The first report of tinea nigra from Iran.

Hortaea werneckii causes Tinea nigra, a rare superficial mycosis. It has not been reported in Iran yet. We report a case of an Iranian boy resident of Amol (Mazandaran, Iran) that developed brown macules on his left palm. Direct microscopic examination and culture confirmed the diagnosis of Tinea nigra.

Neoscytalidium dimidiatum as onychomycosis causative agent in an Iranian patient: a case report and literature review.

A 38-year-old healthy male presented to our medical mycology center with whitish opaque discoloration of the right toenail. He reported a history of some sand scratches subsequent to walking barefoot on the beach two years ago and wearing hard safety shoes for a period of two years. On clinical examination, onycholysis, onychodystrophy, and apparent thickening of the ungual bed in the left big toe were found. The microscopic examination of nail clippings using 15% potassium hydroxide (KOH/) revealed the presence of septate pigmented hyphae. The fungus was identified as Neoscytalidium dimidiatum based on the cultural characteristics, the arrangement of arthroconidia on lactophenol cotton blue (LPCB) staining, blocky-brown pigmented hyphae on serum physiology mounts, and sequencing. Susceptibility of the isolated fungi to amphotericin B, itraconazole, voriconazole, and terbinafine was tested using the standard broth microdilution M38-A2 method developed by the Clinical and Laboratory Standards Institute (CLSI). The minimum inhibitory concentrations (MICs) of the four antifungal drugs used in this study were: amphotericin B: 1 mg/L, itraconazole: 2 mg/L, voriconazole: 0.25 mg/L, and terbinafine: 1 mg/L. The patient underwent terbinafine and clobetasol topical treatments for 6 months.

Subtilisin Gene Activity in Dermatophytes: A study on the Presence of the Subtilisin Gene in Trichophyton verrucosum and Microsporum gypseum in Clinical and Nonclinical Samples in Tehran, Iran.

The keratinolytic activities of dermatophyte species are accompanied by the secretion of enzymes, such as serine proteases, which are coded by the Subtilisin (SUB) genes. This study aimed to determine the presence of the SUB genes in the clinical and nonclinical samples of Trichophyton verrucosum and Microsporum gypseum. Isolation was carried out by direct and laboratory examination. Following that, for the determination of the presence of the SUB gene, polymerase chain reaction with specific primers was conducted. The frequencies of the SUB gene were observed in almost 66% of the isolates. Statistical analysis showed a significant relationship between the presence of the SUB gene and the samples collected from human, animals, and soil (p ˂0.005). The current investigation has been the first study of the presence/absence of the SUB gene in the clinical and nonclinical isolates of T. verrucosum and M. gypseum in Iran which may be a new step to perform further studies.

Comparison of in vitro antifungal activity of novel triazoles with available antifungal agents against dermatophyte species caused tinea pedis.

OBJECTIVE: Dermatophytes are a group of keratinophilic fungi that invade and infect the keratinized tissues and cause dermatophytosis. We investigated effectiveness of novel triazole (luliconazole and lanaconazole) in comparison with available antifungal agents against dermatophyte species isolated from patients with tinea pedis. MATERIAL AND METHODS: A total of 60 dermatophytes species were isolated from the patients with tinea pedis. Identification of species was done by DNA sequencing of the ITS1-5.8S rDNA-ITS2 rDNA region. In vitro antifungal susceptibility testing with luliconazole and lanaconazole and available antifungal agent was done in accordance with the Clinical and Laboratory Standards Institute, M38-A2 document. RESULTS: In all investigated isolates, luliconazole had the lowest minimum inhibitory concentration (MIC) (MIC range=0.0005-0.004µg/mL), while fluconazole (MIC range=0.4-64µg/mL) had the highest MICs. Geometric mean MIC was the lowest for luliconazole (0.0008µg/mL), followed by lanoconazole (0.003µg/mL), terbinafine (0.019µg/mL), itraconazole (0.085 µg/mL), ketoconazole (0.089µg/mL), econazole (0.097µg/mL), griseofulvin (0.351 µg/mL), voriconazole (0.583µg/mL) and fluconazole (11.58µg/mL). CONCLUSION: The novel triazoles showed potent activity against dermatophytes and promising candidates for the treatment of tinea pedis caused by Trichophyton and Epidermophyton species. However, further studies are warranted to determine the clinical implications of these investigations.

A systematic review and meta-analysis on the epidemiology, casual agents and demographic characteristics of onychomycosis in Iran.

Onychomycosis or fungal nail infection is one of the most common fungal infections. Nearly 50% of all nail disorders are caused by fungi. This systematic review and meta-analysis was conducted to determine the prevalence of onychomycosis across Iran. We searched English and Persian databases for studies reporting the epidemiologic features of onychomycosis in Iranian people from January 2000 to December 2018. Literature search revealed 307 studies, of which 24 studies met the eligibility criteria. In order to identifying the existence of publication bias among studies, funnel plots were used. The results of the meta-analysis were visualized as a forest plot representing the prevalence estimates of each study. Heterogeneity was also analyzed using the I2, Chi2, and Tau2 statistics. A high level of I2 and Chi2 was obtained among studies, which provides evidence of notable heterogeneity between studies. The results of current study revealed that the highest prevalence of onychomycosis was related to Mazandaran and Tehran provinces, respectively. As in the literature hypothesized shift in etiologic agents from yeasts to dermatophytes or molds could not be confirmed. Females were affected more frequently than males and in both sexes the highest incidence of infection occurrence was at the ages of >50 years. It seems the highest prevalence of onychomycosis in Mazandaran and Tehran provinces is due to the concentration of specialist doctors and research centers in these two provinces compared with others which leads to more detection and more care of the disease. Therefore, further educational strategies in order to accurate diagnosis in other provinces is necessary to reduce the risk of onychomycosis in Iran.

The frequency of fungi isolated from the skin and hair of asymptomatic cats in rural area of Meshkin-shahr-Iran.

BACKGROUND: Dermatophytosis is a frequent cutaneous infection affecting the keratinized tissues of humans, pets and livestock. Animals can carry dermatophytic elements asymptomatically and are considered to play an important role in the epidemiology of the disease. As exposure to any infected lesion free animals, especially cats, may lead to the development of infection in humans. OBJECTIVES: This study was done to determine the frequency of fungal agents isolated from skin and hair of cats living in rural areas of Meshkin-shahr, Iran. ANIMALS: A total of 103 asymptomatic cats living in rural areas of the region were studied. METHODS: This cross-sectional descriptive study was performed in Medical Mycology Laboratory, School of Public Health, Tehran University of Medical Sciences from February 2015 to July 2016. A total of 103 asymptomatic cats were studied. Mycological analysis including direct examination and culture on SC, SCC and DTM of the collected samples were conducted. For molecular confirmation when needed, panfungal PCR targeting the ITS1 region of the rDNA gene cluster using primers ITS1 and ITS4 were performed. Gender and age were also recorded. RESULTS: None of the 103 cats examined were positive for fungal elements on direct examination. However, 15 (14.5%) cases showed dermatophytes growth. T. verrucosum was the most common etiologic agents of dermatophytosis. Although the gender of the cats had not significant association with dermatophytosis prevalence, age was a significant influential risk factor (P=0.019). Aspergillus spp., Alternaria spp., Rhizopus spp., Penicillium spp.and paecilomyces spp. in descending frequency were the most predominantly identified saprophytic fungi. CONCLUSION: Our findings clearly highlighted the epidemiological role of asymptomatic cats in spreading dermatophytosis to humans and other animals.

Species identification and in vitro antifungal susceptibility testing of Aspergillus section Nigri strains isolated from otomycosis patients.

INTRODUCTION: Aspergillus niger is the most commonly reported etiology of otomycosis based on morphological characteristics. This fungus is a member of Aspergillus section Nigri, a set of morphologically indistinguishable species that can harbor various antifungal susceptibility patterns. The aim of this study was to accurately identify and determine the susceptibility pattern of a set of black aspergilli isolated from otomycosis patients. METHODS: Forty-three black Aspergillus isolates from otomycosis patients were identified by using the PCR-sequencing of the ß-tubulin gene. Furthermore, the susceptibility of isolates to three antifungal drugs, including fluconazole (FLU), clotrimazole (CLT) and nystatin (NS), were tested according to CLSI M38-A2. The data were analyzed using the SPSS software (version 15). RESULTS: The majority of isolates were identified as A. tubingensis (32/43, 74.42%) followed by A. niger (11/43, 25.58%). The lowest minimum inhibitory concentration (MIC) values were observed for NS with geometric means (GM) of 4.65µg/mL and 4.83µg/mL against A. tubingensis and A. niger isolates, respectively. CLT showed wide MIC ranges and a statistically significant inter-species difference was observed between A. tubingensis and A. niger isolates (P<0.05). FLU was inactive against both species with GMs>64µg/mL. CONCLUSION: Species other than A. niger can be more frequent as observed in our study. In addition, considering the low and variable activity of tested antifungal drugs, empirical treatment can result in treatment failure. Accurate identification and antifungal susceptibility testing of isolates is, however, recommended.

Onychomycosis due to dermatophytes species in Iran: Prevalence rates, causative agents, predisposing factors and diagnosis based on microscopic morphometric findings.

OBJECTIVE: Onychomycosis (OM) or fungal nail infection is one of the most common fungal infections, which is increasingly prevalent. OM is caused by dermatophytes spp, yeasts and non-dermatophyte moulds (NDMs). The purpose of this study was to identify and determine the prevalence rates, predisposing factors and causative agents of OM using clinical symptoms and microscopic morphometric findings. MATERIALS AND METHODS: In the present study, 180 patients suspected of OM were evaluated by direct microscopy using KOH 20%, culturing in Mycosel and Sabouraud dextrose agar media and Olysia software for identifying the causative fungi of OM. RESULTS: From 180 referred patients, 118 (65.56%) had OM, of whom 79 (66.94%) were positive for infection with dermatophytes spp. Of the 79 cases, the commonest age group was 61-70 years (21%) with males being 46 (58.23%) and females being 33 (41.77%). Both the fingernail and toenail infections were most prevalent in male patients. Sex, diabetes and age above 60 years were significant predisposing factors for OM development. DLSO was observed as the only clinical pattern of OM and T. rubrum was the commonest dermatophyte isolate (49.34%). CONCLUSION: This study showed that T. rubrum was the most common dermatophyte agent of OM in Iran.

Evaluation of PCR-reverse line blot hybridization assay for simultaneous identification of medically important saprophytic fungi.

BACKGROUND: In immunocompromised patients suffering from invasive fungal infections, rapid identification of fungal species is important since the appropriate treatment is usually related to the responsible species. We describe here, an assay based on combination of PCR and reverse line blot hybridization (PCR/RLB) for differentiation causative agent of fungal infections. MATERIALS AND METHODS: We performed PCR/RLB assay on 10 reference strains, which include Aspergillus species (A. fumigatus, A. flavus, A. niger, A. terreus, and A. clavatus), Mucor circnelloides, Rhizopus oryzae, Alternaria alternata, Cladosporium herbarum, and Fusarium solani. Besides, twenty-two clinical specimens from patients with proven fungal infections were analyzed for the identification of species. The obtained results were then compared with the results of culture and sequence analysis. RESULTS: The fungal species-specific oligonucleotide probes were able to distinguish between all species represented in this study with the exception of cross-reactivity between A. niger and A. fumigatus species. Two specimens, which were represented as mixed fungi in culture, were identified properly by this method. Results of the RLB assay were concordant with the culture and ITS sequencing results. CONCLUSION: Our result demonstrate that the RLB assay potentially is suitable for rapid and simultaneous identification of variety fungal pathogens directly from culture as well as from clinical specimens.

Study of skin and nail Candida species as a normal flora based on age groups in healthy persons in Tehran-Iran.

The skin is the body's largest organ that hosts heterogeneous inhabitants. Until now, the diversity of the cutaneous microbiome was mainly investigated for bacteria and there is a little information about the skin fungal flora. Also, among skin fungal flora, Candida is found as a main member whose distribution is affected by sex, age, climate. In this study, differences in Candida community structure associated with 9 different skin sites of 238 healthy people during 10 months from July to March 2016, are described. These subjects were divided by age into 4 groups: infants, children, adults and geriatrics. The collected samples were examined by culture on Sabouraud Chloramphenicol Agar and CHROM-agar Candida. For precise identification of species ITS1-5. 8S-ITS2 rDNA regions were sequenced where needed. The frequency of Candida species was significantly different between age groups. The most Candida isolations were related to the elderly age group and the fewest in the infants. C. parapsilosis virtually, was the predominant isolated species in all age groups. This study showed no statistically significant effect of the subject's sex on Candida population resident on human skin surface.

In vitro activities of five antifungal agents against 199 clinical and environmental isolates of Aspergillus flavus, an opportunistic fungal pathogen.

Aspergillus flavus is the second leading cause of invasive and non-invasive aspergillosis, as well as the most common cause of fungal sinusitis, cutaneous infections, and endophthalmitis in tropical countries. Since resistance to antifungal agents has been observed in patients, susceptibility testing is helpful in defining the activity spectrum of antifungals and determining the appropriate drug for treatment. A collection of 199 clinical and environmental strains of Aspergillus flavus consisted of clinical (n=171) and environmental (n=28) were verified by DNA sequencing of the partial b-tubulin gene. MICs of amphotericin B, itraconazole, voriconazole, posaconazole, and MEC of caspofungin were determined in accordance with the Clinical and Laboratory Standards Institute M38-A2 document. Caspofungin, followed by posaconazole, exhibited the lowest minimum inhibitory concentrations (MIC). All isolates had caspofungin MEC90 (0.063µg/ml) lower than the epidemiologic cutoff values, and 3.5% of the isolates had amphotericin B MIC higher than the epidemiologic cutoff values. However, their clinical effectiveness in the treatment of A. flavus infection remains to be determined.

In vitro activity of econazole in comparison with three common antifungal agents against clinical Candida strains isolated from superficial infections.

BACKGROUND AND PURPOSE: Candida species are the most common organisms involved in superficial fungal infections, worldwide. Although econazole is among the most frequently used topical formulations for the treatment of candidiasis, no information is available regarding the susceptibility profiles of Candida species in Iran. MATERIALS AND METHODS: In vitro susceptibility of 100 clinical Candida isolates belonging to 6 species from superficial candidiasis of Iran towards to econazole was compared with three other common antifungal agents including itraconazole, fluconazole, and miconazole. Minimum inhibitory concentrations (MICs) values were analyzed according to the Clinical and Laboratory Standards Institute (CLSI) M38-A3 document. All isolates were previously identified to the species level, using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) on ITS region. RESULTS: The MIC of econazole, itraconazole, miconazole, and fluconazole were within the range of 0.016-16, 0.032-16, 0.016-16, and 0.25-64 µg/ml, respectively. In general, econazole and miconazole were more active against Candida isolates, compared to the other two agents. CONCLUSION: The present study demonstrated that for Candida albicans isolates, miconazole and econazole had the best effect, but in non-albicans Candida species, itraconazole and miconazole displayed more activity than other antifungal agents.

Onychomycosis in Tehran: mycological study of 504 patients.

Onychomycosis is a common nail disorder resulting from the invasion of the nail plate by a dermatophyte, yeast or mould species and gives rise to some diverse clinical presentations. The purpose of the present study was to isolate and identify the causative fungi of onychomycosis in the population of Tehran, Iran. Nail samples from 504 patients with prediagnosis of onychomycosis during 2005 were examined both by direct microscopical observation of fungal elements in KOH preparations and in culture for the identification of the causative agent. All samples were inoculated on (i) Sabouraud dextrose agar (SDA, Merck), (ii) SDA with 5% chloramphenicol and cycloheximide in duplicate for dermatophyte and (iii) SDA with 5% chloramphenicol in triplicate for mould isolation. The criteria for the diagnosis of onychomycosis caused by non-dermatophytic moulds were based on microscopical observation of fungal elements, growth of the same mould in all triplicate culture and no growth of a dermatophyte or yeast in all the cultures. Of 504 cases examined, 216 (42.8%) were mycologically proven cases of onychomycosis (144 fingernails, 72 toenails). Among the positive results, dermatophytes were diagnosed in 46 (21.3%), yeasts in 129 (59.7%) and non-dermatophytic moulds in 41 (19%). Trichophyton mentagrophytes was the most common causative agent (n = 22), followed by Trichophyton rubrum (n = 13), Candida albicans (n = 42), Candida spp. (n = 56) and Aspergillus spp. (n = 21). Nearly half of the clinically suspected fungal nail infections are onychomycosis and yeast is responsible for most of the infections in Iran.

Influence of dimethyl formamide pulping of bagasse on pulp properties.

Organosolv pulping of bagasse was conducted following a central composite design using a two-level factorial plan involving three pulping variables (temperature: 190-210 degrees C, time: 120-180 min, organic solvent charge: 40-60% dimethyl formamide). Responses of pulp properties (yield and holocellulose, alpha-cellulose, kappa number, ash and ethanol-dichloromethane extractives contents) and the pH of the resulting wastewater to the process variables were analyzed using statistical software (MINITAB). Main factor analysis revealed that optimum pulp has the following characteristics: 82.7% (yield), 92.9 (kappa number), 95.84% (holocellulose), 83.53% (alpha-cellulose), 1.403% (ash), 2.562% (ethanol-dichloromethane extractives contents) and 6.39 (pH). These results showed that acceptable properties of pulps could be gained at 200-210 degrees C for 150 min and 40-60% DMF. Based on these results, this method could be used for pulping of bagasse equivalent NSSC concerning high yield at a fixed kappa number. In addition, bagasse could be pulped with ease to approximately 55% yield with a kappa number approximately 31. Numerical analyses showed that cooking temperature had the greatest influence on properties of obtained pulps within the DMF concentrations and cooking time as cooking variables.