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1.
Rev. cuba. med. trop ; 72(3): e516, sept.-dic. 2020. tab
Article in Spanish | LILACS, CUMED | ID: biblio-1156536

ABSTRACT

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 , Ecuador
2.
Rev. chil. infectol ; 37(1): 87-88, feb. 2020.
Article in Spanish | LILACS | ID: biblio-1092727

ABSTRACT

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 effects
3.
Braz. J. Pharm. Sci. (Online) ; 56: e18456, 2020. tab, graf
Article in English | LILACS | ID: biblio-1249172

ABSTRACT

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/methods
4.
Braz. j. infect. dis ; 23(6): 468-470, Nov.-Dec. 2019. tab
Article in English | LILACS | ID: biblio-1089315

ABSTRACT

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 fluid
5.
ABCS health sci ; 44(2): 96-102, 11 out 2019. tab, graf
Article in English, Portuguese | LILACS | ID: biblio-1022342

ABSTRACT

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/instrumentation
6.
Rev. cuba. med. trop ; 71(1): e315, ene.-abr. 2019. tab
Article in English | LILACS-Express | LILACS, CUMED | ID: biblio-1093550

ABSTRACT

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/methods
7.
Clinics ; 74: e837, 2019. tab, graf
Article in English | LILACS | ID: biblio-1001824

ABSTRACT

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/methods
8.
Braz. j. biol ; 78(4): 742-749, Nov. 2018. tab, graf
Article in English | LILACS | ID: biblio-951595

ABSTRACT

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/microbiology
9.
Braz. j. biol ; 78(4): 691-696, Nov. 2018. tab
Article in English | LILACS | ID: biblio-951593

ABSTRACT

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 Monoterpenes
10.
Rev. habanera cienc. méd ; 17(4): 603-619, jul.-ago. 2018. tab
Article in Spanish | LILACS | ID: biblio-978555

ABSTRACT

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/methods
11.
Rev. Soc. Bras. Med. Trop ; 51(4): 542-545, July-Aug. 2018. tab
Article in English | LILACS | ID: biblio-1041473

ABSTRACT

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 Aged
12.
Rev. Soc. Bras. Med. Trop ; 51(2): 215-218, Mar.-Apr. 2018. tab
Article in English | LILACS | ID: biblio-1041453

ABSTRACT

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/classification
13.
Rev. chil. infectol ; 35(3): 329-331, 2018. tab
Article in Spanish | LILACS | ID: biblio-1042647

ABSTRACT

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 effects
14.
Rev. Soc. Bras. Med. Trop ; 50(4): 550-553, July-Aug. 2017. tab
Article in English | LILACS | ID: biblio-1041420

ABSTRACT

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.


Subject(s)
Humans , Oxidoreductases/pharmacology , Microbial Sensitivity Tests/methods , Antibiotics, Antitubercular/pharmacology , Mycobacterium tuberculosis/drug effects , Rifampin/pharmacology , Streptomycin/pharmacology , Reproducibility of Results , Drug Resistance, Bacterial , Clinical Enzyme Tests/methods , Ethambutol/pharmacology , Isoniazid/pharmacology , Mycobacterium tuberculosis/isolation & purification
15.
Braz. j. microbiol ; 48(3): 509-514, July-Sept. 2017. tab
Article in English | LILACS | ID: biblio-889143

ABSTRACT

Abstract The production of KPC (Klebsiella pneumoniae carbapenemase) is the major mechanism of resistance to carbapenem agents in enterobacterias. In this context, forty KPC-producing Enterobacter spp. clinical isolates were studied. It was evaluated the activity of antimicrobial agents: polymyxin B, tigecycline, ertapenem, imipenem and meropenem, and was performed a comparison of the methodologies used to determine the susceptibility: broth microdilution, Etest® (bioMérieux), Vitek 2® automated system (bioMérieux) and disc diffusion. It was calculated the minimum inhibitory concentration (MIC) for each antimicrobial and polymyxin B showed the lowest concentrations for broth microdilution. Errors also were calculated among the techniques, tigecycline and ertapenem were the antibiotics with the largest and the lower number of discrepancies, respectively. Moreover, Vitek 2® automated system was the method most similar compared to the broth microdilution. Therefore, is important to evaluate the performance of new methods in comparison to the reference method, broth microdilution.


Subject(s)
Humans , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/metabolism , beta-Lactamases/metabolism , Enterobacteriaceae Infections/microbiology , Klebsiella Infections/microbiology , Klebsiella pneumoniae/drug effects , Microbial Sensitivity Tests/methods , Bacterial Proteins/genetics , beta-Lactamases/genetics , beta-Lactams/pharmacology , Drug Resistance, Bacterial , Enterobacter/drug effects , Enterobacter/genetics , Enterobacter/isolation & purification , Klebsiella pneumoniae/classification , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/isolation & purification , Polymyxin B/pharmacology
16.
Rev. cuba. med ; 56(1): 75-80, ene.-mar. 2017. ilus
Article in Spanish | LILACS | ID: biblio-901266

ABSTRACT

La melioidosis es una enfermedad infecciosa causada por una bacteria gram-negativa intracelular facultativa, Burkholderia pseudomallei, que se encuentra en el suelo y el agua. Es de importancia para la salud pública en las zonas endémicas, sobre todo el sudeste asiático. Se han descrito casos fuera de las regiones endémicas en su mayoría adquiridos por los visitantes a estas áreas. Se presenta el caso de un paciente masculino con sintomatología respiratoria, elementos de sepsis y contracción volumétrica, en el que se diagnosticó diabetes mellitus de debut, neumonía adquirida en la comunidad y finalmente se aisló en hemocultivo la Burkholderia pseudomallei. Se comenzó tratamiento recomendado acorde al antibiograma, observándose evolución satisfactoria del paciente(AU)


Melioidosis is an infectious disease caused by a facultative intracellular gram-negative bacterium, Burkholderia pseudomallei, found in soil and water. It is of importance for public health in endemic areas, especially Southeast Asia. Cases have been described outside the endemic regions, mostly acquired by visitors to these areas. The case is presented of a male patient with respiratory symptoms, elements of sepsis and volumetric contraction, in which the diagnosis of onset diabetes mellitus and community-acquired pneumonia, while Burkholderia pseudomallei was finally isolated in hemoculture. The recommended treatment was started according to the antibiogram, observing the patient's satisfactory evolution(AU)


Subject(s)
Humans , Male , Adult , Melioidosis/diagnosis , Melioidosis/diagnostic imaging , Case Reports , Microbial Sensitivity Tests/methods
17.
São Paulo; s.n; s.n; 2017. 106 p. tab, ilus, graf.
Thesis in Portuguese | LILACS | ID: biblio-883613

ABSTRACT

Seja no meio ambiente, dentro de um hospedeiro ou em outro habitat, bactérias estarão frequentemente enfrentando condições adversas, como exposição a compostos antibacterianos ou carência nutricional. Em situações como essas, as bactérias são capazes de ativar a chamada resposta estringente, modulada pelo alarmônio (p)ppGpp. O acúmulo de (p)ppGpp promove a inibição da transcrição de rRNAs e tRNAs e a supressão do processo de tradução, e a ativação de operons de biossíntese de aminoácidos. Sabe-se também hoje que a resposta estringente está relacionada a outras importantes carências nutricionais em Escherichia coli, como a falta de ácidos graxos, porém não se sabe se o mesmo ocorre em Bacillus subtilis ou em outras Grampositivas. (p)ppGpp atua também direta e indiretamente em vários outros processos celulares, como motilidade, resistência a antibióticos, virulência e persistência, indicando que (p)ppGpp é um regulador central que integra informação metabólica e respostas adaptativas. O presente trabalho buscou estudar a correlação da resposta estringente de B. subtilis com a carência de ácidos graxos e a busca por pequenas moléculas capazes de modular RelA (a principal proteína envolvida na síntese de (p)ppGpp) e impedir o acúmulo de (p)ppGpp. Para a indução da carência de ácidos graxos, foram utilizadas duas estratégias; uso da droga Cerulenina (inibidor de FabF) e mutantes condicionais no gene FabF. Observou-se que mutantes incapazes de ativar a resposta estringente (cepa ppGpp(0) ou RelAD264G) apresentaram grande perda de viabilidade celular durante a carência de ácidos graxos, ao passo que a cepa selvagem manteve sua viabilidade celular. A causa da morte se deu majoritariamente devido ao colapso do potencial de membrana. Apesar de não termos observado aumento de (p)ppGpp nas células selvagens durante a carência de ácidos graxos, observou-se uma redução da razão GTP/ATP, ao passo que na cepa ppGpp(0), a razão GTP/ATP aumentou, devido ao acúmulo de GTP. O uso da droga decoinina, capaz de reduzir os níveis intracelulares de GTP, resgatou parcialmente a viabilidade da cepa e impediu a perda do potencial de membrana, indicando que os níveis de GTP são importantes durante a carência de ácidos graxos em B. subtilis. Para a triagem de pequenas moléculas inibidoras do acúmulo de (p)ppGpp, foi utilizada uma biblioteca de 2320 diferentes compostos químicos, e buscou-se drogas capazes de reverter o fenótipo de crescimento lento de cepas de B. subtilis que acumulam (p)ppGpp (via mutação pontual; mutante RelAH77A e via tratamento com o indutor hidroxamato de arginina) em meio rico. A primeira etapa selecionou 40 moléculas capazes de resgatar o crescimento de células tratadas com arginina-hidroxamato, porém apenas uma, salicilanilida, foi capaz de também resgatar o crescimento da cepa RelAH77A. Todavia, apesar de ser capaz de acelerar o crescimento de B. subtilis esse efeito é limitado. Diversos análogos de salicilanilida foram testados, porém não apresentaram efeito superior a salicilanilida para a reversão do fenótipo de crescimento lento de B. subtilis. Em adição, a droga não foi capaz de aumentar a sensibilidade dos organismos a diversos antibióticos testados, e aparentemente é incapaz de alterar os níveis internos de (p)ppGpp, porém é capaz de causar alterações nos níveis de ATP. Logo, acredita-se que o efeito observado para o crescimento das células seja devido a efeitos indiretos, possivelmente envolvendo alteração de outros nucleotídeos fosforilados


In the environment, inside a host or other habitat, bacteria will always face adverse conditions, as for example exposure to antimicrobials or starvation. In situations like those, bacteria activate the stringent response, modulated by the alarmone (p)ppGpp. (p)ppGpp accumulation promotes inhibition of rRNA and tRNA transcription and suppression of translational process, at the same time that it activates several amino acid biosynthesis operons. It is known also that the stringent response it is related to other starvation stress in Escherichia coli, like lack of fatty acids, but there is no knowledge if the same occurs for Bacillus subtilis or other gram-positive bacteria. ppGpp acts directly and indirectly affecting several other cellular process, as motility, resistance to antibiotics, virulence and persistence, indicating that (p)ppGpp is a central regulator that integrates metabolic information and adaptive responses. This work aimed to study the correlation between the stringent response in B. subtilis with fatty acid starvation, and search for small moleculas capable of modulating RelA (the main enzyme responsible for ppGpp synthesis) and stop (p)ppGpp production. For fatty acid starvation induction, two strategies were used; use of the drug Cerulenin (inhibitor of the FabF protein) and conditional mutants of the FabF gene. We observed that mutants incapable of activating the stringent response (strains ppGpp(0) ou RelAD264G) presented great loss of viability during fatty acid starvation, whereas the wild-type strain keeps its viability. The main cause of death is due membrane rupture in some cells, but mainly due to membrane potential collapse. Although we did not observed increase of (p)ppGpp in wild-type strains during fatty acid starvation, we observed reduction in GTP/ATP ratios, a hallmark of (p)ppGpp production in gram-positive bacteria. In the strain ppGpp(0) GTP/ATP ratio increased, mainly due to GTP increase. Using the drug decoyinine, capable of reducing GTP levels, partially recued viability and protects cells of losing its membrane potential, indicating that GTP levels plays an important role during fatty acid starvation in B. subtilis. For the screening of small molecules capable of inhibit (p)ppGpp production, a library of 2320 different chemical compounds were used, and we looked for drugs capable of reverting the slow growth phenotype of B. subtilis strains with (p)ppGpp accumulation (using a mutant RelAH77A; and using a stringent response inductor, arginine hidroxamate). The first step selected for 40 molecules capable of rescuing the growth of cells treated with arginine hidroxamate, but only one drug, salicilanilyde could also rescue the growth of the strain RelAH77A. Although capable of rescuing growth of B. subtilis that accumulates (p)ppGpp, this rescue is limited. Several analogues of salicilanilyde were tested, but none were stronger than salicilanilyde itself in rescuing growth of slow growing strains of B. subtilis. In addition, the drug was not capable of increasing antibiotic sensibility and it is incapable of changing intracellular (p)ppGpp levels, but it does shifts ATP levels. Therefore, we believe that the observed effects of salicilanilyde is due indirect action, probably involving other phosphorylated nucleotides, rather than modifying (p)ppGpp levels


Subject(s)
Bacillus subtilis/metabolism , Transcription Factor RelA , Cerulenin/administration & dosage , Chromatography, High Pressure Liquid , Fatty Acids/analysis , Microbial Sensitivity Tests/methods , Microscopy, Fluorescence/instrumentation , Salicylanilides/administration & dosage , Triage/statistics & numerical data
19.
Rev. argent. microbiol ; 48(4): 308-312, dic. 2016. tab
Article in Spanish | LILACS | ID: biblio-1041767

ABSTRACT

Dada la importancia de la correcta validación clínica de los aislamientos de Streptococcus bovis, nos planteamos la revisión de su presencia en muestras de orina con el objetivo de conocer su frecuencia relativa y su patrón de sensibilidad antibiótica. Se revisó retrospectivamente la sensibilidad a los antibióticos de 91 aislados de S. bovis recuperados de muestras de orina durante un período de 4 años (2012-2015). La media de la edad de los pacientes fue de 55 años y en su mayoría fueron mujeres (81%). El 37,4% eran pacientes hospitalizados con enfermedades urológicas (61%). La sensibilidad a penicilina, vancomicina y teicoplanina fue del 97,8%. Aunque S. bovis puede ser poco común en los aislamientos de orina, su presencia en sujetos con enfermedades de base justifica la realización de estudios de patogenicidad que demuestren la verdadera capacidad de producir enfermedad de este grupo de bacterias.


Given the relevance of proper clinical validation of Streptococcus bovis, we here consider revising its presence in urine samples in order to determine its relative frequency and the pattern of antibiotic susceptibility. The susceptibility to antibiotics of 91 isolates of S. bovis from urine samples was retrospectively reviewed over a period of 4 years (2012-2015). The mean age of patients was 55 years, 81% of whom were women and 37.4% were hospitalized patients suffering from urological diseases (61%). Susceptibility to penicillin, vancomycin and teicoplanin was 97.8%. Due to the fact that S. bovis can be infrequent in urine isolates and given its presence in patients suffering from urological diseases, further pathogenic studies, showing the true ability of this group of bacteria to produce disease, are required.


Subject(s)
Humans , Male , Female , Streptococcus bovis/isolation & purification , Streptococcus bovis/drug effects , Streptococcus bovis/pathogenicity , Urinary Tract Infections/microbiology , Microbial Sensitivity Tests/methods
20.
Rev. argent. microbiol ; 48(4): 320-324, dic. 2016. ilus, tab
Article in Spanish | LILACS | ID: biblio-1041768

ABSTRACT

En este trabajo se evalúa una prueba rápida in house para la detección de enterobacterias sensibles a cefotaxima, basada en el cambio de pH del rojo fenol debido a la hidrólisis de este antibiótico. Las cepas de enterobacterias procedentes de 1.947 urocultivos se evaluaron mediante los paneles MicroScan y esta prueba in house. Mediante los paneles de MicroScan se estudiaron 499 aislados de enterobacterias, entre los cuales había 27 aislados de Escherichia coli productora de β-lactamasa de espectro extendido (BLEE), 16 de Klebsiella pneumoniae BLEE y una de Klebsiella oxytoca BLEE. La prueba in house mostró una sensibilidad del 98% y una especificidad del 97%, con un valor predictivo negativo del 100% y un valor predictivo positivo del 78%. La prueba in house basada en el cambio de pH es útil en nuestro medio para detectar presuntivamente de forma rápida cepas de enterobacterias con cierta resistencia a cefotaxima.


In this work an "in house" rapid test based on the change in pH that is due to hydrolysis for detecting Enterobacteriaceae susceptible to cefotaxime is evaluated. The strains of Enterobacteriaceae from 1947 urine cultures were assessed using MicroScan panels and the "in house" test. This rapid test includes red phenol solution and cefotaxime. Using MicroScan panels, 499 Enterobacteriaceae isolates were evaluated, which included 27 isolates of Escherichia coli producing extended-spectrum beta-lactamases (ESBL), 16 isolates of Klebsiella pneumoniae ESBL and 1 isolate of Klebsiella oxytoca ESBL. The "in house" test offers the following values: sensitivity 98% and specificity 97%, with negative predictive value 100% and positive predictive value 78%. The "in house" test based on the change of pH is useful in our area for detecting presumptively cefotaxime-resistant Enterobacteriaceae strains.


Subject(s)
Humans , Male , Female , Microbial Sensitivity Tests/methods , Cefotaxime/therapeutic use , Enterobacteriaceae/drug effects , Phenolsulfonphthalein/analysis , beta-Lactamases/analysis , Cefotaxime/pharmacology , Enterobacteriaceae/isolation & purification
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