matched with 30 control patients. Environmental cultures did not reveal a prominent source of S. marcescens. Cultures of specimens from the hands of 100 HCWs revealed colonization of a single HCW with the epidemic strain. Although this HCW instantly went on leave, serial cultures detected prolonged carriage of the epidemic strain on the hands of the HCW for 3 months. The skin of the HCW's hands was psoriatic. The epidemic abruptly ended after the colonized HCW went on leave. Retrospective case-control analysis showed that the patients colonized or infected with S. marcescens received significantly more nursing care from the colonized HCW than did control patients (P<.05). From February 2004 through October 2004, a second cluster of 3 patients was detected with the epidemic strain of S. marcescens. In October 2004, the formerly colonized HCW appeared to have carriage of the epidemic strain on the hands again. CONCLUSIONS: A single HCW with the epidemic strain of S. marcescens on the hands was considered the source of this outbreak.
Assuntos
Portador Sadio/microbiologia , Infecção Hospitalar/epidemiologia , Surtos de Doenças , Mãos/microbiologia , Pessoal de Saúde , Infecções por Serratia/epidemiologia , Serratia marcescens , Centros Médicos Acadêmicos , Antibacterianos/farmacologia , Portador Sadio/transmissão , Estudos de Casos e Controles , Cuidados Críticos , Infecção Hospitalar/microbiologia , Infecção Hospitalar/transmissão , Humanos , Transmissão de Doença Infecciosa do Profissional para o Paciente , Testes de Sensibilidade Microbiana , Países Baixos , Neurocirurgia , Infecções por Serratia/microbiologia , Infecções por Serratia/transmissão , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/enzimologia , Serratia marcescens/crescimento & desenvolvimento , Serratia marcescens/isolamento & purificação , beta-Lactamases/metabolismoRESUMO
Enterococcus faecalis and Enterococcus faecium are among the leading causes of hospital-acquired infections. Reliable and quick identification of E. faecalis and E. faecium is important for accurate treatment and understanding their role in the pathogenesis of infections. Fluorescent in situ hybridization (FISH) of whole bacterial cells with oligonucleotides targeted at the 16S rRNA molecule leads to a reduced time to identification. In clinical practice, FISH therefore can be used in situations in which quick identification is necessary for optimal treatment of the patient. Furthermore, the abundance, spatial distribution and bacterial cell morphology can be observed in situ. This report describes the design of two fluorescent-labelled oligonucleotides that, respectively, detect the 16S rRNA of E. faecalis and the 16S rRNA of E. faecium, Enterococcus hirae, Enterococcus mundtii, Enterococcus villorum and Enterococcus saccharolyticus. Different protocols for the application of these oligonucleotides with FISH in different clinical samples such as faeces or blood cultures are given. Enterococci in a biofilm attached to a biomaterial were also visualized. Embedding of the biomaterial preserved the morphology and therefore the architecture of the biofilm could be observed. The usefulness of other studies describing FISH for detection of enterococci is generally hampered by the fact that they have only focused on one material and one protocol to detect the enterococci. However, the results of this study show that the probes can be used both in the routine laboratory to detect and determine the enterococcal species in different clinical samples and in a research setting to enumerate and detect the enterococci in their physical environment.
Assuntos
Enterococcus faecalis/genética , Enterococcus faecalis/isolamento & purificação , Enterococcus faecium/genética , Enterococcus faecium/isolamento & purificação , Infecções por Bactérias Gram-Positivas/microbiologia , Hibridização in Situ Fluorescente/métodos , Sondas de Oligonucleotídeos , Materiais Biocompatíveis , Biofilmes , Sangue/microbiologia , Cateteres de Demora/microbiologia , DNA Bacteriano/genética , DNA Ribossômico/genética , Fezes/microbiologia , Infecções por Bactérias Gram-Positivas/diagnóstico , Humanos , RNA Bacteriano/genética , RNA Ribossômico 16S/genéticaRESUMO
BACKGROUND: Patients with chronic periodontitis (CP) may yield multiple species of putative periodontal bacterial pathogens that vary in their antibiotic drug susceptibility. This study determines the occurrence of in vitro antibiotic resistance among selected subgingival periodontal pathogens in patients with CP. METHODS: Subgingival biofilm specimens from inflamed deep periodontal pockets were removed before treatment from 400 adults with CP in the United States. The samples were cultured, and selected periodontal pathogens were tested in vitro for susceptibility to amoxicillin at 8 mg/L, clindamycin at 4 mg/L, doxycycline at 4 mg/L, and metronidazole at 16 mg/L, with a post hoc combination of data for amoxicillin and metronidazole. Gram-negative enteric rods/pseudomonads were subjected to ciprofloxacin disk-diffusion testing. RESULTS: Overall, 74.2% of the patients with CP revealed subgingival periodontal pathogens resistant to at least one of the test antibiotics. One or more test species, most often Prevotella intermedia/nigrescens, Streptococcus constellatus, or Aggregatibacter actinomycetemcomitans, were resistant in vitro to doxycycline, amoxicillin, metronidazole, or clindamycin, in 55%, 43.3%, 30.3%, and 26.5% of the patients with CP, respectively. Fifteen percent of patients harbored subgingival periodontal pathogens resistant to both amoxicillin and metronidazole, which were mostly either S. constellatus (45 individuals) or ciprofloxacin-susceptible strains of Gram-negative enteric rods/pseudomonads (nine individuals). CONCLUSIONS: Patients with CP in the United States frequently yielded subgingival periodontal pathogens resistant in vitro to therapeutic concentrations of antibiotics commonly used in clinical periodontal practice. The wide variability found in periodontal pathogen antibiotic-resistance patterns should concern clinicians empirically selecting antibiotic treatment regimens for patients with CP.
Assuntos
Antibacterianos/farmacologia , Periodontite Crônica/microbiologia , Farmacorresistência Bacteriana , Adulto , Idoso , Aggregatibacter actinomycetemcomitans/efeitos dos fármacos , Amoxicilina/farmacologia , Carga Bacteriana , Candida/efeitos dos fármacos , Ciprofloxacina/farmacologia , Clindamicina/farmacologia , Placa Dentária/microbiologia , Doxiciclina/farmacologia , Enterococcus faecalis/efeitos dos fármacos , Feminino , Fusobacterium nucleatum/efeitos dos fármacos , Humanos , Masculino , Metronidazol/farmacologia , Pessoa de Meia-Idade , Bolsa Periodontal/microbiologia , Prevotella intermedia/efeitos dos fármacos , Prevotella nigrescens/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Streptococcus constellatus/efeitos dos fármacosRESUMO
For some microbial species, such as Bacillus anthracis, the etiologic agent of the disease anthrax, correct detection and identification by molecular methods can be problematic. The detection of virulent B. anthracis is challenging due to multiple virulence markers that need to be present in order for B. anthracis to be virulent and its close relationship to Bacillus cereus and other members of the B. cereus group. This is especially the case in environments where build-up of Bacillus spores can occur and several representatives of the B. cereus group may be present, which increases the chance for false-positives. In this study we show the presence of B. anthracis-like bacteria and other members of the B. cereus group in a microbial community within the human environment of the International Space Station and their preliminary identification by using conventional culturing as well as molecular techniques including 16S rDNA sequencing, PCR and real-time PCR. Our study shows that when monitoring the microbial hygiene in a given human environment, health risk assessment is troublesome in the case of virulent B. anthracis, especially if this should be done with rapid, easy to apply and on-site molecular methods.
Assuntos
Bacillus anthracis/isolamento & purificação , Bacillus cereus/isolamento & purificação , Astronave , Medicina Aeroespacial , Antraz/genética , Antraz/microbiologia , Bacillus anthracis/genética , Bacillus anthracis/patogenicidade , Bacillus cereus/genética , Bacillus cereus/patogenicidade , HumanosRESUMO
BACKGROUND: Streptococcus constellatus and Streptococcus intermedius in subgingival dental plaque biofilms may contribute to forms of periodontitis that resist treatment with conventional mechanical root debridement/surgical procedures and may additionally participate in some extraoral infections. Because systemic antibiotics are often used in these clinical situations, and little is known of the antibiotic susceptibility of subgingival isolates of these two bacterial species, this study determined the in vitro susceptibility to six antibiotics of fresh S. constellatus and S. intermedius clinical isolates from human periodontitis lesions. METHODS: A total of 33 S. constellatus and 17 S. intermedius subgingival strains, each recovered from separate patients with severe chronic periodontitis (n = 50) before treatment, were subjected to antibiotic gradient strip susceptibility testing with amoxicillin, azithromycin, clindamycin, ciprofloxacin, and doxycycline on blood-supplemented Mueller-Hinton agar and to the inhibitory effects of metronidazole at 16 mg/L in an enriched Brucella blood agar dilution assay. Clinical and Laboratory Standards Institute and European Committee on Antimicrobial Susceptibility Testing interpretative standards were used to assess the results. RESULTS: Clindamycin was the most active antibiotic against S. constellatus (minimum inhibitory concentration at 90% [MIC90] 0.25 mg/L), and amoxicillin was most active against S. intermedius (MIC90 0.125 mg/L). A total of 30% of the S. constellatus and S. intermedius clinical isolates were resistant in vitro to doxycycline, 98% were only intermediate in susceptibility to ciprofloxacin, and 90% were resistant to metronidazole at 16 mg/L. CONCLUSION: Subgingival S. constellatus and S. intermedius exhibited variable antibiotic susceptibility profiles, potentially complicating empirical selection of periodontitis antibiotic therapy in patients who are species positive.
Assuntos
Antibacterianos/farmacologia , Periodontite Crônica/microbiologia , Farmacorresistência Bacteriana , Streptococcus intermedius/efeitos dos fármacos , Streptococcus/efeitos dos fármacos , Adulto , Idoso , Amoxicilina/farmacologia , Azitromicina/farmacologia , Técnicas Bacteriológicas , Ciprofloxacina/farmacologia , Clindamicina/farmacologia , Doxiciclina/farmacologia , Feminino , Humanos , Masculino , Metronidazol/farmacologia , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Resistência às Penicilinas , Streptococcus/classificação , Resistência a TetraciclinaRESUMO
BACKGROUND: Enterococcus faecalis may contribute to periodontal breakdown in heavily infected subgingival sites, particularly in patients responding poorly to mechanical forms of periodontal therapy. Because only limited data are available on the antimicrobial sensitivity of enterococci of subgingival origin, this study evaluates the in vitro antibiotic susceptibility of E. faecalis isolated from periodontitis patients in the United States. METHODS: Pure cultures of 47 subgingival E. faecalis clinical isolates were each inoculated onto specially prepared broth microdilution susceptibility panels containing vancomycin, teicoplanin, and six oral antibiotics of potential use in periodontal therapy. After incubation in ambient air for 18 to 20 hours, minimal inhibitory drug concentrations were determined using applicable Clinical and Laboratory Standards Institute criteria and interpretative guidelines. The organisms were additionally evaluated for in vitro resistance to metronidazole at 4 µg/mL. RESULTS: Periodontal E. faecalis exhibited substantial in vitro resistance to tetracycline (53.2% resistant), erythromycin (80.8% resistant or intermediate resistant), clindamycin (100% resistant to 2 µg/mL), and metronidazole (100% resistant to 4 µg/mL). In comparison, the clinical isolates were generally sensitive to ciprofloxacin (89.4% susceptible; 10.6% intermediate resistant) and 100% susceptible in vitro to ampicillin, amoxicillin/clavulanate, vancomycin, and teicoplanin. CONCLUSIONS: Tetracycline, erythromycin, clindamycin, and metronidazole revealed poor in vitro activity against human subgingival E. faecalis clinical isolates, and would likely be ineffective therapeutic agents against these species in periodontal pockets. Among orally administered antibiotics, ampicillin, amoxicillin/clavulanate, and ciprofloxacin exhibited marked in vitro inhibitory activity against periodontal E. faecalis, and may be clinically useful in treatment of periodontal infections involving enterococci.
Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Enterococcus faecalis/efeitos dos fármacos , Periodontite/microbiologia , Adulto , Idoso , Combinação Amoxicilina e Clavulanato de Potássio/farmacologia , Ampicilina/farmacologia , Anti-Infecciosos/farmacologia , Técnicas Bacteriológicas , Periodontite Crônica/tratamento farmacológico , Periodontite Crônica/microbiologia , Ciprofloxacina/farmacologia , Clindamicina/farmacologia , Placa Dentária/microbiologia , Enterococcus faecalis/classificação , Eritromicina/farmacologia , Feminino , Gengiva/microbiologia , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Masculino , Metronidazol/farmacologia , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Bolsa Periodontal/tratamento farmacológico , Bolsa Periodontal/microbiologia , Teicoplanina/farmacologia , Resistência a Tetraciclina , Vancomicina/farmacologiaRESUMO
Periodontitis and caries are infectious diseases of the oral cavity in which oral biofilms play a causative role. Moreover, oral biofilms are widely studied as model systems for bacterial adhesion, biofilm development, and biofilm resistance to antibiotics, due to their widespread presence and accessibility. Despite descriptions of initial plaque formation on the tooth surface, studies on mature plaque and plaque structure below the gum are limited to landmark studies from the 1970s, without appreciating the breadth of microbial diversity in the plaque. We used fluorescent in situ hybridization to localize in vivo the most abundant species from different phyla and species associated with periodontitis on seven embedded teeth obtained from four different subjects. The data showed convincingly the dominance of Actinomyces sp., Tannerella forsythia, Fusobacterium nucleatum, Spirochaetes, and Synergistetes in subgingival plaque. The latter proved to be new with a possibly important role in host-pathogen interaction due to its localization in close proximity to immune cells. The present study identified for the first time in vivo that Lactobacillus sp. are the central cells of bacterial aggregates in subgingival plaque, and that Streptococcus sp. and the yeast Candida albicans form corncob structures in supragingival plaque. Finally, periodontal pathogens colonize already formed biofilms and form microcolonies therein. These in vivo observations on oral biofilms provide a clear vision on biofilm architecture and the spatial distribution of predominant species.
Assuntos
Bactérias/crescimento & desenvolvimento , Biofilmes/crescimento & desenvolvimento , Boca/microbiologia , Dente/microbiologia , Actinomyces/genética , Actinomyces/fisiologia , Bactérias/classificação , Bactérias/genética , Aderência Bacteriana , Candida albicans/isolamento & purificação , Cárie Dentária/microbiologia , Placa Dentária/microbiologia , Fusobacterium nucleatum/genética , Fusobacterium nucleatum/fisiologia , Interações Hospedeiro-Patógeno , Humanos , Hibridização in Situ Fluorescente , Lactobacillus/genética , Lactobacillus/fisiologia , Boca/patologia , Periodontite/microbiologia , Filogenia , Streptococcus/crescimento & desenvolvimento , Streptococcus/fisiologiaRESUMO
PURPOSE: Selective digestive microbial decontamination (SDD) is hypothesized to benefit patients in intensive care (ICU) by suppressing Gram-negative potential pathogens from the colon without affecting the anaerobic intestinal microbiota. The purpose of this study was to provide more insight to the effects of digestive tract and oropharyngeal decontamination on the intestinal microbiota by means of a prospective clinical trial in which faecal samples were collected from ICU patients for intestinal microbiota analysis. METHODS: The faecal samples were collected from ICU patients enrolled in a multicentre trial to study the outcome of SDD and selective oral decontamination (SOD) in comparison with standard care (SC). Fluorescent in situ hybridization (FISH) was used to analyze the faecal microbiota. The numbers of bacteria from different bacterial groups were compared between the three regimens. RESULTS: The total counts of bacteria per gram faeces did not differ between regimens. The F. prausnitzii group of bacteria, representing an important group among intestinal microbiota, was significantly reduced in the SDD regimen compared to the SC and SOD. The Enterobacteriaceae were significantly suppressed during SDD compared to both SOD and SC; enterococci increased in SDD compared to both other regimens. CONCLUSIONS: The composition of the intestinal microbiota is importantly affected by SDD. The F. prausnitzii group was significantly suppressed during SDD. This group of microbiota is a predominant producer of butyrate, the main energy source for colonocytes. Reduction of this microbiota is an important trade-off while reducing gram-negative bacteria by SDD.
Assuntos
Descontaminação/métodos , Bactérias Gram-Negativas/efeitos dos fármacos , Unidades de Terapia Intensiva , Intestinos/microbiologia , Metagenoma/efeitos dos fármacos , Orofaringe/microbiologia , Antibioticoprofilaxia , Fezes/microbiologia , Humanos , Pessoa de Meia-Idade , Países BaixosRESUMO
PURPOSE: Although the relevance of cultural factors for antibiotic use has been recognized, few studies exist in Europe. We compared public attitudes, beliefs and knowledge concerning antibiotic use and self-medication between 11 European countries. METHODS: In total, 1101 respondents were interviewed on their attitudes towards appropriateness of self-medication with antibiotics and situational use of antibiotics, beliefs about antibiotics for minor ailments, knowledge about the effectiveness of antibiotics on viruses and bacteria and awareness about antibiotic resistance. To deal with the possible confounding effect of both use of self-medication and education we performed stratified analyses, i.e. separate analyses for users and non-users of self-medication, and for respondents with high and low education. The differences between countries were considered relevant when regression coefficients were significant in all stratum-specific analyses. RESULTS: Respondents from the UK, Malta, Italy, Czech Republic, Croatia, Israel and Lithuania had significantly less appropriate attitudes, beliefs or knowledge for at least one of the dimensions compared with Swedish respondents. The Dutch, Austrian and Belgian respondents did not differ from Swedish for any dimension. CONCLUSIONS: The most pronounced differences were for awareness about resistance, followed by attitudes towards situational use of antibiotics. Awareness about antibiotic resistance was the lowest in countries with higher prevalence of resistance.
Assuntos
Antibacterianos/administração & dosagem , Comparação Transcultural , Conhecimentos, Atitudes e Prática em Saúde , Automedicação/psicologia , Adulto , Idoso , Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana , Europa (Continente) , Feminino , Seguimentos , Pesquisas sobre Atenção à Saúde , História do Século XV , Humanos , Masculino , Pessoa de Meia-Idade , PrevalênciaRESUMO
BACKGROUND: Self-medication with antibiotics may increase the risk of inappropriate use and the selection of resistant bacteria. One of the triggers for using self-medication may be past experience with antibiotics prescribed by health professionals. We examined the association between prescribed use and self-medication with antibiotics. METHODS: A population survey was conducted in 19 European countries, covering 15,548 respondents. Multinomial logistic regression analysis was used to study the relationship between prescribed use and self-medication for all symptoms/diseases and for upper respiratory tract infections (URTIs). RESULTS: The association between prescribed use and self-medication was modified by source of self-medication, region in Europe and education. This association was consistently stronger for self-medication from leftovers than from other sources, primarily directly from a pharmacy. It was stronger also for respondents from Northern/Western Europe than respondents from Eastern Europe and Southern Europe and those with low education. Prescribed use for URTIs (minor ailments such as throat symptom, influenza, etc.) increased the likelihood of self-medication with leftover antibiotics for these symptoms/diseases in all European regions. CONCLUSIONS: Our study shows consistent associations between prescribed use and self-medication with antibiotics from leftovers, but has not been able to support the hypothesis that self-medication from other sources than leftovers is triggered by earlier prescribed use. Preventing leftovers may be one effective way of preventing self-medication. This can be achieved by ensuring that the amount dispensed corresponds to the amount prescribed, by educating patients and by making doctors aware that prescribing for minor ailments may increase the risk of self-medication for such ailments.
Assuntos
Antibacterianos/uso terapêutico , Prescrições de Medicamentos , Automedicação , Estudos Transversais , Humanos , Modelos Logísticos , Educação de Pacientes como Assunto , Infecções Urinárias/tratamento farmacológicoRESUMO
Bacteria play an important role in the initiation and progression of periodontal diseases and are part of a biofilm, which can contain over 100 different species. The aim of the present study was to show the potential of denaturing gradient gel electrophoresis (DGGE) as a tool for the detection of clinically relevant species and to compare the results of detection by DGGE with those by PCR and culturing. Hybridization of the bands from the DGGE profiles with species-specific probes was developed to confirm the band positions in the marker obtained with reference strains. The sensitivities of DGGE compared to those of cultivation for the detection of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, and Tannerella forsythensis were 100, 100, 88, and 100%, respectively; and the sensitivities of DGGE compared to those of PCR were 100, 90, 88, and 96%, respectively. DGGE as a diagnostic tool could easily be extended to other species, as shown for Treponema denticola, which could be detected in 48% of the samples. Three different groups of A. actinomycetemcomitans serotypes could be distinguished by DGGE (i.e., a group comprising serotypes a, d, e, and f; a group comprising serotype b; and a group comprising serotype c). Amplicons from P. gingivalis and T. denticola migrated to the same position in the gel, and P. intermedia produced multiple bands. In the present study we show that the DGGE profiles represent clinically relevant species which can be detected by hybridization with species-specific probes. With DGGE, large numbers of samples can be analyzed for different species simultaneously, and DGGE may be a good alternative in periodontal microbial diagnostics.