Identification of the Trichophyton mentagrophytes complex species using MALDI-TOF mass spectrometry.

Dermatophytes are fungi capable of invading keratinized tissues and are responsible for the most common fungal infection worldwide: dermatophytosis. Identification of these organisms to the species level is often necessary for the correct treatment of these infections, and is always recommended from an epidemiological point of view. Since the identification of dermatophytes is sometimes problematic, we assessed whether Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) could provide a useful tool to identify dermatophytes of the Trichophyton mentagrophytes complex. A reference database was constructed with 17 strains of six different species belonging to this complex. A total of 54 dermatophyte strains of the Belgian co-ordinated collections of micro-organisms, Scientific Institute of Public Health, Brussels, Belgium (BCCM™/IHEM) collection were used to challenge this database; 89% of the tested strains (not used as reference strains in the database) could readily be identified. When incorrect identifications were encountered, the confusion was always between phylogenetically closely related taxa which indicates that observations made by MALDI-TOF MS correlate with phylogenetic data. To assess this observation, a dendrogram outlining the similarities between the obtained spectra was constructed. Strikingly, the relationships found in this dendrogram were highly similar to the ones observed in the phylogenetic tree recently reported by Beguin and co-workers. In conclusion, MALDI-TOF MS is a fast and reliable tool for the identification of dermatophytes, since it can even discriminate between the closely related species of the T. mentagrophytes complex. Moreover, our data indicate that the data obtained by MALDI-TOF MS correlate with phylogenetic data.

Is Trichophyton simii endemic to the Indian subcontinent?

Trichophyton simii is considered to be prevalent only in the Indian subcontinent where it was isolated from soil, as well as from infections of humans and animals. We have investigated a case of onychomycosis caused by this exotic dermatophyte, not traceable to endemic areas. This case, as in others due to this fungus in man or animals, that have been previously and sporadically reported worldwide, suggests infections caused by T. simii might be underestimated, especially outside its primary geographic areas. Indeed, there are isolates that do not show species-specific morphology, as in our case isolate, and as a result may be misidentified by classical methods. By checking the identity of some strains preserved in the collection BCCM/IHEM, we found several that proved to be T. simii, originating from non-endemic areas (Belgium, France and Ivory Coast). Therefore, the natural distribution of T. simii is probably not as restricted as has previously been proposed.

The taxonomic status of Trichophyton quinckeanum and T. interdigitale revisited: a multigene phylogenetic approach.

Trichophyton quinckeanum, known as the causative agent of mouse favus, has been a subject of controversy since its discovery, 125 years ago. The purpose of this study was to examine the phylogenetic relationships between this fungus and related taxa. To achieve this objective, the ITS rDNA region, as well as actin and ß-tubulin gene regions of various isolates were sequenced. Bayesian inference and maximum likelihood analyses were conducted with T. rubrum as outgroup. Our study showed that strains identified as T. quinckeanum and others identified as T. schoenleinii are part of the complex T. mentagrophytes, and that their genotype cannot be confused with any other dermatophytes. Furthermore, this study demonstrates that the choice of the neotype of T. mentagrophytes was inappropriate. The beta-tubulin topology also revealed that isolates of T. interdigitale form a genetically distinct population from the type strains of Arthroderma vanbreuseghemii. Therefore, contrary to what is generally accepted, the anthropophilic species T. interdigitale cannot be considered as the anamorph associated with the latter.

Unusual Aspergillus species in patients with cystic fibrosis.

Poorly sporulating Aspergillus isolates from patients with cystic fibrosis (CF) are generally identified in routine procedures as Aspergillus spp. In this study, we identified and characterized 11 isolates belonging to two unusual Aspergillus species of the section Fumigati (A. lentulus and Neosartorya pseudofischeri) recovered from four different patients. Aspergillus lentulus was found occasionally during a 10-year follow-up study of one CF patient colonized by A. fumigatus. Neosartorya pseudofischeri was isolated from three patients followed in different European hospitals. This species was recovered from two sputum samples of one patient, and from four successive samples of the two other patients, suggesting that it may be responsible for chronic colonization. Both species were isolated together with A. fumigatus. Isolates from both species did not grow at 50°C, and DNA sequence analysis, together with further morphological observations permitted identification at the species level. Growth at different temperatures and antifungal susceptibility were also investigated. All the isolates of N. pseudofischeri exhibited a very low susceptibility to voriconazole (VRZ) whereas a very low susceptibility to VRZ and amphotericin B was seen with the A. lentulus isolates.

Tritirachium egenum, a thiamine- and siderophore-auxotrophic fungal species isolated from a Penicillium rugulosum.

Tritirachium egenum sp. nov., a biotrophic mycosymbiont, was found growing in association with a Penicillium rugulosum. This new species was unable to grow in axenic culture on traditional semi-synthetic culture media unless the growth medium was supplemented with a fraction of the culture filtrate of its host. The hot water extract of Alternaria alternata, containing the 'growth factor' of several contact mycosymbionts (biotrophic contact mycoparasites) also supported the growth of T. egenum. Signs, particularly the functional equivalence of this extract and ferrichrome on the T. egenum growth, suggested that this growth factor, referred to in the literature as mycotrophein, could actually be a hydroxamate-type siderophore. Moreover, it was shown that this siderophore-dependent organism had a deficient metabolism requiring, in addition, an exogenous source of thiamine or a precursor molecule of thiamine. Among mycosymbionts of fungi, the nutrient acquisition strategy of T. egenum is new, because it did not live in close association with the cytoplasm of its host. Indeed, it is neither a haustorial mycosymbiont nor a contact mycosymbiont, the two groups into which the biotrophic mycosymbionts of fungi are classified. Moreover, an interfungal association based on the utilization of siderophores has not yet been reported.

Zygomycosis in the immunocompromised patient: a case report.

Zygomycosis is a highly aggressive infection observed in immunocompromised patients, such as those with haematological malignancies. The sites most frequently involved are the sinuses and the lungs. New diagnostic tools and new antifungal treatments are essential in order to diagnose early and treat efficiently infections due to moulds. We report a case of sinusitis due to Absidia corymbifera occurring during chemotherapy-induced bone marrow aplasia in a patient with acute leukaemia. The sinusitis was successfully treated with AmBisome, and surgical debridement.

Cutaneous phaeohyphomycosis due to Alternaria infectoria.

Here we report a case of cutaneous alternariosis in a 74-year-old man treated by corticotherapy for myasthenia, and presenting with papular, crusted lesions on the left elbow and the right knee. Histological examination of the biopsy specimens showed fungal hyphae associated with round-shaped cells which were highly suggestive of alternariosis. Mycological culture allowed the isolation of a dematiaceous fungus which was identified as a member of the Alternaria infectoria species-group. This was confirmed by PCR amplification and sequencing of the internal transcribed spacer domain of the gene encoding nuclear ribosomal DNA and of the mitochondrial small subunit ribosomal DNA domain. The fungus was therefore referred to the Scientific Institute of Public Health where it was identified as Alternaria infectoria, on the basis of its very small 1 or 2-celled conidia often arranged in long chains and presenting with very long secondary conidiophores. Corticotherapy was stopped and a local antifungal treatment with ketoconazole was initiated, allowing the stabilisation of the cutaneous lesions within 2 months.

Airway colonization by Acrophialophora fusispora in patients with cystic fibrosis.

Acrophialophora fusispora is a thermotolerant soil fungus which is very unusual in clinical samples. Here we report four cases of transient or chronic airway colonization by A. fusispora in patients with cystic fibrosis (CF). However, the prevalence of this fungus in CF patients may be underestimated due to the currently poor knowledge of this fungus in most of the medical mycology laboratories. In addition, its clinical importance regarding CF remains to be evaluated.