RESUMEN
Periodontitis is an infectious disease characterized by the destruction of supporting tissues. Antimicrobial photodynamic therapy (aPDT) has been proposed as an improved method for eliminating microorganisms. Its efficiency depends on the correct use of physical and chemical parameters. Thus, these parameters and their relations were evaluated in this study with the purpose of establishing lethal conditions for combating bacterial agents. Diode lasers and light-emitting diodes (LEDs) were characterized to evaluate the absorption profile and resonance of methylene blue (MB) and toluidine blue O (TBO). The relations between light energy density and photosensitizer absorption were determined. Two methodologies were used to evaluate the effects of aPDT against Aggregatibacter actinomycetemcomitans. LED light exhibited a broad emission spectrum with a peak light wavelength of 637 nm and 99% purity. The resonance intensity of MB was higher with diode laser irradiation, and TBO showed higher resonance intensity with LED irradiation. There was no difference in the absorption profile of photosensitizers using diode lasers or LEDs, and variations in power density did not result in an increasing or decrease in light absorption. A. actinomycetemcomitans was susceptible to photodynamic processes. Emission spectra and peak light wavelengths of light sources combined with the absorption profiles of photosensitizers were the main parameters involved in determining the efficiency of photodynamic effects. Power density did not alter the light absorption of photosensitizers. The association between adequate irradiation characteristics and photosensitizer absorption results in complete inactivation of A. actinomycetemcomitans. In addition, the bactericidal effect was not altered by an increase in energy densities.
Asunto(s)
Antiinfecciosos , Fotoquimioterapia , Aggregatibacter actinomycetemcomitans , Fármacos Fotosensibilizantes/farmacología , Cloruro de TolonioRESUMEN
The aim of this study was to phenotypically and genotypically identify coagulase-negative Staphylococcus (CoNS) recovered from the nostrils of waste workers and from dental waste; 135 strains were recovered and S. epidermidis was the prevailing species. Genetic similarity (100%) was observed between the two S. epidermidis isolated from different employees on the same shift and 85% similarity between the S. epidermidis recovered from an employee's nostril and from waste. The mecA gene was found in 20 CoNS, and 20% were also found to possess the vanA gene. The blaZ gene was detected in 46.7%, and the icaA (34.8%), B and C genes (11.8% each). Our findings emphasized the biological risk to which waste workers are exposed and unprecedently confirms that it was possible to recover genetically identical bacterial species from waste and from workers' nostrils. It is important to highlight that this risk is raised by the detection of relevant antimicrobial resistance genes. The results also suggest that effective measures to correctly manage waste and promote the rational use of antimicrobials should be adopted.
Asunto(s)
Antiinfecciosos , Infecciones Estafilocócicas , Antibacterianos/farmacología , Coagulasa/genética , Farmacorresistencia Bacteriana/genética , Humanos , Pruebas de Sensibilidad Microbiana , Residuos Sólidos , Infecciones Estafilocócicas/microbiología , Staphylococcus/genéticaRESUMEN
OBJECTIVE: To quantify Enterococcus faecalis density in root canal dentin after chemomechanical preparation (CMP) using alternated irrigating regimen. METHODOLOGY: Root canals (RC) were contaminated with E. faecalis (ATCC 19433) for 3 weeks and evident biofilms were obtained. After initial sampling (S1), the CMP was aided by irrigants: saline solution (control; n=12), a conventional regimen (CR) (group 1; n=12) using 5.25% NaOCl and a final rinse with 17% EDTA, and an alternating regimen (AR) of intercalated use of NaOCl and EDTA (group 2, n=12), followed by a second sampling (S2). After 2 weeks, S3 was obtained. Two roots were analyzed by scanning electron microscopy. Each root was divided into cervical, mild, and apical segments and sampling of the superficial (n=90) and deep (n=90) dentin layers was obtained using Gates-Glidden burs. The E. faecalis density (CFU/mg) in log10 was categorized as residual (0 > 0.2), moderate (0.2 ≥ 0.5), or elevated (> 0.5). The prevalence of positive samples in BHI and BHI-A was analyzed by Pearson's chi-square test. The data were normalized by a log10 transformation of CFU and were analyzed by one-way ANOVA and Tukey's tests. RESULTS: Biofilms were observed only in the control root canal walls. Topographically, the controls and CR showed similar distributions of E. faecalis in the dentin. Microbiologically positive root canals harbored much E. faecalis in the adjacent dentin (p < 0.05). Irrigating saline provided moderate density of E. faecalis in the dentin while CR and AR resulted in a residual density of microorganisms (p < 0.05). CONCLUSIONS: The Enterococcus faecalis density in dentin was influenced by the irrigating regimen and the microbiological status of the root canal. The CMP aided by the alternating regimen interfered with the recolonization of the root canal and topographic distribution of Enterococcus in root dentin.
Asunto(s)
Biopelículas/crecimiento & desarrollo , Dentina/microbiología , Enterococcus faecalis/crecimiento & desarrollo , Irrigantes del Conducto Radicular/farmacología , Biopelículas/efectos de los fármacos , Humanos , Hipoclorito de Sodio/farmacologíaRESUMEN
Healthcare wastes are those generated inside healthcare services, including dental clinics. Workers coming into close proximity to hazardous healthcare waste are potentially at risk. In an attempt to assess the knowledge and attitudes of workers dealing with infectious waste, a questionnaire was administered. The biological risk was investigated by evaluating the microbial load and screening some clinically relevant micro-organisms in the nasal mucosa, hands and coats of these workers. The results showed that 66.6% of the study population had incomplete primary education. Only two workers have had their blood tested to confirm anti-HBs levels. Microbial load evaluation on hand surfaces of morning workers showed statistically significant lower microbial loads after the workday when compared with the beginning of the work period. It is important to highlight that some clinically relevant bacteria, such as Pseudomonas aeruginosa and Klebsiella pneumoniae were isolated from worker's hands. This study revealed the need for more training programmes regarding awareness of safe waste disposal protocols and also the necessity of discussing vaccination and its implications. Data regarding microbial loads of the worker's hands, mostly at the beginning of the workday when handwashing is recommended worldwide, emphasise that hygiene measures should receive more attention during training exercises.
Asunto(s)
Residuos Dentales , Eliminación de Residuos Sanitarios , Brasil , Residuos Peligrosos , Conocimientos, Actitudes y Práctica en Salud , Personal de Salud , Humanos , Riesgo , Residuos SólidosRESUMEN
UNLABELLED: Acanthamoeba polyphaga mimivirus (APMV) is a giant virus from the Mimiviridae family. It has many unusual features, such as a pseudoicosahedral capsid that presents a starfish shape in one of its vertices, through which the â¼ 1.2-Mb double-stranded DNA is released. It also has a dense glycoprotein fibril layer covering the capsid that has not yet been functionally characterized. Here, we verified that although these structures are not essential for viral replication, they are truly necessary for viral adhesion to amoebae, its natural host. In the absence of fibrils, APMV had a significantly lower level of attachment to the Acanthamoeba castellanii surface. This adhesion is mediated by glycans, specifically, mannose and N-acetylglucosamine (a monomer of chitin and peptidoglycan), both of which are largely distributed in nature as structural components of several organisms. Indeed, APMV was able to attach to different organisms, such as Gram-positive bacteria, fungi, and arthropods, but not to Gram-negative bacteria. This prompted us to predict that (i) arthropods, mainly insects, might act as mimivirus dispersers and (ii) by attaching to other microorganisms, APMV could be ingested by amoebae, leading to the successful production of viral progeny. To date, this mechanism has never been described in the virosphere. IMPORTANCE: APMV is a giant virus that is both genetically and structurally complex. Its size is similar to that of small bacteria, and it replicates inside amoebae. The viral capsid is covered by a dense glycoprotein fibril layer, but its function has remained unknown, until now. We found that the fibrils are not essential for mimivirus replication but that they are truly necessary for viral adhesion to the cell surface. This interaction is mediated by glycans, mainly N-acetylglucosamine. We also verified that APMV is able to attach to bacteria, fungi, and arthropods. This indicates that insects might act as mimivirus dispersers and that adhesion to other microorganisms could facilitate viral ingestion by amoebae, a mechanism never before described in the virosphere.
Asunto(s)
Acanthamoeba/virología , Glicoproteínas/metabolismo , Mimiviridae/fisiología , Proteínas Virales/metabolismo , Acoplamiento Viral , Acanthamoeba/fisiología , Acanthamoeba/ultraestructura , Acetilglucosamina/metabolismo , Análisis de Varianza , Manosa/metabolismo , Microscopía Electrónica de Transmisión , Especificidad de la Especie , Replicación Viral/fisiologíaRESUMEN
Diarrhoeal disease is still considered a major cause of morbidity and mortality among children. Among diarrhoeagenic agents, Shigella should be highlighted due to its prevalence and the severity of the associated disease. Here, we assessed Shigella prevalence, drug susceptibility and virulence factors. Faeces from 157 children with diarrhoea who sought treatment at the Children's Hospital João Paulo II, a reference children´s hospital in Belo Horizonte, state of Minas Gerais, Brazil, were cultured and drug susceptibility of the Shigella isolates was determined by the disk diffusion technique. Shigella virulence markers were identified by polymerase chain reaction. The bacterium was recovered from 10.8% of the children (88.2% Shigella sonnei). The ipaH, iuc, sen and ial genes were detected in strains isolated from all shigellosis patients; set1A was only detected in Shigella flexneri. Additionally, patients were infected by Shigella strains of different ial, sat, sen and set1A genotypes. Compared to previous studies, we observed a marked shift in the distribution of species from S. flexneri to S. sonnei and high rates of trimethoprim/sulfamethoxazole resistance.
Asunto(s)
Diarrea/microbiología , Disentería Bacilar , Shigella/patogenicidad , Factores de Virulencia/genética , Enfermedad Aguda , Ampicilina/farmacología , Antibacterianos/farmacología , Brasil/epidemiología , Preescolar , Diarrea/prevención & control , Pruebas Antimicrobianas de Difusión por Disco , Disentería Bacilar/diagnóstico , Disentería Bacilar/tratamiento farmacológico , Disentería Bacilar/microbiología , Heces/microbiología , Femenino , Genotipo , Humanos , Lactante , Masculino , Reacción en Cadena de la Polimerasa , Prevalencia , Shigella/clasificación , Shigella/efectos de los fármacos , Combinación Trimetoprim y Sulfametoxazol/farmacologíaRESUMEN
Bacteriocins are antibacterial, proteinaceous substances that mediate microbial dynamics. Bacteriocin production is a highly disseminated property among all major lineages of bacteria, including Shigella. In this paper, we addressed the purification and characterisation of a bacteriocin produced by a Shigella sonnei strain (SS9) isolated from a child with acute diarrhoea. The substance was purified through ammonium-sulphate precipitation and sequential steps of chromatography. The intracellular fraction obtained at 75% ammonium sulphate maintained activity following exposure to pH values from 1-11 and storage at -80ºC for more than two years and was inactivated by high temperatures and proteases. The molecular mass of the purified bacteriocin was determined by mass spectrometry to be 18.56 kDa. The N-terminal sequence of the bacteriocin did not match any other antibacterial proteins described. A putative new bacteriocin produced by S. sonnei has been detected. This bacteriocin may represent a newly described protein or a previously described protein with a newly detected function. Considering that SS9 expresses antagonism against other diarrhoeagenic bacteria, the bacteriocin may contribute to S. sonnei virulence and is potentially applicable to either preventing or controlling diarrhoeal disease.
Asunto(s)
Bacteriocinas/aislamiento & purificación , Shigella sonnei/química , Enfermedad Aguda , Secuencia de Aminoácidos , Bacteriocinas/química , Bacteriocinas/metabolismo , Cromatografía de Fase Inversa , Diarrea/microbiología , Humanos , Espectrometría de Masas , Shigella sonnei/crecimiento & desarrolloRESUMEN
The biological activity of antimicrobial peptides and proteins is closely related to their structural aspects and is sensitive to certain post-translational modifications such as glycosylation, lipidation and PEGylation. However, PEGylation of protein and peptide drugs has expanded in recent years due to the reduction of their toxicity. Due to their size, the PEGylation process can either preserve or compromise the overall structure of these biopolymers and their biological properties. The antimicrobial peptide LyeTx I-bcys was synthesized by Fmoc strategy and coupled to polyethylene glycol 2.0 kDa. The conjugates were purified by HPLC and characterized by MALDI-ToF-MS analysis. Microbiological assays with LyeTx I-bcys and LyeTx I-bPEG were performed against Staphylococcus aureus (ATCC 33591) and Escherichia coli (ATCC 25922) in liquid medium. MIC values of 2.0 and 1.0 µM for LyeTx I-bcys and 8.0 and 4.0 µM for LyeTx I-bPEG were observed against S. aureus and E. coli, respectively. PEGylation of LyeTx I-bcys (LyeTx I-bPEG) decreased the cytotoxicity determined by MTT method for VERO cells compared to the non-PEGylated peptide. In addition, structural and biophysical studies were performed to evaluate the effects of PEGylation on the nature of peptide-membrane interactions. Surface Plasmon Resonance experiments showed that LyeTx I-b binds to anionic membranes with an association constant twice higher than the PEGylated form. The three-dimensional NMR structures of LyeTx I-bcys and LyeTx I-bPEG were determined and compared with the LyeTx I-b structure, and the hydrodynamic diameter and zeta potential of POPC:POPG vesicles were similar upon the addition of both peptides. The mPEG-MAL conjugation of LyeTx I-bcys gave epimers, and it, together with LyeTx I-bPEG, showed clear α-helical profiles. While LyeTx I-bcys showed no significant change in amphipathicity compared to LyeTx I-b, LyeTx I-bPEG was found to have a slightly less clear separation between hydrophilic and hydrophobic faces. However, the similar conformational freedom of LyeTx I-b and LyeTx I-bPEG suggests that PEGylation does not cause significant structural changes. Overall, our structural and biophysical studies indicate that the PEGylation does not alter the mode of peptide interaction and maintains antimicrobial activity while minimizing tissue toxicity, which confirmed previous results obtained in vivo. Interestingly, significantly improved proteolytic resistance to trypsin and proteinase K was observed after PEGylation.
RESUMEN
Background: Asthma is a chronic pulmonary disease that affects about 300 million people worldwide. Previous studies have associated antimicrobial use with allergies, but the real impact of antibiotics on asthma is still elusive. We investigated the potential impact of amoxicillin (Amox), trimethoprim/sulfamethoxazole (TMP/SMX), and metronidazole (Metro) in a murine model of OVA-induced allergic airway inflammation. Methods: BALB/c mice received three cycles of 7 days of antibiotics in drinking water followed by 7 days washout and were sensitized i.p. with OVA/Alum at days 0 and 14. After the end of the last antibiotic washout, the mice were challenged with aerosolized OVA. Pulmonary parameters were evaluated, and serum, BAL, and feces were collected for analysis. Results: Amox- and TMP/SMX-treated animals displayed more severe allergic airway inflammation parameters with increased airway hyperresponsiveness, reduced lung alveolar volume, and increased levels in BAL of IL-4 and IL-6. In contrast, Metro-treated mice showed preserved FEV-50, decreased lung inflammation, and higher levels of butyrate and propionate in their feces. Metro treatment was associated with increased OVA-specific IgA in serum. BAL microbiota was abundant in allergic groups but not in nonallergic controls with the Amox-treated group displaying the increased frequency of Proteobacteria, while Metro and TMP/SMX showed increased levels of Firmicutes. In the gut, we observed the enrichment of Akkermansia muciniphila associated with reduced airway inflammation phenotype in the Metro group, even after the recovery period. Conclusion: Our data suggest that different antibiotic treatments may impact the course of experimental allergic airway inflammation in diverse ways by several mechanisms, including modulation of short-chain fat acids production by intestinal microbiota.
Asunto(s)
Asma , Hipersensibilidad , Microbiota , Animales , Antibacterianos/uso terapéutico , Asma/tratamiento farmacológico , Humanos , Hipersensibilidad/tratamiento farmacológico , Inflamación/tratamiento farmacológico , Pulmón , Ratones , Combinación Trimetoprim y SulfametoxazolRESUMEN
In the continuing search for novel antibiotics, antimicrobial peptides are promising molecules, due to different mechanisms of action compared to classic antibiotics and to their selectivity for interaction with microorganism cells rather than with mammalian cells. Previously, our research group has isolated the antimicrobial peptide LyeTx I from the venom of the spider Lycosa erythrognatha. Here, we proposed to synthesize three novel shortened derivatives from LyeTx I (LyeTx I mn; LyeTx I mnΔK; LyeTx I mnΔKAc) and to evaluate their toxicity and biological activity as potential antimicrobial agents. Peptides were synthetized by Fmoc strategy and circular dichroism analysis was performed, showing that the three novel shortened derivatives may present membranolytic activity, like the original LyeTx I, once they folded as an alpha helix in 2.2.2-trifluorethanol and sodium dodecyl sulfate. In vitro assays revealed that the shortened derivative LyeTx I mnΔK presents the best score between antimicrobial (↓ MIC) and hemolytic (↑ EC50) activities among the synthetized shortened derivatives, and LUHMES cell-based NeuriTox test showed that it is less neurotoxic than the original LyeTx I (EC50 [LyeTx I mnΔK] â EC50 [LyeTx I]). In vivo data, obtained in a mouse model of septic arthritis induced by Staphylococcus aureus, showed that LyeTx I mnΔK is able to reduce infection, as demonstrated by bacterial recovery assay (â¼10-fold reduction) and scintigraphic imaging (less technetium-99m labeled-Ceftizoxime uptake by infectious site). Infection reduction led to inflammatory process and pain decreases, as shown by immune cells recruitment reduction and threshold nociception increment, when compared to positive control group. Therefore, among the three shortened peptide derivatives, LyeTx I mnΔK is the best candidate as antimicrobial agent, due to its smaller amino acid sequence and toxicity, and its greater biological activity.
Asunto(s)
Péptidos Catiónicos Antimicrobianos/farmacología , Animales , Péptidos Catiónicos Antimicrobianos/síntesis química , Péptidos Catiónicos Antimicrobianos/aislamiento & purificación , Bacterias/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Dicroismo Circular , Eritrocitos/efectos de los fármacos , Eritrocitos/metabolismo , Hongos/efectos de los fármacos , Humanos , Inflamación/patología , Ratones , Pruebas de Sensibilidad Microbiana , Nocicepción/efectos de los fármacos , ConejosRESUMEN
INTRODUCTION: This randomized clinical study compared the in vivo antibacterial efficacy of Reciproc Blue (RB), XP-endo Shaper (XP-S), and XP-endo Shaper associated with XP-endo Finisher (XP-F) systems in infected oval-shaped root canals with primary apical periodontitis. METHODS: In this study, 28 human teeth with a single root and a single canal were randomly assigned to 2 groups according to the instrumentation technique: group 1, RB (n = 14) and group 2, XP-endo (XP-S and XP-F, n = 14). The single-rooted teeth were prepared by reciprocating and rotary nickel-titanium instruments with 5.25% sodium hypochlorite irrigation. Samples were collected from the canal at the baseline (S1), after chemomechanical preparation (S2), and after XP-F instrumentation (S3). The DNA extracts were subjected to quantitative analysis for total bacterial counts by quantitative real-time polymerase chain reaction. The data were analyzed using the analysis of variance test, and the level of significance was set at 5%. RESULTS: All samples tested positive for the presence of bacteria at baseline, and the bacterial counts substantially reduced after treatment procedures (P < .01). The results showed no statistical difference between RB and XP-S instrumentation with respect to the bacterial reduction (P > .05). A marked bacterial reduction was observed after the use of the XP-F instrument (P < .01). CONCLUSIONS: The XP-S and RB systems sharply reduced the bacterial load in oval-shaped root canals with primary apical periodontitis. XP-F used as a supplementary instrument to chemomechanical preparation promoted a significantly higher bacterial reduction.
Asunto(s)
Periodontitis Periapical , Preparación del Conducto Radicular , Carga Bacteriana , Cavidad Pulpar , Humanos , Irrigantes del Conducto Radicular , Hipoclorito de SodioRESUMEN
OBJECTIVES: The polymerization of adhesive systems is incomplete and the residual monomers that have been released have a cytotoxic capacity. Some teeth develop into pulp necrosis after composite resin restorations. Considering frequent pulpal inflammation in response to cariogenic bacteria, substances released from the patches could affect the cells of the inflammatory infiltrate and interfere with the mechanisms of defense against microorganisms and protection of pulpal tissue. The aim of this study was to evaluate the effect of substances released by different resinous adhesive systems on cell viability and cytokine expression by human monocytes stimulated in vitro with Streptococcus mutans. DESIGN: Peripheral blood mononuclear cells from 10 healthy subjects were stimulated with S. mutans and then incubated with supernatants obtained from the Single Bond Universal (SBU) or Clearfil SE Bond (CSEB) adhesive systems for eight hours. Staining with Annexin V and 7AAD for analysis of apoptosis were performed and detection of monocytes expressing cytokines IL-1α, IL-6, IL-8, IL-10, IL-12 and TNF-α were performed by flow cytometry. RESULTS: No treatment significantly affected apoptosis in monocytes. SBU supernatant increased the frequency of monocytes expressing IL-8 and decreased the monocytes expressing IL-10. Considering S. mutans-stimulated cells, while SBU increased the frequency of IL-8+ monocytes, CSEB reduced the frequency of IL-6 and TNF-αâ¯positive monocytes. CONCLUSIONS: Products released from SBU seem to induce proinflammatory effects on monocytes while those from CSEB show an anti-inflammatory outcome. These effects may interfere in the control of cytokine-mediated immunoinflammatory pulp reactions, both in the presence and absence of stimulation by cariogenic bacteria.
Asunto(s)
Monocitos , Streptococcus mutans , Resinas Compuestas , Citocinas , Cementos Dentales , Humanos , Leucocitos Mononucleares , Factor de Necrosis Tumoral alfaRESUMEN
OBJECTIVE: to evaluate the potential contamination of enzymatic detergent from its reuse and to identify the microbiological profile in the solution used to clean gastrointestinal endoscopic devices. METHOD: cross-sectional study based on microbiological analysis of 76 aliquots of 19 different enzymatic detergent solutions used to clean endoscopic devices. The aliquots were homogenized, subjected to Millipore® 0.45 µm membrane filtration and the presumptive identification of microorganisms was performed by biochemical-physiological methods according to previously established specific bacterial groups that are of clinical and epidemiological relevance. RESULTS: the mean values, as well as the standard deviation and the median, of the enzymatic detergent microbial load increased as the solution was reused. There was a significant difference between the means of after first use and after fifth reuse. A total of 97 microorganisms were identified, with predominance of the coagulase-negative Staphylococcus, Pseudomonas spp., Klebsiella spp., Enterobacter spp. genus, and Escherichia coli species. CONCLUSION: the reuse of the enzymatic detergent solution is a risk to the safe processing of endoscopic devices, evidenced by its contamination with pathogenic potential microorganisms, since the enzymatic detergent has no bactericidal property and can contribute as an important source for outbreaks in patients under such procedures.
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Detergentes/efectos adversos , Contaminación de Equipos , Gastroscopios/efectos adversos , Gastroscopios/microbiología , Carga Bacteriana , Estudios Transversales , Detergentes/farmacología , Transmisión de Enfermedad Infecciosa , Bacterias Gramnegativas/clasificación , Bacterias Gramnegativas/aislamiento & purificación , Bacterias Grampositivas/clasificación , Bacterias Grampositivas/aislamiento & purificación , Humanos , Control de InfeccionesRESUMEN
BACKGROUND: The failure of endodontic treatment is linked to the presence of Enterococcus faecalis in the root canals. The scope of this study was to evaluate the influence of the energy dose and frequency of photodynamic therapy (PDT cycles), as well as the volume of bacterial suspensions (BS) in the elimination of Enterococcus faecalis in planktonic form. METHODS: In four successive assays BS of Enterococcus faecalis ATCC 19433 were irradiated with a diode laser (40â¯mW) using the photosensitizer (PS) methylene blue (MB) (0.005⯵g/mL). Group 1 - Effect of energy dose: 100⯵â¯L of BS and 100⯵â¯L of PS were irradiated by 1, 2.5, 5, 7.5 and 10â¯minuteâ¯s. Group 2 - Effect of PDT cycles: The BS received 1, 2, 3 or 4 PDT cycles (in each cycle 100â¯mâ¯L of PS was added and irradiated by 2.5â¯minutes). Group 3 - Effect of energy dose and bacterial suspension volume: 10⯵â¯L of BS and 10⯵â¯L of PS were irradiated similar to group 1. Group 4 - Effect of energy dose, bacterial suspension volume and PDT cycles: 10⯵â¯L of BS and 10⯵L of PS were irradiated according to group 2. The laser source and MB isolated represented the controls. RESULTS: The mean log reduction after separate applying laser light and MB were 0.01 and 0.07, respectively. It was found that wells with 100⯵â¯L of BS irradiated with 2.4 to 24â¯J of energy did not cause significant bacterial elimination (pâ¯>â¯0.05), on the other hand PDT cycles above 12â¯J increased significantly bacterial elimination (pâ¯<â¯0.05). In 10⯵â¯L wells irradiation from 12â¯J of energy provided higher bacterial elimination (pâ¯<â¯0.05) which combined with PDT cycle resulted in the logarithmic elimination of E. faecalis (pâ¯<â¯0.05). CONCLUSIONS: The energy dose, the volume of the bacterial suspension and, especially, the PDT cycles optimized the bacterial elimination of Enterococcus faecalis in planktonic form.
Asunto(s)
Enterococcus faecalis/efectos de los fármacos , Azul de Metileno/farmacología , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/farmacología , Plancton/efectos de los fármacos , Láseres de Semiconductores/uso terapéuticoRESUMEN
Trying to widen the discussion on the risks associated with dental waste, this study proposed to investigate and genetically compare yeast isolates recovered from dental solid waste and waste workers. Three samples were collected from workers' hands, nasal mucosa, and professional clothing (days 0, 30, and 180), and two from dental waste (days 0 and 180). Slide culture, microscopy, antifungal drug susceptibility, intersimple sequence repeat analysis, and amplification and sequencing of internal transcribed spacer regions were performed. Yeast strains were recovered from all waste workers' sites, including professional clothes, and from waste. Antifungal susceptibility testing demonstrated that some yeast recovered from employees and waste exhibited nonsusceptible profiles. The dendrogram demonstrated the presence of three major clusters based on similarity matrix and UPGMA grouping method. Two branches displayed 100% similarity: three strains of Candida guilliermondii isolated from different employees, working in opposite work shifts, and from diverse sites grouped in one part of branch 1 and cluster 3 that included two samples of Candida albicans recovered from waste and the hand of one waste worker. The results suggested the possibility of cross-contamination from dental waste to waste workers and reinforce the need of training programs focused on better waste management routines.
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Antifúngicos/farmacología , Candida/clasificación , Candida/efectos de los fármacos , Residuos Dentales , Residuos Sólidos , Instalaciones de Eliminación de Residuos , Secuencia de Bases , Candida/genética , ADN de Hongos/genética , ADN Intergénico/genética , Farmacorresistencia Fúngica/genética , Variación Genética/genética , Humanos , Pruebas de Sensibilidad Microbiana , Análisis de Secuencia de ADNRESUMEN
Rotavirus has been considered the main agent of infectious diarrhea especially among younger children. We addressed the prevalence of rotavirus-associated diarrhea and the diversity of circulating electropherotypes by immunochromatography and RNA electrophoresis. Stool samples were taken from 391 children (267 with diarrhea) from the lower socioeconomic stratum who sought treatment in the Hospital Infantil João Paulo II/Belo Horizonte, during 2005 and 2006. Rotavirus was detected in 79/20.2% of subjects, 64/24.0% with diarrhea and 15/12.1% with no diarrhea. The virus was strongly associated with diarrhea (p = 0.003). A total of 76/19.4% and 69/17.6% rotavirus-positive children were identified by immunochromatography and electrophoresis, respectively. Rotavirus-associated diarrhea was more frequently detected in dry months (p < 0.001) and almost exclusively in children aged up to three years. Long profile strains prevailed (54/78.3%) but a shift toward short electropherotype was identified. Despite the decrease seen in 2006, rotavirus infection is still very common in our area. Although viral RNA electrophoresis is useful as a typing method, it should not be used exclusively in the diagnosis of rotavirus infection. We confirmed a shift from long to short profile strains, as already described for other South American countries.
Asunto(s)
Diarrea/virología , Infecciones por Rotavirus/virología , Rotavirus/genética , Brasil/epidemiología , Niño , Preescolar , Diarrea/epidemiología , Electroforesis en Gel de Poliacrilamida , Heces/virología , Humanos , Prevalencia , ARN Viral/análisis , Rotavirus/clasificación , Infecciones por Rotavirus/epidemiología , Estaciones del Año , Factores SocioeconómicosRESUMEN
O presente estudo objetivou avaliar o perfil de susceptibilidade antimicrobiana e a ocorrência de patotipos diarreiogênicos (E. coli entoropatogênica/EPEC, E. coli enterotoxigênica/ETEC, E. coli produtora de shiga toxina/STEC, E. coli enteroinvasiva/EIEC e E. coli enteroagregativa/EAEC) em amostras de Escherichia coli, isoladas de diversos alimentos destinados ao consumo humano e naqueles envolvidos em surtos de Doença de Transmissão Alimentar (DTA), em Minas Gerais, Brasil. O perfil de susceptibilidade antimicrobiana das amostras de E. coli foi avaliado por meio da técnica de Concentração Mínima Inibitória e os patotipos de E. coli diarreiogênica pela Reação em Cadeia de Polimerase, utilizando-se os marcadores de virulência eae, bfpA, eltB, estA, st1, stx1, stx2, ipaH e aatA. Dentre as 220 amostras de E. coli analisadas, 60 (27,3%) apresentaram-se resistentes ou com resistência intermediária. Entre estas, oito (13,3%) foram consideradas multirresistentes. Os fármacos amicacina e gentamicina foram os mais eficazes, diferentemente da ampicilina, que apresentou o maior percentual de amostras resistentes (10,9%). Não foram identificados os patotipos de EPEC, STEC, EIEC e EAEC. Duas amostras de E. coli (0,9%), isoladas de um salgado (pão de queijo) e de uma refeição mista (galinhada), foram classificadas como ETEC. Ambas albergavam os marcadores de virulência eltB, estA e st1. Estas foram caracterizadas como pertencentes sorotipos O9:H10 e O9:H33. Conclui-se que entre as amostras de E. coli, isoladas de alimentos destinados ao consumo humano em Minas Gerais e naqueles envolvidos em surtos de DTA, existem de amostras consideradas multirresistentes e diarreiogênicas.
This study aimed to evaluate the profile of antimicrobial susceptibility and the occurrence of diarrheagenic pathotypes (E. colientoropatogênic/EPEC, E. coli enterotoxigenic/ETEC, E. coli producing shiga toxin/STEC, E. coli enteroinvasive/EIEC and E. colienteroaggregative/EAEC) in Escherichia coli isolated from different foods intended for human consumption and those involved in foodborne disease outbreaks, in Minas Gerais, Brazil. The antimicrobial susceptibility of strains of E. coli was evaluated by the Minimum Inhibitory Concentration technique and pathotypes of E. coli diarrheagenic by polymerase chain reaction, using the virulence markers eae, bfpA, eltB, st1, stx1, stx2, ipaH and aatA. Among the 220 strains of E. coli analyzed, 60 (27.3%) were resistant or intermediate resistance. Among these, eight (13.3%) were considered multiresistant. The amikacin and gentamicin drugs were the most effective, unlike ampicillin, which had the highest percentage of resistant strains (10.9%). Pathotypes of EPEC, STEC, EIEC and EAEC were not identified. Two strains of E. coli (0.9%) isolated from a salty (cheese bread) and a mixed meal (chicken meal) were classified as ETEC. Both harbored the virulence markers eltB, estA and st1. These were characterized as serotypes O9:H10 and O9:H33. We conclude that among samples of E. coli, isolated from foods for human consumption in Minas Gerais and those involved in outbreaks of DTA, there are samples considered multiresistant and diarrheogenic.
Asunto(s)
Amicacina , GentamicinasRESUMEN
This 3.5-year prospective study was conducted to ascertain the level of attaching and effacing Escherichia coli (AEEC) associated diarrhoea in children from Teresina, a northeastern state of Brazil. Passed faecal specimens from 400 patients (250 with and 150 without diarrhoea) up to 60 months of age attending from 2004 to 2007 at two public hospitals were investigated. Conventional microbiology methods and PCR were employed. Escherichia coli was isolated from 390 children, 240 of them with diarrhoea. A total of 117 AEEC strains were cultivated from specimens from 63 children, 37 with and 26 without diarrhoea. No association between AEEC and diarrhoea was observed. Atypical enteropathogenic E. coli (a-EPEC) (79.4%) was more commonly found than typical EPEC (t-EPEC). Association between EPEC and EPEC subtypes and diarrhoea was not detected. Mixed infection by t-EPEC and a-EPEC and infection by Shiga toxin-producing E. coli (STEC) were rare. Enteropathogenic E. coli was more common in males and in children aged less than 12 months. Correlation between serotyping and PCR results was 0.19. High resistance rates of AEEC to ampicillin, cephalotin, and trimethoprim-sulfamethoxazole were found. In conclusion, EPEC is very common in children with diarrhoea and controls in the population we studied, with a-EPEC predominating. This diarrhoeagenic E. coli (DEC) pathotype is more common in infant males and is resistant to drugs frequently used in clinical practice.
Asunto(s)
Diarrea/microbiología , Infecciones por Escherichia coli/epidemiología , Escherichia coli/aislamiento & purificación , Heces/microbiología , Toxina Shiga/aislamiento & purificación , Enfermedad Aguda , Adhesinas Bacterianas/genética , Adhesinas Bacterianas/aislamiento & purificación , Adhesión Bacteriana/genética , Brasil/epidemiología , Diarrea/epidemiología , Escherichia coli/genética , Escherichia coli/metabolismo , Infecciones por Escherichia coli/complicaciones , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/aislamiento & purificación , Femenino , Proteínas Fimbrias/genética , Proteínas Fimbrias/aislamiento & purificación , Humanos , Lactante , Masculino , Estudios ProspectivosRESUMEN
ABSTRACT Antimicrobial photodynamic therapy (aPDT) involves the association of a photosensitizing agent with a light source with the goal of causing apoptosis or microbial lysing. The use of compounds with natural active principles is gaining prominence throughout the world. Several studies from groups that are linked to the development of innovations in the pharmaceutical market have used natural dyes, such as curcumin, the efficacy of which has been demonstrated in aPDT trials. Difficulties related to physicochemical stability, solubility and cell penetration are some of the challenges associated with this field. The present work aimed to prepare, investigate the characteristics and improve the photodynamic activity of PLGA-based nanoparticles loaded with curcumin for use in aPDT therapy. Using the simple technique of emulsion during the evaporation of a solvent, the particles were built, characterized and tested against microorganisms with importance for medicine and dentistry. The results revealed that the particles were able to protect the curcumin against degradation and eliminate some microorganism species at nanomolar concentrations.
Asunto(s)
Curcumina/análisis , Nanopartículas/análisis , Fotoquimioterapia/efectos adversos , Composición de MedicamentosRESUMEN
Shigella is a common agent of diarrhoea, a worldwide major health problem. The bacterium produces bacteriocins; however, the role of these substances as a virulence factor is completely unknown. With the aim to search for colicin production by Shigella sonnei, to evaluate the influence of culture conditions on bacteriocin expression, and to characterize the substance partially, 16 S. sonnei strains isolated from children with diarrhoea were tested for antagonism against members of the intestinal microbiota or agents of diarrhoea. Nine strains exhibited isoantagonism and heteroantagonism against S. flexneri and diarrhoeagenic Escherichia coli. Autoantagonism and antagonism against the intestinal microbiota were not detected. Culture medium and incubation conditions influenced antagonism expression. Antagonism resulting from bacteriophages, low pH, fatty acids, hydrogen peroxide, and chloroform was excluded. The activity of the intracellular fraction obtained with 75% ammonium sulphate was preserved at pH 1.0-11.0, and was found to be reduced by organic solvents and affected by high temperatures and proteases. The antagonistic spectrum and the in vitro conditions for better antagonism expression suggest that the role of colicin in S. sonnei virulence, if any, would be expressed prior to infection, and may regulate population density of enteropathogens by helping in organism transmission.