RESUMEN
Streptococcus salivarius is a common, harmless, and prevalent member of the oral microbiota in humans. In the present study, the safety of S. salivarius UBSS-01 was evaluated using in silico methods and preclinical and clinical studies. In an acute toxicity study, rats were administered with 5 g/kg (500 × 109 CFU) S. salivarius UBSS-01. The changes in phenotypic behaviors and hematological, biochemical, electrolytes, and urine analyses were monitored. No toxicity was observed at 14 days post-treatment. The no observable effects limit (NOEL) of S. salivarius UBSS-01 was >5 g/kg in rats. In a 28-day repeat dose toxicity study, rats were administered S. salivarius UBSS-01 once daily at doses of 0.1, 0.5, and 1 g/kg (10, 50, and 100 billion CFU/kg, respectively) body weight. S. salivarius UBSS-01 did not influence any of the hematology parameters and clinical chemistry parameters in plasma and serum samples after 28-day repeated administration. No structural abnormality was observed in the histological examination of organs. Whole genome analysis revealed the absence of virulence factors or genes that may transmit antibiotic resistance. In the double-blind study with 60 human participants (aged 18-60 years), consumption of S. salivarius UBSS-01 for 30 days was found to be safe and results were comparable with placebo treatment These findings indicate that S. salivarius UBSS-01 may be safe for human consumption.
Asunto(s)
Streptococcus salivarius , Adulto , Animales , Femenino , Humanos , Masculino , Persona de Mediana Edad , Ratas , Adulto Joven , Voluntarios Sanos , Nivel sin Efectos Adversos Observados , Probióticos/toxicidad , Ratas Sprague-Dawley , Streptococcus salivarius/efectos de los fármacos , Pruebas de Toxicidad AgudaRESUMEN
This study aimed to evaluate the antimicrobial effect of Thymoquinone (TQ) on four different oral microorganisms. Minimum Bactericidal Concentration (MBC), Minimum Inhibition Concentration (MIC), Broth microdilution, and Well diffusion tests were used to determine the optimum antimicrobial concentrations of TQ against Streptococcus salivarius, Streptococcus oralis, Streptococcus mutans, and Staphylococcus aureus over 1, 3, 6, 12 and 24 h. Chlorhexidine 0.12% was selected as a positive control. The inhibitory effect of TQ on bacterial growth was most noticeable with S. salivarius, while the least affected was S. aureus. TQ's MBC and MIC for S. oralis and S. aureus were comparable 2 mg/mL and 3 mg/mL, respectively. S. salivarius was most resistant to TQ and displayed a value of 5 mg/mL and 4 mg/mL for MIC and MBC, respectively. The viable count of different strains after exposure to TQ's MBC values was most noticeable with S. aureus followed by S. oralis and S. mutans, while S. salivarius was least affected. This study emphasized the promising antimicrobial effect of TQ against the four main oral microorganisms. It has a potential preventive effect against dental caries as well as other oral diseases.
Asunto(s)
Antibacterianos/farmacología , Benzoquinonas/farmacología , Staphylococcus aureus/efectos de los fármacos , Streptococcus mutans/efectos de los fármacos , Streptococcus oralis/efectos de los fármacos , Streptococcus salivarius/efectos de los fármacos , Antibacterianos/química , Benzoquinonas/química , Pruebas de Sensibilidad MicrobianaRESUMEN
The biofilm formation by oral bacteria on the implant surface is one of the most remarkable factors of peri-implant infections, which may eventually lead to bone resorption and loss of the dental implant. Therefore, the elimination of biofilm is an essential step for the successful therapy of implant-related infections. In this work we created a basic in vitro model to evaluate the antibacterial effect of three widely used antiseptics.Commercially pure (CP4) titanium sample discs with sand blasted, acid etched, and polished surface were used. The discs were incubated with mono-cultures of Streptococcus mitis and Streptococcus salivarius. The adhered bacterial biofilms were treated with different antiseptics: chlorhexidine-digluconate (CHX), povidone-iodine (PI), and chlorine dioxide (CD) for 5 min and the control discs with ultrapure water. The antibacterial effect of the antiseptics was tested by colorimetric assay.According to the results, the PI and the CD were statistically the most effective in the elimination of the two test bacteria on both titanium surfaces after 5 min treatment time. The CD showed significant effect only against S. salivarius.Based on our results we conclude that PI and CD may be promising antibacterial agents to disinfecting the peri-implant site in the dental practice.
Asunto(s)
Clorhexidina/análogos & derivados , Compuestos de Cloro/farmacología , Desinfectantes Dentales/farmacología , Óxidos/farmacología , Periimplantitis/prevención & control , Povidona Yodada/farmacología , Streptococcus mitis/efectos de los fármacos , Streptococcus salivarius/efectos de los fármacos , Antibacterianos/farmacología , Antiinfecciosos Locales/farmacología , Adhesión Bacteriana/efectos de los fármacos , Biopelículas/efectos de los fármacos , Clorhexidina/farmacología , Implantes Dentales/microbiología , Humanos , Periimplantitis/microbiología , Streptococcus mitis/crecimiento & desarrollo , Streptococcus salivarius/crecimiento & desarrollo , TitanioRESUMEN
Antimicrobials have become a mainstay of healthcare in the past century due to their activity against pathogens. More recently, it has become clear that they can also affect health via their impact on the microbiota and inflammation. This may explain some of their clinical benefits despite global increases in antimicrobial resistance (AMR) and reduced antimicrobial effectiveness. We showed in a randomized controlled trial of stopping versus continuing cotrimoxazole prophylaxis among HIV-positive Zimbabwean children taking antiretroviral therapy (ART), that continuation of cotrimoxazole persistently suppressed gut-resident viridans group streptococcal species (VGS) that were associated with intestinal inflammation. In this addendum, we provide a broader overview of how antibiotics can shape the microbiota and use high read-depth whole metagenome sequencing data from our published study to investigate whether (i) the impact of cotrimoxazole on gut VGS and (ii) VGS associated inflammation, is attributable to strain-level variability. We focus on S. salivarius, the VGS species that was most prevalent in the cohort and for which there was sufficient genome coverage to differentiate strains. We demonstrate that suppression of S. salivarius by cotrimoxazole is not strain specific, nor did stool concentration of the pro-inflammatory mediator myeloperoxidase vary by S. salivarius strain. We also show that gut-resident S. salivarius strains present in this study population are distinct from common oral strains. This is the first analysis of how cotrimoxazole prophylaxis used according to international treatment guidelines for children living with HIV influences the gut microbiome at the strain-level. We also provide a detailed review of the literature on the mechanisms by which suppression of VGS may act synergistically with cotrimoxazole's anti-inflammatory effects to reduce gut inflammation. A greater understanding of the sub-clinical effects of antibiotics offers new insights into their responsible clinical use.
Asunto(s)
Profilaxis Antibiótica , Microbioma Gastrointestinal/efectos de los fármacos , Infecciones por VIH/microbiología , Streptococcus salivarius/efectos de los fármacos , Combinación Trimetoprim y Sulfametoxazol/uso terapéutico , Estreptococos Viridans/efectos de los fármacos , Antibacterianos/uso terapéutico , Fármacos Anti-VIH/uso terapéutico , Niño , Heces/microbiología , Infecciones por VIH/tratamiento farmacológico , Humanos , Inflamación/microbiología , Inflamación/prevención & control , Intestinos/inmunología , Intestinos/microbiología , Especificidad de la Especie , Streptococcus salivarius/clasificación , Streptococcus salivarius/fisiología , Estreptococos Viridans/clasificación , Estreptococos Viridans/fisiología , ZimbabweRESUMEN
Constantly surrounded by kin or alien organisms in nature, eukaryotes and prokaryotes developed various communication systems to coordinate adaptive multi-entity behavior. In complex and overcrowded environments, they require to discriminate relevant signals in a myriad of pheromones to execute appropriate responses. In the human gut commensal Streptococcus salivarius, the cytoplasmic Rgg/RNPP regulator ComR couples competence to bacteriocin-mediated predation. Here, we describe a paralogous sensor duo, ScuR and SarF, which circumvents ComR in order to disconnect these two physiological processes. We highlighted the recurring role of Rgg/RNPP in the production of antimicrobials and designed a robust genetic screen to unveil potent/optimized peptide pheromones. Further mutational and biochemical analyses dissected the modifiable selectivity toward their pheromone and operating sequences at the subtle molecular level. Additionally, our results highlight how we might mobilize antimicrobial molecules while silencing competence in endogenous populations of human microflora and temper gut disorders provoked by bacterial pathogens.
Asunto(s)
Proteínas Bacterianas/metabolismo , Bacteriocinas/metabolismo , Competencia de la Transformación por ADN/efectos de los fármacos , Microbioma Gastrointestinal , Microbiota , Feromonas/metabolismo , Streptococcus salivarius/metabolismo , Antibacterianos/metabolismo , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Redes Reguladoras de Genes/efectos de los fármacos , Humanos , Streptococcus salivarius/efectos de los fármacos , Streptococcus salivarius/genética , Streptococcus salivarius/crecimiento & desarrolloRESUMEN
This study compared the outcome of photosensitization on the viability of four different cariogens in planktonic form as well as biofilms in human dentine. Photodynamic therapy was carried out with a gallium aluminium arsenide laser (670 nm wavelength) using Toluidine blue O (TBO) as the photosensitizer. Cariogenic bacteria ( Streptococcus mutans , Lactobacillus casei , Streptococcus salivarius and Actinomyces viscosus ) were exposed to TBO and then to the laser for 1 minute in planktonic suspension. Then, tooth slices previously incubated for 24 hours with broth cultures of broth culture of the four cariogenic organisms were exposed to antimicrobial photosensitization. The control samples consisted of planktonic and sessile cells that were exposed to TBO alone, laser alone and the bacterial cells that were not treated with TBO or laser. The results showed significant reductions in the viability of S. mutans , L. casei and A. viscosus in both planktonic form (to 13%, 30%, and 55%, respectively) and sessile form hosted in dentinal tubules (to 19%, 13% and 52%, respectively), relative to the controls. S. salivarius was the least affected in planktonic (94% viability) and sessile form (86% viability). In conclusion, sensitivity to photosensitization is species-dependent and sessile biofilm cells are affected to the same extent as their planktonic counterparts.This study compared the outcome of photosensitization on the viability of four different cariogens in planktonic form as well as biofilms in human dentine. Photodynamic therapy was carried out with a gallium aluminium arsenide laser (670 nm wavelength) using Toluidine blue O (TBO) as the photosensitizer. Cariogenic bacteria (Streptococcus mutans, Lactobacillus casei, Streptococcus salivarius and Actinomyces viscosus) were exposed to TBO and then to the laser for 1 minute in planktonic suspension. Then, tooth slices previously incubated for 24 hours with broth cultures of broth culture of the four cariogenic organisms were exposed to antimicrobial photosensitization. The control samples consisted of planktonic and sessile cells that were exposed to TBO alone, laser alone and the bacterial cells that were not treated with TBO or laser. The results showed significant reductions in the viability of S. mutans, L. casei and A. viscosus in both planktonic form (to 13%, 30%, and 55%, respectively) and sessile form hosted in dentinal tubules (to 19%, 13% and 52%, respectively), relative to the controls. S. salivarius was the least affected in planktonic (94% viability) and sessile form (86% viability). In conclusion, sensitivity to photosensitization is species-dependent and sessile biofilm cells are affected to the same extent as their planktonic counterparts.
Asunto(s)
Biopelículas/efectos de los fármacos , Biopelículas/efectos de la radiación , Caries Dental/microbiología , Fármacos Fotosensibilizantes/farmacología , Cloruro de Tolonio/farmacología , Diente/microbiología , Adulto , Humanos , Lacticaseibacillus casei/efectos de los fármacos , Lacticaseibacillus casei/efectos de la radiación , Viabilidad Microbiana/efectos de los fármacos , Viabilidad Microbiana/efectos de la radiación , Fotoquimioterapia , Streptococcus mutans/efectos de los fármacos , Streptococcus mutans/efectos de la radiación , Streptococcus salivarius/efectos de los fármacos , Streptococcus salivarius/efectos de la radiación , Adulto JovenRESUMEN
BACKGROUND: Streptococcus salivarius is an abundant isolate of the human oral microbiota. Since both pH and glucose availability fluctuate frequently in the oral cavity, the goal of this study was to investigate regulation by CodY, a conserved pleiotropic regulator of Gram positive bacteria, in response to these two signals. The chemostat culture system was employed to precisely control the growth parameters, and the transcriptomes of wild-type S. salivarius 57.I and its CodY-null derivative (ΔcodY) grown at pH 7 and 5.5, with limited and excessive glucose supply were determined. RESULTS: The transcriptomic analysis revealed that CodY was most active at pH 7 under conditions of glucose limitation. Based on whether a CodY binding consensus could be located in the 5' flanking region of the identified target, the transcriptomic analysis also found that CodY shaped the transcriptome via both direct and indirect regulation. Inactivation of codY reduced the glycolytic capacity and the viability of S. salivarius at pH 5.5 or in the presence of H2O2. Studies using the Galleria mellonella larva model showed that CodY was essential for the toxicity generated from S. salivarius infection, suggesting that CodY regulation was critical for immune evasion and systemic infections. Furthermore, the CodY-null mutant strain exhibited a clumping phenotype and reduced attachment in biofilm assays, suggesting that CodY also modulates cell wall metabolism. Finally, the expression of genes belonging to the CovR regulon was affected by codY inactivation, but CodY and CovR regulated these genes in opposite directions. CONCLUSIONS: Metabolic adaptation in response to nutrient availability and growth pH is tightly linked to stress responses and virulence expression in S. salivarius. The regulation of metabolism by CodY allows for the maximal utilization of available nutrients and ATP production. The counteractive regulation of the CovR regulon could fine tune the transcriptomes in response to environmental changes.
Asunto(s)
Proteínas Bacterianas/metabolismo , Glucosa/farmacología , Streptococcus salivarius/crecimiento & desarrollo , Streptococcus salivarius/metabolismo , Factores de Transcripción/metabolismo , Relación Dosis-Respuesta a Droga , Glucólisis/efectos de los fármacos , Concentración de Iones de Hidrógeno , Estrés Oxidativo/efectos de los fármacos , Streptococcus salivarius/efectos de los fármacosRESUMEN
BACKGROUND: The aim of this study was to examine effectiveness of oral probiotic Streptococcus salivarius K12 in preventing group A streptococcus pharyngitis in 5- to 14-year-old children at high risk of acute rheumatic fever. New Zealand has high rates of acute rheumatic fever among Maori and Pacific children. Children were already enrolled in a school-based Ministry of Health throat swabbing and treatment program. Children self-identified and reported sore throats daily and were swabbed twice weekly. METHODS: A total of 1314 children were quasirandomized (based on odd or even birthdates) to receive either K12 (2.5 × 10(9) cfu per lozenge) or placebo lozenges and continued observed daily treatment (in the school week, during school time) for one school year. RESULTS: A total of 801 children (61.0%) reported a sore throat on one or more occasions resulting in 2927 pharyngeal swabs. Of these swabs, 1525 (52.1%) were taken from 411 children receiving K12 and 119 (7.8%) of these were positive for group A streptococcus on routine culture. In addition, 1402 (47.8%) swabs were taken from 390 children receiving placebo and 124 (8.8%) were positive. Overall there was a nonsignificant 11.2% relative reduction in positive swabs among children receiving K12. This relative reduction was greater for older children, 7-9 years of age, 15.6%, and for children 10 years and older, 30.2%. CONCLUSIONS: S. salivarius K12 had modest nonsignificant effects on culture-positive sore throats when given at school, during the school day. Based on our pragmatic trial, the routine use of this probiotic in the prevention of pharyngitis associated with GAS detection is not supported.
Asunto(s)
Antibacterianos/administración & dosificación , Faringitis/terapia , Probióticos/administración & dosificación , Fiebre Reumática/prevención & control , Infecciones Estreptocócicas/prevención & control , Enfermedad Aguda/epidemiología , Administración Oral , Adolescente , Niño , Preescolar , Femenino , Humanos , Masculino , Nueva Zelanda/epidemiología , Faringitis/epidemiología , Faringitis/microbiología , Fiebre Reumática/epidemiología , Infecciones Estreptocócicas/epidemiología , Streptococcus pyogenes , Streptococcus salivarius/efectos de los fármacosRESUMEN
In this study, the essential oils of Orthosiphon stamineus Benth and Ficus deltoidea Jack were evaluated for their antibacterial activity against invasive oral pathogens, namely Enterococcus faecalis, Streptococcus mutans, Streptococcus mitis, Streptococcus salivarius, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Fusobacterium nucleatum. Chemical composition of the oils was analyzed using gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The antibacterial activity of the oils and their major constituents were investigated using the broth microdilution method (minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC)). Susceptibility test, anti-adhesion, anti-biofilm, checkerboard and time-kill assays were also carried out. Physiological changes of the bacterial cells after exposure to the oils were observed under the field emission scanning electron microscope (FESEM). O. stamineus and F. deltoidea oils mainly consisted of sesquiterpenoids (44.6% and 60.9%, respectively), and ß-caryophyllene was the most abundant compound in both oils (26.3% and 36.3%, respectively). Other compounds present in O. stamineus were α-humulene (5.1%) and eugenol (8.1%), while α-humulene (5.5%) and germacrene D (7.7%) were dominant in F. deltoidea. The oils of both plants showed moderate to strong inhibition against all tested bacteria with MIC and MBC values ranging 0.63-2.5 mg/mL. However, none showed any inhibition on monospecies biofilms. The time-kill assay showed that combination of both oils with amoxicillin at concentrations of 1× and 2× MIC values demonstrated additive antibacterial effect. The FESEM study showed that both oils produced significant alterations on the cells of Gram-negative bacteria as they became pleomorphic and lysed. In conclusion, the study indicated that the oils of O. stamineus and F. deltoidea possessed moderate to strong antibacterial properties against the seven strains pathogenic oral bacteria and may have caused disturbances of membrane structure or cell wall of the bacteria.
Asunto(s)
Antibacterianos/farmacología , Ficus/química , Boca/microbiología , Aceites Volátiles/farmacología , Orthosiphon/química , Sesquiterpenos/farmacología , Aggregatibacter actinomycetemcomitans/efectos de los fármacos , Aggregatibacter actinomycetemcomitans/crecimiento & desarrollo , Aggregatibacter actinomycetemcomitans/aislamiento & purificación , Amoxicilina/farmacología , Antibacterianos/química , Antibacterianos/aislamiento & purificación , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Sinergismo Farmacológico , Enterococcus faecalis/efectos de los fármacos , Enterococcus faecalis/crecimiento & desarrollo , Enterococcus faecalis/aislamiento & purificación , Fusobacterium nucleatum/efectos de los fármacos , Fusobacterium nucleatum/crecimiento & desarrollo , Fusobacterium nucleatum/aislamiento & purificación , Humanos , Pruebas de Sensibilidad Microbiana , Boca/patología , Aceites Volátiles/química , Aceites Volátiles/aislamiento & purificación , Sesquiterpenos Policíclicos , Porphyromonas gingivalis/efectos de los fármacos , Porphyromonas gingivalis/crecimiento & desarrollo , Porphyromonas gingivalis/aislamiento & purificación , Sesquiterpenos/química , Sesquiterpenos/aislamiento & purificación , Streptococcus mitis/efectos de los fármacos , Streptococcus mitis/crecimiento & desarrollo , Streptococcus mitis/aislamiento & purificación , Streptococcus mutans/efectos de los fármacos , Streptococcus mutans/crecimiento & desarrollo , Streptococcus mutans/aislamiento & purificación , Streptococcus salivarius/efectos de los fármacos , Streptococcus salivarius/crecimiento & desarrollo , Streptococcus salivarius/aislamiento & purificaciónRESUMEN
Alterations of gut bacterial metabolism play an important role in their host metabolism, and can result in diseases such as obesity and diabetes. While many factors were discovered influencing the gut bacterial metabolism, exposure to ultrafine particles (UFPs) from engine combustions were recently proposed to be a potential risk factor for the perturbation of gut bacterial metabolism, and consequentially to obesity and diabetes development. This study focused on evaluation of how UFPs from diesel engine combustions impact gut bacterial metabolism. We hypothesize that UFPs from different type of diesel (petrodiesel vs. biodiesel) will both impact bacterial metabolism, and the degree of impact is also diesel type-dependent. Targeted metabolic profiling of 221 metabolites were applied to three model gut bacteria in vitro, Streptococcus salivarius, Lactobacillus acidophilus and Lactobacillus fermentum. UFPs from two types of fuels, petrodiesel (B0) and a biodiesel blend (B20: 20% soy biodiesel/80% B0 by volume), were exposed to the bacteria and their metabolic changes were compared. For each bacterial strain, metabolites with significantly changed abundance were observed in both perturbations, and all three strains have increased number of altered metabolites detected from B20 UFPs perturbation in comparison to B0 UFPs. Multivariate statistical analysis further confirmed that the metabolic profiles were clearly different between testing groups. Metabolic pathway analyses also demonstrated several important metabolic pathways, including pathways involves amino acids biosynthesis and sugar metabolism, were significantly impacted by UFPs exposure.
Asunto(s)
Biocombustibles/toxicidad , Gasolina/toxicidad , Lactobacillus acidophilus/efectos de los fármacos , Limosilactobacillus fermentum/efectos de los fármacos , Metaboloma/efectos de los fármacos , Material Particulado/toxicidad , Streptococcus salivarius/efectos de los fármacos , Emisiones de Vehículos/toxicidad , Biocombustibles/análisis , Cromatografía Líquida de Alta Presión , Gasolina/análisis , Tracto Gastrointestinal/microbiología , Humanos , Lactobacillus acidophilus/metabolismo , Limosilactobacillus fermentum/metabolismo , Metabolómica , Tamaño de la Partícula , Material Particulado/análisis , Streptococcus salivarius/metabolismo , Espectrometría de Masas en Tándem , Emisiones de Vehículos/análisisRESUMEN
Most people have heard of the placebo effect, while relatively few have heard of nocebo, even within the circles of medical staff. Placebo effect means positive results by treatment via pharmacological inert substances. In contrast to placebo, by nocebo effect, due to negative beliefs and expectations, opposite results are achieved. Said in a more formal manner, what a sick person expects, unfortunately, he/she most often gets. It is a fact that a high number of medical staff is still uninterested in placebo and nocebo effects, although they would benefit from them. Maybe this is because the treatment would not seem "scientific enough". However, the newest scientific evidence undoubtedly shows that placebo and nocebo effects arise out of very active neurobiological processes intervened by psychological mechanisms such as expectations and conditions. Regardless of whether or not the doctor or the patient are aware of this, placebo and nocebo effects are extremely powerful and represent a significant part of the treatment process, in treatment by methods of ancient cultures, as well as in modern medicine. Of course medicines hold their role, but understanding how the human mind processes information is also very important.
Asunto(s)
Antiinfecciosos/farmacología , Placa Dental/prevención & control , Antisépticos Bucales/farmacología , Probióticos/farmacología , Pastas de Dientes/química , Candida albicans/efectos de los fármacos , Humanos , Técnicas In Vitro , Lactobacillus acidophilus , Lacticaseibacillus paracasei , Pruebas de Sensibilidad Microbiana , Streptococcus salivarius/efectos de los fármacos , Pastas de Dientes/farmacologíaRESUMEN
Magnetite nanoparticles (NPs) decorated with silver (magnetite/Ag) are intensively investigated due to their application in the biomedical field. We demonstrate that the increase of silver content on the surface of nanoparticles improves the adsorptivity of antibiotic rifampicin as well as antibacterial properties. The use of ginger extract allowed to improve the silver nucleation on the magnetite surface that resulted in an increase of silver content. Physicochemical and functional characterization of magnetite/Ag NPs was performed. Our results show that 5%-10% of silver content in magnetite/Ag NPs is already sufficient for antimicrobial properties against Streptococcus salivarius and Staphylococcus aureus. The rifampicin molecules on the magnetite/Ag NPs surface made the spectrum of antimicrobial activity wider. Cytotoxicity evaluation of the magnetite/Ag/rifampicin NPs showed no harmful action towards normal human fibroblasts, whereas the effect on human embryonic kidney cell viability was time and dose dependent.
Asunto(s)
Antibacterianos/farmacología , Nanopartículas de Magnetita/química , Rifampin/farmacología , Plata/farmacología , Zingiber officinale/química , Adsorción/efectos de los fármacos , Antibacterianos/química , Línea Celular , Supervivencia Celular/efectos de los fármacos , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Células HEK293 , Humanos , Nanopartículas de Magnetita/ultraestructura , Pruebas de Sensibilidad Microbiana , Extractos Vegetales/química , Extractos Vegetales/farmacología , Rifampin/química , Plata/química , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/crecimiento & desarrollo , Streptococcus salivarius/efectos de los fármacos , Streptococcus salivarius/crecimiento & desarrolloRESUMEN
Emerging antibiotic resistance in the oropharyngeal microbiota, of which Streptococcus salivarius is a prominent species, represents a challenge for treating paediatric populations. In this study, we investigated the role of Streptococcussalivarius as a reservoir for antibiotic resistance genes (ARG) in the oral microbiota by analysing 95 Streptococcussalivarius isolates from 22 healthy infants (2-16 months of age). MICs of penicillin G, amoxicillin, erythromycin, tetracycline, doxycycline and streptomycin were determined. ARG profiles were assessed in a subset of 21 strains by next-generation sequencing of genomes, followed by searches of assembled reads against the Comprehensive Antibiotic Resistance Database. Strains resistant to erythromycin, penicillins and tetracyclines were isolated from 83.3, 33.3 and 16.6 %, respectively, of infants aged 2 to 8 months with no prior antibiotic treatment. These percentages were100.0, 66.6 and 50.0 %, by 13 to 16 months of age. ARG or polymorphisms associated with antibiotic resistance were the most prevalent and involved genes for macrolide efflux (mel, mefA/E and macB), ribosomal protection [erm(B), tet(M) and tet(O)] and ß-lactamase-like proteins. Phylogenetically related strains showing multidrug-resistant phenotypes harboured multidrug efflux ARG. Polymorphic genes associated with antibiotic resistance to drugs affecting DNA replication, folate synthesis, RNA/protein synthesis and regulators of antibiotic stress responses were detected. These data imply that Streptococcussalivarius strains established during maturation of the oral microbiota harbour a diverse array of functional ARG, even in the absence of antibiotic selective pressures, highlighting a potential role for this species in shaping antibiotic susceptibility profiles of oropharyngeal communities.
Asunto(s)
Antibacterianos/farmacología , Farmacorresistencia Bacteriana Múltiple , Boca/microbiología , Streptococcus salivarius/efectos de los fármacos , Streptococcus salivarius/genética , Antibacterianos/clasificación , Proteínas Bacterianas/genética , Proteínas Portadoras/genética , Farmacorresistencia Bacteriana Múltiple/genética , Femenino , Genes Bacterianos , Genes MDR , Genoma Bacteriano , Genotipo , Voluntarios Sanos , Humanos , Lactante , Masculino , Proteínas de la Membrana/genética , Pruebas de Sensibilidad Microbiana , Fenotipo , Análisis de Secuencia de ADN , Streptococcus salivarius/clasificación , Streptococcus salivarius/fisiologíaRESUMEN
Recently, it has been reported that eriC and crcB are involved in bacterial fluoride resistance. However, the fluoride-resistance mechanism in oral streptococci remains unclear. BLAST studies showed that two types of eriCs (eriC1 and eriC2) and two types of crcBs (crcB1 and crcB2) are present across 18 oral streptococci, which were identified in ≥ 10% of 166 orally healthy subjects with ≥ 0.01% of the mean relative abundance. They were divided into three groups based on the distribution of these four genes: group I, only eriC1; group II, eriC1 and eriC2; and group III, eriC2, crcB1, and crcB2. Group I consisted of Streptococcus mutans, in which one of the two eriC1s predominantly affected fluoride resistance. Group II consisted of eight species, and eriC1 was responsible for fluoride resistance, but eriC2 was not, in Streptococcus anginosus as a representative species. Group III consisted of nine species, and both crcB1 and crcB2 were crucial for fluoride resistance, but eriC2 was not, in Streptococcus sanguinis as a representative species. Based on these results, either EriC1 or CrcBs play a role in fluoride resistance in oral streptococci. Complementation between S. mutans EriC1 and S. sanguinis CrcB1/CrcB2 was confirmed in both S. mutans and S. sanguinis. However, neither transfer of S. sanguinis CrcB1/CrcB2 into wild-type S. mutans nor S. mutans EriC1 into wild-type S. sanguinis increased the fluoride resistance of the wild-type strain. Co-existence of different F- channels (EriC and CrcB) did not cause the additive effect on fluoride resistance in oral Streptococcus species.
Asunto(s)
Fluoruros/farmacología , Canales Iónicos/fisiología , Streptococcus/efectos de los fármacos , Farmacorresistencia Bacteriana/genética , Genes Bacterianos/genética , Humanos , Canales Iónicos/genética , Boca/microbiología , Streptococcus/genética , Streptococcus anginosus/efectos de los fármacos , Streptococcus anginosus/genética , Streptococcus gordonii/efectos de los fármacos , Streptococcus gordonii/genética , Streptococcus intermedius/efectos de los fármacos , Streptococcus intermedius/genética , Streptococcus mutans/efectos de los fármacos , Streptococcus mutans/genética , Streptococcus oralis/efectos de los fármacos , Streptococcus oralis/genética , Streptococcus salivarius/efectos de los fármacos , Streptococcus salivarius/genética , Streptococcus sobrinus/efectos de los fármacos , Streptococcus sobrinus/genéticaRESUMEN
BACKGROUND: The rise in disease incidence, increased resistance of pathogenic bacteria to currently used antibiotics and chemotherapeutics, opportunistic infections in immunocompromised individuals, and financial considerations in developing countries necessitates alternate preventive and treatment strategies for oral diseases. OBJECTIVE: The objective of the study is to assess the antimicrobial efficacy of triple and quadruple combinations of Acacia nilotica (AN), Murraya koenigii (Linn.) (MKL) Sprengel, Eucalyptus (Euca), and Psidium guajava (PS) on primary plaque colonizers. MATERIALS AND METHODS: The phytochemicals in four plants were extracted using Soxhlet apparatus. The dried extracts were diluted with dimethyl sulfoxide (DMSO) to prepare stock solutions (100 mg/ml) of each plant. The triple and quadruple combinations were prepared after mixing equal quantities of stock solutions from each plant extracts. The antimicrobial efficacy testing was done on Streptococcus mutans, Streptococcus sanguis, and Streptococcus salivarius using agar well diffusion method. Chlorhexidine of 0.2% composition and DMSO were used as positive and negative controls, respectively. The mean diameter of inhibition zone between different categories was compared using one-way analysis of variance. RESULTS: The combination of AN + MKL Sprengel + Euca + PS produced the highest mean diameter of inhibition zone (23.5 ± 2.17 mm) against S. mutans. The combination of AN + MKL Sprengel + Euca produced the maximum antimicrobial efficacy against S. sanguis (19.83 ± 1.33). CONCLUSION: All the triple and quadruple combinations of the plant extracts offered antimicrobial benefits either superior or comparable to 0.2% chlorhexidine against S. mutans, S. sanguis, and S. salivarius.
Asunto(s)
Acacia , Placa Dental/tratamiento farmacológico , Placa Dental/microbiología , Eucalyptus , Murraya , Extractos Vegetales/farmacología , Psidium , Antiinfecciosos/farmacología , Combinación de Medicamentos , Técnicas In Vitro , Pruebas de Sensibilidad Microbiana , Streptococcus mutans/efectos de los fármacos , Streptococcus salivarius/efectos de los fármacos , Streptococcus sanguis/efectos de los fármacosRESUMEN
The study aimed to determine the effects of a single-dose antibiotic prophylaxis on normal oral microflora. A single dose of 2 g amoxicillin was given to 29 healthy volunteers. Saliva was collected before antibiotic administration (day 1), and again on days 2, 5, 10, 17 and 24 and subjected to culturing and antibiotic sensitivity analysis. Twenty-one per cent (6/29) of the individuals carried penicillin-V- and amoxicillin-resistant viridans streptococci before antibiotic administration. After a single dose of amoxicillin there was a significant reduction in Streptococcus salivarius on days 2 and 5, a significant reduction in other viridans streptococci on day 2 and the proportion of viridans streptococci with reduced susceptibility to amoxicillin was significantly increased on days 2 and 5. A single dose of amoxicillin can cause an ecological disturbance and induce selection of resistant strains in the oral microflora.
Asunto(s)
Amoxicilina/administración & dosificación , Antibacterianos/administración & dosificación , Saliva/microbiología , Streptococcus salivarius/efectos de los fármacos , Adulto , Amoxicilina/farmacología , Antibacterianos/farmacología , Profilaxis Antibiótica , Farmacorresistencia Bacteriana , Femenino , Humanos , Masculino , Pruebas de Sensibilidad Microbiana , Adulto JovenRESUMEN
Oral infections such as periodontitis and tooth decay are the most common diseases of humankind. Oleoresins from different copaifera species display antimicrobial and anti-inflammatory activities. Copaifera reticulata is the commonest tree of this genus and grows abundantly in several Brazilian states, such as Pará, Amazonas, and Ceará. The present study has evaluated the chemical composition and antimicrobial potential of the Copaifera reticulata oleoresin (CRO) against the causative agents of tooth decay and periodontitis and has assessed the CRO cytotoxic potential. Cutting edge analytical techniques (GC-MS and LC-MS) aided the chemical characterization of CRO. Antimicrobial assays included determination of the Minimum Inhibitory Concentration (MIC), determination of the Minimum Bactericidal Concentration (MBC), determination of the Minimum Inhibitory Concentration of Biofilm (MICB50), Time Kill Assay, and Checkerboard Dilution. Conduction of XTT assays on human lung fibroblasts (GM07492-A cells) helped to examine the CRO cytotoxic potential. Chromatographic analyses revealed that the major constituents of CRO were ß-bisabolene, trans-α-bergamotene, ß-selinene, α-selinene, and the terpene acids ent-agathic-15-methyl ester, ent-copalic acid, and ent-polyalthic acid. MIC and MBC results ranged from 6.25 to 200 µg/mL against the tested bacteria. The time-kill assay conducted with CRO at concentrations between 50 and 100 µg/mL showed bactericidal activity against Fusobacterium nucleatum (ATCC 25586) and Streptococcus mitis (ATCC 49456) after 4 h, Prevotella nigrescens (ATCC 33563) after 6 h, Porphyromonas gingivalis (ATCC 33277) and Lactobacillus casei (clinical isolate) after 12 h, and Streptococcus salivarius (ATCC 25975) and Streptococcus mutans (ATCC 25175) after 18 h. The fractional inhibitory concentration indexes (FICIs) revealed antagonistic interaction for Lactobacillus casei (clinical isolate), indifferent effect for Porphyromonas gingivalis (ATCC 33277), Fusobacterium nucleatum (ATCC 25586), Prevotella nigrescens (ATCC 33563), and Streptococcus salivarius (ATCC 25975), and additive effect for Streptococcus mutans (ATCC 25175) and Streptococcus mitis (ATCC 49456). Treatment of GM07492-A cells with CRO demonstrated that concentrations up to 39 µg/mL significantly reduced cell viability as compared to the negative control, being IC50 equal to 51.85 ± 5.4 µg/mL. These results indicated that CRO plays an important part in the search for novel sources of agents that can act against oral pathogens.
Asunto(s)
Antibacterianos/farmacología , Fabaceae/química , Extractos Vegetales/farmacología , Porphyromonas gingivalis/efectos de los fármacos , Prevotella nigrescens/efectos de los fármacos , Antibacterianos/química , Antibacterianos/aislamiento & purificación , Compuestos Bicíclicos con Puentes/aislamiento & purificación , Línea Celular , Supervivencia Celular/efectos de los fármacos , Caries Dental/microbiología , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Humanos , Lacticaseibacillus casei/efectos de los fármacos , Lacticaseibacillus casei/crecimiento & desarrollo , Lacticaseibacillus casei/aislamiento & purificación , Pruebas de Sensibilidad Microbiana , Sesquiterpenos Monocíclicos , Periodontitis/microbiología , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Porphyromonas gingivalis/crecimiento & desarrollo , Porphyromonas gingivalis/aislamiento & purificación , Prevotella nigrescens/crecimiento & desarrollo , Prevotella nigrescens/aislamiento & purificación , Sesquiterpenos/aislamiento & purificación , Streptococcus mutans/efectos de los fármacos , Streptococcus mutans/crecimiento & desarrollo , Streptococcus mutans/aislamiento & purificación , Streptococcus salivarius/efectos de los fármacos , Streptococcus salivarius/crecimiento & desarrollo , Streptococcus salivarius/aislamiento & purificación , Terpenos/aislamiento & purificaciónRESUMEN
Antibacterial surfaces have been in the focus of research for years, driven by an unmet clinical need to manage an increasing incidence of implant-associated infections. The use of silver has become a topic of interest because of its proven broad-spectrum antibacterial activity and track record as a coating agent of soft tissue implants and catheters. However, for the time being, the translation of these technological achievements for the improvement of the antibacterial property of hard tissue titanium (Ti) implants remains unsolved. In our study, we focused on the investigation of the photocatalysis mediated antibacterial activity of silver (Ag), and Ti nanoparticles instead of their pharmacological effects. We found that the photosensitisation of commercially pure titanium discs by coating them with an acrylate-based copolymer that embeds coupled Ag/Ti nanoparticles can initiate the photocatalytic decomposition of adsorbed S. salivarius after the irradiation with an ordinary visible light source. The clinical isolate of S. salivarius was characterised with MALDI-TOF mass spectrometer, while the multiplication of the bacteria on the surface of the discs was followed-up by MTT assay. Concerning practical relevance, the infected implant surfaces can be made accessible and irradiated by dental curing units with LED and plasma arc light sources, our research suggests that photocatalytic copolymer coating films may offer a promising solution for the improvement of the antibacterial properties of dental implants.