Aspergillus species as emerging causative agents of onychomycosis.

BACKGROUND: Onychomycosis is a common nail infection caused by dermatophytes, non-dermatophyte molds (NDM), and yeasts. Aspergillus species are emerging as increasing causes of toenail onychomycosis. The purpose of this study was species delineation of Aspergillus spp. isolated from patients with onychomycosis. METHODS: During a period of one year (2012-2013), nail samples were collected from patients clinically suspected of onychomycosis and subjected to microscopic examination and culture. Species identification was performed based on macro- and micro-morphology of colonies. For precise species identification, PCR-amplification and sequencing of the beta-tubulin gene followed by BLAST queries were performed where required. RESULTS: A total of 463/2,292 (20.2%) tested nails were diagnosed with onychomycosis. Among the positive specimens, 154 cases (33.2%) were identified as saprophytic NDM onychomycosis, 135 (29.2%) of which were attributable to Aspergillus. Aspergillus species isolated from the infected nails included Aspergillus flavus (77.3%, n=119), Aspergillus niger (n=4), Aspergillus tubingensis (n=4), Aspergillus terreus (n=3), Aspergillus sydowii (n=2), Aspergillus spp. (n=2), and Aspergillus candidus (n=1). Among the patients diagnosed with onychomycosis due to Aspergillus (average patient age, 47.4 years), 40 had fingernail and 95 toenail involvement. The large toenails were most commonly affected. CONCLUSIONS: This study identified a markedly high occurrence of A. flavus, and this fungus appears to be an emerging cause of saprophytic onychomycosis in Iran. The study moreover highlights the necessity of differentiating between dermatophytic and non-dermatophytic nail infections for informed decisions on appropriate therapy.

A comparative study on morphological versus molecular identification of dermatophyte isolates.

OBJECTIVE: Dermatophytes are taxonomically classified in the genera Trichophyton, Microsporum, and Epidermophyton. Pleomorphism, cultural variability, slow growth and sporulation, and the need for additional physiological tests make dermatophytes notoriously difficult to identify. The present study aimed to compare the results of morphological and molecular identification of certain groups of clinical isolates of dermatophytes with a view to evaluating the accuracy of molecular methods. PATIENTS AND METHODS: For each sample, the ITS1-5.8S-ITS2 rDNA region was amplified using the primers ITS1 and ITS4. PCR products were subjected to restriction fragment length polymorphism (RFLP) analysis using the enzyme MvaI and isolate identification was performed by comparing the electrophoretic RFLP patterns with reference profiles obtained previously. Finally, paired comparative analyses of molecular and conventional methods were performed. RESULTS: While morphology results from routine daily reports of the laboratories indicated that 18 (6.8%) and 136 (52.10%) of the isolates were T. rubrum and T. interdigitale, respectively, PCR-RFLP results suggested that T. rubrum was the most common etiological agent of ringworm accounting for 94 (36.01%), followed by T. interdigitale accounting for 71 (27.20%). Interestingly, 80.8% out of the 94 isolates identified as T. rubrum by molecular testing had been identified by morphological examination as belonging to different species, such as T. interdigitale (75.5%), E. floccosum (2.1%) and M. canis, T. verrucosum, and T. tonsurans (each 1.06%). Ten strains out of 261 (T. interdigitale, n=8; E. floccosum, n=2) had been defined as unknown species by morphological tests. CONCLUSION: An unexpected high percent of isolates identified as T. interdigitale by conventional methods were in effect T. rubrum shown by PCR-RFLP, and regarding the necessity of correct identification of dermatophytes recovered from different clinical forms of the infection, we highly recommend ITS-sequencing or ITS-RFLP of the isolates, particularly for epidemiological research studies.

Using SCC8, SCF27 and VMC7f2 markers in grapevine breeding for seedlessness via marker assisted selection.

We used molecular markers associated with seedlessness in grapes, namely SCC8, SCF27 and VMC7f2, to improve the efficiency of seedless grapevine breeding via marker assisted selection (MAS). DNA from 372 F1 hybrid progeny from the cross between seeded "Alphonse Lavallée" and seedless "Sultani" was amplified by PCR using three markers. After digestion of SCC8 marker amplification products by restriction enzyme BgIII, 40 individuals showed homozygous SCC8+/SCC8+ alleles at the seed development inhibitor (SdI) locus. DNA from 80 of the progeny amplified with the SCF27 marker produced bands; 174 individuals had 198-bp alleles of the VMC7f2 marker associated with seedlessness. In the second year, based on MAS, 183 F1 hybrids were designated as seedless grapevine candidates because they were positive for a minimum of one marker. Twenty individuals were selected as genetic resources for future studies on seedless grapevine breeding because they carried alleles for the three markers associated with seedlessness. The VMC7f2 SSR marker was identified as the marker most associated with seedlessness.

Prevalence of fungal infection among Iranian patients with chronic sinusitis.

Chronic sinusitis is a major cause of morbidity today. Regional variations in the incidence of this disease have been reported. The aim of this study was to evaluate the incidence of fungal infection as the causative agent of chronic sinusitis among Iranian patients. A cross sectional hospital based study was designed; the patients underwent paranasal sinus washing and maxillary sinus biopsy. All specimens were studied by light microscopy. Fungal culturing was employed to confirm diagnosis. The patients underwent Computed Tomography for sinus evaluation. Of 162 participants, 12 samples from patients showed fungal elements, 2 of them Aspergillus fulvous (1.2%), 9 of them Alternaria species (5.56%) and 1 of them Psilomysis (0.6%). All patients presented radiologic evidence of sinusitis, ranging from mucosal thickening to total opacity. In conclusion, results obtained showed a low prevalence of fungal sinusitis among Iranian patients with chronic sinusitis. Findings also showed that Alternaria is the most causative agent.

Antifungal activity of the methanolic extract and alkaloids of Glaucium oxylobum.

The methanolic extract and total alkaloids of the aerial parts of Glaucium oxylobum exhibited good activity against Microsporum gypseum, Microsporum canis, Trichophyton mentagrophytes and Epidermophyton floccosum. Four alkaloids, dicentrine, glaucine, protopine, and alpha-allocryptopine, were identified as the compounds responsible for the antifungal activity of this plant.

Novel therapeutic approach by culture filtrate of Cryptococcus neoformans var. gattii (CneF) in experimental immune complex glomerulonephritis.

The present study was undertaken to determine the therapeutic effect of the culture filtrate of Cryptococcus neoformans var. gattii (CneF) in experimental immune complex glomerulonephritis. Bovine serum albumin (BSA) nephritis was induced in rats by a subcutaneous immunization and daily intravenous administration of BSA. CneF solution at three different doses (36, 54, and 90 mg/kg based on carbohydrate concentration) was administered intraperitoneally at regular 72-h intervals for 4 weeks. Onset of treatment was day 65, and urinary protein was measured at different intervals. Animals were euthanized on day 107. Serum and urine determinants were measured at the time of sacrifice and kidney specimens were examined. Results of this experiment showed that CneF therapy could significantly reduce the urinary protein excretion, blood urea nitrogen (BUN), plasma concentration of triglyceride, and increase the serum HDL cholesterol in treated rats vs. nontreated controls. Moreover, there was significant difference in glomerular changes between treated and nontreated groups. These observations show that the beneficial effect of CneF may be related to decreased number of glomerular leukocytes. Our findings suggest that treatment with CneF as a new antiinflammatory compound can reduce proteinuria, suppress the development of glomerular lesions, and exert lipid-lowering property in a rat model of immune complex glomerulonephritis.

The first survey of Hendersonula toruloidea as a human pathogen in Iran.

The possible presence of Hendersonula toruloidea in the environment and its role as a human pathogen in Iran was studied. Samples were collected both from trees likely to be infected and from patients who presented with suspected fungal infections of the hands and feet. The samples were mainly collected in areas of southern Iran where the climate is similar to that of areas where H. toruloidea has been found previously to infect humans. H. toruloidea type B was isolated from plant samples (eucalyptus trees) for the first time in Iran but it could not be isolated as a human pathogen.