Sensibilidad de Escherichia coli frente a fosfomicina, en infecciones urinarias provenientes de personas con discapacidad que acudieron a la Secretaría Nacional por los Derechos Humanos de las Personas con Discapacidad-SENADIS

Introducción: Fosfomicina es un antimicrobiano de amplio espectro utilizado para el tratamiento de las infecciones urinarias bajas; tiene actividad sobre bacilos gramnegativos y cocos grampositivos, así también sobre microorganismos multirresistentes, además de ofrecer una alternativa terapéutica de administración vía oral en dosis única, alcanzando una efectividad de 90%. Objetivo: Conocer la sensibilidad in vitro de Escherichia coli frente a fosfomicina, en infecciones urinarias provenientes de personas con discapacidad. Material y Método: Estudio observacional, descriptivo, prospectivo, en el que se incluyó un total de 273 muestras de urocultivo, de pacientes de ambos sexos que acudieron a SENADIS, y que en el momento de la consulta presentaban síntomas de infección del tracto urinario, por lo que se les solicitó el análisis de orina simple y cultivo. De las muestras procesadas en el laboratorio de microbiología, que fueron positivas con crecimiento bacteriano significativo, se procedió a la identificación bacteriana y a la realización del antibiograma según las recomendaciones de CLSI. Resultados: De estas 273 muestras, 91 fueron positivas para diferentes uropatógenos, 62/91 (68%) resultaron ser E. coli. De estas cepas de E. coli, 59/62 (95%) mostraron sensibilidad in vitro a fosfomicina. Comentario: Aunque el número de muestra obtenido es pequeño y no extrapolable ampliamente, pretendemos extender el trabajo por un tiempo más para compararlo más adelante. Conclusiones: Se observa que fosfomicina presenta buena actividad in vitro frente a cepas de E. coli aisladas de urocultivo, pudiendo representar una buena alternativa terapéutica a ser utilizada en la población en estudio.

Background: Fosfomycin is a broad-spectrum antibiotic used for the treatment of lower urinary tract infections, it is active against gramnegative bacilli and grampositive cocci, as well as against multi-resistant microorganism, in addition to offering a therapeutic alternative for oral administration in a single dose, reaching an effectiveness of 90%. Aim: To study the susceptibility of Escherichia coli to fosfomycin in urinary tract infections, of isolated strains obtained from patients with disabilities. Methods: It is an observational, descriptive, prospective study in which a total of 273 urine culture samples of patients of both sexes who attended the SENADIS were included, and who at the time of the consultation presented symptoms of urinary tract infection. The urine positive cultures with significant bacterial growth were performed to determine its bacterial identification and the antibiogram according to CLSI recommendations. Results: Of these 273 samples, 91 samples were positive for different uropathogens, with 62/91 (68%) being positive for E. coli. Of these E. coli strains, 59/62 (95%) showed in vitro susceptibility to fosfomycin. Comment: Although the number of samples obtained is small and it cannot be extrapolated, we pretend to extend the work for a while longer to be able to compare it later. Conclusion: Fosfomycin has good activity in vitro against E. coli isolated from urine culture in our institution, representing a good alternative to be used in our study population

Biofilm forming capability and antifungal susceptibility profile of Candida spp. from blood

Although Candida albicans remains the most frequent Candida species; however other species have emerged as important causes of candidiasis. In this work, we evaluated the in vitro susceptibility profile of C. albicans, C. parapsilosis, and C. tropicalis biofilms isolated from patients with candidemia to fluconazole, voriconazole, amphotericin B, and caspofungin. Differences between the biofilm ultrastructure of the three species were also determined. The isolates were phenotypically determined by growth on a ChromagarTM medium and assimilation profile on ID32C. The Scanning Electron Microscopy method (SEM) on biofilm was performed using polyurethane strips. For the in vitro susceptibility profile a microdilution in broth was used. Sessile cells were resistant to fluconazole, voriconazole and caspofungin. The resistance to amphotericin B was less pronounced and more variable between the tested isolates. In the SEM, slight differences in ultrastructural morphology for each species in biofilms were observed. Our results verified biofilm formation. Low susceptibility to the drugs in the three researched species confirmed the higher virulence of them.

Invasive fungal infection in patients with hematologic disorders in a Brazilian tertiary care hospital

ABSTRACT INTRODUCTION: Invasive fungal infections (IFIs) are an important complication in immunocompromised individuals, particularly neutropenic patients with hematological malignancies. In this study, we aimed to verify the epidemiology and diagnosis of IFIs in patients with hematologic problems at a tertiary hospital in Goiânia-GO, Brazil. METHODS: Data from 117 patients, involving 19 cases of IFIs, were collected. The collected data included diagnosis methods, demographics, clinical characteristics, and in vitro susceptibility to different antifungal agents. Among the 19 cases, 12 were classified as proven IFI and 7 as probable invasive aspergillosis with detection of galactomannan in blood and presence of lung infiltrates in radiographic images. Logistic regression analysis showed that the proven and probable IFIs were associated with increased risk of death. Statistical analysis demonstrated that age, sex, and underlying disease were not independently associated with risk of death in IFI patients. RESULTS: Most bloodstream isolates of Candida spp. exhibited low minimum inhibitory concentrations (MICs) to all antifungal agents tested. Voriconazole and amphotericin had the lowest MICs for Aspergillus spp. and Fusarium spp., but Fusarium spp. showed the least susceptibility to all antifungals tested. Amphotericin B, fluconazole, and itraconazole were found to be inactive in vitro against Acremonium kiliense; but this fungus was sensitive to voriconazole. CONCLUSIONS: Considering the high number of IFI cases, with crude mortality rate of 6%, we could conclude that IFIs remain a common infection in patients with hematological malignancies and underdiagnosed ante mortem. Thus, IFIs should be monitored closely.

Successful therapy in unusual generalized Dermatophilus congolensis infection in a calf based on modified in vitro disk diffusion test / Sucesso no tratamento de infecção generalizada em bezerra por Dermatophilus congolensis baseado em teste in vitro modificado de difusão com discos

Bovine dermatophilosis is a dermatitis characterized by typical focal or localized lesions with "paintbrush" aspect and occasionally as disseminated cutaneous disease. We report the case of a one-year-old Nelore female with history of chronic cutaneous disseminated lesions that appeared immediately after a rainfall period. Serous to purulent exudates, hair with tufted appearance, hyperkeratotic, non-pruritic, hardened, yellowish to brown, and coalescent crusty lesions were observed distributed all over its body. Removal of the crusts revealed ulcerated or hemorrhagic areas, with irregular elevated crusts like "paintbrush". Microbiological diagnosis enabled the identification of a microorganism, the Dermatophilus congolensis. Despite disseminated and chronic lesions, we obtained a successful therapy with parenteral therapy using long-acting tetracycline based on modified in vitro disk diffusion test. The present report highlights success therapy in uncommon generalized bovine dermatophilosis with selection of first-choice drugs based on modified in vitro susceptibility test, and need of responsible use of antimicrobials in livestock.(AU)

A dermatofilose bovina é uma dermatite caracterizada por lesões focais ou localizadas com aspecto de "pincel" e, ocasionalmente, como lesão cutânea disseminada. Relata-se o caso de uma fêmea bovina de um ano de idade, que foi atendida apresentando história de lesões cutâneas crônicas imediatamente após um período de alta pluviosidade. Ao exame clínico, lesões serosas a purulentas, com hiperqueratose, coalescentes, não pruriginosas, ressecadas, de coloração amarelada à acinzentada foram observadas distribuídas de modo generalizado pelo animal. A remoção das crostas revelou áreas ulceradas ou hemorrágicas, com crostas irregulares e elevadas semelhantes a "pincel". O diagnóstico microbiológico possibilitou a identificação do micro-organismo Dermatophilus congolensis. Apesar das lesões disseminadas e crônicas, a cura do animal foi obtida com tratamento parenteral usando oxitetraciclina de longa duração, baseado em teste in vitro de sensibilidade microbiana modificado. O presente relato ressalta o sucesso no tratamento de caso incomum de lesões generalizadas de dermatofilose bovina com respaldo de teste in vitro de sensibilidade modificado, bem como a necessidade do uso responsável de antimicrobianos em animais de produção.(AU)

Susceptibility and molecular characterization of Candida species from patients with vulvovaginitis

Abstract Vulvovaginal candidiasis affects women of reproductive age, which represents approximately 15–25% of vaginitis cases. The present study aimed to isolate and characterize yeast from the patients irrespective of the presentation of clinical symptoms. The isolates were subjected to in vitro susceptibility profile and characterization by molecular markers, which intended to assess the distribution of species. A total of 40 isolates were obtained and identified through the CHROMagar, API20aux and by ITS and D1/D2 regions sequencing of DNAr gene. Candida albicans strains were genotyped by the ABC system and the isolates were divided into two genotypic groups. The identity of the C. albicans, C. glabrata, C. guilliermondii, C. kefyr and Saccharomyces cerevisiae isolates was confirmed by the multilocus analysis. The strains of Candida, isolated from patients with complications, were found to be resistant to nystatin but sensitive to fluconazole, amphotericin B and ketoconazole, as observed by in vitro sensitivity profile. The isolates from asymptomatic patients, i.e., the colonized group, showed a dose-dependent sensitivity to the anti-fungal agents, fluconazole and amphotericin B. However, the isolates of C. albicans that belong to distinct genotypic groups showed the same in vitro susceptibility profile.

Frecuencia y perfil de sensibilidad in vitro de aislamientos del Complejo Candida parapsilosis provenientes de pacientes con candidemias / Frequency and in vitro susceptibility profile of Candida parapsilosis Complex isolates from patients with candidemia

El objetivo de este trabajo fue conocer la frecuencia y el perfil de sensibilidad in vitro de aislamientos del Complejo Candida parapsilosis provenientes de casos de candidemias. Se estudiaron 754 cepas (Periodo 2008-2011), de la Red de Vigilancia de Candidemia del Instituto Nacional de Higiene “Rafael Rangel”. La identificación de las cepas se realizó por pruebas fenotípicas. La sensibilidad in vitro a los antifúngicos se evaluó por el método de Etest® y se determinó la concentración mínima inhibitoria a anfotericina B (AB), caspofungina (CS), fluconazol (FZ), y voriconazol (VZ). Se calcularon los puntos de corte epidemiológicos (PCE) y los rangos de cepas salvajes (PS) para cada antifúngico. El 43,6% de las cepas (n=328) fueron identificadas como Complejo C. parapsilosis; todas fueron sensibles a AB y presentaron bajos porcentajes de resistencia a FZ (4,3%), VZ (1,2%) y CS (0,6%). Los PCE y los rangos de PS (en µg/mL) fueron: FZ: 2/0,03-2; VZ y AB: 0,06/0,002-0,06 y CS: 0,5/0,002-0,5 respectivamente. Los resultados de este estudio aportaron información importante sobre el comportamiento del Complejo C. parapsilosis frente a los antifúngicos más utilizados en el tratamiento de las candidemias.

The aim of this study was to determine the frequency and in vitro susceptibility profile of Candida parapsilosis Complex isolates from patients with candidemia. Seven hundred and fifty four (754) strains (Period 2008-2011), from the Candidemia Surveillance Network of the Instituto Nacional de Higiene “Rafael Rangel” were studied. The strains identification was performed by phenotypic methods. In vitro antifungal susceptibility was evaluated by the Etest® method and minimum inhibitory concentration for amphotericin B (AB), caspofungin (CS), fluconazole (FZ), and voriconazole (VZ) was determined. Epidemiological cut off values (ECV) and ranges for wild type strains (WT) were also calculated for each antifungal. Forty three point six (43.6%) of the isolates (n=328) belonged to C. parapsilosis Complex; all of them were susceptible to AB and showed low resistance percentages to FZ (4.3%), VZ (1.2%) and CS (0.6%). The ECV and WT strains ranges (in mcg/mL) were: FZ: 2/0.03-2; VZ and AB: 0.06/0.002-0.06 and CS: 0.5/0.002-0.5 respectively. The results of this study provided important information about the behavior of the C. parapsilosis Complex against the most commonly antifungal agents used for the treatment of candidemias.

Susceptibilidad in vitro a cinco antifúngicos de aislamientos del Complejo Fusarium solani provenientes de úlceras corneales / In vitro susceptibility to five antifungals of Fusarium solani Complex isolates obtained from corneal ulcers

El Complejo Fusarium solani (CFS) se encuentra distribuido en la naturaleza, causando un amplio espectro de infecciones en los humanos, desde superficiales, como la queratitis, hasta infecciones fúngicas invasoras, caracterizándose por su resistencia a los antimicóticos. El objetivo de esta investigación fue determinar la susceptibilidad in vitro del CFS frente a cinco antifúngicos. Se utilizaron 30 aislados obtenidos de úlceras corneales provenientes de la colección de cultivos del Departamento de Micología del Instituto Nacional de Higiene “Rafael Rangel” y se siguió el protocolo descrito en el documento M38-A2 del Instituto de Estándares Clínicos y de Laboratorio (CLSI), determinando las Concentraciones Mínimas Inhibitorias (CMI) por microdilución en caldo para anfotericina B, itraconazol, posaconazol, voriconazol y fluconazol. En general, todas las drogas presentaron CMI elevadas, siendo voriconazol y anfotericina B los antifúngicos que exhibieron mejor actividad, mientras que itraconazol, posaconazol y fluconazol mostraron actividad nula. Los resultados de este estudio aportaron información importante sobre el comportamiento del CFS frente a los antifúngicos de uso común en la práctica clínica por primera vez en Venezuela. Es imprescindible que el médico conozca la actividad de estas drogas para poder tomar decisiones y orientar una conducta terapéutica adecuada.

The Fusarium solani Complex (FSC) is distributed in nature, producing a wide spectrum of infections in humans, from superficial ones such as keratitis, to invasive fungal infections, characterized by their resistance to antimycotics. The purpose of this investigation was to determine the in vitro susceptibility of the FSC to five antifungals. We used 30 isolates obtained from corneal ulcers kept at the culture collection of the Instituto Nacional de Higiene “Rafael Rangel” and we followed the protocol described in the M38-A2 document of the Clinical and Laboratory Standards Institute (CLSI) determining the Minimal Inhibitory Concentrations (MIC) by broth microdilution for amphotericin B, itraconazole, posaconazole, voriconazole and fluconazole. In general, all the drugs presented high MICs, voriconazole and amphotericin B being the antifungals which showed the best activity, while itraconazole, posaconazole and fluconazole showed a null activity. The results of this study provided, for the first time in Venezuela, important information about the behavior of the FSC towards commonly used antifungals. It is mandatory that physicians know the activity of these drugs in order to be able to take decisions and devise an guide an appropriate therapeutic management.

Risk factors associated with the antimicrobial resistance of Staphylococcus aureus isolated from bovine mastitis / Fatores de risco associados com a resistência antimicrobiana de Staphylococcus aureus isolados da mastite bovina

The objective of this study was to evaluate herd management practices and mastitis treatment procedures as risk factors associated with Staphylococcus aureus antimicrobial resistance. For this study, 13 herds were selected to participate in the study to evaluate the association between their management practices and mastitis treatment procedures and in vitro antimicrobial susceptibility. A total of 1069 composite milk samples were collected aseptically from the selected cows in four different periods over two years. The samples were used for microbiological culturing of S. aureus isolates and evaluation of their antimicrobial susceptibility. A total of 756 samples (70.7%) were culture-positive, and S. aureus comprised 27.77% (n=210) of the isolates. The S. aureus isolates were tested using the disk-diffusion susceptibility assay with the following antimicrobials: ampicillin 10mg; clindamycin 2μg; penicillin 1mg; ceftiofur 30μg; gentamicin 10mg; sulfa-trimethoprim 25μg; enrofloxacin 5μg; sulfonamide 300μg; tetracycline 30μg; oxacillin 1mg; cephalothin 30μg and erythromycin 5μg. The variables that were significantly associated with S. aureus resistance were as follows: the treatment of clinical mastitis for ampicillin (OR=2.18), dry cow treatment for enrofloxacin (OR=2.11) and not sending milk samples for microbiological culture and susceptibility tests, for ampicillin (OR=2.57) and penicillin (OR=4.69). In conclusion, the identification of risk factors for S. aureus resistance against various mastitis antimicrobials is an important information that may help in practical recommendations for prudent use of antimicrobial in milk production.

Objetivou-se com este estudo avaliar os fatores de risco associados às práticas de manejo e tratamento de mastite e a resistência aos antimicrobianos de Staphylococcus aureus isolados de vacas com mastite. Foram selecionados para o presente estudo 13 rebanhos localizados na região de Pirassununga/SP. Foi aplicado um questionário contendo informações para o levantamento de fatores de risco relacionados à resistência aos antimicrobianos e às práticas de manejo e tratamento de mastite. Após a seleção dos rebanhos e aplicação dos questionários, foram utilizados 210 isolados de S. aureus de amostras compostas de leite coletadas durante 24 meses, em quatro períodos, para realização dos testes de resistência. Os antimicrobianos testados foram: ampicilina 10µg, clindamicina 2µg, penicilina 1µg, eftiofour 30µg, gentamicina 10µg, sulfatrimetropin 25µg, enrofloxacina 5µg, sulfonamida 300µg, tetraciclina 30µg, oxacilina 1µg, cefalotina 30µg e eritromicina 5µg. As variáveis que foram significativamente associadas à resistência de S. aureus foram: o tratamento da mastite clínica para ampicilina (OR = 2,18), o tratamento da vaca seca para enrofloxacina (OR=2,11), e o não envio de amostras de leite para a cultura microbiológica e testes de sensibilidade, para ampicilina (OR=2,57) e penicilina (OR=4,69). Em conclusão, a identificação dos fatores de risco para a resistência S. aureus frente aos principais agentes antimicrobianos, utilizados para tratamento da mastite, pode auxiliar o estabelecimento do uso prudente de antimicrobianos na produção de leite.

In vitro antimalarial activity of tigecycline against Plasmodium falciparum culture-adapted reference strains and clinical isolates from the Brazilian Amazon

Introduction: We evaluated the in vitro antimalarial activity of tigecycline as an alternative drug for the treatment of severe malaria. Methods: A chloroquine-sensitive Plasmodium falciparum reference strain, a chloroquine-resistant reference strain, and three clinical isolates were tested for in vitro susceptibility to tigecycline. A histidine-rich protein in vitro assay was used to evaluate antimalarial activity. Results: The geometric-mean 50% effective concentration (EC50%) of tigecycline was 535.5 nM (confidence interval (CI): 344.3-726.8). No significant correlation was found between the EC50% of tigecycline and that of any other tested antimalarial drug. Conclusions: Tigecycline may represent an alternative drug for the treatment of patients with severe malaria. .

Molecular analysis and dimorphism of azole-susceptible and resistant Candida albicans isolates / Análise molecular e dimorfismo de isolados de Candida albicans suscetíveis e resistentes aos azólicos

INTRODUCTION: Candida albicans is responsible for superficial or systemic infections known as candidiasis, which may be found in infected tissue as unicellular budding yeasts, hyphae, or pseudohyphae. In this study, the effects of both fluconazole and itraconazole antifungal agents on the hyphal formation and genotypic characterization of C. albicans isolates classified as either susceptible or resistant were investigated. METHODS: The hyphal production of five C. albicans isolates under the action of antifungal agents was investigated by culturing yeast on growth medium and on hyphal induction medium. The genotypic characterization was carried out for 13 isolates of C. albicans using the random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) method. RESULTS: The dimorphism analysis showed that the hyphal formation was higher in resistant than in the susceptible isolates to both azoles. The RAPD-PCR method identified the formation of two different groups. In group A, four resistant and two susceptible isolates were clustered, and in group B, one resistant and six susceptible isolates were clustered. CONCLUSIONS: Considering that hyphal formation was higher in resistant isolates in the presence of azole drugs, we confirmed that the hyphal production is closely related to susceptibility to azoles. These drugs may affect the morphogenesis of C. albicans depending on their susceptibility to these drugs. In relation to RAPD-PCR, most resistant isolates classified in group A and susceptible isolates in group B demonstrated that this method presented a similar standard between the two groups, suggesting that by this technique, a strong correlation between genotypes and fluconazole-resistant samples may be found.

INTRODUÇÃO: Candida albicans é responsável por infecções superficiais ou sistêmicas conhecidas como candidíase, encontrada em tecidos infectados na forma de leveduras brotantes unicelulares, hifas ou pseudohifas. Neste estudo, os efeitos de agentes antifúngicos como o fluconazol e o itraconazol sobre a formação de hifas e caracterização genotípica de isolados de C. albicans suscetíveis ou resistentes foram investigados. MÉTODOS: A produção de hifas de cinco isolados de C. albicans, sob a ação de antifúngicos foi investigada pelo cultivo da levedura em meios de crescimento e de indução de hifas. A caracterização genotípica foi realizada para 13 isolados de C. albicans pelo método de RAPD-PCR. RESULTADOS: A análise do dimorfismo mostrou que a formação de hifas foi maior nos isolados resistentes do que nos suscetíveis aos antifúngicos. O método de RAPD-PCR identificou a formação de dois diferentes grupos. No grupo A, foram agrupados quatro isolados resistentes e dois suscetíveis e no grupo B um resistente e seis suscetíveis. CONCLUSÕES: Considerando que a formação hifal foi maior em isolados resistentes na presença de azólicos, concluimos que a produção hifal está muito relacionada a suscetibilidade a estes fámacos. Estes antifúngicos podem alterar a morfologia de C. albicans em dependência da sua suscetibilidade. No método de RAPD-PCR, o encontro da maioria dos isolados resistentes classificados como pertencentes ao grupo A e suscetíveis ao grupo B demonstrou que este método apresentou um padrão semelhante entre os dois grupos, sugerindo que por este método pode ser detectado uma estreita correlação entre genótipos e amostras resistentes ao fluconazol.

Polymyxins: Antimicrobial susceptibility concerns and therapeutic options.

The increasing prevalence of multidrug-resistant nosocomial pathogens such as Acinetobacter baumannii, Pseudomonas aeruginosa and Klebsiella pneumoniae poses a great challenge to the treating physicians. The paucity of newer effective antimicrobials has led to renewed interest in the polymyxin group of drugs, as a last resort for treatment of gram-negative bacterial infections. There is a dearth of information on the pharmacological properties of colistin, leading to difficulties in selecting the right dose, dosing interval, and route of administration for treatment, especially in critically-ill patients. The increasing use of colistin over the last few years necessitates the need for accurate and reliable in vitro susceptibility testing methods. Development of heteroresistant strains as a result of colistin monotherapy is also a growing concern. There is a compelling need from the clinicians to provide options for probable and possible colistin combination therapy for multidrug-resistant bacterial infections in the ICU setting. Newer combination drug synergy determination tests are being developed and reported. There are no standardized recommendations from antimicrobial susceptibility testing reference agencies for the testing and interpretation of these drug combinations. Comparison and analysis of these reported methodologies may help to understand and assist the microbiologist to choose the best method that produces accurate results at the earliest. This will help clinicians to select the appropriate combination therapy. In this era of multidrug resistance it is important for the microbiology laboratory to be prepared, by default, to provide timely synergistic susceptibility results in addition to routine susceptibility, if warranted. Not as a favour or at request, but as a responsibility.

Estudio comparativo de medición entre lecturas visuales y espectrofotométricas en pruebas de susceptibilidad in vitro de aislamientos de candida / Comparative study of measure visual and spectrophotometer lectures in a in vitro antifungal susceptibility testing in candida isolates

Se realizó un estudio de susceptibilidad in vitro frente a itraconazol, ketoconazol y clotrimazol de 144 cepas de Candida, conservadas y previamente identificadas, aisladas de la cavidad oral de pacientes infectados por el virus de inmunodeficiencia humana (VIH) con cuadros clínicos de candidiasis orofaríngea (COF). El estudio se llevó a cabo mediante dos metodologías; la primera, utilizando los requerimientos del Clinical and Laboratory Standard Institute (CLSI), en el cual está establecida la lectura visual para determinar los patrones de susceptibilidad; y la segunda, mediante la propuesta de la European Committee on Antibiotic Susceptibility Testing, el cual tiene fijado la lectura espectrofotométrica para eliminar las posibles subjetividades de la metodología del CLSI. Los resultados obtenidos mediante ambas lecturas no mostraron diferencias mayores a dos diluciones en los valores de concentración mínima inhibitoria, y demostraron que ambos métodos se correlacionan y es importante para aquellos laboratorios de pocos recursos económicos.

A study of in vitro susceptibility was realized opposite to itraconazol, ketoconazol and clotrimazol of 144 strains of Candida, conserved and previously identified, isolated of the oral cavity of patients infected by the virus of human immunodeficiency (VIH) with clinical pictures of oropharingeal candidiasis (COF). The study was realized by means of two methodologies; the first one, using the requests of the Clinical and Laboratory Standard Institute (CLSI), in which the visual reading is established to determine the patterns of susceptibility; and the second one, by means of the proposal of the European Committee on Antibiotic Susceptibility Testing, which has fixed the spectrophotometric to eliminate the possible subjectivities of the methodology of the CLSI. The results obtained by means of both readings did not show differences bigger than two dilutions in the values of minimal inhibitory concentration, demonstrating that both methods are correlated and it is important for those laboratories of few economic resources.

Caracterización fenotípica y factores de virulencia en cepas de Aeromonas aisladas de pacientes con enfermedad diarreica aguda en Cuba / Phenotypic characteristics and virulence factors in Aeromonas strains isolated from patients with diarrheic disease in Cuba

Fifty four strains of Aeromonas spp were isolated from patients with acute diarrheic episodes by using Aerokey II and Aeroesquema methods. In vitro antimicrobial susceptibility and virulence factors were analyzed. The most frequently isolated specie was Aeromonas caviae. Over 75 percent of strains exhibited resistance to penicillins and ce-phalosporins; for the other antibiotic groups resistance was under 20 percent. Twenty six strains (48.1 percent) were multiresist-ant. At least one virulence factor among those evaluated in the study was present in 53 (98.1 percent) of the 54 strains.

Se identificaron 54 cepas de Aeromonas aisladas de pacientes con enfermedad diarreica aguda mediante los métodos Aerokey II y Aeroesquema. Se determinó la susceptibilidad antimicrobiana y algunos factores de virulencia. La especie encontrada en mayor frecuencia fue Aeromonas caviae. Se observaron valores de resistencia por sobre 75 por ciento para penicilinas y cefalotina; para el resto de los antimicrobianos estos valores se encontraron bajo 20 por ciento>; 26 cepas (48,1 por ciento) presentaron multi-resistencia. Al menos un factor de virulencia de los investigados estuvo presente en 53 (98,1 por ciento) de las 54 cepas analizadas.

Susceptibilidad a la nistatina de aislamientos bucales de Candida y su correlación con la respuesta al tratamiento / Susceptibility to nystatin of oral Candida isolates and its correlation with the response to treatment

INTRODUCCIÓN: la candidiasis bucofaríngea es un marcador precoz de progresión a sida en los pacientes seropositivos al VIH sin tratamiento antirretroviral, así como un indicador de no adherencia al tratamiento antirretroviral o su posible fracaso. OBJETIVO: determinar la susceptibilidad frente a nistatina de las cepas de Candida aisladas de la cavidad bucal de pacientes seropositivos al VIH, y correlacionar los resultados con la respuesta al tratamiento. MÉTODOS: se realizó un estudio de susceptibilidad in vitro frente a nistatina para 104 cepas aisladas de lesiones de candidiasis bucofaríngea de 97 pacientes VIH/sida que habían participado, como voluntarios, en un ensayo clínico para evaluar la respuesta a cuatro drogas. De los 104 aislamientos, 58 fueron obtenidos antes de iniciar los esquemas de tratamientos y 46 una vez finalizado estos. Se procedió según el micrométodo el documento M27-A3 del Clinical and Laboratory Standards Institute. RESULTADOS: todas las cepas se comportaron como sensibles a la nistatina, con valores de concentración mínima inhibitoria (CMI) menores que 16 µg/mL, independientemente del momento de su aislamiento y de la droga administrada a los pacientes. En el grupo de pacientes tratados con nistatina, los rangos de CMI80 (1-2 µg/mL), las medias geométricas y los valores acumulativos de la droga frente a los aislamientos de Candida albicans obtenidos antes de iniciar el tratamiento, resultaron similares a los obtenidos frente a las cepas recobradas al finalizar este. De todas las especies identificadas, C. lusitaniae fue la que alcanzó el valor absoluto de CMI más elevado, 4 µg/mL. CONCLUSIONES: la correlación entre la evolución clínica de los pacientes y los resultados de los estudios de susceptibilidad de sus aislamientos in vitro, fue buena para los pacientes con evolución favorable, pero no para predecir posibles fracasos terapéuticos.

INTRODUCTION: oropharyngeal candidiasis is an early marker of progression to AIDS in HIV-positive patients and an indicator of non-adherence of treatment or possible failure in patients undergoing anti-retroviral therapy. OBJECTIVE: to determine the in vitro susceptibility to nystatin in Candida strains isolated from the oral cavity of HIV-positive patients, and to correlate the results with the response to treatment. METHODS: a study of in vitro susceptibility to nystatin was conducted in 104 oral isolates from 97 HIV/AIDS patients, who participated as volunteers in a clinical trial to evaluate the response to four antifungal drugs. Fifty-eight of the 104 isolates were obtained before starting treatment and the remaining 46 at the end of therapy. Broth microdilution method was performed according to the document M27-A3 of the Clinical and Laboratory Standards Institute. RESULTS: one hundred percent of the strains were susceptible to nystatin, with minimum inhibitory concentration (MIC) values of <16 µg/mL, regardless of the time of isolation and of the drug administered to patients. In the group of patients treated with nystatin, the MIC80 ranges (1-2 µg/mL), geometric mean and cumulative values of the drug against Candida albicans isolates before starting treatment were similar to those obtained against strains recovered at the end. Among the identified species, C. lusitaniae reached the highest absolute MIC value (4 µg/mL). CONCLUSIONS: the correlation between the clinical evolution and the results of in vitro susceptibility tests was good in patients with a favorable outcome; however, it did not allow predicting possible treatment failure.

Aspectos micológicos e suscetibilidade in vitro de leveduras do gênero Candida em pacientes HIV-positivos provenientes do Estado de Mato Grosso / Mycological aspects and susceptibility in vitro the yeast of the genus Candida from HIV-positive patients in the State of Mato Grosso

INTRODUÇÃO: A candidíase é uma das infecções fúngicas mais frequentes entre os pacientes infectados pelo vírus da imunodeficiência humana. O presente estudo objetivou a caracterização das leveduras do gênero Candida de distintas amostras clínicas, provenientes de pacientes HIV - positivos, assim como a determinação do perfil de suscetibilidade in vitro a cinco drogas antifúngicas. MÉTODOS: A caracterização dos isolados de Candida sp foi realizada através da metodologia clássica, testes bioquímicos (zimograma e auxanograma) e morfológicos (prova do tubo germinativo e microcultivo em lâmina). Também, foram realizadas a técnica genotípica (PCR) e identificação pelo método comercial API 20C AUX (BioMeriéux). Para a determinação do perfil de suscetibilidade in vitro, foram utilizadas cinco drogas antifúngicas (cetoconazol, fluconazol, itraconazol, voriconazol e anfotericina B), através do método comercialmente disponível - Etest. RESULTADOS: Foram identificados 105 isolados de leveduras do gênero Candida provenientes de 102 pacientes infectados pelo vírus HIV. Destes, foram caracterizadas 82 (78,1 por cento) Candida albicans, 8 (7,6 por cento) Candida parapsilosis, 8 (7,6 por cento) Candida tropicalis, 4 (3,8 por cento) Candida krusei, 2 (1,9 por cento) Candida glabrata e 1 (1 por cento) Candida guilliermondii. CONCLUSÕES: Considerando o perfil geral de sensibilidade, 60 por cento dos isolados foram suscetíveis a todos os antifúngicos testados, porém as espécies C. tropicalis e C. krusei demonstraram uma tendência a valores mais elevados de CIMs para os azóis do que os encontrados paraC. albicans, sugerindo resistência.

INTRODUCTION: Candidiasis is one of the most common fungal infections among patients infected by human immunodeficiency virus. The present study aimed to characterize yeasts of the genus Candida from distinct clinical samples from HIV-positive patients and determine the in vitro susceptibility profile to five antifungal drugs. METHODS: Characterization of Candida sp was achieved using the classic methodology: biochemical (zymogram and auxanogram) and micromorphology (germinative tube growth test and slide microculture) tests. Genotypic technique (PCR) and identification by the commercial method API 20C AUX (Biomeriéux) were also performed. To determine the in vitro susceptibility profile, five antifungal drugs were used (ketoconazole, fluconazole, itraconazole, voriconazole and amphotericin-B) following a commercially available method, the Etest. RESULTS: The procedure isolated 105 yeasts of the genus Candida from 102 HIV-infected patients. Of these, 82 (78.1 percent) were characterized as Candida albicans, 8 (7.6 percent) as C. parapsilosi s, 8 (7.6 percent) C. tropicalis, 4 (3.8 percent) C. krusei, 2 (1.9 percent) C. glabrata, and 1 (1 percent) as C. guiilliermondii. CONCLUSIONS: Considering the general profile of sensitivity, 60 percent of isolates were susceptible to all the antifungal drugs tested; however, the species C. tropicalis and C. krusei showed a tendency toward higher MICs to azoles than those obtained for C. albicans, suggesting resistance.

Comparison of in vitro activity of five antifungal agents against dermatophytes, using the agar dilution and broth microdilution methods / Comparação da atividade in vitro de cinco agentes antifúngicos para dermatófitos, usando os métodos de diluição em ágar e microdiluição em caldo

The purpose of this study was to compare the agar dilution and broth microdilution methods for determining the minimum inhibitory concentration (MIC) of fluconazole, itraconazole, ketoconazole, griseofulvin and terbinafine for 60 dermatophyte samples belonging to the species Trichophyton rubrum, Trichophyton mentagrophytes and Microsporum canis. The percentage agreement between the two methods, for all the isolates with < 2 dilutions that were tested was 91.6 percent for ketoconazole and griseofulvin, 88.3 percent for itraconazole, 81.6 percent for terbinafine and 73.3 percent for fluconazole. One hundred percent agreement was obtained for Trichophyton mentagrophytes isolates evaluated with ketoconazole and griseofulvin. Thus, until a reference method for testing the in vitro susceptibility of dermatophytes is standardized, the similarity of the results between the two methods means that the agar dilution method may be useful for susceptibility testing on these filamentous fungi.

O propósito do presente trabalho foi comparar os métodos de diluição em ágar e diluição em caldo para a determinação de concentração inibitória mínima de fluconazol, itraconazol, cetoconazol, griseofulvina e terbinafina para 60 amostras de dermatófitos pertencentes às espécies, Trichophyton rubrum, Trichophyton. mentagrophytes e Microsporum canis. A porcentagem de acordo entre os dois métodos para todos os isolados testados considerando-se valores < 2 diluições, foram de 91,6 por cento para cetoconazol e para griseofulvina, de 88,3 por cento para itraconazol, de 81,6 por cento para terbinafina e de 73,3 por cento para fluconazol. Uma concordância de 100 por cento foi obtido para isolados de Trichophyton mentagrophytes avaliados com cetoconazol e griseofulvina. Desta forma, até que um método de referência seja padronizado para testar a suscetibilidade in vitro para os dermatófitos, os resultados semelhantes encontrados para os dois métodos fazem com que o método de diluição em ágar possa ser útil no teste de suscetibilidade para estes fungos filamentosos.

Susceptibilidad in vitro frente a fluconazol y voriconazol de cepas de Cryptococcus aisladas en Cuba / In vitro susceptibility of isolated Cryptococcus strains to fluconazole and voriconazole

Las infecciones fúngicas invasoras se han incrementado como consecuencia de los avances de la medicina en el ßrea de la transplantología, el tratamiento del cßncer, las enfermedades autoinmunes y el sida, fundamentalmente. La criptococosis, antes infrecuente, ocupa ahora el tercer lugar en la incidencia de micosis graves en este último grupo de pacientes y constituye además la de mayor letalidad. Hasta el momento existen pocos reportes sobre el desarrollo de resistencia de Cryptococcus a los antifúngicos, pero se prevé que el amplio uso de fluconazol pudiera favorecer la emergencia de cepas menos sensibles. Dada la importancia de mantener la vigilancia de la susceptibilidad a los antifúngicos y de poder evaluar nuevas opciones terapéuticas, se determinó la actividad in vitro del fluconazol y el voriconazol frente a un amplio número de cepas de Cryptococcus aisladas a partir de muestras clínicas y ambientales mediante el método de Etest. Se determinó la susceptibilidad in vitro mediante Etest frente a fluconazol y voriconazol de 159 cepas de Cryptococcus (117 obtenidas a partir de muestras clínicas y 42 de fuentes ambientales de la región centro-occidental de Cuba). Los aislamientos estudiados mostraron valores bajos de concentraciones mínimas inhibitorias, sobre todo frente a voriconazol. Los rangos, las medias geométricas y las modas fueron de 0,031-128, 4,631 y 3 mg/mL para fluconazol y 0,003-8, 0,02 y 0,016 mg/mL para voriconazol, respectivamente. Aunque de forma general los valores de concentraciones mínimas inhibitorias fueron mßs elevados en los aislamientos ambientales, no se encontraron diferencias estadísticamente significativas (p= 0,05) entre las cepas de acuerdo con el origen (clínica/ambiental) o la procedencia geográfica. Se señala la importancia de continuar este tipo de estudio para la detección de la aparición de resistencia a las drogas más empleadas en la terapéutica anticriptocósica, así como en la búsqueda de nuevas estrategias para...

Invasive fungal infections have increased as a result of medical advances in the sphere of transplantation, cancer treatment, autoimmune diseases and AIDS fundamentally. Cryptococcus disease, infrequent before, holds the third place in the incidence of severe mycosis in AIDS patients and it is also the most lethal. Up to the present, there exist few reports on the development of Cryptococcus resistance to antifungal products, however, it is foreseen that wide use of fluconazole might encourage the emergence of less susceptible strains. Given the importance of surveillance of antifungal susceptibility and of the evaluation of new therapeutical options, in vitro action of fluconazole and voriconazole on a high number of isolated Cryptococcus strains was determined through Etest method on clinical and environmental samples. In vitro susceptibility to fluconazole and voriconazole of 159 Cryptococcus strains (117 from clinical samples and 42 from environmental sources of Central and Western regions in Cuba) was determined through Etest method. The studied isolates showed low MIC values, mainly susceptibility to voriconazole. Ranges, geometric means and modes were 0.031-128, 4.631 y 3 mg/mL for fluconazole and 0.003-8, 0.02 y 0.016 mg/mL for voriconazole, respectively. Although MIC values were generally higher than those of environmental isolates, there were no statistically significant differences (p= 0.05) among strains according to their origin (clinical/environmental) or to their geographical location. This paper points out the importance of continuing this type of study for the detection of resistance to the most used drugs in anticryptococcus disease therapy as well as in the search of new strategies to treat this infection.

Sensibilidad de cepas de Pseudomonas aeruginosa frente a beta-lactámicos / Beta-lactams susceptibility in Pseudomonas aeruginosa strains

Los pacientes portadores de enfermedad fibroquística presentan manifestaciones respiratorias a repetición o crónicas, las cuales se acompañan de infecciones bacterianas. Pseudomonas aeruginosa es la especie bacteriana que se aísla con mayor frecuencia en estos pacientes a partir de muestras de origen respiratorio. Teniendo en cuenta que las infecciones por esta bacteria son difíciles de tratar y que la terapia usualmente empleada es la combinación de un aminoglucósido con un antibiótico beta-lactámico, se analizó el comportamiento por el método de microdilución en caldo de 54 cepas aisladas de pacientes con fibrosis quística frente a cuatro beta-lactámicos (carbenicilina, azlocilina, cefotaxima, ceftazidima), y se observó que el 85% de las cepas fueron sensibles a ceftazidima, 83% a azlocilina, 37% sensibles a carbenicilina, y solamente un 20% de las cepas estudiadas fueron sensibles a cefotaxima. Los resultados evidencian que ceftazidima continúa siendo un beta-lactámico de elección en el tratamiento de las infecciones causadas por Pseudomonas aeruginosa en pacientes con fibrosis quística.

Repetitive or chronic respiratory infections are common in cystic fibrosis patients, usually due to Pseudomonas aeruginosa. The recommended therapy is the combination of one aminoglycoside with a beta-lactam antibiotic. For this reason, the susceptibility of 54 Pseudomonas aeruginosa isolated obtained from the respiratory tract of cystic fibrosis patients to four beta-lactams was analysed. The study was performed by broth microdilution. Ceftazidime was the most active agent against Pseudomonas aeruginosa (85% of the total strains being susceptible).. Eighty three percent of the strains were inhibited by azlocillin and 37% by carbenicillin. Only 20% of all studied strains were susceptible to cefotaxime.

Suscetibilidade in vitro a antibióticos de cepas de Staphylococcus spp e Micrococcus spp isoladas a partir de mucosa oral de macacos-pregos (Cebus apella) mantidos em cativeiro / In vitro susceptibility to antibiotics in Staphylococcus spp e Micrococcus spp strains isolated from oral mucosae of captive capuchin monkeys (Cebus apella)

O estudo foi realizado com 29 macacos-pregos (Cebus apella). Foram colhidas 50 amostras de suabe da mucosa oral, junto à transição muco-gengival maxilar, com auxílio de zaragatoas esterilizadas, embebidas em caldo Brain Heart Infusion (BHI). Todos os animais foram submetidos a exame clínico para avaliação periodontal. As amostras obtidas foram cultivadas em meios apropriados: caldo simples, caldo BHI, e ágar sangue para o isolamento de cocos Gram-positivos aeróbios da família Micrococcaceae. Para sua classificação utilizou-se as provas de catalase, Staphy-test (teste rápido para caracterização de Staphylococcus aureus) e sensibilidade à bacitracina. Foram identificados 73,1 por cento de Staphylococcus spp; 15,4 por cento de Staphylococcus aureus; e 11,5 por cento Micrococcus spp. As cepas isoladas foram testadas em relação à sua suscetibilidade a antibióticos pela técnica de difusão em ágar. Verificou-se para as cepas de Staphylococcus spp, 94,7 por cento de sensibilidade a cefalotina e resistência de 89,5 por cento à penicilina, 97,4 por cento à oxacilina, 55,3 por cento à tetraciclina, 57,9 por cento à clindamicina e 63,2 por cento à amoxicilina. Os dados obtidos demonstraram que a cefalotina foi o antibiótico para o qual as amsotras de Staphylococcus spp estudadas apresentaram, in vitro, maior grau de sensibilidade.

Twenty-nine capuchin monkeys (Cebus apella) were used in this study. Fifty samples of oral mucous membrane were collected in area next to their muco-gingival-maxilar transition using sterilized swabs soaked in Brain Heart Infusion (BHI). All animals were clinically examined for periodontal evaluation. The samples were cultivated in appropriate media, namely: simple broth, BHI broth and, blood agar in order to get aerobic Gram positive cocos, from the Micrococcaceae family, isolated. Catalase test, Staphy-test (a quick-test for Staphylococcus aureus characterization) and bactracin-sensitivity test were the tools employed for their classification. Data were follows: 73.1 percent of them were Staphylococcus spp; 15.4 percent Staphylococcus aureus; and, 11.5 percent Micrococcus spp. The isolated were strains tested for their in vitro susceptible to antibiotics by the agar diffusion technique. Concerning the Staphylococcus spp strains, 94.7 percent were susceptibility to cephalotin; however, 89.5 percent of them were resistant to penicillin; 97.4 percent to oxacilin; 55.3 percent to tetracicline; 57.9 percent to clindamicine; and 63.2 percent to amoxiciline. Staphylococcus spp strains studied presented the highest in vitro sensitivity degree to cephalotin.