Autochthonous and dormant Cryptococcus gattii infections in Europe.

Until recently, Cryptococcus gattii infections occurred mainly in tropical and subtropical climate zones. However, during the past decade, C. gattii infections in humans and animals in Europe have increased. To determine whether the infections in Europe were acquired from an autochthonous source or associated with travel, we used multilocus sequence typing to compare 100 isolates from Europe (57 from 40 human patients, 22 from the environment, and 21 from animals) with 191 isolates from around the world. Of the 57 human patient isolates, 47 (83%) were obtained since 1995. Among the 40 patients, 24 (60%) probably acquired the C. gattii infection outside Europe; the remaining 16 (40%) probably acquired the infection within Europe. Human patient isolates from Mediterranean Europe clustered into a distinct genotype with animal and environmental isolates. These results indicate that reactivation of dormant C. gattii infections can occur many years after the infectious agent was acquired elsewhere.

Cryptococcus gattii infection in a Spanish pet ferret (Mustela putorius furo) and asymptomatic carriage in ferrets and humans from its environment.

A domestic ferret (Mustela putorius furo) was presented with lymphadenopathy and acute bilateral blindness. Cytologic evaluation and biopsy of an affected lymph node revealed pyogranulomatous lymphadenitis with intralesional yeast consistent with Cryptococcus sp. Subsequent studies demonstrated Cryptococcus gattii serotype B VGI/AFLP4 as the causative agent. The ferret was treated with fluconazole and prednisone. After one month of therapy, an improvement of the clinical symptoms was detected although blindness persisted. Seven months after presentation, the disease progressed to a severe neurologic condition, and it was euthanized. Postmortem exam revealed disseminated cryptococcosis with prominent neurologic involvement. Nasal swabs of other ferrets and humans from the same household revealed that two ferrets and two humans to be asymptomatic carriers of the same strain of cryptococcus as the necropsied ferret. These findings stress the importance of veterinary diagnostic work with pets and epidemiological investigations for disease prevention in them and in their owners.

Pyrenochaeta keratinophila sp. nov., isolated from an ocular infection in Spain.

The new coelomycete Pyrenochaeta keratinophila, isolated from corneal scrapings of a case of keratitis in Spain, is described and illustrated. This fungus is morphologically characterized by grey-olivaceous to greenish olivaceous colonies, scarce pycnidial setae placed mainly near the ostiole and production of phialoconidia from the aerial mycelium. The latter feature is unknown in any other species of the genus Pyrenochaeta. Sequencing of the ITS rDNA region of this clinical strain confirmed this proposal and revealed its close genetic relationship with the Leptosphaeriaceae.

New pyrenochaeta species causing keratitis.

We report a new fungus as an agent of fungal keratitis in a diabetic woman. The fungal etiology was established by classic microbiology and PCR following 3 months of antibacterial therapy. The morphological features of the isolate and sequence analysis of the internal transcribed spacer region indicate a new species of Pyrenochaeta (Coelomycetes).

[Pigeons and other birds as a reservoir for Cryptococcus spp]. / La paloma y otras aves como reservorio de Cryptococcus spp.

In the last 25 years, the cases of human and animal cryptococcosis have increased significantly. This is mostly due to the improvement in the survival of immunocompromised patients. The disease is frequently related to the exposure of this type of patients to avian droppings. Among birds, pigeon, Columba livia, is undoubtedly the most important reservoir for the Cryptococcus species. Nevertheless, the study of a large number of bird's species demonstrated that pigeons are not the only Cryptococcus spp. carriers. The suspicion of the birds being the source for the infection is now becoming a demonstrable fact thanks to the use of molecular typing methods. These methods allow the comparison between strains from birds to patients living around them, with high level of discrimination.

Polymerase chain reaction and DNA typing for diagnosis of infectious crystalline keratopathy.

A 63-year-old man developed infectious crystalline keratopathy (ICK) in his right eye 1 year after phacoemulsification. The white peripheral lesion was adjacent to the corneal phacoemulsification incision. Infiltrates in the form of creamy-white, midstromal branching, needle-like opacities without evidence of inflammatory cells were noted. A corneal biopsy by double lamellar flap was done and studied by 3 techniques: microbiological culture, stain, and polymerase chain reaction (PCR). Fungal and bacterial PCR were positive. A second sample was necessary to obtain a positive stain and culture. The DNA sequence analysis showed Candida parapsilosis and Staphylococcus aureus as the causal agents of the crystalline keratopathy. Treatment was started with amphotericin B 1% and cefazolin 6 times a day, and systemic voriconazole was recommended. This is the first reported case of ICK after cataract surgery. Polymerase chain reaction amplification and subsequent DNA typing were useful tools in detecting and identifying the ocular pathogens involved in this case.

First case of human cryptococcosis due to Cryptococcus neoformans var. gattii in Spain.

We report the first case of human cryptococcosis due to Cryptococcus neoformans var. gattii described in our country, which was presented as brain cryptococcoma in an immunocompetent patient. An extensive sampling of the patient's environment was carried out to find the source of infection.

Biología molecular en el diagnóstico de la candidiasis profunda en el paciente crítico no neutropénico / Molecular biology in the diagnosis of deep-seated candidiasis in the critically ill non-neutropenic patient

Las infecciones por Candida son una importante causa de mortalidad y morbilidaden pacientes críticos. La detección rápida de la presencia de la levadura ensangre y otros tejidos es un objetivo que recientemente se intenta abordar aplicandodistintos métodos moleculares de diagnóstico. Analizamos la sensibilidady especificidad de los métodos aplicados a la detección de Candida en muestrasclínicas de pacientes críticos y otros susceptibles de padecer candidiasis profunda.Los métodos basados en la reacción en cadena de la polimerasa (PCR),especialmente los que tienen como diana las secuencias de los genes ribosomalesy sus espaciadores internos, muestran claramente una mayor sensibilidadque el cultivo y una especificidad equiparable a este. Todavía no hay propuestasclaras en cuanto a la posible estandarización de la extracción y purificación delDNA y de los sistemas de lectura de productos, aunque la técnica de PCR atiempo real con emisión de fluorescencia, parece la propuesta más interesantepor la ventaja añadida de permitir la cuantificación de la carga fúngica y minimizarla manipulación de las muestras y por tanto el riesgo de falsos positivos

Candida infections are an important cause of morbidity and mortality in criticallyill patients. Rapid detection of the yeast in blood and other tissues by molecularbiology methods has been the goal of some recent studies. An analysis of thesensitivity and specificity of these methods assayed in clinical specimens fromcritically ill and other patients is carried out. PCR amplification of ribosomalgenes and their internal spacers showed a higher sensitivity than culture basedmethods. A standardization of most of the methodological steps in molecularmethods is needed. Real time PCR with fluorescent probes seems to be themost interesting proposal. It has the advantage of the possible quantification offungal presence in tissues and minimizes the samples’ contamination risk