ABSTRACT
Introducción: En la actualidad las infecciones por hongos afectan entre el 20 y el 25 por ciento de la población. Objetivo: Determinar la concentración mínima inhibitoria de dos lotes de OLEOZON® almacenados a temperaturas de 5 y 30°C en envases de vidrio y polietileno de alta densidad, durante 24 meses como parte del estudio de estabilidad del OLEOZON® tópico. Métodos: Mediante el método dilución en agar fueron evaluadas cinco concentraciones del producto frente a los dermatofitos trichophyton rubrum, trichophyton mentagrophytes y epidermophyton floccosum. Resultados: Se obtuvo que el 8,9 mg/mL fue el valor de la CMI para las cepas evaluadas en el estudio vida de estante; se observó en el estudio acelerado el mismo valor frente a las cepas de trichophyton mientras que para epidermophyton floccosum fue de 17,8 mg/mL a excepción del envase frasco de vidrio del lote 803295 donde se obtuvo 8,9 mg/mL. El análisis estadístico, tanto del estudio acelerado como en vida de estante, mostró que existe diferencia estadísticamente significativa entre el primer y el último mes de ensayo, estas son las diferencias más apreciables en los lotes almacenados en frasco de vidrio. El OLEOZON® tópico almacenado en frasco de polietileno de alta densidad presentó mejores valores de actividad frente a los dermatofitos. Conclusiones: Todos los valores de la concentración mínima inhibitoria encontrados, independiente del tipo de envase, el tiempo o la temperatura de almacenamiento, muestran que el producto mantiene su actividad antifúngica. Se evidenció una similitud entre las cepas del género trichophyton en comparación con el género epidermophyton(AU)
Introduction: Currently fungal infections affect between 20 and 25percent of the population. Objective: To determine the minimum inhibitory concentration of two batches of OLEOZON® stored at temperatures of 5 and 30 0C in glass and high-density polyethylene containers, for 24 months as part of the stability study of topical OLEOZON®. Methods: Using the agar dilution method, five concentrations of the product were evaluated against the dermatophytes Trichophyton rubrum, Trichophyton mentagrophytes and Epidermophyton floccosum. Results: It was found that 8.9 mg/mL was the MIC value for the strains evaluated in this shelf life study; and the same value was observed in the accelerated study against the trichophyton strains, while for epidermophyton floccosum it was 17.8 mg/mL with the exception of the glass bottle container of batch 803295 where 8.9 mg/mL was obtained. The statistical analysis, both in the accelerated study and in shelf life, showed that there is a statistically significant difference between the first and the last month of the trial, these are the most appreciable differences in the batches stored in glass jars. Topical OLEOZON® stored in a high-density polyethylene bottle presented better activity values against dermatophytes. Conclusions: All the values of the minimum inhibitory concentration found, regardless of the type of container, the time or the storage temperature, show that the product maintains its antifungal activity during the months of study. A similarity was apparent between the strains of the genus trichophyton compared to the genus epidermophyton(AU)
Subject(s)
Humans , Male , Female , Microbial Sensitivity Tests/methods , Reference DrugsABSTRACT
Abstract The present study was aimed at conducting phytochemical analysis and evaluating the in vitro antifungal and antioxidant activities of the essential oil obtained from the fruits of J. oxycedrus L. Hydro-distillation was used to extract the essential oil from the fruits of Juniper oxycedrus. The essential oil was analyzed using gas chromatography with a flame ionization detector (GC-FID) and gas chromatography coupled with mass spectrometry (GC/MS). The antioxidant activity of the essential oil against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals was determined in vitro using varying concentrations of the essential oil and vitamin C as a standard antioxidant compound. A disc diffusion test was employed to evaluate the antifungal activity of the essential oil against two test fungal strains, Penicillium citrinum, and Aspergillus niger. The results revealed that 49 constituents were identified in fruit oil, representing 91.56% of the total oil and the yield was 1.58%. Juniper fruit oil was characterized by having high contents of ß-pinene (42.04%), followed by limonene (15.45%), sabinene (9.52%), α-pinene (5.21%), (E)-caryophyllene (3.77%), ρ-cymene (1.56%), caryophyllene oxide (2.02%), and myrcene (1.02%). The radical scavenging activity (% inhibition) of the essential oil was highest (81.87± 2.83%) at a concentration of 200 µg/mL. The essential oil of J. oxycedrus exhibited antifungal activity against A. niger and P. citrinum with minimum inhibitory concentration values (MIC) ranging from 2.89 to 85.01 µl/mL. The findings of the study reveal that the antioxidant and antifungal properties of J. oxycedrus essential oil and their chemical composition are significantly correlated
Subject(s)
Oils, Volatile/analysis , Juniperus/adverse effects , Phytochemicals/analysis , Fruit/classification , Morocco/ethnology , Antioxidants/pharmacology , Mass Spectrometry/methods , In Vitro Techniques/methods , Microbial Sensitivity Tests/methods , Chromatography, Gas/methods , Antifungal Agents/pharmacologyABSTRACT
Introduction: Staphylococcus aureus and Escherichia coli are pathogens that cause chronic infections due to antibiotic resistance mechanisms and their ability to adhere to surfaces and to form biofilms. The search for new agents from natural resources to counter microbial biofilms is an urgent priority in healthcare. Objetive: chemical composition, antibacterial and anti-biofilm activity of Plectranthus amboinicus essential oil (PAEO) and carvacrol were investigated against E. coli and S. aureus. Methodology: PAEO was chemically analyzed using gas chromatography coupled to a mass spectrometer. Antimicrobial activity was assessed by the disc diffusion method and broth microdilution method to determine minimum inhibitory concentrations (MICs). Antibiofilm activity was investigated using 96-well plates with a crystal violet assay. Results: carvacrol (85.25%) was the major component of PAEO. The disc diffusion test confirmed the ability of PAEO and carvacrol in inhibiting bacteria in their planktonic form. The MICs of PAEO against S. aureus and E. coli were 0.31 and 1.25%, respectively, with bactericidal effect. Carvacrol demonstrated a significant antibacterial property (MIC = 0.31%), exhibiting bacteriostatic effects against S. aureus and bactericidal effects against E. coli. Carvacrol considerably inhibited E. coli biofilm formations (58.9%). Moreover, carvacrol inactivated the mature biofilms. Conclusion: the data obtained are promising, and facilitates the development of new therapeutic alternatives. These results indicate the potential of carvacrol in treating diseases caused by E. coli and S. aureus
Introdução: Escherichia coli e Staphylococcus aureus são patógenos que podem causar infecções persistentes devido aos mecanismos de resistência e à sua capacidade de aderir e formar biofilme. A busca por novos agentes a partir de recursos naturais para combater o biofilme é uma prioridade na área da saúde. Objetivo: a composição química, a atividade antibacteriana e anti-biofilme do óleo essencial de Plectranthus amboinicus (PAEO) e do carvacrol foram investigadas frente a S. aureus e E. coli. Metodologia: a análise química do PAEO foi realizada por cromatografia gasosa acoplada a espectrômetro de massa. A atividade antimicrobiana foi avaliada por difusão em disco e microdiluição em caldo para determinar as concentrações inibitórias mínimas (CIMs). As atividades do anti-biofilme foram investigadas usando placas de 96 poços pelo ensaio de cristal violeta. Resultados: o carvacrol (85,25%) foi o principal componente do PAEO. A capacidade do PAEO e do carvacrol para inibir bactérias planctônica foi confirmada por difusão em disco. A CIM do PAEO para S. aureus e E. coli foi de 0,31 e 1,25%, respectivamente, com efeito bactericida. O carvacrol demonstrou uma propriedade antibacteriana significativa (CIM=0,31%), exibindo efeito bacteriostático frente a S. aureus e efeito bactericida para E. coli. O carvacrol inibiu significativamente a formação de biofilme de E. coli (58,9%) e promoveu a desestabilização do biofilme maduro. Conclusão: os dados obtidos são promissores, possibilitando o desenvolvimento de novas alternativas terapêuticas e revelam a potencialidade do carvacrol como fonte para o tratamento de doenças causadas por E. coli e S. aureus.
Subject(s)
Staphylococcus aureus/drug effects , Oils, Volatile/pharmacology , Biofilms/drug effects , Plectranthus/chemistry , Escherichia coli/drug effects , Anti-Bacterial Agents/pharmacology , Microbial Sensitivity Tests/methodsABSTRACT
Introducción: El tratamiento de las infecciones por Klebsiella pneumoniae productora de carbapenemasa tipo KPC es complicado debido a las escasas opciones terapéuticas existentes, lo cual obliga a optimizar los esquemas terapéuticos disponibles. Objetivo: Determinar la concordancia de la tarjeta AST-N272 del Sistema Vitek 2 Compact y las tiras M.I.C.ETM Evaluator con la dilución en agar para la determinación de la concentración mínima inhibitoria del meropenem en Klebsiella pneumoniae productora de carbapenemasa tipo KPC. Métodos: Se estudiaron 53 aislados de K. pneumoniae bla KPC positivas no clonales, provenientes de hisopados rectales recolectados en diferentes unidades hospitalarias de Guayaquil, Ecuador, entre enero a junio de 2016. Se determinó la concentración mínima inhibitoria de meropenem por dilución en agar (método de referencia), así como por el sistema Vitek 2 Compact (AST-N272) y las tiras M.I.C.ETM. Se determinó la CMI 50, CMI 90 y la concordancia esencial. Resultados: El rango de la CMI de meropenem de los aislados estudiados fue de 1 a ≥ 32 µg/mL, con una CMI50= 4 µg/mL y una CMI90= ≥ 32 µg/mL. El 86,79 por ciento (n= 46) de los aislados tuvo una CMI≤ 8 µg/mL. Se observó un 94,33 por ciento de concordancia esencial con las tiras M.I.C.ETM, mientras que la tarjeta AST-N272 mostró una concordancia esencial inferior al 50 por ciento. Conclusiones: Los resultados sugieren posibles implicaciones en el tratataminto del paciente, pues reduce opciones terapéuticas en contextos de difícil manejo. Además, resaltan la necesidad de la confirmación de la resistencia a carbapenémicos mediante el método de Kirby Bawer en aquellos laboratorios que tienen métodos automatizados para estudios de susceptibilidad(AU)
Introduction: The treatment for KPC carbapenemase-producing Klebsiella pneumoniae infections is complicated, due to the scant therapeutic options available, which forces us to optimize the therapies at hand. Objective: Determine the agreement between the AST-N272 card of the Vitek 2 Compact system and the M.I.C.E.TM Evaluator strips, and the agar dilution method for determination of the minimum inhibitory meropenem concentration in KPC carbapenemase-producing Klebsiella pneumoniae. Methods: A study was conducted of 53 positive non-clonal K. pneumoniae bla KPC isolates from rectal swabs collected at several hospitals in Guayaquil, Ecuador, from January to June 2016. Minimum inhibitory meropenem concentration was determined by agar dilution (reference method), the Vitek 2 Compact system (AST-N272) and M.I.C.E.TM strips. Determination was made of MIC 50, MIC 90 and essential agreement. Results: The meropenem MIC range for the isolates studied was 1 to ≥ 32 µg/ml, with MIC50= 4 µg/ml and MIC90= ≥ 32 µg/ml. In 86.79 percent (n= 46) of the isolates MIC was ≤ 8 µg/ml. Essential agreement was 94.33 percent with the M.I.C.E.TM strips and under 50 percent with the AST-N272 card. Conclusions: The results obtained suggest potential implications for the treatment of patients, since therapeutic options are reduced in difficult management contexts. They also highlight the need for confirmation of carbapenem resistance by the Kirby-Bauer procedure in laboratories equipped with automated methods for susceptibility studies(AU)
Subject(s)
Humans , Microbial Sensitivity Tests/methods , Enterobacteriaceae Infections/drug therapy , Meropenem/therapeutic use , Klebsiella pneumoniae , EcuadorABSTRACT
Resumen Utilizando cepas clínicas de bacilos gramnegativos multi-resistentes (MDR), comparamos las CIM obtenidas de la microdilución en caldo, el método de referencia y el método de elución de sensidiscos. Encontramos que, con la excepción de A. baumannii, los resultados fueron muy similares. El método de elución de sensidiscos podría ser una buena alternativa y confiable para la determinación de la resistencia a colistín.
Abstract Using clinical strains of multidrug resistant (MDR) Gram negative bacilli, we compared MICs obtained from both broth microdilution, the reference method, and sensi-disk elution method. We found that, with A. baumannii exception, results were very similar. Sensi-disk elution method could be a good and reliable alternative for colistin resistance determination.
Subject(s)
Microbial Sensitivity Tests/methods , Microbial Sensitivity Tests/standards , Colistin/pharmacology , Gram-Negative Bacteria/drug effects , Anti-Bacterial Agents/pharmacology , Drug Resistance, Multiple, Bacterial , Acinetobacter baumannii/drug effectsABSTRACT
The aim of this study was evaluate the in vitro antifungal activity of crude extracts from Eugenia uniflora, Libidibia ferrea and Psidium guajava. The extracts were obtained by turbo-extraction using water (AQ) or acetone-water (AC-W) (7:3, v/v) as solvents and lyophilized to obtain the crude extracts (CE). The CE were characterized by UV-Vis, TLC and HPLC. The activity of CEs was investigated against clinical isolates of Candida spp. and the Minimum Inhibitory Concentration (MIC), MIC50 and MIC90 were determinated. The analysis by TLC showed that all CEs presented polyphenols (flavonoids and tannins). The CEs from E. uniflora showed higher amount of polyphenols (30.35 ± 2.15%, AC-W) and the HPLC analysis revealed the tannins in all extracts. The CEs of E. uniflora showed MIC range from 1.9 to 500.0 µg/mL, and lower values of MIC50 and MIC90 against non-albicans Candida isolates. Regarding L. ferrea and P. guajava, the results showing MIC from 3.9 to 1000.0 µg/mL (CE-AQ) against C. albicans. The results demonstrate antifungal performance from CE against various species of Candida spp., suggesting that the herbal species may be use as new potential antifungal agents. Additionally, the polyphenol content can play a pivotal role in the antifungal properties of CE.
Subject(s)
In Vitro Techniques/methods , Plant Extracts/adverse effects , Polyphenols/analysis , Phytochemicals , Antifungal Agents/administration & dosage , Flavonoids/pharmacokinetics , Microbial Sensitivity Tests/methods , Chromatography, High Pressure Liquid/methodsABSTRACT
ABSTRACT The precise diagnosis of bacterial meningitis is essential. Cytological and biochemical examination of cerebrospinal fluid (CSF) are not specific. Conventional methods for bacterial meningitis lack sensitivity or take too long for a final result. Therefore, other methods for rapid and accurate diagnosis of central nervous system infections are required. FilmArray meningitis/encephalitis (ME) panel is a PCR multiplex for simultaneous and rapid identification of 14 pathogens, including 6 bacteria, 7 viruses, and Cryptococcus. We evaluated 436 CSF samples submitted to FilmArray ME Panel. Among them, 25 cases were positive for bacteria, being Streptococcus pneumonia the most frequent (48 %). Among positive cases for bacteria, 60 % were positive only with FilmArray. All the bacterial meningitis cases in which the only positive test was FilmArray had CSF findings suggestive of bacterial meningitis, including neutrophilic pleocytosis, increased CSF protein and lactate, and decreased CSF glucose. These findings suggest that FilmArray may increase the diagnostic sensitivity for bacterial meningitis.
Subject(s)
Humans , Cerebrospinal Fluid/microbiology , Cerebrospinal Fluid/virology , Meningitis, Bacterial/diagnosis , Multiplex Polymerase Chain Reaction/methods , Bacteria/isolation & purification , Viruses/isolation & purification , Microbial Sensitivity Tests/methods , Sensitivity and Specificity , Meningitis, Bacterial/cerebrospinal fluidABSTRACT
INTRODUÇÃO: A automação laboratorial é cada vez mais utilizada em microbiologia, no entanto, poucos estudos avaliam desfechos clínicos em comparação aos métodos tradicionais. No Brasil, nenhum estudo com esse objetivo foi detectado. OBJETIVO: Analisar os impactos clínicos e microbiológicos após implantação de método fenotípico automatizado em um serviço de microbiologia. MÉTODOS: Realizamos estudo observacional e retrospectivo no laboratório de microbiologia referente a exame de hemocultura de pacientes da Unidade de Terapia Intensiva (UTI). Os dados foram coletados de pacientes internados entre janeiro/2014 a dezembro/2015. Analisou-se o tempo de internação, número de terapias empíricas, óbitos e dados relacionados ao isolamento microbiológico. A amostra foi obtida por conveniência. Para a comparação entre os desfechos foram empregados os testes t de Student e Qui-quadrado de Pearson. O programa empregado foi o Stata release, versão 11, sendo considerados significativos valores de p<0,05. RESULTADOS: Foram avaliados 472 pacientes. Não houve redução na prescrição empírica de antimicrobianos (54,7% vs 45,3%; p=0,33), tempo de internação na UTI (14,5 dias vs 15,8 dias p=0,78) e na taxa de óbitos (54,4% vs 45,6%; p=0,36). Similarmente, o perfil de agentes isolados em ambos os métodos não parece ser discrepante, no entanto, houve um aumento de 44,7% no número de isolados microbianos (76 vs 110) com melhor caracterização dos mesmos. CONCLUSÃO: A automação do laboratório de microbiologia não impactou no tempo de internação, mortalidade na UTI e no número de terapias empíricas. No entanto, a identificação e o isolamento de microrganismos melhoraram.
INTRODUCTION: Automation is increasingly used in microbiology laboratory, however, few studies assessed clinical outcomes compared to traditional methods. In Brazil, no studies with this objective were detected. OBJECTIVE: To analyze the clinical and microbiological impacts after implantation of an automated phenotypic method in a microbiology service. METHODS: Observational and retrospective study carried out on the microbiology laboratory involving blood culture test from intensive care unit (ICU) patients. Data were collected from hospitalized patients between January 2014 and December 2015. The length of hospitalization, number of empirical therapies, deaths and information related to microbiological isolation were analyzed. The sample was obtained by convenience. Pearson's Chisquare and Student's t-tests were used to compare outcomes. The program used was the Stata release, version 11, being considered significant values of p<0.05. RESULTS: A total of 472 patients were evaluated. There was no reduction in the empirical prescription of antimicrobials (54.7% vs 45.3%; p=0.33), ICU stay (14.5 days vs 15.8 days; p=0.78) and mortality (54.4% vs 45.6%; p=0.36). Similarly, profile of isolated agents in both methods did not appear to be discrepant, however, there was an increase of 44.7% in the number of microbial isolates (76 vs 110) and a better characterization of them. CONCLUSION: The microbiology laboratory automation did not modify the length of stay, ICU mortality and the number of empirical therapies. However, identification and isolation of microorganisms was improved.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Microbial Sensitivity Tests/instrumentation , Microbial Sensitivity Tests/methods , Automation, Laboratory/instrumentation , Automation, Laboratory/methods , Blood Culture/instrumentation , Blood Culture/methods , Microbiology/instrumentationABSTRACT
Resumen Introducción. Las infecciones por Staphylococcus aureus y Staphylococcus coagulasa negativa multirresistentes a los antibióticos y asociadas con la atención en salud tienen un gran impacto epidemiológico por su alta morbimortalidad; además, se han relacionado con la formación de biopelículas, lo cual también se asocia con la resistencia a los antimicrobianos. Objetivo. Determinar la resistencia a la meticilina y cuantificar la producción de biopelículas para establecer su posible relación con los aislamientos clínicos de S. aureus y Staphylococcus coagulasa negativa. Materiales y métodos. Se estudiaron 11 cepas de S. aureus y 12 de Staphylococcus coagulasa negativa. La resistencia a la meticilina se determinó con discos de cefoxitina tomando como valores de referencia los estándares del Clinical Laboratory Standards Institute (CLSI) de 2018. La producción de biopelícula se cuantificó con cristal violeta. Los genes mecA e icaADBC se identificaron mediante reacción en cadena de la polimerasa (PCR), y se hizo un análisis bivariado con la prueba de ji al cuadrado y el coeficiente V de Cramér, utilizando el programa SPSS™, versión 20.0. Resultados. Nueve cepas de S. aureus fueron resistentes a la meticilina (SARM) y dos fueron sensibles. Ocho cepas de Staphylococcus coagulasa negativa fueron resistentes y cuatro fueron sensibles. El genotipo mecA se encontró en ocho de las nueve cepas de S. aureus y en seis de las ocho de Staphylococcus coagulasa negativa resistentes a meticilina. Todas las cepas formaron biopelícula. Diez cepas de S. aureus y 11 de Staphylococcus coagulasa negativa presentaron el genotipo icaADCB. No se encontró asociación entre la resistencia a meticilina y la formación de biopelícula. Conclusiones. La cefoxitina es suficiente para determinar el fenotipo resistente a meticilina y se asoció con el genotipo mecA. Las cepas resistentes a la meticilina y poseedoras del gen mecA pueden presentar un mecanismo de resistencia alterno. Los dos grupos de cepas formadoras de biopelícula se relacionaron con la presencia del operón icaADCB. La formación de biopelícula y la resistencia a la meticilina se expresaron como características independientes en los dos grupos de cepas.
Abstract Introduction: Infections associated with health care caused by S. aureus and coagulase- negative Staphylococci multi-resistant to antibiotics cause a high epidemiological impact due to their high morbidity and mortality. Biofilm formation, which has been associated with antimicrobial resistance, can also occur. Objectives: To determine methicillin resistance and to quantify the biofilm production to establish if there is a relationship in clinical isolates of S. aureus and coagulase-negative Staphylococci. Material and methods: A total of 11 strains of S. aureus and 12 of coagulase-negative Staphylococci were studied. Methicillin resistance was determined with cefoxitin discs and the Clinical Laboratory Standards Institute (CSLI), 2018 reference values. Biofilm production was quantified by the crystal violet method. The mecA and icaADBC genes were identified by PCR. A bivariate analysis was performed with chi-square (c2) and Cramér's V statistical tests, using SPSS™, version 20.0 software. Results: Nine S. aureus strains were methicillin-resistant and two were sensitive. Eight coagulase-negative Staphylococci strains were resistant and four were sensitive. The mecA genotype was found in eight of the nine S. aureus resistant strains and six of eight resistant coagulase-negative Staphylococci. All strains formed biofilms. Ten strains of S. aureus and 11 of coagulase-negative Staphylococci presented the icaADCB genotype. No association was found between methicillin-resistance and biofilm formation. Conclusions: Cefoxitin is enough to define the resistance phenotype and is associated with the mecA genotype. All strains formed biofilms and were related to the presence of the icaADCB operon. Biofilm formation and methicillin resistance were independent features in both groups of strains.
Subject(s)
Humans , Staphylococcus/drug effects , Staphylococcus/physiology , Staphylococcus aureus/drug effects , Staphylococcus aureus/physiology , Methicillin Resistance , Biofilms/growth & development , Oxacillin/pharmacology , Staphylococcal Infections/microbiology , Staphylococcus/enzymology , Staphylococcus/genetics , Staphylococcus aureus/genetics , Bacterial Proteins/genetics , DNA, Bacterial/isolation & purification , Microbial Sensitivity Tests/methods , Cefoxitin/pharmacology , Methicillin Resistance/genetics , Coagulase , Penicillin-Binding Proteins/genetics , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/genetics , Genes, Bacterial , Mexico , Anti-Bacterial Agents/pharmacologyABSTRACT
According to the World Health Organization, diarrheal infections cause 525 000 deaths of children under five years of age every year, and shigellosis. Shigellosis is a relevant cause of dysentery, which increases the morbidity and mortality in pediatric patients. Therefore, emergingthe emergence of antimicrobial resistant strains of Shigella is a concerningworrisome problem worldwide. We report the case of a 7-year-old patient with acute dysentery caused by CTX-M Type ESBL Producing Shigella flexneri, being. This was the first case treated in the Specialties Hospital of Specialties of the Armed Forces N°1, in Quito, Ecuador. The antibiogram demonstrated sensibilityshowed sensitivity to ampicillin-sulbactam. As a result, after five days of microbiologically directed treatment, the patient improved his condition without relapse. Proper clinical diagnoses and accurate laboratory studies like stool culture and antibiogram are crucial to givingindicate an appropriate therapy in infections caused by Shigella and other enteric bacilli(AU)
Según la Organización Mundial de la Salud, las infecciones diarreicas provocan 525 000 muertes de niños menores de cinco años de edad cada año. La shigelosis es una causa importante de disentería que aumenta la morbilidad y mortalidad de los pacientes pediátricos. Es por eso que el surgimiento de cepas de Shigella resistentes a los antibióticos es un preocupante problema a nivel mundial. Presentamos el caso de un paciente de 7 años de edad con disentería aguda provocada por Shigella flexneri productora de BLEE tipo CTX-M. Se trata del primer caso tratado en el Hospital de Especialidades de las Fuerzas Armadas Nº 1, en Quito, Ecuador. El antibiograma mostró sensibilidad a la combinación ampicilina/sulbactam. Al cabo de cinco días de tratamiento microbiológico, el paciente mejoró su estado y no se produjeron recaídas. Un diagnóstico clínico correcto, así como estudios precisos de laboratorio como los cultivos de heces y los antibiogramas, son vitales para indicar una terapia apropiada en las infecciones causadas por Shigella y otros bacilos entéricos(AU)
Subject(s)
Humans , Male , Child , Clinical Diagnosis , Dysentery/prevention & control , Dysentery, Bacillary/drug therapy , Microbial Sensitivity Tests/methodsABSTRACT
OBJECTIVE: To report our experience using conventional culture methods (CM) and pediatric blood culture bottles (PBCBs) for vitreous sample culture of acute postoperative endophthalmitis. METHODS: A retrospective study was conducted at the Department of Ophthalmology, Hospital das Clinicas, HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo, Sao Paulo, BR, from January 2010 to December 2015, and it included 54 patients with clinically suspected acute postoperative endophthalmitis. Vitreous samples were obtained by vitreous tap or vitrectomy. Samples from January 2010 to December 2011 were cultivated in CM, whereas samples from January 2012 to December 2015 were inoculated in PBCBs. The measured outcome was the yield of positive cultures. RESULTS: Twenty cases were included in the CM group, and 34 cases were included in the PBCB group. The yield of positive cultures in PBCBs (64.7%) was significantly higher than that in conventional CM (35%, p=0.034). Staphylococcus epidermidis and Streptococcus viridans were the two most commonly found agents. CONCLUSION: PBCBs can be used successfully in clinically suspected endophthalmitis. The method showed a higher yield of positive cultures than the conventional method. This technique appears to have several advantages over the traditional method: it saves time, as only one medium needs to be inoculated; transportation to a laboratory is easier than in the traditional method, and there is no need to maintain a supply of fresh agar media. The use of PBCBs may be recommended as the primary method for microbiological diagnosis and is especially suitable for office settings and remote clinics.
Subject(s)
Humans , Child , Postoperative Complications/diagnosis , Staphylococcus epidermidis/isolation & purification , Endophthalmitis/diagnosis , Culture Media/standards , Viridans Streptococci/isolation & purification , Blood Culture/instrumentation , Vitreous Body/microbiology , Microbial Sensitivity Tests/methods , Acute Disease , Retrospective Studies , Blood Culture/methodsABSTRACT
Abstract Although invasive infections and mortality caused by Candida species are increasing among compromised patients, resistance to common antifungal agents is also an increasing problem. We analyzed 60 yeasts isolated from patients with invasive candidiasis using a PCR/RFLP strategy based on the internal transcribed spacer (ITS2) region to identify different Candida pathogenic species. PCR analysis was performed from genomic DNA with a primer pair of the ITS2-5.8S rDNA region. PCR-positive samples were characterized by RFLP. Restriction resulted in 23 isolates identified as C. albicans using AlwI, 24 isolates as C. parapsilosis using RsaI, and 13 as C. tropicalis using XmaI. Then, a group of all isolates were evaluated for their susceptibility to a panel of previously described killer yeasts, resulting in 75% being susceptible to at least one killer yeast while the remaining were not inhibited by any strain. C. albicans was the most susceptible group while C. tropicalis had the fewest inhibitions. No species-specific pattern of inhibition was obtained with this panel of killer yeasts. Metschnikowia pulcherrima, Pichia kluyveri and Wickerhamomyces anomalus were the strains that inhibited the most isolates of Candida spp.
Resumo Embora as infecções invasivas e a mortalidade causada por espécies de Candida estejam aumentando entre pacientes comprometidos, a resistência a agentes antifúngicos comuns também é um problema crescente. Analisamos 60 leveduras isoladas de pacientes com candidíase invasiva utilizando como estratégia PCR/RFLP baseada na região espaçadora transcrita interna (ITS2) para identificar diferentes espécies patogênicas de Candida. A análise por PCR foi realizada a partir de ADN genómico com um par de iniciadores da região ITS2-5.8S rDNA. As amostras PCR-positivas foram caracterizadas por RFLP. A restrição resultou em 23 isolados identificados como C. albicans usando AlwI, 24 isolados como C. parapsilosis usando RsaI e 13 como C. tropicalis usando XmaI. Em seguida, avaliou-se o grupo de todos os isolados quanto à sua susceptibilidade a um painel de leveduras killer previamente descritas, resultando em 75% sendo suscetíveis a pelo menos uma levedura killer, enquanto que as restantes não foram inibidas por qualquer cepa. C. albicans foi o grupo mais suscetível enquanto C. tropicalis teve o menor número de inibições. Não se obteve um padrão de inibição específico da espécie com este painel de leveduras killer. Metschnikowia pulcherrima, Pichia kluyveri e Wickerhamomyces anomalus foram as cepas que inibiram a maioria dos isolados de Candida spp.
Subject(s)
Humans , Adult , Candida/drug effects , Candidiasis, Invasive/drug therapy , Antifungal Agents/pharmacology , Polymorphism, Restriction Fragment Length , Candida/genetics , Microbial Sensitivity Tests/methods , Polymerase Chain Reaction/methods , Candidiasis, Invasive/microbiologyABSTRACT
Abstract This study aimed to evaluate the in vitro antibacterial activity of the phytochemicals thymol, linalool, and citronellol against Streptococcus mutans, Streptococcus salivarius and Streptococcus oralis. Disk diffusion screening on solid medium and measurement of the diameter of the bacterial growth inhibition halos was the technique utilized. The Minimum Inhibitory Concentration (MIC) of the substances was determined using serial substance dilutions and microdilution technique in Brain Heart Infusion culture medium. After incubation for 24 hours in an oven at 37 °C, plate reading was completed and confirmed by visual method using 2,3,5 triphenyl tetrazolium chloride dye. The Minimum Bactericidal Concentration (MBC) was determined from MIC subcultures. Assays were performed in triplicate, and chlorhexidine was used as a positive control. The diameters in mm of the growth inhibition halos ranged between 7.3 and 10.7 for S. mutans, 7.3 and 10.0 for S. oralis, and 8.2 and 9.8 for S. salivarius. The MIC and MBC values obtained converged, ranging from maximum values in the presence of Linalool (1,250.0 mg/mL, 2,500.0 mg/mL and 2,500.0 mg/mL, respectively, for S. mutans, S. oralis, and S. salivarius); and minimum values with Thymol (312.5 μg/ml, 156.2 μg/mL and 156.2 μg/ml, respectively for S. mutans, S. oralis, and S. salivarius). All the tested phytochemicals displayed antibacterial activity, thus representing substances with potential applications in preventing tooth decay.
Resumo Este estudo objetivou avaliar a atividade antibacteriana in vitro dos fitoquímicos timol, linalol e citronelol sobre Streptococcus mutans, Streptococcus salivaris e Streptococcus oralis. Utilizou-se a técnica de discos de difusão em meio sólido e medição do diâmetro dos halos de inibição. A concentração inibitória mínima (CIM) das substâncias foi determinada utilizando diluições em série das substâncias e técnica de microdiluição em meio de cultura de Brain Heart Infusion. Após incubação durante 24 horas em estufa a 37 °C, a leitura da placa foi confirmada pelo método visual usando o corante 2,3,5 trifenil cloreto de tetrazólio. A concentração bactericida mínima (CBM) foi determinada a partir de subculturas de MIC. Os ensaios foram realizados em triplicata, e clorexidina foi usada como um controle positivo. Os diâmetros dos halos de inibição do crescimento variaram entre 7,3 e 10,7 por S. mutans, 7,3 e 10,0 por S. oralis, e 8,2 e 9,8 para S. salivaris. Os valores de CIM e CBM obtidos variaram de valores máximos na presença de linalol (1.250,0 mg/mL, 2.500.0 mg/mL e 2.500.0 mg/mL, respectivamente, para o S. mutans, S oralis e S. salivaris); a valores mínimos com timol (312,5 μg/ml, 156,2 μg/mL e 156,2 μg/ml, respectivamente para S. mutans, S. oralis e S. salivaris). Todos os fitoquímicos testados apresentaram atividade antibacteriana, representando, assim, substâncias com potencial de aplicações na prevenção da cárie dentária.
Subject(s)
Streptococcus/drug effects , Thymol/pharmacology , Monoterpenes/pharmacology , Anti-Bacterial Agents/pharmacology , Microbial Sensitivity Tests/methods , Dental Caries/microbiology , Phytochemicals/pharmacology , Acyclic MonoterpenesABSTRACT
Introducción: La resistencia bacteriana amenaza la efectividad de todos los antibióticos disponibles en la actualidad. Como parte de las acciones para mejora del uso de antimicrobianos, la lectura interpretada del antibiograma surge como una herramienta clave en la optimización del uso de antimicrobianos. Objetivo: Exponer las ideas fundamentales tras la lectura interpretada del antibiograma, orientadas hacia los médicos de asistencia clínica de centros que solo dispongan de microbiología básica en sus laboratorios. Material y métodos: Se realizó una búsqueda sistemática en la base de datos MEDLINE de información biomédica a través del motor PubMed, además del Google Académico. Para artículos publicados en revistas cubanas, se revisó la biblioteca virtual electrónica ScIELO. Resultados: Se describe el enfoque de la lectura interpretada del antibiograma en dos grupos de bacterias: los cocos Gram positivos y los bacilos Gram negativos, con énfasis en aquellos resultados que se pueden obtener con los recursos disponibles en la mayoría de los laboratorios en el país. Se señalan además los casos en que es de extrema importancia la determinación de la concentración mínima inhibitoria para la obtención de resultados satisfactorios respecto a la evolución del paciente. Son mencionadas adicionalmente algunas alternativas terapéuticas para gérmenes multirresistentes. Conclusiones: Convertir la lectura interpretada del antibiograma en un acto cotidiano es una necesidad impostergable, que requiere además de los conocimientos básicos, una estrecha interrelación entre el clínico y el laboratorio de microbiología y la disponibilidad de recursos mínimos, como los discos de antibióticos marcadores(AU)
Introduction: Bacterial resistance threatens the effectiveness of all available antibiotics at present. Interpretive reading of the antibiogram emerges as a key element for an optimal use of these unique drugs as part of the actions for the improvement of antimicrobial use. Objective: To present basic ideas directed to attending physicians from centers with basic microbiology resources at their disposal, after an interpretive reading of the antibiogram. Material and Methods: A systematic search on MEDLINE database was conducted using PubMed and Google Scholar search engines. A search on the Scientific Electronic Virtual Library Online - SciELO was used for the articles published in Cuban magazines. Results: The review is focused mainly to two groups of bacteria: Gram-positive cocci and Gram-negative bacilli, making an emphasis on the results that can be obtained with the resources, which are available in most laboratories of the country. Cases where the determination of minimal inhibitory concentration is of extreme importance to obtain satisfactory results regarding the patients´ evolution are also pointed out. In addition, some therapeutic alternatives for multi-resistant germs are presented. Conclusions: To turn the interpretive reading of the antibiogram into a daily act is an extremely urgent need that requires, besides the basic knowledge, a close relationship between the clinician and the microbiology lab, as well as a group of basic resources such as key antibiotics discs(AU)
Subject(s)
Humans , Male , Female , Microbial Sensitivity Tests/methods , Microbiological Techniques/methods , Drug Resistance, Bacterial , Clinical Laboratory Techniques/methodsABSTRACT
Abstract INTRODUCTION This study aims to evaluate the antifungal susceptibility of different species of Candida isolated from diabetic patients against eight antifungal agents. METHODS Susceptibility testing of 111 clinical isolates of Candida species was performed against 8 antifungals using the M27-A3 protocol of the Clinical and Laboratory Standards Institute (CLSI). RESULTS Voriconazole, lanoconazole, and caspofungin showed the highest in vitro activity against all the isolates of C. albicans. Resistance against the tested antifungals was only observed in the C. albicans isolates. CONCLUSIONS Our finding revealed that resistance against amphotericin B, itraconazole, ketoconazole, posaconazole, and fluconazole can be observed in C. albicans.
Subject(s)
Humans , Male , Female , Adult , Aged , Aged, 80 and over , Candida/drug effects , Diabetes Mellitus/microbiology , Antifungal Agents/pharmacology , Candida/isolation & purification , Candida/classification , Microbial Sensitivity Tests/methods , Middle AgedABSTRACT
Abstract INTRODUCTION: Bloodstream infections can be fatal, and timely identification of the etiologic agent is important for treatment. METHODOLOGY: An alternative method, consisting of direct identification and susceptibility testing of blood culture bottles using the automated VITEK 2® system, was assessed. RESULTS: All 37 of the Gram-negative bacilli (GNB) identifications and 57.1% of the 28 Gram-positive cocci (GPC) identifications matched those obtained with standard methods. In susceptibility testing, the agreement was greater than 90%. CONCLUSIONS: This alternative methodology may assist in the early identification and susceptibility testing of GNB. Further research is necessary to develop appropriate methods for GPC.
Subject(s)
Humans , Microbial Sensitivity Tests/methods , Bacteremia/microbiology , Gram-Positive Cocci/drug effects , Gram-Negative Bacteria/drug effects , Anti-Bacterial Agents/pharmacology , Prospective Studies , Bacteriological Techniques , Bacteremia/diagnosis , Gram-Positive Cocci/classification , Qualitative Research , Gram-Negative Bacteria/classificationABSTRACT
Objective: To evaluate the diagnostic accuracy of line probe assays for drug- resistant tuberculosis (TB) in China. Methods: Chinese databases (CNKI, Wanfang, SinoMed, VIP Information) and English databases (PubMed, Embase, Cochrane Library) were used to retrieve the literatures regarding the accuracy of line probe assays in the diagnosis of drug-resistant tuberculosis in China between January 1, 2000 and September 1, 2017. Quality Assessment of Diagnostic Accuracy Studies-2 was used to evaluate the quality of the included studies. Sensitivity and specificity in different studies (using drug sensitivity test or gene sequencing as gold standard) were combined by Meta-analysis using bivariate or univariate model. In addition, subgroup analysis (GenoType MTBDRplus, GenoType MTBDRsl and Reverse dot blot hybridization) and sensitivity analysis were also carried out. Results: A total of 24 literatures involving 82 studies were included in the final analysis. The sensitivity and specificity of line probe assays for rifampicin resistant TB were 0.91(0.88-0.94) and 0.98 (0.97-0.99), respectively. The sensitivity and specificity of line probe assays for isoniazid resistant TB were 0.80 (0.77-0.83) and 0.98 (0.96-0.99), respectively. The sensitivity and specificity of line probe assays for multidrug-resistant TB were 0.81 (0.76-0.85) and 0.99 (0.99-1.00), respectively. The sensitivity and specificity of line probe assays for quinolone resistant TB were 0.92(0.88-0.95) and 0.94 (0.91-0.97), respectively. The sensitivity and specificity of line probe assays for second-line injectable drug resistant TB (including kanamycin, Capreomycin, amikacin) were 0.79(0.58-0.91) and 0.98 (0.90-1.00), respectively. The sensitivity and specificity of line probe assays for extensively drug-resistant TB were 0.46 (0.19-0.75) and 1.00 (0.98-1.00), respectively. Subgroup analysis showed that the overall diagnostic accuracy of GenoType MTBDRplus and GenoType MTBDRsl was higher than that of Reverse dot blot hybridization. According to the results of sensitivity analysis, the results of this study were robust. Conclusion: The diagnostic accuracy of line probe assays for drug-resistant TB is high.
Subject(s)
Humans , Antitubercular Agents/therapeutic use , Biological Assay/methods , China , Isoniazid/pharmacology , Microbial Sensitivity Tests/methods , Mycobacterium tuberculosis/isolation & purification , Rifampin/pharmacology , Sensitivity and Specificity , Tuberculosis, Multidrug-Resistant/drug therapyABSTRACT
Resumen Actualmente se recomienda el uso de cefazolina para determinar la susceptibilidad a cefalosporinas orales de primera generación en cepas de enterobacterias en ITU no complicada. Nuestro objetivo fue establecer la susceptibilidad a cefalosporinas orales en cepas urinarias según puntos de corte para cefalotina o cefazolina y la correlación de susceptibilidad entre cefazolina y cefadroxilo. Se estudió la concordancia entre cefalotina y cefazolina en 52 cepas por método de Kirby-Bauer y Vitek XL. En Escherichia coli fue de 0% para VitekXL y 50% para Kirby-Bauer. La concordancia entre cefazolina y cefadroxilo fue 95,6%. En el laboratorio debiera usarse cefazolina para determinar susceptibilidad a cefalosporinas orales de primera generación. La concordancia entre cefazolina y cefadroxilo sugiere que cefazolina podría predecir susceptibilidad para cefadroxilo.
Currently, the use of cefazolin is recommended to determine the susceptibility to first-generation oral cephalosporins in strains of enterobacteria in uncomplicated UTI. We determined susceptibility differences to oral cephalosporins in urinary strains according to cefazolin or cefalotin breakpoints and the correlation of susceptibility between cefazolin and cefadroxil. We studied 52 strains with cefalotin and cefazolin by disk-diffusion and MIC (Kirby-Bauer and Vitek XL) and a subgroup by disk-diffusion for cefadroxil. Agreement among different methods was 100% for K. pneumoniae and Proteus spp. In Escherichia coli, agreement for Vitek and disk-diffusion were 0 and 50% respectively. Susceptibility to first generation cephalosporins in E. coli should be determined with cefazolin. Agreement between cefazolin and cefadroxil suggests that cefazolin could also predict the susceptibility of cefadroxil.
Subject(s)
Humans , Cephalosporins/pharmacology , Enterobacteriaceae/drug effects , Anti-Bacterial Agents/pharmacology , Proteus/drug effects , Urinary Tract Infections/microbiology , Microbial Sensitivity Tests/methods , Cefadroxil/pharmacology , Cefazolin/pharmacology , Cephalosporins/classification , Cephalothin/pharmacology , Enterobacteriaceae/classification , Enterobacteriaceae Infections/microbiology , Escherichia coli/drug effects , Klebsiella pneumoniae/drug effectsABSTRACT
Currently, there is a controversy in how to determine the minimal inhibitory concentration (MIC) of colistin against Acinetobacter baumannii. We compared three methods, concluding that the addition of Tween-80 (0.002%) to Müller-Hinton broth in the microdilution method could improve MIC determination and it could reduce false resistance.
Subject(s)
Humans , Microbial Sensitivity Tests/methods , Colistin/pharmacology , Acinetobacter baumannii/drug effects , Anti-Bacterial Agents/pharmacologyABSTRACT
Abstract INTRODUCTION: This study aimed to evaluate a new commercial kit, Kit SIRE Nitratase-PlastLabor, for testing the drug susceptibility of clinical Mycobacterium tuberculosis isolates. METHODS: The accuracy of the Kit SIRE Nitratase was evaluated by examining the susceptibility (streptomycin, isoniazid, rifampicin, and ethambutol) of 40 M. tuberculosis isolates, using the proportion method with Lowenstein-Jensen medium or the BACTEC MGIT 960 system. RESULTS: The detection accuracy for streptomycin, isoniazid, rifampicin, and ethambutol was 95%, 97.5%, 100%, and 80%, respectively. CONCLUSIONS: The exceptional accuracy demonstrated by Kit SIRE Nitratase for isoniazid and rifampicin makes the kit an attractive option for screening M. tuberculosis strain resistance.