Direct identification of microorganisms from thioglycolate broth by MALDI-TOF MS.

We developed an easy MALDI-TOF MS-based assay to identify microorganisms directly from thioglycolate broth. A total of 101 positive thioglycolate broths inoculated with 15 different kinds of samples were evaluated. In 91 samples (90.1%), direct MALDI-TOF MS identifications were the same as those obtained after conventional laboratory procedures including subcultures. In 10 samples misidentified by direct processing, yeasts or mixed cultures grew in the thioglycolate subcultures, or high cellular debris hampered a correct analysis. This rapid method can provide a fast, clinically- relevant species-level identification without disturbing the daily workflow in clinical microbiology laboratories.

First Report of Acute Postoperative Endophthalmitis Caused by Rothia Mucilaginosa after Phacoemulsification.

We aimed at reporting the first case of rapidly progressive acute postoperative endophthalmitis after phacoemulsification cataract surgery in an immunocompetent patient caused by Rothia mucilaginosa. An immunocompetent patient manifested endophthalmitis signs 48 hours after an uncomplicated cataract surgery by phacoemulsification. A bacteria of the family Micrococcaceae was cultured in the vitreous biopsy, namely R. mucilaginosa. The patient did not show a favorable clinical response after vitrectomy and systemic, intravitreal, and topical fortified antibiotics. The patient's eye was very painful, and consequently, it deemed necessary to perform an evisceration. R. mucilaginosa may be an aggressive etiologic agent for postoperative endophthalmitis. Although the isolated R. mucilaginosa was susceptible to empirical treatment, it was impossible to control the infection with standard treatment, probably due to its ability to create a biofilm around the intraocular lens.

Candida nivariensis as a New Emergent Agent of Vulvovaginal Candidiasis: Description of Cases and Review of Published Studies.

Candida nivariensis is a new emergent agent related to human infections in the vaginal tract and other localizations, but the phenotypic characteristics are very similar to Candida glabrata and can be misidentified and underdiagnosed. We described four cases of vulvovaginitis identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and confirmed the results with PCR amplification and sequencing of the entire ITS genomic region (ITS1, ITS2 and 5.8 rRNA). We reinforce the need for new diagnostic tools for the correct identification of yeast infections.

Aneurisma infeccioso de aorta abdominal infrarrenal por Eikenella corrodens / Aortic abdominal aneurysm infection due to Eikenella corrodens

No disponible

Diversity of bla genes and low incidence of CTX-M in plasmid-mediated AmpC-producing Escherichia coli clinical isolates.

The aim of this study was to characterize plasmid-mediated AmpC (pAmpC)-producing Escherichia coli clinical isolates. A total of 101 strains with AmpC-susceptibility pattern were prospectively included. All isolates were tested by multiplex PCR to detect different bla genes. Phylogenetic groups were determined by a multiplex PCR assay. Antimicrobial susceptibility was tested by a microdilution commercial method. Presence of blapAmpC was detected in 79 (78.2%) of the strains; in these pAmpC-producing isolates, blaTEM was detected in 41 (51.9%) strains, blaSHV in 5 (6.3%) strains, blaOXA in 3 (3.8%) strains, and blaCTX-M in 3 (3.8%) strains. blaVIM and blaKPC were detected in one strain. Sixteen strains belonged to phylogroup A, 27 to B1, 20 to B2, and 16 to D. As conclusion, the majority of the strains of E. coli with AmpC-susceptibility pattern are pAmpC positive, although the association of extended-spectrum beta-lactamases (ESBL) and pAmpC is unusual.

Evaluation of the Sensititre Yeast One microdilution method for susceptibility testing of Candida species to anidulafungin, caspofungin, and micafungin

Introducción. Las equinocandinas representan un nuevo grupo de antifúngicos con gran actividad frente a especies de Candida. El propósito de este estudio fue evaluar el método Sensititre Yeast One para determinar la actividad in vitro de anidulafungina, micafungina y caspofungina frente a especies de Candida aisladas de muestras clínicas. Métodos. Un total de 131 cepas de Candida identificadas como: 42 C. albicans, 36 C. glabrata, 21 C. parapsilosis, 12 C. tropicalis, 10 C. krusei, 3 C. guilliermondii, 2 C. famata, 3 C. kefyr, 1 C. lusitaniae, 1 C. zeylanoides y 1 C. lipolytica, fueron ensayadas mediante el método colorimétrico de microdilución Sensititre Yeast One. Se consideraron sensibles las cepas inhibidas por concentraciones <=2 mg/L de anidulafungina, caspofungina o micafungina. Resultados. El 80,1% de las cepas fueron inhibidas por concentraciones <=0,25 mg/L de anidulafungina y micafungina. La actividad de caspofungina fue ligeramente inferior (78,6% de las cepas inhibidas por concentraciones <=0,25 mg/L). El 96,9% de las cepas resultaron sensibles frente a las tres equinocandinas. Dos cepas de C. parapsilosis (9,5%), 1 de C. guilliermondii y 2 de C. famata no mostraron sensibilidad a una o más equinocandinas. Conclusiones. En nuestra serie, anidulafungina, micafungina y caspofungina fueron efectivas frente a C. albicans, C. glabrata, C. tropicalis, C. krusei, C. kefyr, C. lusitaniae y C. lipolytica. El 96,9% de las cepas fueron sensibles a las tres equinocandinas. Se puede afirmar que las equinocandinas tienen una excelente actividad frente a las especies de Candida más frecuentes en infecciones humanas, excepto Candida parapsilosis(AU)

Introduction. Echinocandins represent a new antifungal group with potent activity against Candida species. The purpose of our study was to evaluate the utility of the Sensititre Yeast One method to determine the in vitro activity of anidulafungin, micafungin, and caspofungin against Candida species isolated from clinical specimens. Methods. A total of 131 Candida strains were tested using Sensititre Yeast One colorimetric microdilution method. They belonged to the following species: 42 C. albicans, 36 C. glabrata, 21 C. parapsilosis, 12 C. tropicalis, 10 C. krusei, 3 C. guilliermondii, 2 C. famata, 3 C. kefyr, 1 C. lusitaniae, 1 C. zeylanoides, and 1 C. lipolytica. For being considered susceptible the strains had to be inhibited by concentrations <=2 mg/L of anidulafungin, caspofungin or micafungin. Results. The 80.1% of the strains tested were inhibited by concentrations <=0.25 mg/L of anidulafungin and micafungin. The activity of caspofungin was slightly lower (78.6% of strains inhibited by concentrations <=0.25 mg/L). The 96.9% of strains turned out susceptible to concentrations <=2 mg/L against the three echinocandins. Two strains of C. parapsilosis (9.5%), one of C. guilliermondii, and two of C. famata showed non-susceptible to one or more echinocandins. Conclusions. In our series, anidulafungin, micafungin, and caspofungin were effective against C. albicans, C. glabrata, C. tropicalis, C. krusei, C. kefyr, C. lusitaniae and C. lipolytica. The 96.9% of strains were susceptible to all three echinocandins. Thus, echinocandins are proved to exhibit excellent activity to the Candida species most frequently involved in human infections, except Candida parapsilosis(AU)

Sensibilidad a fluconazol de levaduras de interés clínico: nuevos puntos de corte / Susceptibility to fluconazole of clinical interest yeasts: new breakpoints

Introducción. Recientemente, Pfaller y colaboradores (Drug Resist Updat 2010; 13:180-95), han propuesto nuevos puntos de corte para determinar la sensibilidad in vitro a fluconazol de Candida albicans, C. parapsilosis y C. tropicalis. El objetivo de este trabajo ha sido establecer las variaciones de sensibilidad de estas especies al aplicar estos puntos de corte, en relación con los del Clinical and Laboratory Standards Institute (CLSI). Métodos. Analizamos 112 cepas de Candida: 49 C. albicans, 40 C. parapsilosis y 23 C. tropicalis. La sensibilidad a fluconazol se ensayó por el método Sensititre YeastOne. Los puntos de corte para categorizar la concentración mínima inhibitoria (CMI) fueron los del CLSI y los propuestos por Pfaller y colaboradores. Resultados. Según los criterios del CLSI, todas las cepas fueron sensibles a fluconazol. Las CMI50 y CMI90 fueron 0,5 mg/L y 2 mg/L para C. albicans y C. parapsilosis, 0,5 mg/L y 1 mg/L para C. tropicalis. Con los nuevos criterios, 109 (97%) cepas fueron sensibles. Solamente se apreciaron variaciones en C. albicans (6% sensibles dosis dependientes). Conclusiones. Al aplicar los puntos de corte recomendados por Pfaller y colaboradores, y los del EUCAST, el número de cepas sensibles a fluconazol disminuye en relación con los criterios del CLSI, especialmente de C. albicans(AU)

Introduction. Recently, Pfaller et al (Drug Resist Update 2010; 13:180-95), have proposed new breakpoints for determining the in vitro susceptibility to fluconazole of Candida albicans, C. parapsilosis and C. tropicalis. The aim of this study was to establish the variations in sensitivity of these species applying these breakpoints, in relation to those of the Clinical and Laboratory Standards Institute (CLSI). Methods. We analyzed 112 strains of Candida: 49 C. albicans, 40 C. parapsilosis and 23 C. tropicalis. Susceptibility to fluconazole was performed by the method Sensititre YeastOne. The breakpoints used to determine the minimum inhibitory concentration (MIC) were identified by CLSI and the ones proposed by Pfaller et al. Results. According to the CLSI criteria, all isolates were susceptible to fluconazole. MIC50 and MIC90 were 0.5 mg/L and 2 mg/L for C. albicans and C. parapsilosis, 0.5 mg/L and 1 mg/L for C. tropicalis. With the new criteria, 109 (97%) strains were susceptible. Variations were seen in C. albicans, with 3 strains (6%) susceptible dose-dependent. Conclusions. When applying the breakpoints recommended by Pfaller et al, and EUCAST, the number of fluconazole-susceptible strains decreased according to the CLSI criteria, especially C. albicans(AU)

Evaluation of the Sensititre Yeast One microdilution method for susceptibility testing of Candida species to anidulafungin, caspofungin, and micafungin.

INTRODUCTION: Echinocandins represent a new antifungal group with potent activity against Candida species. The purpose of our study was to evaluate the utility of the Sensititre Yeast One method to determine the in vitro activity of anidulafungin, micafungin, and caspofungin against Candida species isolated from clinical specimens. METHODS: A total of 131 Candida strains were tested using Sensititre Yeast One colorimetric microdilution method. They belonged to the following species: 42 C. albicans, 36 C. glabrata, 21 C. parapsilosis, 12 C. tropicalis, 10 C. krusei, 3 C. guilliermondii, 2 C. famata, 3 C. kefyr, 1 C. lusitaniae, 1 C. zeylanoides, and 1 C. lipolytica. For being considered susceptible the strains had to be inhibited by concentrations ≤2 mg/L of anidulafungin, caspofungin or micafungin. RESULTS: The 80.1% of the strains tested were inhibited by concentrations ≤0.25 mg/L of anidulafungin and micafungin. The activity of caspofungin was slightly lower (78.6% of strains inhibited by concentrations ≤0.25 mg/L). The 96.9% of strains turned out susceptible to concentrations ≤2 mg/L against the three echinocandins. Two strains of C. parapsilosis (9.5%), one of C. guilliermondii, and two of C. famata showed non-susceptible to one or more echinocandins. CONCLUSIONS: In our series, anidulafungin, micafungin, and caspofungin were effective against C. albicans, C. glabrata, C. tropicalis, C. krusei, C. kefyr, C. lusitaniae and C. lipolytica. The 96.9% of strains were susceptible to all three echinocandins. Thus, echinocandins are proved to exhibit excellent activity to the Candida species most frequently involved in human infections, except Candida parapsilosis.

[Susceptibility to fluconazole of clinical interest yeasts: new breakpoints]. / Sensibilidad a fluconazol de levaduras de interés clínico: nuevos puntos de corte.

INTRODUCTION: Recently, Pfaller et al (Drug Resist Update 2010; 13:180-95), have proposed new breakpoints for determining the in vitro susceptibility to fluconazole of Candida albicans, C. parapsilosis and C. tropicalis. The aim of this study was to establish the variations in sensitivity of these species applying these breakpoints, in relation to those of the Clinical and Laboratory Standards Institute (CLSI). METHODS: We analyzed 112 strains of Candida: 49 C. albicans, 40 C. parapsilosis and 23 C. tropicalis. Susceptibility to fluconazole was performed by the method Sensititre YeastOne. The breakpoints used to determine the minimum inhibitory concentration (MIC) were identified by CLSI and the ones proposed by Pfaller et al. RESULTS: According to the CLSI criteria, all isolates were susceptible to fluconazole. MIC50 and MIC90 were 0.5 mg/L and 2 mg/L for C. albicans and C. parapsilosis, 0.5 mg/L and 1 mg/L for C. tropicalis. With the new criteria, 109 (97%) strains were susceptible. Variations were seen in C. albicans, with 3 strains (6%) susceptible dose-dependent. CONCLUSIONS: When applying the breakpoints recommended by Pfaller et al, and EUCAST, the number of fluconazole-susceptible strains decreased according to the CLSI criteria, especially C. albicans.

Otomycosis due to filamentous fungi.

Otomycosis is common throughout the world but barely studied in Spain. Our objective was to determine the microbiological and epidemiological characteristics of this pathology in Cadiz (Spain) between 2005 and 2010. Samples from patients with suspicion of otomycosis underwent a direct microscopic examination and culture on different media for fungi and bacteria. Mycological cultures were incubated at 30°C for at least seven days. Identification of fungi was based on colonial morphology and microscopic examination of fungal structure. From a total of 2,633 samples, microbial growth was present in 1,375 (52.2%) and fungal isolation in 390 (28.4%). We identified 228 yeasts and 184 filamentous fungi (13.4% of positive cultures and 47.2% of otomycosis), associated with yeasts in 22 cases (5.6%). The most frequent species were Aspergillus flavus (42.4%), A. niger (35.9%), A. fumigatus (12.5%), A. candidus (7.1%), A. terreus (1.6%), and Paecilomyces variotii (0.5%). Infection was predominant in men (54.9%) and patients beyond 55 years old (46.8%). The most common clinical symptoms were itching (98.9%), otalgia (59.3%), and hypoacusis (56.0%). Fall season reported the lowest number of cases (20.1%). Incidence of otomycosis and fungi producing otomycosis vary within the distinct geographical areas. In Cadiz, this infection is endemic due to warm temperatures, high humidity, sea bathing, and wind, which contributes to disseminate the conidia. Despite Aspergillus niger has been reported as the main causative agent, A. flavus is predominant in Cadiz. Although infection is usually detected in warm months, we observed a homogeneous occurrence of otomycosis in almost all the seasons.

Urethritis due to Corynebacterium glucuronolyticum.

We describe a case of urethritis in a young man caused by Corynebacterium glucuronolyticum. This bacterium is a synonym of Corynebacterium seminale, a known agent of non-gonococcal urethritis, that cannot be regarded as commensal flora in the urogenital region when is isolated in a symptomatic clinical context. Accuracy in diagnosis and correct treatment is important for avoiding probable complications, for example prostatitis. Bacterial isolation is convenient, and an antimicrobial susceptibility test should be conducted to discover antimicrobial resistance. In our case the patient was successfully treated with fluoroquinolones and was symptom-free in 2 weeks.

Emergence of CTX-M-15-producing Klebsiella pneumoniae of multilocus sequence types 1, 11, 14, 17, 20, 35 and 36 as pathogens and colonizers in newborns and adults.

OBJECTIVES: To characterize the population structure and resistance mechanisms of Klebsiella pneumoniae isolates that are highly resistant to third-generation cephalosporins, collected from five Spanish hospitals. METHODS: A total of 162 K. pneumoniae isolates from five hospitals located in three geographical areas of Spain were characterized. The number of isolates from each hospital ranged from 3 to 82. The genetic relationship between isolates was established by PFGE and multilocus sequence typing (MLST). bla(ESBL) types and other antibiotic resistance genes were analysed by PCR and sequencing. Plasmids were classified according to their incompatibility group by a PCR-based replicon-typing scheme. RESULTS: All 162 isolates carried the bla(CTX-15) gene. Fifty-eight isolates (35.8%) caused clinical infections and 104 (64.2%) were colonizers. Sixty-nine (42.6%) isolates were collected from newborns and 93 (57.4%) from adults. Using PGFE, the 162 isolates were grouped into seven clusters that were further identified as members of the MLST types 1, 11, 14, 17, 20, 35 and 36. Two hospitals each had two different clones and the remaining three hospitals had a single CTX-M-15-producing K. pneumoniae clone. All clones carried different antibiotic resistance genes, including bla(OXA-1), aac(3)-IIa, aac(6')-Ib-cr, qnrS1 and qnrB. In four of the seven (57.1%) clones the bla(CTX-M-15) gene was transferred by conjugation; in all cases plasmids of the incompatibility group IncF were identified by PCR. CONCLUSIONS: This study shows that multiresistant K. pneumoniae producing CTX-M-15 of MLST types 1, 11, 14, 17, 20, 35 and 36 are spreading as pathogens and colonizers among newborns and adult patients in Spain.

[Evaluation of an applied program for the prevention and control of tuberculosis]. / Valoración de la aplicación de un programa de prevención y control de tuberculosis.

BACKGROUND: To evaluate the efficacy of a tuberculosis prevention and control program applied in the health care area of Cadiz (Spain). MATERIALS AND METHODS: Tuberculosis cases diagnosed by microbiological criteria (Ziehl-Neelsen/auramine smear and/or culture in Löwenstein-Jensen medium) from 1996 to 2000 were analyzed in relation to epidemiologic characteristics of the patients. Susceptibility studies were performed from 1997 onwards using the proportion method described by Canetti, Rist and Grosset. RESULTS: The rate of tuberculous infections gradually dropped from 38.4 cases/100,000 inhabitants in 1996 to 16.0 cases/100,000 inhabitants in 2000. Along the study period, the percentage of men affected by the disease ranged from 81.2% in 1996 to 70.0% in 2000. In 1996, 26.0% of tuberculous patients were HIV-positive, and in 2000, 27.5%. The sensitivity of microbiologic diagnosis increased considerably from 60.4% in 1996 to 67.5% in 2000. Multi-drug resistance (9.2% en 1997) has virtually disappeared in our area since 1998 (1.6%).CONCLUSIONS. Implementation of a plan for the prevention and control of tuberculosis in our geographic area was effective, resulting in a decrease in the infection rate to less than 25 cases/100,000 inhabitants, and improvements in microbiologic diagnosis and compliance with treatment. Tuberculous disease has affected an increasingly higher proportion of women, regardless of HIV status. Antimicrobial resistance, particularly multi-drug resistance is now low in our area.

Valoración de la aplicación de un programa de prevención y control de tuberculosis / Evaluation of an applied program for the prevention and control of tuberculosis

ANTECEDENTES. Evaluar la aplicación de un programa de prevención y control de la tuberculosis en el área sanitaria de Cádiz. MATERIAL Y MÉTODOS. Se analizan los casos de tuberculosis diagnosticados microbiológicamente (tinción de Ziehl-Neelsen/auramina y/o cultivo en medio de Löwenstein-Jensen) desde 1996 a 2000, considerando las características epidemiológicas de los pacientes. Se realizó estudio de sensibilidad a partir de 1997 por el método de las proporciones de Canetti, Rist y Grosset. RESULTADOS. La tasa de infección tuberculosa desciende de manera progresiva desde 38,4 casos/100.000 habitantes en 1996, hasta 16,0 casos/100.000 habitantes en el año 2000. El porcentaje de hombres afectados por la enfermedad osciló a lo largo del estudio desde el 81,2 por ciento en 1996 al 70,0 por ciento en 2000. El 26,0 por ciento de los pacientes con tuberculosis eran positivos para el virus de la inmunodeficiencia humana (VIH) en 1996, y el 27,5 por ciento en el año 2000. La sensibilidad del diagnóstico microbiológico aumentó de forma importante desde el 60,4 por ciento en 1996 al 67,5 por ciento en 2000. La multirresistencia (9,2 por ciento en 1997) prácticamente desaparece de nuestra área a partir de 1998 (1,6 por ciento). CONCLUSIONES. La implantación del Programa de Prevención y Control de la Tuberculosis en nuestra área sanitaria se ha mostrado eficaz, al conseguir una disminución de la tasa de infección por debajo de 25 casos/100.000 habitantes y una mejora en el rendimiento del diagnóstico microbiológico y en la adherencia de los pacientes al tratamiento. La enfermedad tuberculosa afecta cada vez más a mujeres, con independencia de si están infectadas por el VIH o no. En el momento actual, la resistencia observada en nuestra zona geográfica es escasa, sobre todo la multirresistencia (AU)