Genotypes analysis of Candida albicans species complex from healthy individual saliva in Ahvaz, Iran.

Although Candida albicans is a part of the mycoflora of healthy individuals, it is causing mild to severe forms of candidiasis in patients with underlying diseases. The recent molecular investigations were classified three genotypes, A, B and C for C. albicans. The aim of the present study was to detect different genotypes of C. albicans complex species in a normal population in Iran. Saliva was randomly collected from a normal population, homogenized and cultured on CHROMagar Candida. Classical and molecular methods were used for the detection of isolates. Candida 25S rDNA gene was amplified using the primer pairs of CA-INT-L and CA-INT-R for ABC genotyping of C. albicans. Candida albicans complex was recovered from 103/194 (53·1%) normal papulation. Genotype A with a frequency of 41·7% was the most common isolate, followed by genotype C (34%), genotype B (20·4%) and genotype D (3·9%). Genotyping of C. albicans species complex from healthy individuals showed the presence of three A, B and C genotypes of C. albicans and one D genotype of C. dubliniensis.

Genotypic diversity and antifungal susceptibility pattern of Candida albicans species isolated from hospitalized paediatric patients with urinary tract infection in Iran.

AIMS: The present study aimed to determine the microsatellite length polymorphism (MLP) genotypic patterns and antifungal susceptibility profiles of Candida albicans isolated from patients with candiduria. METHODS AND RESULTS: DNA of 50 C. albicans isolates was used for molecular identification based on the ITS1 -5.8s-ITS2 region. MLP assays were performed to amplify three loci (EF3, CDC3 and HIS3), and PCR products were used for fragment analysis. Antifungal susceptibility tests were performed according to CLSI M27 4th ed guidelines. In all, 38 different genotypes were detected with the three polymorphic loci among C. albicans isolates, and only one genotype was homozygous. In comparison to other countries, our genotypes were divided into three clusters, two of which were linked to France and a third of which was linked to Austria. The genetic structures of the 50 C. albicans isolates revealed varied heterozygosity and significant Hardy-Weinberg equilibrium at the EF3 locus. Only one (2%) and four (8%) of isolates showed resistance to fluconazole and itraconazole, respectively. In C. albicans genotype G25, one (2%) of the isolates showed cross-resistance and non-wild-type resistance to posaconazole, itraconazole and fluconazole. CONCLUSION: MLP typing is a useful tool to analyse the genetic structure of C. albicans isolates. High genetic diversity (38 genotypes) was detected in the isolates tested here. Compared to isolates in other countries, the ones from our patients had a clear relationship with French and Austrian isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: Iranian isolates of C. albicans have a distinct genotype and show similarities only with French and Austrian isolates.

Monitoring of mycoflora in outdoor air of different localities of Ahvaz, Iran.

INTRODUCTION: Airborne fungi are considered important causes of respiratory allergy and diseases. The knowledge of these fungi in a city or region is important for the ecological diagnosis and specific treatment of respiratory manifestations induced by inhalation of fungal agents. Therefore, in recent years bioaerosols and their density in the air is highly regarded. Identifying fungi with common mycological methods as well as molecular about 5% of people are prone to allergic symptoms of the respiratory tract caused by molds during their lives. Fungi are being considered as one of the most common triggers of asthma, allergic rhinitis and other respiratory problems. AIMS: We aimed at the investigation of the airborne fungi of Ahvaz, Khuzestan province, Iran, during spring and autumn months. MATERIALS AND METHODS: A total of 224 Petri dishes with Sabouraud dextrose agar medium containing chloramphenicol were exposed at seven different regions in the city. Two hundred and twenty-four samples (112 samples per season) were taken from outdoor air seven areas of Ahvaz using microbial sampler (Quick Take-30, SKC, USA) with Debbi 14.3L/min during 5minutes directly on Sabouraud dextrose agar medium with Chloramphenicol and Dechlorane. The effects of environmental factors such as temperature, humidity and wind velocity on bioaerosol concentrations were studied. RESULTS: The dishes exposed yielded 1240 fungal colonies of 24 genera. The most predominant were: Cladosporium spp. (37.1%), Alternaria spp. (25.7%), Aspergillus spp. (13.5%), Penicillium spp. (5.9%), Drechslera spp. (3.46%) and Stemphylium spp. (3.46%). The maximum and minimum numbers of airborne fungi were isolated from humid (32.3%) and industrial (17%) environments, respectively. In addition, positive correlation was observed between the number of fungi and relative humidity (r=0.476, P=0.000) in spring and autumn seasons. CONCLUSION: This study indicated that the incidence of airborne fungi with clinical significance had a direct relationship with the variation of environmental conditions.

Luliconazole, a new antifungal against Candida species isolated from different sources.

OBJECTIVE: Luliconazole is an inhibitor for sterol 14-α-demethylase in fungal cells with a broad-spectrum antifungal activity against dermatophytes, Candida albicans, Malassezia species, dematiaceous and hyaline hyphomycetes. Furthermore, luliconazole has been clinically used for the treatment of pityriasis versicolor, dermatophytosis, onychomycosis, cutaneous and mucocutaneous candidiasis. In the present study, we aimed to evaluate in vitro antifungal activity of luliconazole against several strains of Candida species recovered from different clinical materials. MATERIALS AND METHODS: In the present study, 104 strains of Candida species including, 34 isolates from vaginitis, 23 isolates from AIDS patients with vaginal candidiasis, 24 isolates from neutropenic patients and 24 isolates from tracheal tubes, were examined for susceptibility tests. A serial dilution of luliconazole (4-0.008µg/mL) was tested against different strains of Candida species recovered from different sources. RESULTS: The minimum inhibitory concentration (MIC) range and MIC90 of vaginal isolates (HIV-) were 1-0.063 and 1µg/mL. Furthermore, the most of strains (50%) had a MIC of 0.5µg/mL. The MIC ranges were similar (2-0.016µg/mL) for both vaginal (HIV+) and neutropenic patients isolates, whereas, MIC90 for them were 0.5 and 1µg/mL, respectively. All tracheal tubes strains were inhibited at the range of 2-0.008µg/mL with MIC90=1µg/mL. Totally, the lowest MIC50 (MIC=0.015µg/mL), MIC90 (MIC=1µg/mL) and MICGM (MIC=0.05µg/mL) are correlated to C. glabrata, a non-albicans species. CONCLUSION: It is concluded that, luliconazole could be an alternative anti-Candida agent, however, in vivo studies must be confirmed usefulness of drug for clinical usage.

The efficacy of gaseous ozone against different forms of Candida albicans.

BACKGROUND AND PURPOSE: Ozone is an inorganic molecule with effective antimicrobial properties. Clinical treatment of ozonated water was used for the elimination of Candidaalbicans, Enterococcus faecalis, endotoxins, and biofilms from root canals. In addition, its therapeutic effects for tinea pedis, ulcers, and leishmaniasis were investigated. The purpose of the present study was to evaluate the fungicidal effects of ozone on different forms of C. albicans. In addition, antifungal susceptibility profile of strains was assessed before and after exposure to ozone. MATERIALS AND METHODS: Fifty strains of C. albicans were exposed to gaseous ozone at different times. Furthermore, biofilm formation and germ tube production were evaluated when yeast suspensions were exposed to ozone. In addition, antifungal susceptibility of ozone resistant colonies was investiagted as compared to controls. RESULTS: Ozone was highly effective in killing C.albicans in yeast form and inhibition of germ tube formation during 210 and 180 s, respectively. Although with increasing exposure time biofilm production was considerably decreased, resistance to ozone was much higher among vaginal and nail isolates even after 60 min. All the strains were sensitive to fluconazole, caspofungin, and terbinafine pre- and post-ozone exposure. Resistance to amphotericin B was significantly enhanced after exposure to ozone. CONCLUSION: Although ozone was highly effective on the yeast form of C.albicans and it can inhibit the formation of germ tubes in C.albicans, the complete removal of biofilms did not happen even after 60 min. It seems that ozone therapy induces resistance to amphotericin B.

Luliconazole, an alternative antifungal agent against Aspergillus terreus.

Aspergillus terreus is the fourth leading cause of invasive and non-invasive aspergillosis and one of the causative agents of morbidity and mortality among immunocompromised and high-risk patients. A. terreus appears to have increased as a cause of opportunistic fungal infections from superficial to serious invasive infections. Although, invasive aspergillosis is often treated empirically with amphotericin B, most A. terreus isolates are resistant both in vivo and in vitro to some antifungal drugs. In this study, we aimed to evaluate antifungals susceptibility profiles of the different strains of A. terreus against amphotericin B, caspofungin, fluconazole, voriconazole, posaconazole and luliconazole. Forty A. terreus strains originating from environmental sources (air and soil) were identified using by macroscopic and microscopic features. Six antifungals including, amphotericin B, caspofungin, fluconazole, voriconazole, posaconazole and luliconazole were applied for susceptibility tests. Our results show that tested isolates had different susceptibility to antifungals. The lowest MICGM related to luliconazole (0.00236µg/ml), followed by posaconazole (0.18621µg/ml), voriconazole (0.22925µg/ml), caspofungin (0.86µg/ml), fluconazole (8µg/ml) and amphotericin B (11.12µg/ml). This study demonstrated that luliconazole had an excellent in vitro activity against all tested isolates of A. terreus, with MICGM 0.00236µg/mL than other tested antifungals. As a result, luliconazole could be a possible alternative antifungal for the treatment of aspergillosis due to A. terreus.

Lamisil, a potent alternative antifungal drug for otomycosis.

BACKGROUND AND PURPOSE: Otomycosis is an acute, subacute or chronic fungal infection of the pinna, the external auditory meatus and the ear canal caused mainly by several species of saprophytic fungi. Lamisil (Terbinafine) is an allylamine antifungal agent, that is used both in the topical and oral administration for the treatment of dermatophytosis, cutaneous candidiasis, and the pityriasis versicolor. We investigated the in vitro activity of clotrimazole, miconazole, nystatin, and Lamisil against the agents of otomycosis. MATERIALS AND METHODS: Fifteen clinically obtained isolates from otomycosis (Aspergillus species; n=13, and Candida species, n=2) and 8 environmental isolates of Aspergillus were tested. The disk diffusion method was employed to detect susceptibility. In the present study, the in vitro activity of the terbinafine with clotrimazole, miconazole, and nystatin against several isolates of Aspergillus and Candida with different sources were compared. RESULTS: Out of 23 isolates of Aspergillus, Candida 4(17.4%) and 1(4.4%) were resistant to nystatin and miconazole, respectively. In addition, all tested organisms were sensitive to clotrimazole and terbinafine. Statistical analysis has shown that there are no significant differences on the effects of clotrimazole, miconazole and, terbinafine on saprophytic (environmental) and pathogenic isolates of A. niger, A. flavus, and A. terreus (P value= 0.85). In addition, all tested organisms were found to be highly susceptible to terbinafine (P< 0.04). CONCLUSION: This is a new approach for the possible use of Lamisil for the treatment of otomycosis.

Candiduria in hospitalized patients in teaching hospitals of Ahvaz.

BACKGROUND AND OBJECTIVES: Nosocomial infections are usually acquired during hospitalization. Fungal infection of the urinary tract is increasing due to predisposing factors such as; antibacterial agents, indwelling urinary catheters, diabetes mellitus, long hospitalization, immunosuppressive agents, use of IV catheters, radiation therapy, malignancy. The aim of our study was to determine the prevalence of candiduria and urinary tract infection in patients admitted in Golestan and Emam Khomeini hospitals of Ahvaz, Iran. MATERIALS AND METHODS: During 14 months, a total of 744 urine samples were collected and transferred to medical mycology laboratory immediately. Ten µl of uncentrifuged sample was cultured on CHROM agar Candida plates and incubated at 37°C for 24-48h aerobically. Candida species were identified based on colony morphology on CHROM agar Candida, germ tube production and micro-morphology on corn meal agar including 1% Tween 80. RESULTS: In the present study, 744 hospitalized patients were sampled (49.5%, female; 50.5%, male). The prevalence of candiduria in subjects was 16.5% that included 65.1% female and 34.9% male. The most common isolates were C. albicans (53.3%), followed by C. glabrata (24.4%), C. tropicalis (3.7%), C. krusei (2.2%), and Geotrichum spp. (0.7%) Urine cultures yielded more than 10,000 yeast colonies in 34.1% of cases, and the major predisposing factor associated with candiduria was antibiotic therapy (69.1%). CONCLUSION: Candiduria is relatively common in hospitalized patients in educational hospitals of Ahvaz. In addition, there is a strong correlation between the incidence of candiduria in hospitalized patients and broad-spectrum antibiotics therapy.

Interactions between Entomopathogenic Fungus, Metarhizium anisopliae and Sublethal Doses of Spinosad for Control of House Fly, Musca domestica.

BACKGROUND: Metarhizium anisopliae strain IRAN 437C is one of the most virulent fungal isolates against house fly, Musca domestica. The objective of this study was to determine the interaction of this isolate with sublethal doses of spinosad against housefly. METHODS: In adult bioassay, conidia of entomopathogenic fungus were applied as inoculated bait at 10(5) and 10(7) spore per gram and spinosad at 0.5, 1 and 1.5 µg (A.I.) per gram bait. In larval bioassay, conidia were applied as combination of spore with larval bedding at 10(6) and 10(8) spore per gram and spinosad at sublethals of 0.002, 0.004 and 0.006 µg (AI) per gram medium. RESULTS: Adult mortality was 48% and 72% for fungus alone but ranged from 66-87% and 89-95% in combination treatments of 10(5) and 10(7) spore/g with sublethal doses of spinosad respectively. The interaction between 10(5) spore/g with sublethals exhibited synergistic effect, but in combination of 10(7) spore in spite of higher mortality, the interaction was additive. There was significant difference in LT(50) among various treatments. LT(50) values in all combination treatments were smaller than LT(50) values in alone ones. Larval mortality was 36% and 69% for fungus alone but ranged from 58%-78% and 81%-100% in combination treatments of 10(6) and 10(8) spore/g medium with sublethals of spinosad respectively. The interaction was synergistic in all combination treatments of larvae. CONCLUSION: The interaction between M. anispliae and spinosad indicated a synergetic effect that increased the house fly mortality as well as reduced the lethal time.