Azole resistance in Aspergillus is emerging in European and Asian countries. As azoles are mainstay of therapy in the management of aspergillosis, azole resistance has serious implications in patient management. We report the emergence of resistance to triazoles in environmental Aspergillus fumigatus isolates in Iran. The TR34/L98H mutation was the only resistance mechanism. Overall 3.3% of the A. fumigatus isolates from hospital surroundings in Sari and Tehran had the same TR34/L98H STRAf genotype and were related to some resistant clinical and environmental TR34/L98H isolates from the Netherlands and India. It is emphasised that routine resistance surveillance studies focusing on environmental and clinical samples are warranted to yield the true prevalence of azole resistance in A. fumigatus in Iran.
The dermatophyte fungus Trichophyton rubrum often produces arthroconidia in vivo, and these cells are thought to be involved in pathogenesis, and, in shed skin scales, to act as a source of infection. The purpose of this study was (i) to examine the environmental and iatrogenic factors which affect arthroconidiation in T. rubrum in vitro, (ii) to look at arthroconidia formation in a large number of clinical isolates of T. rubrum and (iii) to develop a new model for the study of arthroconidia formation in nail tissue. Arthroconidia production was studied in T. rubrum grown on a number of media and under conditions of varying pH, temperature and CO 2 concentration. The effect of the presence of antifungals and steroids on arthroconidia formation was also examined. Nail powder from the healthy toenails of volunteers was used as a substrate for arthroconidial production. On Sabouraud dextrose agar in the presence of 10 % CO 2 plus air, arthroconidial formation occurred optimally at 37 6C and pH 7?5, and was maximal at 10 days. Most isolates of T. rubrum showed a similar level of arthroconidial production, and only two out of 50 strains were unable to produce arthroconidia. Subinhibitory levels of some antifungals and betamethasone resulted in the stimulation of arthroconidia formation. Arthroconidial production in ground nail material also occurred under the same optimal conditions, but took longer to reach maximal levels (14 days). These in vitro and ex vivo results provide a useful basis for the understanding of arthroconidium formation in vivo in infected tissues such as nails.
Phylogenetic studies highlight Candida africana as an atypical variant within Candida albicans species complex which is dominantly recovered from vaginal specimens. This study aimed to characterize C. africana isolates from patients with vulvovaginal candidiasis (VVC) by molecular methods and in vitro susceptibilities. One hundred and fifty-six (48.44%) Candida strains were collected from 322 patients diagnosed with VVC. Of these, 114 (73.07%) were germ tube positive and presented green color on the chromogenic medium, thus classified as C. albicans species complex. One hundred and nine (95.61%) out of 114 isolates were identified as C. albicans, while five (4.38%) isolates were identical with C. africana based on hwp1 PCR. C. africana appeared to be highly susceptible to the tested antifungals. For all strains of C. africana, fluconazole MIC was 2-log2-dilution steps less active than amphotericin B, which in turn was 2-log2-dilution steps and 3-log2-dilution steps less active than other azoles and echinocandin agents, respectively. In conclusion, among the C. albicans species complex, C. albicans predominantly and C. africana rarely occur in vaginal mucosa. Due to limited information on molecular epidemiology of this novel yeast, more studies using molecular methods are needed to elucidate the inter- and intraspecific genomic variations of C. africana isolates.
The genus Cladophialophora comprises etiologic agents of disease in immunocompetent patients, ranging from mild cutaneous colonization to cerebral encephalitis, in addition to saprobic species. Due to the high degree of phenotypic similarity between closely related species of the genus, identification problems are imminent. In the present study, we described rapid and sensitive rolling circle amplification (RCA) method based on species-specific padlock probes targeted for the internal transcribed spacer regions of rDNA. ITS regions of 12 Cladophialophora species were sequenced, and subsequently, 10 specific padlock probes were designed for the detection of single nucleotide polymorphisms. The majority of circularizable padlock probes were designed based on single nucleotide polymorphisms (SNPs), while for C. bantiana, C. immunda and C. devriesii were characterized by two or more nucleotides. Individual species-specific probes correctly identified in all ten Cladophialophora species correctly by visualization on 1.2 % agarose gels used to verify specificity of probe-template binding; no cross-reactivity was observed. Simplicity, sensitivity, robustness and low costs provide RCA a distinct place among isothermal techniques for DNA diagnostics. However, restriction and specificity and sensitivity should be lowered and increased, respectively, to be useful for a wide variety of clinical applications.
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