Rare and new etiological agents revealed among 178 clinical Aspergillus strains obtained from Czech patients and characterized by molecular sequencing.

A collection of 178 Aspergillus isolates, recovered from Czech patients, mostly from 2007-2011, was subjected to multilocus DNA sequence typing using the ITS region, ß-tubulin, and calmodulin genes. An unusually wide spectrum of etiologic agents that included 36 species of Aspergillus is discussed in the context of recent taxonomic and clinical reports. Invasive aspergillosis (IA), onychomycosis, and otitis externa were the predominant clinical entities. Five cases due to species newly proven as etiologic agents of human mycoses, as well as cases with unique clinical manifestations caused by unusual agents are discussed in more detail. Three species (i.e., A. insulicola, A. westerdijkiae and A. tritici) were identified as the confirmed etiologic agents of non-dermatophytic onychomycosis. Emericella rugulosa was recovered from a premature newborn with a fatal necrotising disseminated infection and is reported for only the second time as the cause of IA. Furthermore, we document the first infection due to A. calidoustus in a patient with chronic granulomatous disease. The infection manifested as a latent brain aspergilloma with an unusual clinical-laboratory finding. In addition to the well-known agents of human mycosis, several rarely isolated or poorly documented species were identified. An undescribed cryptic species related to A. versicolor was found to be common among isolates linked to proven and probable onychomycosis. An isolate representing A. fresenii, or an unnamed sister species, were causal agents of otomycosis. Three well defined, and tentative new species belonging to section Cervini, Candidi and Aspergillus (Eurotium spp.), were associated with cases of probable onychomycosis.

Development of rapid and specific molecular discrimination methods for pathogenic emericella species.

Aspergillosis is an important mycosis caused primarily by Aspergillus fumigatus and its relatives. The genus Emericella is a teleomorph related to the Aspergillus section Nidulantes. The typical anamorphic stage species in this genus is Aspergillus nidulans, which is sometimes a significant agent in chronic granulomatous disease (CGD) patients. The mortality rate of osteomyelitis in CGD patients due to A. nidulans ( E. nidulans ) is very high compared to that due to A. fumigatus. Moreover, two Emericella species ( E. nidulans and E. quadrilineata ) from clinical specimens exhibit different sensitivities against several antifungal drugs. In aspergillosis, correct species identification is important for antifungal therapy. We attempted to develop rapid and specific molecular discrimination by polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) methods in the principal pathogenic Emericella species, and succeeded in establishing species-specific primers corresponding to the hydrophobin gene. These primers discriminate E. nidulans and E. quadrilineata rapidly and specifically. These methods and primers make it possible to diagnose etiological agents in aspergillosis quickly and easily.

Reportes clínicos: infección probada por Aspergillus flavus y probable infección pulmonar por Emericella nidulans en pacientes inmunodeprimidos / Clinical reports: tested rhinosinusal infection by Aspergillus flavus and probable pulmonary infection by Emericella nidulans in immunodepressed patients

En los últimos 20 años la incidencia de aspergilosis invasiva ha aumentado entre 5 y 10 veces, presentando una mortalidad que varía desde un 60 a 98 por ciento dependiendo de su localización, enfermedad de base (inmunodepresión), diagnóstico y tratamiento precoz. A pesar que se ha logrado un importante avance en estos dos últimos decenios, aproximadamente un 50 por ciento de los casos se diagnostican post mortem. Se reportan dos casos de aspergilosis (probada y probable) en pacientes inmunodeprimidos de 21 y 44 años respectivamente. El primer caso corresponde a una infección rinosinusal en una paciente con leucemia linfoblástica, con un cuadro clínico caracterizado por aumento de volumen facial izquierdo, compromiso progresivo del estado general y equimosis en región periorbitaria. La endoscopia nasal detectó abundante secreción y tejido friable amarillento sugerente de infección micótica. Las muestras de tejidos enviadas al laboratorio para examen directo y cultivos arrojaron presencia de hifas en la mucosa y crecimiento abundante de Aspergillus flavus. Se inició terapia con itraconazol, cambiándose posteriormente a voriconazol, con respuesta favorable y disminución del dolor y volumen facial. Sin embargo, la paciente fallece al 10º día del ingreso debido a su mal estado general, avance de la falla medular y posterior insuficiencia respiratoria. El segundo caso corresponde a una probable infección pulmonar en una paciente con cáncer de mama en tratamiento quimioterápico. La paciente ingresó con el diagnóstico de neutropenia febril y probable neumonía de etiología bacteriana, fue tratada con antimicrobianos tanto en el hospital comunal como en el de base. Sin embargo, presentó una evolución tórpida, cursando posteriormente con hemoptisis, insuficiencia respiratoria y compromiso de conciencia. Se tomaron muestras de secreción endotraqueal las que fueron enviadas para estudio bacteriológico(TBC) y micológico, además de hemo y urocultivos. Los resultados de los...

In the lately twenty years the incidence of invasive spergillosis has increased five and ten times, causing a mortality rate ranging from 60 to 98 percent, depending on its location, nature of disease (immunodepression), diagnosis and forward treatment. In spite of the fact that there has been a significant progress in this lately period of twenty years, about 50 percent of cases were detected post mortem. Two cases of spergillosis (tested and probable) in 21 and 44 aged immunodepressed patients respectively are herein reported. The first case involves a rhinosinusal infection in a female patient diagnosed with lymphoblastic leukemia, whose clinical symptoms were an increase in the left volume of her face, a progressivecompromise in her overall state of health and ecchymosis in her periorbitary region. Nasal endoscopy revealed abundant secretion and yellowish friable tissues which suggested a mycotic infection. Samples of tissues sent to the lab for direct exam and cultures revealed the presenceof hypha in the mucus and an abundant growth of Aspergillus flavus. The first therapy was with itraconazol, changing later on to voriconazol what resulted in adecrease of pain and face volume. However, the patient dies on the tenth day of her ingress due to her bad state ofhealth, progress in glandular failure and ultimate respiratory insufficiency. The second case involves a probable pulmonary infection in a patient diagnosed with breast cancer and receiving chemotherapy treatment. She was ingressed with a febrile neutropenia and a probable bacterial etiology pneumonia and she was treated with antimicrobial medication both in the community hospital and in the base building. However, she showed a torpid evolution followed furthermore by hemoptisis, respiratory insufficiencyand consciousness compromise. Samples of endotraqueal secretion were taken in order to submit them to bacteriological and mycological (TBC) studies, together with hemo...