Characterization of clinically important dermatophytes in North of Iran using PCR-RFLP on ITS region.

OBJECTIVE: Dermatophytosis is considered as one of the major public health problems in the world. Accurate identification of the dermatophytes species is important for recognition of the environment and animal sources of infection. In this survey, 1500 clinical specimens taken from outpatients suspected of dermatophytosis and they were examined by direct microscopy and culture. MATERIALS AND METHODS: In total, three hundred and twelve dermatophytes were isolates recovered in culture, they were identified using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method based on the ribosomal DNA internal transcribed spacer (ITS) regions. RESULTS: Tinea pedis was the most common clinical form (32.1%) followed by tinea cruris (24.4%), tinea manuum (16.4%), tinea unguium (8.3%), tinea corporis (7.3%), tinea faciei (6.4%), and tinea capitis (5.1%). Trichophyton interdigitale was the most frequent isolate (38.2%), followed by Trichophyton rubrum (29.8%), Epidermophyton floccosum (16.6%), Trichophyton tonsurans (14.8%) and Microsporum canis (0.6%). The frequency of dermatophytosis was higher in males than in females and in the age-group of 21-30 years. CONCLUSION: Our finding indicated that the incidence of dermatophytosis caused by anthropophilic dermatophytes in Mazandaran province is increasing. Also, this study provides valuable data for the prevention and control of dermatophytosis in the southern coast of the Caspian Sea.

Development of a high-resolution melting analysis assay for rapid and high-throughput identification of clinically important dermatophyte species.

Accurate identification of dermatophyte species is important both for epidemiological studies and for implementing antifungal treatment strategies. Although nucleic acid amplification-based assays have several advantages over conventional mycological methods, a major disadvantage is their high cost. The aim of this study was to develop a rapid and accurate real-time PCR-based high-resolution melting (HRM) assay for differentiation of the most common dermatophyte species. The oligonucleotide primers were designed to amplify highly conserved regions of the dermatophyte ribosomal DNA. Analysis of a panel containing potentially interfering fungi demonstrated no cross reactivity with the assay. To evaluate the performance characteristics of the method, a total of 250 clinical isolates were tested in comparison with the long-established PCR-RFLP method and the results were reassessed using DNA sequencing, as the reference standard method. The assay is able to type dermatophytes using normalised melting peak, difference plot analysis or electrophoresis on agarose gel methods. The results showed that, in comparison to PCR-RFLP, the developed HRM assay was able to differentiate at least 10 common dermatophytes species with a higher speed, throughput and accuracy. These results indicate that the HRM assay will be a useful sensitive, high throughput and cost-effective method for differentiating the most common dermatophyte species.

Clinical features, diagnosis, and outcomes of rhino-orbito-cerebral mucormycosis- A retrospective analysis.

BACKGROUND AND PURPOSE: Rhino-orbito-cerebral mucormycosis (ROCM) is a rare disease with acute and fulminant manifestation. This infection is associated with high morbidity and mortality rates. Herein, we reviewed the manifestations, underlying conditions, medical treatments, and surgical interventions in ROCM patients admitted to a tertiary referral center in northern Iran over a seven-year period. MATERIALS AND METHODS: In a retrospective analysis, 15 cases of ROCM were identified from 2007 to 2013 in Bu Ali Sina Hospital, Sari, Iran. All the ROCM cases were clinically diagnosed and confirmed by histopathological and/or mycological examination. The relevant demographic data, clinical, ophthalmic, and neurologic manifestations, underlying conditions, medical treatments, and surgical interventions were recorded and analyzed. RESULTS: The mean age of the patients was 54±11 years (age range: 28-70 years); 26.7% of the patients were male and 73.3% female (male: female ratio of 1: 2.7). Uncontrolled diabetes was noted in at least 86.7% (13/15) of the cases. The maxillary sinuses were the most frequently involved sites (66.7% of the cases) followed by the ethmoid sinus. Amphotericin B in combination with surgical debridement was used in the treatment of 80% of the cases. Furthermore, 73.3% of the patients who were diagnosed early and underwent medical and extensive surgical debridement of the infected tissues survived. CONCLUSION: Uncontrolled diabetes mellitus is considered to be the main predisposing factor for ROCM. To prevent and reduce mortality rate of this acute disease, early diagnosis based on clinical findings and biopsy is recommended.

In vitro activity of five antifungal agents against Candida albicans isolates, Sari, Iran.

BACKGROUND AND PURPOSE: Candidaalbicans is the most common causative agent of candidiasis. Candidiasis management is dependent on the immune status of the host, severity of disease, and the choice of antifungal drug. Antifungals, specifically triazoles, are widely administered for the treatment of invasive fungal infections. Herein, we aimed to evaluate the invitro susceptibility of C.albicans isolates to fluconazole (FLZ), itraconazole (ITZ), voriconazole (VRZ), amphotericin B (AMB), and Caspofungin (CAS). MATERIALS AND METHODS: A total of 44 clinical strains of C.albicans were collected from 36 patients admitted to four hospitals in Mazandaran Province, Iran. The invitro antifungal susceptibility testing was performed based on the Clinical and Laboratory Standards Institute methods. RESULTS: Generally, 34 isolates were susceptible to all the five antifungal drugs, while four isolates were susceptible or susceptible dose-dependent (SDD) and six isolates were SDD or resistant to these antifungals. The lowest minimum inhibitory concentration (MIC; 0.016 µg/ml) belonged to AMB and the highest MIC was for FLZ )16 µg/ml). The lowest MIC (50 0.063 µg/ml) was related to ITZ and the lowest MIC (90 0.25 µg/ml) pertained to CAS, in addition , the highest MIC (50 1 µg/ml) and MIC (90 4 µg/ml) were for FLZ. Four of the isolates showed resistance to both FLZ and VRZ, separately, and five isolates were resistant to ITZ. Caspofungin showed potent activity against more than %95 of the C.albicans isolates. CONCLUSION: Overall, we reported %9.1 resistance to FLZ and VRZ ,%11.3 resistance to ITZ and AMB, and %4.6 resistance to caspofungin .Our finding is in agreement with previous observations proposing that C.albicans isolates develop resistance to some antifungal drugs such as FLZ since they are widely used as prophylaxis.

Prevalence of otomycosis in Khouzestan Province, south-west Iran.

BACKGROUND: This study aimed to investigate the prevalence of otomycosis and aetiological agents in Khouzestan province, south-west Iran. METHODS: This cross-sectional study examined and cultured 881 swabs from suspected external otitis cases, collected from throughout Khouzestan province. Fungal agents were identified by slide culture and complementary tests when necessary. RESULTS: The mean patient age was 37 years. The 20-39 year age group had the highest prevalence of otomycosis: 293 cases, comprising 162 (55.3 per cent) women and 131 (44.7 per cent) men. The seasonal distribution of cases was: summer, 44.7 per cent; autumn, 28.7 per cent; winter, 14.7 per cent; and spring, 11.9 per cent. The fungal agents isolated were Aspergillus niger (67.2 per cent), Aspergillus flavus (13 per cent), Candida albicans (11.6 per cent), Aspergillus fumigatus (6.2 per cent) and penicillium species (2 per cent). CONCLUSION: Fungal otomycosis is still one of the most important external ear diseases. In this study in south-west Iran, Aspergillus niger was the predominant aetiological agent. However, clinicians should be cautious of candidal otomycosis, which has a lower rate of incidence but is more prevalent among 20-39 year olds.

Fungal Contamination of Indoor Public Swimming Pools, Ahwaz, South-west of Iran.

BACKGROUND: Using public swimming pools during different seasons and poor health behavior could be responsible in transmission of fungal disease through pool water and its environment. Therefore, this research was conducted to investigate fungal agents of indoor public swimming pools of Ahwaz, capital city of Khouzestan Province, south-west of Iran. METHODS: Ten indoor swimming pools of Ahwaz were investigated during two seasons for 6 months. Water specimens were collected by pump and environment samples including shower-bath area, margin of pool walls, dressing rooms, and slippers, by sterile carpet pieces. All specimens were cultured in SC and SCC culture media and fungal agents identification were based on macroscopic, microscopic characteristic and complement tests when it was necessary. Data analyzing was performed using SPSS version 13 for descriptive analyzing. RESULTS: A total of 593 samples were collected from different parts of pools. Interestingly in 13 samples from environmental places, dermatophytes were isolated as follows: Trichophyton mentagrophytes, T. rubrum, T. verrucosum and Epidermophyton floccosum 5, 4, 3 and 1 cases respectively. Ten cases of dermatophytes were isolated from floor of dressing area. Three hundred seventy two saprophytic fungi species and 32 yeasts were recovered from water and environment surfaces samples. Aspergillus, Penicillium and Mucor were the most common isolated saprophytic fungi. CONCLUSION: Existence of saprophytic fungi and yeast in pools water seems to be an indicator of their resistance to detergent agents. In addition, yeast water contamination could be from swimmers. Dermatophytes isolation from pools environment areas and foot washing sink, reveals the importance of public swimming pools in disease transmission. Because dressing places are being used by all of the swimmers, take care of hygienic discipline in these places should be noted by health policy markers.