Bahram Ahmadi

A case of onychomycosis caused by Aspergillus candidus

Based on epidemiological studies, Aspergillus candidus has been demonstrated as an emerging fungal agent of toenail onychomycosis. Here we report a case of a toenail infection caused by A. candidus in a healthy 60-year-old woman. Based on macroscopic and microscopic characteristics of the culture as well as nucleotide sequencing of 28S region, the causative agent was identified as A. candidus.

Translation elongation factor 1-α gene as a potential taxonomic and identification marker in dermatophytes

Intra- and interspecies variations of the translation elongation factor 1-α (Tef-1α) gene were evaluated as a new identification marker in a wide range of dermatophytes, which included 167 strains of 30 species. An optimized pan-dermatophyte primer pair was designed, and the target was sequenced. Consensus sequences were used for multiple alignment and phylogenetic tree analysis and the levels of intra- and interspecific nucleotide polymorphism were assessed. Between species, the analyzed part of the Tef-1α gene varied in length from 709 to 769 nucleotides. Significant numbers of species including Trichophyton rubrum, T. tonsurans, T. schoenleinii, T. concentricum, T. violaceum, Epidermophyton floccosum, Microsporum ferrugineum, M. canis, M. audouinii, T. equinum, T. eriotrephon, and T. erinacei were invariant in Tef-1α and had sufficient barcoding distance with neighboring species. Although overall consistency was found between ITS phylogeny as the current molecular marker of dermatophytes and Tef-1α, a higher discriminatory power of Tef-1α appeared particularly useful in some clades of closely related species such as the A. vanbreuseghemii, T. rubrum, A. benhamiae, and A. otae complexes. Nevertheless, we stress that a single gene can not specify species borderlines among dermatophytes and multiple lines of evidence based on a multilocus inquiry may ascertain an incontrovertible evaluation of kinship.

Microsporum fulvum, an Ignored Pathogenic Dermatophyte: A New Clinical Isolation from Iran

Misidentifying with Microsporum gypseum has for a long time been accounted for less prevalence of the geophilic species, Microsporum fulvum in human dermatophytosis. We describe a new case of infection with the species in an Iranian young man. Direct examination of skin scrapings revealed a tinea corporis, and morphological study of the recovered isolate from the culture resulted in the identification of M. gypseum. However, PCR amplification of ITS1-5.8S rDNA-ITS2 region and subsequent ITS-RFLP and sequencing were indicative of M. fulvum as the true causative agent. To recognize M. fulvum in human infections and to validate the morphologically distinguished isolates of M. gypseum, the genetic-based identification is strongly recommended.

First case of disseminated phaeohyphomycosis in an immunocompetent individual due to Alternaria malorum

A 27-year-old Iranian, previously healthy male presented with sub-cutaneous necrotic lesions with a localized dermatosis affecting the anterior chest, neck and face. These lesions consisted of singular, well-defined verrucous plaques which gradually developed and disseminated over time. The dermatosis was followed by the development of necrotic swollen lesions localized on the hard palate. The patient did not recall any history of trauma or puncture at any of the sites of infection. While histopathological examination of periodic acid-Schiff (PAS) stained material revealed irregular, unbranched, septate hyphae, direct examination (KOH 10%) of lesion samples demonstrated the presence of septate indistinct brownish hyphae. Alternaria malorum was isolated (CBS 126589) and its identity was confirmed by sequencing of the internal transcribed spacer (ITS rDNA). Since the palate lesion reoccurred after 10 years and the patients condition did not improve with amphotericin B combination therapy, the lesion was surgical excised and he underwent antifungal therapy with amphotericin B and itraconazole. There was no dehiscence or fistula formation or any evidence of relapse of fungal infection during a one year follow-up and the patient was successfully cured. In vitro antifungal susceptibility tests revealed that the MIC values for those antifungals employed in this case were amphotericin B (0.125 μg/ml), fluconazole (32 μg/ml), itraconazole (0.125 μg/ml), voriconazole (1 μg/ml), and posaconazole (0.063 μg/ml). The MECs for caspofungin and anidulafungin were 0.25 μg/ml and 0.016 μg/ml, respectively. However, treatment of A. malorum infections with the latter agents remains to be evaluated.

Phylogenetic analysis of dermatophyte species using DNA sequence polymorphism in calmodulin gene

Use of phylogenetic species concepts based on rDNA internal transcribe spacer (ITS) regions have improved the taxonomy of dermatophyte species; however, confirmation and refinement using other genes are needed. Since the calmodulin gene has not been systematically used in dermatophyte taxonomy, we evaluated its intra- and interspecies sequence variation as well as its application in identification, phylogenetic analysis, and taxonomy of 202 strains of 29 dermatophyte species. A set of primers was designed and optimized to amplify the target followed by bilateral sequencing. Using pairwise nucleotide comparisons, a mean similarity of 81% was observed among 29 dermatophyte species, with inter-species diversity ranging from 0 to 200 nucleotides (nt). Intraspecies nt differences were found within strains of Trichophyton interdigitale, Arthroderma simii, T. rubrum and A. vanbreuseghemii, while T. tonsurans, T. violaceum, Epidermophyton floccosum, Microsporum canis, M. audouinii, M. cookei, M. racemosum, M. gypseum, T. mentagrophytes, T schoenleinii, and A. benhamiae were conserved. Strains of E. floccosum/M. racemosum/M. cookei, A. obtosum/A. gertleri, T. tonsurans/T. equinum and a genotype of T. interdigitale had identical calmodulin sequences. For the majority of the species, tree topology obtained for calmodulin gene showed a congruence with coding and non-coding regions including ITS, BT2, and Tef-1α. Compared with the phylogenetic tree derived from ITS, BT2, and Tef-1α genes, some species such as E. floccosum and A. gertleri took relatively remote positions. Here, characterization and obtained dendrogram of calmodulin gene on a broad range of dermatophyte species provide a basis for further discovery of relationships between species. Studies of other loci are necessary to confirm the results.