RESUMEN
In order to know the epidemiology and the changes of antifungal resistance in invasive candidiasis (IC) we carried out this prospective study of Candida strains belonging to patients admitted to 26 Spanish hospitals from June 2011 to June 2012 diagnosed with IC. Clinical information and the identity of the Candida species were collected and antifungal susceptibility was tested using broth microdilution in five agents: amphotericin B, fluconazole, voriconazole, caspofungin and anidulafungin. A total of 705 cases-isolates were documented. Most of the patients suffered from candidemia and several underlying diseases and more than half of them were neutropenic or under immunosuppressive therapy, factors associated with higher mortality. Thirty percent of global mortality was documented. C. albicans was the most frequently isolated species, although an increase of non-C. albicans species was observed. Resistance to fluconazole was of 3.4%, lower than in previous years (6.3%). C. parapsilosis presented a higher MIC90 of echinocandins compared to other species.
Asunto(s)
Antifúngicos/farmacología , Candida/efectos de los fármacos , Candidiasis Invasiva/epidemiología , Candidiasis Invasiva/microbiología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Candida/aislamiento & purificación , Candida albicans/efectos de los fármacos , Candida albicans/aislamiento & purificación , Candidiasis Invasiva/tratamiento farmacológico , Candidiasis Invasiva/mortalidad , Niño , Preescolar , Farmacorresistencia Fúngica/efectos de los fármacos , Femenino , Hospitales , Humanos , Lactante , Recién Nacido , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Estudios Prospectivos , Vigilancia de Guardia , España/epidemiología , Adulto JovenRESUMEN
Since two new species phylogenetically related to Candida glabrata with slightly different phenotypes and antifungal susceptibility profiles have been described, it seems to be necessary from clinical point of view, to develop a rapid and accurate identification system in order to distinguish between these three fungal species. We studied the performance of denaturing high performance liquid chromatography (dHPLC) as a faster (less than 7 min) and alternative novel technique for simultaneous analysis of Candida species in different biological matrices. The analyses show the good low limit of detection (LLOD) in all biological matrices studied (5.16-9.56 ngµL(-1), 4.14-4.70 ng µL(-1) and 3.99-4.66 ng µL(-1) for Candida bracarensis, Candida nivariensis and C. glabrata, respectively). 180 Candida isolates were analyzed in order to demonstrate the method suitability for screening analysis to identify C. glabrata and its cryptic species (C. bracarensis and C. nivariensis) in clinical routine.
Asunto(s)
Candida glabrata/clasificación , Candida/clasificación , Cromatografía Líquida de Alta Presión/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Sangre/microbiología , Candida/genética , Candida/aislamiento & purificación , Candida glabrata/genética , Candida glabrata/aislamiento & purificación , Cromatografía Líquida de Alta Presión/normas , ADN Bacteriano/química , ADN Intergénico/química , Humanos , Límite de Detección , Desnaturalización de Ácido Nucleico , Reacción en Cadena en Tiempo Real de la Polimerasa/normas , Reproducibilidad de los Resultados , Esputo/microbiología , Orina/microbiologíaRESUMEN
Candidemia studies have documented geographic differences in rates and epidemiology, underscoring the need for surveillance to monitor trends. We conducted prospective candidemia surveillance in Spain to assess the incidence, species distribution, frequency of antifungal resistance, and risk factors for acquiring a Candida infection. Prospective laboratory-based surveillance was conducted from June 2008 to June 2009 in 40 medical centers located around the country. A case of candidemia was defined as the isolation of a Candida species from a blood culture. Incidence rates were calculated per 1,000 admissions. Antifungal susceptibility tests were performed by using broth microdilution assay according to the guidelines of the Clinical and Laboratory Standards Institute. We detected 984 cases, for an overall incidence of 1.09 cases per 1,000 admissions. The crude mortality was 20.20%. Candida albicans was the most common species (49.08%), followed by C. parapsilosis (20.73%), C. glabrata (13.61%), and C. tropicalis (10.77%). Overall, decreased susceptibility to fluconazole occurred in 69 (7.01%) incident isolates. Antifungal resistance was rare, and a moderate linear correlation between fluconazole and voriconazole MICs was observed. This is the largest multicenter candidemia study conducted to date and shows the substantial morbidity and mortality of candidemia in Spain.
Asunto(s)
Candida/aislamiento & purificación , Candidiasis/epidemiología , Fungemia/epidemiología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Antifúngicos/farmacología , Candida/clasificación , Candida/efectos de los fármacos , Niño , Preescolar , Farmacorresistencia Fúngica , Femenino , Fungemia/microbiología , Hospitales , Humanos , Incidencia , Lactante , Recién Nacido , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Estudios Prospectivos , Vigilancia de Guardia , España/epidemiología , Adulto JovenRESUMEN
Stir bar soptive extraction (SBSE) coupled to thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS) and membrane-assisted solvent extraction (MASE) coupled to large volume injection-programmed temperature vaporisation-GC-MS (LVI-PTV-GC-MS) were optimised for the simultaneous determination of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), phthalate esters (PEs), nonylphenols (NPs), polybrominated biphenyls (PBBs) and polybrominated diphenyl ethers (PBDEs) in water samples. In the case of SBSE-TD, variables affecting the extraction (extraction time, addition of sodium chloride or methanol and sample volume) and desorption (cryofocusing temperature, desorption time and temperature, vent pressure and desorption flow) were fitted for the simultaneous determination. The extraction solvent nature (n-hexane, cyclohexane, n-heptane, ethyl acetate, toluene, dichloromethane or cyclohexane:ethyl acetate mixtures), as well as the addition of methanol (0-30%) and sodium chloride (0-20%), the extraction temperature (30-60 degrees C), shaking speed (250-750 rpm) and extraction time (5-150 min) were studied for the simultaneous membrane-assisted preconcentration. Finally, PTV-LVI variables such as injection volume (100-600 microL), injection speed (10-40 microL s(-1)), vent pressure (0-12.7 psi), vent time (0.05-0.8 min), vent flow (30-80 mL min(-1)), cryofocusing temperature (20-70 degrees C), split flow (20-100 mL min(-1)) and split time (1-5 min) were optimised. The optimisation was carried out by means of experimental design approaches in most of the cases. Precision (approximately 3-19% for both SBSE-TD and MASE-LVI-PTV), accuracy (approximately 80-120% for both SBSE-TD and MASE-LVI-PTV), limits of detection (LoDs) (0.1-222 ng L(-1) for MASE-LVI-PTV and 0.03-20.4 ng L(-1) for SBSE-TD in dependence of substance) and linearity (from 25 ng L(-1) up to at least 500 ng L(-1) for both procedures) were established for both procedures. Finally, the developed methods were applied to the determination of the free concentrations of PAHs, PCBs, PEs, NPs, PBBs and PBDEs in natural water samples (estuarine water and sea water) from the Bilbao estuary (Northern Spain) and comparable results were obtained with both procedures.