molecular epidemiological survey of clinically important dermatophytes in Iran based on specific RFLP profiles of betatubulin gene

Surveillance of dermatophytosis is essential to determine the likely changes in etiological trends and distribution profile of this infection. In this study beta tubulin gene [BT2], was used as the first time in a PCR-RFLP format to clarify the distribution of dermatophytosis agents in some parts of Iran. A total of 603 clinical isolates was obtained from 500 patients in Tehran, Isfahan, Mazandaran and Guilan provinces. The isolates were identified using macro/micro-morphological criteria and electrophoretic patterns of PCR amplicons of BT2 after digestion with each of the restriction enzymes. FatI, Hpy CH4V, MwoI and Alw21L. Among the patients, 59.2% were male and 40.8% female. The most prevalent clinical form was tinea pedis [42.4%], followed by tinea cruris [24.2%], tinea unguium [12.3%], tinea corporis [10.8%], tinea faciei [4%], tinea manuum [3.14%], tinea capitis [3%] and tinea barbae [0.16%], respectively. Trichophyton interdigitale ranked the first, followed by T.rubrum, Epidermophyton floccosum, Microsporum canis, T. tonsurans, T. erinacei and T. violaceum [each 0.49%] and the less frequent species were T. schoenleinii, M. gypseum and T. anamorph of Arthroderma benhamiae [each 0.16%]. A case of scalp infection by E. floccosum was an exceptional event in the study. No case of T.verrucosum was found. Trichophyton species and E. floccosum are yet the predominant agents of infection in Iran, while Microsporum species are decreasing. T.interdigitale and Tinea pedis remain as the most causal agent and clinical form of dermatophytosis, respectively. it seems that BT2 can be a useful genetic marker for epidemiological survey of common pathogenic dermatophytes

[ study on fungal colonization of respiratory tract in patients admitted to intensive care units of Sari and Babol hospitals]

Fungi are considered as a life threatening in immunocompromised patients and respiratory tract is the main involvement location. Critically ill patients who are admitted to intensive care units [ICU] may also be susceptible to these infections, because of their conditions. Fungal colonization in respiratory tract maybe consider as a probable source for infection. Therefore, in the present study we evaluatedfungal flora of respiratory tract in patients admitted to ICU. Bronchoalveolar lavage samples were collected by bronchoscope from 45 patients with underlying predisposing conditions for invasive fungal infection twice a week. Samples were homogenated by pancreatin for performance of direct microscopic examination and cultured on Sabouraud's dextrose agar. The grown fungi on culture media were identified by standard mycological procedures. The main underlying predisposing conditions were COPD [22.2%], hematologic malignancy [20.3%] and prolonged stay in the ICU [16.9%]. The mean length of ICU stay was 19.6 days. Overall, 80 samples had positive result in direct examination [76.2%] and culture [71.2%], respectively. The most frequent isolated fungi were Candida [64.7%], Aspergillus [19.3%] and Penicillium [7.9%]. Among Candida and Aspergillus species, C. albicans and A. flavus were most common. In 48.4% of patients, similar fungal species were isolated in both sampling times. The results of our study showed that the ICU patients were susceptible to fungal resistant colonization especially Candida and Aspergillus as two life threatening fungal agents. So we emphasize the control procedures to reduce the fungal infection possibility among ICU patients

[Identification of malassezia species isolated from patients with pityriasis versicolor and seborrhoeic dermatitis by PCR-RFLP]

Lipophilic yeast of the genus Malassezia are members of normal human cutaneous micro flora which are also associated with several skin diseases. It is strongly suspected that Malassezia species are responsible for pityriasis versicolor [PV], and seborrhoeic dermatitis [SD]. Considering various sensitivities among Malassezia species to antifungal, accurate species identification will facilitate the treatment of relevant diseases. Malassezia species can be identified through their morphological features and bio-chemical characteristics. However, these phenotypic methods are usually time consuming, and lack sufficient discriminatory power. Development of DNA- based methods for detection and identification of Malassezia species provide helpful alternatives for solving problems. The aim of this study was to examine the distribution of Malassezia species in patients with pityriasis versicolor and seborrhoeic dermatitis, using molecular methods. A total of 63 clinical isolates of Malassezia spp, 30 strains isolated from patients with PV and 33 strains isolated from patients with SD, were studied. To investigate the strains at molecular level, genomic DNA of Malassezia isolates were extracted and amplified within the ITS1 region [located between 18S and 5.8S rDNA] by polymerase chain reaction [PCR] assay. DNA sequencing of ITS1 of rDNA in Malassezia spp was performed to type the species. Restriction fragment length polymorphism [RFLP] analysis of ITS1 PCR product with two restrictive enzymes CFOI and BSTF5I, was used in subsequent species identification. In this study, 37 patients with PV [20 females, 17 males; 2 to 64 years old], 81.1% [30 case] yielded growth of Malassezia in culture, while the frequency of isolation of M. globosa was 53.3% [16 case], M .furfur 40% [12 case] and M. sympodialis 6.7% [2 case]. Of the 41 patients with SD [22 females, 19 males; 1 to 52 years old], 80.5% [33 case] yielded growth of Malassezia in culture, while the frequency of M. furfur was 42.4% [14 case], M.globosa 39. 4% [13 case], M. restrict 15.2% [5 case] and M. sympodialis 3% [1 case]. This PCR-RFLP Profile allows us to clearly identify important Malassezia species. The results of the PCR-RFLP analyses of clinical isolates were in complete agreement with those from DNA sequencing, morphological features and bio-chemical characterization. In patients with PV, the most frequently isolated species were M. globosa, followed by M .furfur. However, in patients with SD, the most frequently isolated species were M. furfur, followed by M. globosa. The PCR-RFLP system applied for the ITS1 fragment of the rDNA is a reliable, simple and rapid method for identification of the most important Malassezia species, but further work will be necessary in applying these techniques to additional patients

[ effect of ketoconazole 2% solution in comparison with ketoconazole 2% shampoo on clinical signs and malassezia yeasts in seborrhoeic dermatitis patients]

Seborrhoeic dermatitis [SD] is a common skin disorder. Malassezia yeasts have an important role in the etiology of SD. Since anti-fungal agents, especially in azoles are effective for treating SD, in this study, the effect of ketoconazole 2% solution on clinical signs and Malassezia in SD patients were assayed. 100 patients with SD were enrolled in this study. Patients were scored in regard to the severity of lesions at the initial evaluation and every 2 weeks for a 1 month period. Microscopic examination and culture of patients scale in days 0 and 28 were used for isolation and identification of Malassezia species. Patients were divided into two groups [ketoconazole 2% solution and shampoo] and followed after 14 and 28 days, and then clinical response was graded. 58% of patients showed lesions on their heads. In day 0, 51% of patients showed > 7 yeasts in each microscopic field. 77% of scale samples were positive to Malassezia spp. Growth and M. globosa [57.1%] had the most frequency. In day 28, 89.6% and 82.6% of treated patients with solution and shampoo showed 1-3 yeast in within entire smear, respectively. 94.8% and 82.6% of scale samples were negative to Malassezia spp growth, respectively. In day 0, patients with moderate SI had the most prevalence, whereas in day 28, patients with mild SI were predominant. Statistical test showed the correlation is significance only between SI and treatment with solution. The results of our study showed that according to decrease of yeast load and increase of improvement of SD signs after treatment with ketoconazole 2% solution, compared with ketoconazole 2% shampoo, 2% ketoconazole solution can be considered as an appropriate agent in treatment of Sd

[ survey on myco-flora of air, book and cabinet of Mazandaran univerity of medical sciences libraries]

Grown fungi on books can be a risk factor for occupants as well as its known agents of bio-deterioration. Therefore, in this study, we surveyed the myco-flora of air, book and cabinets at Mazandaran University of Medical Sciences Libraries. Opened plates [containing Sabouraud's dextrose agar with chloramphenicole media [SC] were used for the isolation of fungi in the air of indoor environment of libraries. Pleated carpet sterile fragments were used for survey of cabinets and books contamination. Then, these fragments were cultured on SC in laboratory. A total of 939 colonies with 17 genera of fungi were identified from the environment of 4 school libraries at the Mazandaran University of Medical Sciences. The most common fungi isolated were: Penicillium [62.0%], Yeast [13.6%], sterile hyphae [7.6%] and Candida [5.6%]. The most number of colonies were isolated from the air. Penicillium, Aspergillus, Alternaria and Stachybotrys were isolated from the libraries. They are considered toxigenic, allergenic, infective and also, as book deterioration agents

Airborne fungi in indoor and outdoor of asthmatic patients' home, living in the city of Sari

The aim of this study was identification of fungi in indoor and outdoor of asthmatic patients´ home environment. Opened plates [containing of Malt extract agar media] were used for isolation of fungi in the air of indoor [n=360] and outdoor [n=180] of 90 asthmatic patients? home living in the city of Sari at the level of breathing height. Plates were incubated in room temperature for 7- 14 days. Then grown fungi were identified by standard mycological techniques. A total of 1876 colonies with 31 and 1692 colonies with 27 genera of fungi were identified from indoor and outdoor of asthmatic patients' home respectively. The most common fungi isolated were Cladosporium, Aspergillus and Penicillium. Stachibotyris, Oedocephalum, and Stemphillium showed the least frequencies among the isolated fungi. Cladosporium, Aspergillus, Penicillium, and Alternaria as the most common allergenic moulds had the most frequencies in indoor air of the houses of asthmatic patients