Toxicities of the copper and zinc oxide nanoparticles on Marshallagia marshalli (Nematoda: Trichostrongylidae): evidence on oxidative/nitrosative stress biomarkers, DNA damage and egg hatchability.

This study investigated the in vitro anthelmintic activity of copper oxide (CuO) and zinc oxide (ZnO) nanoparticles (NPs) against Marshallagia marshalli. The in vitro study was based on an egg hatch assay, adult and larvae motility inhibition assays, DNA damage, intensity protein profile along with several oxidative/nitrosative stress biomarkers including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), protein carbonylation (PCO), malondialdehyde (MDA), total antioxidant status (TAS) and nitric oxide (NO) content. Different concentrations of CuO-NPs and ZnO-NPs (1, 4, 8, 12 and 16 ppm) were used to assess anthelmintic effects on three stages of M. marshalli life cycle - that is, eggs, larvae and adult parasites for 24 h. The results indicated that CuO-NPs and ZnO-NPs played a significant role as anthelminthics, and the effect was dependent on time and concentration. The concentrations of 12 and 16 ppm of CuO-NPs and 16 ppm of ZnO-NPs resulted in the induction of oxidative/nitrosative stress (decreased SOD, GSH-Px and CAT, and increased MDA, PCO and NO), increased DNA damage, inhibition of adult and larval motility, egg hatch and low intensity of protein bands following sodium dodecyl sulphate-polyacrylamide gel electrophoresis, compared to control. It was concluded that CuO-NPs and ZnO-NPs could be utilized as novel and potential agents for the control and treatment of M. marshalli infection, and they have the pharmacological potential to be studied in vivo for further utilization in treating parasitic infections.

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.

Molecular identification, phylogenetic analysis and antifungal susceptibility patterns of Aspergillusnidulans complex and Aspergillusterreus complex isolated from clinical specimens.

OBJECTIVE: Aspergillus sections Terrei and Nidulantes are the less common causes of invasive aspergillosis and pulmonary aspergillosis (PA) in immunocompromised patients when compared to A. fumigatus and A. flavus. Identifying these fungi as the infectious agent is crucial because of the resistance to amphotericin B (AMB) and increased lethality. The aim of this study was to identify the molecular status, evaluate the genetic diversity and examine the antifungal susceptibility profile of the uncommon Aspergillus species. Forty-five uncommon Aspergillus species were identified based on the microscopic and macroscopic criteria. Then, the molecular identification was performed using the sequencing beta tubulin (benA) gene. In vitro antifungal susceptibility to amphotericin B (AMB), itraconazole (ITC), ravuconazole (RAV), voriconazole (VRC), caspofungin (CFG) isavuconazole (ISA) and posaconazole (POS) test was performed according to the CLSI M38-A2 guidelines. RESULTS: A. terreus was the most species detected, followed by A. nidulans, A. latus, A.ochraceus, and A. citrinoterreus, respectively. The analysis of the benA gene showed the presence of 12 distinct genotypes among the A. terreus isolates. The other species did not show any intraspecies variation. CFG exhibited the lowest MEC50/MIC50 (0.007µg/mL), followed by POS (0.125µg/mL), VRC, ITC, ISA (0.25µg/mL), RAV (0.5µg/mL), and AMB (8µg/mL). Among all the isolates, only 15.5% (7/45) were susceptible to AMB. CONCLUSION: Antifungal susceptibility pattern of the uncommon Aspergillus species is useful to improve patient management and increase knowledge concerning the local epidemiology. Moreover, this information is necessary when an outbreak dealing with drug-resistant infections occurs.

Comparative in vitro activities of seven antifungal drugs against clinical isolates of Candida parapsilosis complex.

OBJECTIVE: Candida parapsilosis species complex, an important set of non-albicans Candida species, is known to cause candidaemia particularly in neonates and infants. However, the incidence has increased in recent years, owing to higher numbers of at individuals at risk for these infections. Our objective was to evaluate the in vitro susceptibility of clinical isolates of C. parapsilosis complex isolates from Iran to seven antifungal drugs. MATERIAL AND METHODS: One hundred-one clinical isolates of C. parapsilosis species complex cultured from humans were included. Species identification had been previously confirmed by combined phenotypic characteristics, matrix-assisted laser desorption ionization-time of flight mass spectrometry-based assay and reconfirmed by DNA sequence analysis of the ITS rDNA region and D1/D2 gene. Minimum inhibitory concentrations (MICs) for amphotericin B, fluconazole, itraconazole, voriconazole, posaconazole, micafungin and anidulafungin were determined against well-characterized isolates by broth microdilution susceptibility testing according to the CLSI M27-A3 guideline. RESULTS: Species identifications were performed on 101 isolates, of which 88 (87.2%) C. parapsilosis sensu stricto and 13 (12.8%) C. orthopsilosis. Amphotericin B and posaconazole were the most active drugs with 100% of isolates being wild-type (WT). Voriconazole and micafungin, 99% of isolates were WT. The low activity was recorded for fluconazole and itraconazole with 93.1% and 89.1% of isolates being WT, respectively. At the species level, all Candida parapsilosis sensu stricto isolates were WT to amphotericin B and posaconazole and all Candida orthopsilosis isolates were WT to amphotericin B, voriconazole, posaconazole, anidulafungin and micafungin. In contrast, the highest rate of non-WT was observed in C. orthopsilosis to itraconazole (4 of 13, 30.8%). CONCLUSIONS: Although almost all of the tested drugs demonstrated potent activity against C. parapsilosis species complex, it seems that more especially C. orthopsilosis isolates had decreased susceptibility to itraconazole. Further studies are needed to determine how these findings may switch into in vivo efficacy.

Experiences of the mothers of infants hospitalized in the neonatal intensive care unit (NICU).

BACKGROUND: The mother-newborn relationship is more important in neonates hospitalized in the NICU than in healthy neonates. This study was conducted to explore the experiences of the mothers of infants hospitalized in the NICU. MATERIALS AND METHODS: This qualitative study was done in 2016 by adopting a conventional content analysis approach. Thirty-five mothers in the NICUs, Imam Hossein Hospital and Fatemieh Hospital were selected. Their experiences were assessed using in-depth individual semi-structured interviews. Sampling was purposive and was continued until reaching data saturation. RESULTS: Two hundred and nine primary codes were extracted. After removing duplicates and overlaps, 95 primary codes were categorized in 8 subcategories, 2 accessory categories and 1 main category based on their appropriateness, agreement, and similarity. The accessory categories of "mothers' worries" and "mothers' hopes" were merged into a more general, abstract category named "dual feelings about the baby". CONCLUSIONS: The nurses' awareness of the mothers' experiences can help design interventions to promote the quality of care for mothers and infants in the critical period of the NICU admission.

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.

Comparison of biofilm-producing ability of clinical isolates of Candida parapsilosis species complex.

OBJECTIVE: Candida parapsilosis is one of the main emerging non-Candida albicans species leading to superficial and systemic fungal infections in humans. Candida has the ability to produce biofilms associated with pathogenesis. The aim of the study was to estimate biofilm-producing ability of clinical isolates of C. parapsilosis sp. complex. METHODS: Clinical samples of C. parapsilosis complex have been analyzed. Crystal violet (CV) staining and tetrazolium reduction assay (MTT) have been used to analyze the clinical isolates ability to produce biofilms. The biofilm's structural characteristics have been assessed by using scanning electron microscopy. RESULTS: All 65 isolates were able to form biofilm. In addition, no significant difference was found between biofilm quantification based on two assays at different time intervals (24h, 48h, 72h, 96h) (P>0.05), with the exception of Candida orthopsilosis, which exhibited higher metabolic activity at 24h time point (P<0.05). Moreover, metabolic activity and production of biofilm biomass demonstrated statistically significant correlation (r=0.685, P<0.01). According to microscopic observations, the investigated clinical strains formed the similar surface topography with the slight differences in morphology; in addition, there was no statistically significant difference between efficiency of two assays to quantify biofilm. CONCLUSION: It was shown that, similar to C. parapsilosissensu stricto, two cryptic identified species (C. orthopsilosis and Candida metapsilosis) obtained from different clinical samples, were biofilm producers, while C. parapsilosissensu stricto exhibited the highest biofilm production.

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.

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.

Isolation of keratinophilic fungi from the soil of Greater Tunb, Abu-Musa, and Sirri islands in the Persian Gulf, Iran.

BACKGROUND AND PURPOSE: Keratinophilic fungi are among the important groups of fungi living in the soil. This study aimed to isolate and identify keratinophilic fungi from the soil of three Iranian islands, namely Greater Tunb, Abu Musa, and Sirri, located in the Persian Gulf using morphological and molecular (polymerase chain reaction) methods. MATERIALS AND METHODS: In this study, a total of 60 soil samples were collected from the three islands of Greater Tunb, Abu Musa, and Sirri. The samples were analyzed for the presence of the keratinophilic fungi using a hair baiting technique. Furthermore, the identification of keratinophilic fungi was accomplished through the employment of molecular and sequencing techniques. RESULTS: A total of 130 fungal isolates, including 11 genera with 24 species, were collected. Accordingly, Chrysosporium tropicum (24;18.5%), C. keratinophilum (17; 13.1%), Chrysosporium species (15; 11.5%), Aspergillus species ( 8;6.1%), Aspergillus flavus (8; 6.1%), Penicillium species (8;6.1%), Alternaria spp ( 6; 4.6%), Phoma species (5; 3.8%), Aphanoascus verrucosus (4;3.1%), Fusarium chlamydosporum (4; 3.1%), Aspergillustrreus (4;3.1%), Acremonium species (4; 3.1%), and other fungi( 23; 17.8 %) isolates were identified . All isolates of keratinophilic fungi were isolated from the soils with the pH range of 7-9. CONCLUSION: The results of this study contributed towards a better conceptualization of the incidence pattern of keratinophilic fungi in the regions of Iran. Given that no study has investigated this issue, the findings of the present study can be beneficial for the management of public health surveillance, physicians, and epidemiologists.

First case of superficial infection due to Naganishia albida (formerly Cryptococcus albidus) in Iran: A review of the literature.

BACKGROUND AND PURPOSE: Naganishia albida (formerly Cryptococcus albidus) is a non-neoformans cryptococcal species rarely isolated as a human pathogen. CASE REPORT: Herein, we present the case of a 26-year-old Iranian man with a superficial cutaneous lesion in the axilla. The initial treatment for pityriasis versicolor by clotrimazole was unsuccessful. We performed skin sampling based on the standard protocol and conducted further investigations by the conventional laboratory tests and molecular analysis of the skin samples. All the mentioned analyses revealed N.albida as the causative agent of infection. The minimum inhibitory concentration (MIC) analysis was carried out for the isolated agent, and the patient was treated using 100 mg daily of oral itraconazole. CONCLUSION: N. albida can be the causative agent of some superficial infections. This is the first report on the successful detection and treatment of a superficial skin infection due to N. albida by oral itraconazole.

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 antifungal susceptibility of clinical species belonging to Aspergillus genus and Rhizopus oryzae.

OBJECTIVE: Among filamentous fungal pathogens, Aspergillus spp. and zygomycetes account for highest rates of morbidity and mortality among immunocompromised patients. Recently developed antifungal drugs offer the potential to improve management and therapeutic outcomes of fungal infections. The aim of this study was to analyse the in vitro activities of voriconazole, itraconazole, amphotericin B and caspofungin against clinical isolates of Aspergillus spp. and Rhizopus oryzae. MATERIAL AND METHODS: The in vitro antifungal susceptibility of 54 isolates belonging to different clinical isolates of Aspergillus spp. and R. oryzae was tested for four antifungal agents using a microdilution reference method (CLSI, M38-A2). All isolates were identified by typical colony and microscopic characteristics, and also characterized by molecular methods. RESULTS: Caspofungin (MEC range: 0.008-0.25 and MEC50: 0.0023µg/mL) was the most active drug in vitro against Aspergillus spp., followed by voriconazole (MIC range: 0.031-8 and MIC50: 0.5µg/mL), itraconazole (MIC range: 0.031-16 and MIC50: 0.25µg/mL), and amphotericin B (MIC range: 0.125-4 and MIC50: 0.5µg/mL), in order of decreasing activity. The caspofungin, voriconazole, and itraconazole demonstrated poor in vitro activity against R. oryzae isolates evaluated, followed by amphotericin B. CONCLUSION: This study demonstrates that caspofungin had good antifungal activity and azole agents had better activity than amphotericin B against Aspergillus species. Although, azole drugs are considered ineffective against R. oryzae. This result is just from a small scale in vitro susceptibility study and we did not take other factors into consideration.

Pseudohyphae formation in Candida glabrata due to CO2 exposure.

BACKGROUND AND PURPOSE: Formation of pseudohyphae is considered a virulence factor in Candida species. Generally, Candida glabrata grows as budding yeast cells; however, reports illustrated that C. glabrata could form pseudohyphal cells in response to some stimuli. In this study, we provided insight into the ability of C. glabrata in forming pseudohyphal cells under different levels of carbon dioxide (CO2). MATERIALS AND METHODS: Candida glabrata reference strain (ATCC 90030) was used in this study. Yeast samples were cultured on Sabouraud dextrose broth (SDB) medium and incubated under 3%, 5%, and 10% CO2 levels for 24, 48 and 72 h. Control cultures were prepared without CO2 pressure for three days. The possibility of pseudohyphae and mycelium formation in C. glabrata was investigated. RESULTS: The results of this study revealed that the most branching filament-like cells were obtained at high CO2 pressure (10%) after 72 h. After three days of low CO2 pressure (3%), only yeast and budding cells were observed without any pseudohyphae formation. CONCLUSION: CO2 could act as a stimulus and induced formation of pseudohyphae in Candida glabrata yeast cells.

PCR - RFLP patterns for the differentiation of the Fusarium species in virtue of ITS rDNA.

BACKGROUND AND PURPOSE: The Fusarium species are among the most important fungi in the medical, veterinary and agricultural fields. MATERIALS AND METHODS: In the present study, 172 strains of these fungi have been analyzed. The high molecular weight DNAs were extracted from 23 reference strains as well as from 149 isolated Fusarium species. Using the designed nucleotide primers from rDNA of Fusarium species, PCR analysis was performed for the amplification of ITS regions. Afterwards, the location of the effective endonuclease enzymes has been evaluated within approximately 930 bp of rDNA sequence. RESULTS: Through the selected enzymes including; HhaI, MspI, TaqI and FaqI, the mentioned Fusarium species have been divided into 33 groups. The first three enzymes were able to classify Fusarium species into 23 groups of which 19 groups included one member, one group included two members and three groups included three members of the Fusarium species. This study also revealed the possibility in the identification of F. semitectum, F. solani complex, F. pseudograminearum, F. nisikadoi, F. coeruleum and F. acuminatum species by one unique enzyme. In addition, our study indicated the ability of the differentiation of F. Compactum from F. equiseti. CONCLUSION: As Compared to previous studies with more endonuclease enzymes and with limited in identifications, the ITS-RFLP patterns reported here an attempted to evaluate most of the Fusarium species successfully.

Effects of thiamine on growth, aflatoxin production, and aflr gene expression in A.parasiticus.

BACKGROUND AND PURPOSE: Mycotoxins are secondary fungal metabolites with a very high diversity that are produced by some species of Aspergillus which frequently leads to contaminate food and agricultural products. Recently, elimination of aflatoxin contamination in food and feed has been considered by scientists worldwide. Although, the antibacterial and antifungal effects of vitamins as natural compounds have been proven, the mechanism of vitamins effect on Aspergillus parasiticus growth and aflatoxin production is not yet clear. In this study, the effect of thiamine (vitamin B1) was studied on Aspergillus parasiticus growth, aflatoxins production and the afIR gene expression. MATERIALS AND METHODS: A standard strain of Aspergillus parasiticus was applied for performing antifungal susceptibility test in different concentrations of thiamine. Antifungal susceptibility test was performed according to CLSI M38-A2 document. The concentration of aflatoxin was determined by HPLC. Moreover, the quantitative changes in the aflR gene expression were analyzed by Real Time PCR method. RESULTS: The minimum inhibitory concentration was yielded as > 500 mg/ml. However, HPLC analysis results showed that aflatoxin production reduced in samples treated with 500 mg/ml of thiamine. In addition, the level of afIR gene expression was significantly reduced after treating with 500 and 250 mg/ml of vitamin B1. CONCLUSION: Based on the obtained results, thiamine could not inhibit the fungal growth completely. However, the rate of afIR gene expression and aflatoxin production was significantly reduced after fungal treating with thiamine. Consequently, using natural compounds such as vitamins may be regarded as potential antitoxic agent in food industry and the industries related to agriculture.

Development of RFLP-PCR method for the identification of medically important Aspergillus species using single restriction enzyme MwoI.

In this study we attempted to modify the PCR-RFLP method using restriction enzyme MwoI for the identification of medically important Aspergillus species. Our subjects included nine standard Aspergillus species and 205 Aspergillus isolates of approved hospital acquired infections and hospital indoor sources. First of all, Aspergillus isolates were identified in the level of species by using morphologic method. A twenty four hours culture was performed for each isolates to harvest Aspergillus mycelia and then genomic DNA was extracted using Phenol-Chloroform method. PCR-RFLP using single restriction enzyme MwoI was performed in ITS regions of rDNA gene. The electrophoresis data were analyzed and compared with those of morphologic identifications. Total of 205 Aspergillus isolates included 153 (75%) environmental and 52 (25%) clinical isolates. A. flavus was the most frequently isolate in our study (55%), followed by A. niger 65(31.7%), A. fumigatus 18(8.7%), A. nidulans and A. parasiticus 2(1% each). MwoI enabled us to discriminate eight medically important Aspergillus species including A. fumigatus, A. niger, A. flavus as the most common isolated species. PCR-RFLP method using the restriction enzyme MwoI is a rapid and reliable test for identification of at least the most medically important Aspergillus species.

Development of RFLP-PCR method for the identification of medically important Aspergillus species using single restriction enzyme MwoI

In this study we attempted to modify the PCR-RFLP method using restriction enzyme MwoI for the identification of medically important Aspergillus species. Our subjects included nine standard Aspergillus species and 205 Aspergillus isolates of approved hospital acquired infections and hospital indoor sources. First of all, Aspergillus isolates were identified in the level of species by using morphologic method. A twenty four hours culture was performed for each isolates to harvest Aspergillus mycelia and then genomic DNA was extracted using Phenol-Chloroform method. PCR-RFLP using single restriction enzyme MwoI was performed in ITS regions of rDNA gene. The electrophoresis data were analyzed and compared with those of morphologic identifications. Total of 205 Aspergillus isolates included 153 (75%) environmental and 52 (25%) clinical isolates. A. flavus was the most frequently isolate in our study (55%), followed by A. niger 65(31.7%), A. fumigatus 18(8.7%), A. nidulans and A. parasiticus 2(1% each). MwoI enabled us to discriminate eight medically important Aspergillus species including A. fumigatus, A. niger, A. flavus as the most common isolated species. PCR-RFLP method using the restriction enzyme MwoI is a rapid and reliable test for identification of at least the most medically important Aspergillus species.