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Background: Cholestasis is described as a disease in which bile flow from the liver is reduced or stopped, and due to its oxidative effects, irreversible consequences may occur. Objectives: Due to the remarkable antioxidant properties of Oliveria decumbens (OD) and the contribution of oxidants to the progression of bile duct ligation (BDL)-induced cholestasis, this research aimed to examine how the OD ethanolic extract affected liver damage and oxidant-antioxidant balance markers in BDL-induced cholestasis. Methods: Forty male Wistar rats weighing 200 - 250 g were used. Cholestasis was induced using the BDL approach. The rats were categorized into four groups: Group 1, sham-control (SC); group 2, cholestatic; group 3, SC + OD; and group 4, cholestatic + OD. A dose of OD ethanolic extract was administered orally (500 mg/kg/day) to rats for seven days. Seven days following surgery, the rats’ blood samples were collected; after sacrifice, a part of the liver tissue was isolated. A histopathological examination was performed, while the rest was stored at -70°C in liquid nitrogen. Heparin-containing tubes were used to gather blood samples. In plasma and hepatic tissue, biochemical tests, histopathological evaluations, and oxidative stress markers staining levels were performed. Results: Our findings showed that OD could effectively reduce liver injury by reducing the activity of liver function enzymes (AST and ALP). At the same time, it did not affect total bilirubin and protein. Bile duct ligation-induced hepatic markers of protein oxidation (PCO) and reactive nitrogen species (NO) were significantly decreased by OD, and it also promoted liver antioxidant capacity by enhancing superoxide dismutase (SOD) activities. Moreover, OD treatment prevented liver bile duct proliferative changes in histopathologic analysis. Conclusions: Our study confirmed that OD exerts substantial hepatoprotective activities against BDL-induced cholestasis by improving liver damage markers and regulating oxidative stress. It may be a beneficial therapeutic agent for managing cholestasis. Bioassay-guided isolation and identification of bioactive OD secondary metabolites can further direct the discovery of potential natural-based drug candidates.

Background The current study aimed to determine the seroprevalence of Toxoplasma gondii infection among HIV-positive patients and healthy subjects. Methods This study was carried out on HIV-positive patients and healthy individuals in Southwest Iran. Five millilitres of venous blood samples were collected aseptically from each individual. Sera and buffy coats were isolated from each sample and evaluated for anti-Toxoplasma antibodies and T. gondii DNA using ELISA kit and real-time PCR, respectively. Statistical analysis was performed using SPSS 18 software. Results Of 64 AIDS/HIV-positive patients, six (9.3%, 95% CI 7.2 to 11.3%) were seropositive for only IgG and five (7.8%, 95% CI 6.0 to 9.5%) were seropositive for both IgG and IgM. Moreover, among 64 healthy controls, 10 (15.6%, 95% CI 12.1 to 19.0%) were seropositive for only IgG and 2 (3.1%, 95% CI 2.4 to 3.7%) were seropositive for both IgG and IgM. Toxoplasma gondii DNA was detected in six samples (9.3%, 95% CI 7.2 to 11.3%) in the AIDS/HIV-positive patients group and eight samples (5.95%, 95% CI 4.6 to 7.2%) in the control group using real-time PCR. Consumption of undercooked meat was documented as an associated risk factor for T. gondii seropositivity in AIDS patients (OR 4.06, 95% CI 0.966 to 17.09; p=0.045). Conclusions Our findings showed a lower prevalence of Toxoplasma infection in AIDS/HIV-positive patients vs healthy controls; however, a considerable number of AIDS/HIV-positive patients were also seen to be at risk of toxoplasmosis. Based on the findings, screening and prophylaxis for toxoplasmosis should be implemented for all AIDS/HIV-positive patients in Southwest Iran.

Background Species from the Trichophyton benhamiae complex are mostly zoophilic dermatophytes which cause inflammatory dermatophytosis in animals and humans worldwide. Objectives This study was purposed to (a) to identify 169 reference and clinical dermatophyte strains from the T benhamiae complex species by molecular method and adhering to the newest taxonomy in the complex (b) to evaluate the in vitro antifungal susceptibility profile of these strains against eight common and new antifungal agents that may be used for the treatment of dermatophytosis. Methods All isolates, mainly originated from Europe but also from Iran, Japan and USA, were subjected to ITS-rDNA sequencing. The in vitro antifungal susceptibility profiles of eight common and new antifungal drugs against the isolates were determined by CLSI M38-A2 protocol and according to microdilution method. Results Based on the ITS-rDNA sequencing, T benhamiae was the dominant species (n = 102), followed by T europaeum (n = 29), T erinacei (n = 23), T japonicum (n = 10), Trichophyton sp (n = 4) and T eriotrephon (n = 1). MIC ranges across all isolates were as follows: luliconazole: 0.0002-0.002 µg/ml, terbinafine: 0.008-0.125 µg/ml, efinaconazole: 0.008-0.125 µg/ml, ciclopirox olamine: 0.03-0.5 µg/ml, itraconazole: 0.06-2 µg/ml, griseofulvin: 0.25-4 µg/ml, amorolfine hydrochloride: 0.125-4 µg/ml and tavaborole: 1-16 µg/ml. Conclusion Luliconazole, efinaconazole and terbinafine were the most potent antifungals against T benhamiae complex isolates, regardless of the geographic locations where strains were isolated. These data might help dermatologists to develop effective therapies for successful treatment of infections due to T benhamiae complex species.

The ability of dermatophytes to develop biofilm, as one of the virulence factors in fungal infections which contribute to antifungal resistance, is an outstanding aspect of dermatophytosis that has been noted recently. Because of the paucity of data about the biofilm formation by dermatophytes and their susceptibility to antifungal drugs, this study evaluated the biofilm formation by clinical isolates of dermatophytes and antibiofilm activity of common antifungals widely used to manage dermatophytosis.The ribosomal DNA internal transcribed spacer (ITS) regions sequencing for species identification of 50 clinical dermatophyte isolates was performed. The ability of isolates to form biofilm and inhibitory activity of itraconazole, terbinafine, and griseofulvin against biofilm formation was assayed by the crystal violet staining method. Optical microscopy and scanning electron microscopy (SEM) were applied for the visualization of the biofilm structures.Trichophyton (T.) mentagrophytes (n: 14; 28%) and T. rubrum (n: 13;26%) were included in more than half of the dermatophyte isolates. Biofilm formation was observed in 37 out of 50 (74%) isolates that were classified as follows: nonproducers (n: 13; 26%), weak producers (n: 4; 8%), moderate producers (n: 16; 32%), and strong producers (n: 17; 34%) by comparison of the absorbance of biofilms produced by clinical strains with control. The mean IC50 values for terbinafine, griseofulvin, and itraconazole were 2.42, 3.18, and 3.78 μg/ml, respectively.The results demonstrated that most of the clinical dermatophyte isolates are capable to form biofilm in vitro with variable strength. Moreover, terbinafine can be suggested as the first-line choice for the treatment of biofilm-formed dermatophytosis.

Introduction. Aspergillus sections Flavi and Nigri comprise clinically relevant and cryptic species that differ significantly in drug susceptibility, meaning that effective treatment depends on correct species identification. Hypothesis/Gap Statement. There are no comprehensive data for molecular identification and antifungal susceptibility testing (AFST) of clinically relevant and cryptic species of Aspergillus sections Flavi and Nigri as the main agents of invasive and non-invasive aspergillosis in Iran. We aimed to perform molecular identification and AFST of 213 clinical Aspergillus isolates belonging to sections Flavi and Nigri. Molecular identification of isolates was performed using sequencing of the β-tubulin gene and in vitro AFST was conducted according to the Clinical and Laboratory Standards Institute (CLSI) M38-A3 guidelines. Results. The most common isolates in sections Flavi and Nigri were Aspergillus flavus (110/113, 97.3 %) and Aspergillus tubingensis (49/100, 49.0 %), respectively. A total of 62/213 (29.1 %) isolates belonging to cryptic species were identified; among them, A. tubingensis was the most prevalent (49/62, 79.0%). Aspergillus flavus and A. niger isolates that responded to the minimum inhibitory concentrations (MICs) of itraconazole above the epidemiological cutoff values were the most frequently detected: 8/110 (7.3 %) and 3/41 (7.3 %), respectively. In section Flavi, Aspergillus alliaceus responded to amphotericin B at a high MIC (>16 µg mL−1) and in section Nigri, one of the three Aspergillus luchuensis/awamori isolates responded to itraconazole at an MIC >16 µg ml−1. Interestingly, for all Aspergillus welwitschiae isolates, the MIC50 and MIC90 of itraconazole were both 16 µg ml−1. Conclusion. A considerable presence of A. flavus and A. niger isolates showing non-wild-type responses to azoles in clinical cases of aspergillosis indicates the importance of classifying clinical Aspergillus isolates at the species level and performing antifungal susceptibility testing on the isolates, which would ensure appropriate treatment.

Candida glabrata infections are responsible for deaths of people globally. Fluconazole is known to be less effective against C. glabrata, which developed many strategies to evade being destroyed by fluconazole. To achieve enhanced efficacy of fluconazole against C. glabrata, the interaction of fluconazole with sodium bicarbonate was investigated using the CLSI guidelines. The efficacy of fluconazole alone and in combination with sodium bicarbonate was evaluated using the time-kill and phospholipase production assays. Eventually, the expression of PLB was assessed using semi-quantitative RT-PCR to investigate the inhibitory properties of fluconazole alone and in combination with sodium bicarbonate against C. glabrata. The fluconazole/sodium bicarbonate combination displayed synergistic and antagonistic effects (FICI= 0.375-4.25). In C. glabrata ATCC, SN 152, and SN 164, the fluconazole/sodium bicarbonate combination exhibited a significant fungicidal activity (p< 0.05) but antagonistic effect in the case of SN 283. With exception of SN 283, a significant reduction was noted in phospholipase production in clinical isolates of C. glabrata treated with fluconazole/sodium bicarbonate combination. The PLB was down-regulated significantly by 0.168-0.515 fold in C. glabrata treated with fluconazole/sodium bicarbonate. The results suggested fluconazole/sodium bicarbonate to have a potential synergistic interaction in C. glabrata, and the underlying mechanism may be associated with phospholipase gene.

Background and Purpose: Invasive aspergillosis (IA) of the central nervous system (CNS) is a devastating complication which is rarely reported in immunocompromised children. In this case presentation, we reported a rare and fatal IA with spinal cord involvement in a 10-year-old child with X-linked chronic granulomatosis disease (CGD). Case report: The child had a previous history of pulmonary tuberculosis. A cervical spine X-ray revealed the involvement of cervical vertebrae (T4/T5) and ribs causing spinal cord compression and epidural abscess. The patient underwent a decompressive laminectomy and mass removal. The histopathology and culture results suggested IA. Despite the aggressive and prolonged therapy, he died within one year. Aspergillus nidulans was identified as the causative agent based on morphological and molecular studies. Conclusion: This synopsis represents the aggressive behavior of infection caused by A. nidulans in the CGD patient.

Otomycosis is a superficial infection of the external ear caused by fungal pathogens. The genera Aspergillus and Candida are considered the main fungal causative agents, with the predominance of Aspergillus section Nigri. The present study aimed to evaluate the clinical symptoms of patients with otomycosis and predisposing factors and to identify fungal etiological agents using molecular approaches. We also present an overview of published papers on tympanic membrane perforation (TMP) secondary to otomycosis.An otorhinolaryngologist collected specimens from external ear canals of patients with suspected otomycosis based on the patient's history and clinical examinations. The specimens were collected using sterile swabs. Fungal isolates were confirmed in clinical specimens by direct microscopy and culture methods. Fungal isolates were identified based on molecular approaches.In total, specimens from 211 patients with suspected otomycosis were examined. The presence of fungi was confirmed in about 51% of patients based on fungal elements in direct microscopy and culture-positive fungi. Aspergillus tubingensis was the most commonly isolated species (52.77%), followed by Aspergillus niger (25.92%). Otomycosis due to infection with Candida species was observed in 16% of cases. Of note, in 36.11% of cases, otomycosis was associated with TMP.A mycological examination is indispensable for a correct diagnosis in patients with otitis extern. TMP should be considered in patients with otomycosis, as it appears to be relatively common in this population.

Pathogens from the Trichophyton benhamiae complex are one of the most important causes of animal mycoses with significant zoonotic potential. In light of the recently revised taxonomy of this complex, we retrospectively identified 38 Trichophyton isolates that could not be resolved into any of the existing species. These strains were isolated from Iranian and Czech patients during molecular epidemiological surveys on dermatophytosis and were predominantly associated with highly inflammatory tinea corporis cases, suggesting possible zoonotic etiology. Subsequent phylogenetic (4 markers), population genetic (10 markers), and phenotypic analyses supported recognition of two novel species. The first species, Trichophyton persicum sp. nov., was identified in 36 cases of human dermatophytosis and one case of feline dermatophytosis, mainly in Southern and Western Iran. The second species, Trichophyton spiraliforme sp. nov., is only known from a single case of tinea corporis in a Czech patient who probably contracted the infection from a dog. Although the zoonotic sources of infections summarized in this study are very likely, little is known about the host spectrum of these pathogens. Awareness of these new pathogens among clinicians should refine our knowledge about their poorly explored geographic distribution. IMPORTANCE In this study, we describe two novel agents of dermatophytosis and summarize the clinical manifestation of infections. These new pathogens were discovered thanks to long-term molecular epidemiological studies conducted in Czechia and Iran. Zoonotic origins of the human infections are highly probable, but the animal hosts of these pathogens are poorly known. Further research is needed to refine our knowledge about these new dermatophytes.

Background: Aspergillus and Candida species are the most commonly identified fungal pathogens in otomycosis. However, we usually encounter some difficulties in its treatment because many patients show resistance to antifungal agents and present a high recurrence rate. Objectives: The current research was conducted to compare the in vitro activities of luliconazole (LUL), and efinaconazole (EFN) and the nine comparators on Aspergillus and Candida strains isolated from otomycosis. Methods: The in vitro activities of nine common antifungal drugs (amphotericin B (AMB), voriconazole (VRC), fluconazole (FLU), itraconazole (ITC), ketoconazole (KTO), clotrimazole (CLO), nystatin (NYS), terbinafine (TRB), and caspofungin (CAS)) and two novel new azoles (LUL and EFN) against 108 clinical isolates of Aspergillus and Candida species obtained from otomycosis were assessed according to the CLSI broth microdilution document. Results: The LUL and EFN had the geometric mean minimum inhibitory concentrations (GM MICs) of 0.098 and 0.109 μg/mL against all Aspergillus strains, respectively. Furthermore, the GM MICs of all Candida isolates for LUL, EFN, CAS, CLO, VRC, AMB, ITC, KTO, FLU, NYS, and TRB were calculated to be 0.133, 0.144, 0.194, 0.219, 0.475, 0.537, 0.655, 1.277, 4.905, 9.372, and 13.592 μg/mL, respectively. Additionally, 6 (35.29%), 2 (11.7%), and 1 (5.88%) Candida isolates were resistant to FLU, CAS, and VRC, respectively. Conclusions: As the findings indicated, LUL and EFN showed the lowest GM MIC values against the examined species. Accordingly, these novel imidazole and triazole antifungal agents can be regarded as proper candidates for the treatment of otomycosis caused by Aspergillus and Candida strains.

Candidemia is the most common systemic infection in hospitalized patients and causing high mortality. Hence, the diagnosis of this infection in the early-stage with appropriate antifungal therapy has been attributed to the lowest mortality. The aims of this study are molecular identification of Candida species isolated from candidemia patients and evaluated the in vitro antifungal susceptibility patterns of these strains to fluconazole, amphotericin B, and caspofungin. In the present study, 800 hospitalized patients who suspected candidemia were sampled. Candida species were isolated and identified based on morphological and PCR-sequencing of the ITS1-5.8S-ITS2 region. Antifungal susceptibility tests for fluconazole, amphotericin B, and caspofungin was performed according to the Clinical and Laboratory Standards Institute M27-A3. Also, clinical data were recorded from patient's records. Overall, 27 candidemia patients were detected among hospitalized patients. 33.3% of candidemia patients were treated with amphotericin B, however, the mortality rate was 14.8%. The majority of patients (59%) were from the neonatal intensive care unit and premature born was the most underlying condition. C. albicans (n = 18; 66.6%) was the most common species isolated from blood cultures, followed by C.parapsilosis (n = 7; 25.9%), C.pelliculosa ( n = 1;3.7% ) and C.tropicalis (n = 1;3.7%). Only one C. albicans isolate were resistance to fluconazole (MIC = 32 µg/mL). Generally, C. albicans has been the most frequent causative agent of candidemia. Resistance to antifungal drugs among candidemia agents was rare. Also, the identification of Candida isolates at the species level with in-vitro antifungal susceptibility tests can manage and decrease the mortality rate among candidemia patients.

A molecular epidemiology study was conducted between 2016 and 2017 by a network of collaborators from 12 provinces in the Islamic Republic of Iran. A total of 1484 soil samples from different habitats were screened for the presence of dermatophytes by using the hair baiting technique. The primary identification of isolates was carried out by amplification and MvaI restriction fragment length polymorphism (RFLP) of the internal transcribed spacers regions of ribosomal DNA (ITS-rDNA). The identifications, especially in the cases of isolates with unknown RFLP patterns, were confirmed by sequencing of the ITS-rDNA region. As a result, 256 isolates were recovered. The isolation rate was higher in soils with pH range 7.1–8.0, collected from animal habitats (n = 78; 34%) and parks and gardens (n = 75; 32%), geographically from Mazandaran Province (n = 115; 49.5%) and seasonally in the spring (n = 129; 50.4%), all of which were statistically significant (p < 0.05). The dermatophytes comprising five species of the two genera, viz., Nannizzia fulva (n = 214), N. gypsea (n = 34), Arthroderma quadrifidum (n = 5), A. gertleri (n = 2) and A. tuberculatum (n = 1), were isolated. The geophilic dermatophytes occurred in various soils from different parts of Iran; however, surprisingly, N. fulva emerged as the dominant species, outnumbering the common geophilic species of N. gypsea. For the definitive identification of soil inhabitant dermatophytes, DNA-based identification is strongly recommended.

Aspergillus clavatus is a common environmental species known to cause occupational allergic disease in grain handlers. We have recently observed azole-resistant isolates of this fungus as a cause o...

Purpose Otomycosis is a mycotic infection of the external auditory canal and can be caused by a wide range of fungal species. In this study, we aimed to identify fungal isolates from patients suspected of otomycosis. Methodology External ear canal samples were taken from patients referred to the outpatient department of Shahid-Mofatteh Clinic in the city of Yasuj, Iran, and examined by direct microscopy and culture. DNA of the isolated fungi was tested by internal transcribed spacer PCR restriction fragment length polymorphism analysis for identification of yeasts and β-tubulin sequencing for identification of Aspergillus species. Results Among 275 patients suspected of otomycosis, 144 cases (83 female and 61 male) were confirmed with otomycosis. For 89% (n=128) of positive cultures, microscopy was also positive, while there were no cases with a microscopy-positive and culture-negative result. The predominant predisposing factor was self-cleaning of the external ear using unhygienic tools, and the main risk occupation was 'housewife'. The most common isolated fungi were typically Aspergillus (n=120), including 73 isolates of Aspergillus section Nigri, 43 of section Flavi, 3 of section Terrei and 1 of section Fumigati. After sequencing, 44 out of 73 strains primarily identified as Aspergillus niger turned out to be Aspergillus tubingensis. Thirty-five isolates were identified as Candida, including Candida parapsilosis (n=22), Candida albicans (n=12) and Candida tropicalis (n=1). Conclusion Aspergillus tubingensis was the most common species involved in otomycosis. This work corroborates the difficulty of precise identification of species within the black Aspergilli by morphological characteristics.

Orthodontic removable appliances can facilitate the accumulation of certain microorganisms and microbial plaque on tooth surfaces and appliance components. Since long-term use of chemical agents is not recommended, an alternative option would be medicinal plants for disinfection of the oral cavity.The aim of this study was to assess the efficacy ofIn this 2-arm parallel controlled clinical trial, forty-four patients whose removable orthodontic appliances were contaminated with C. albicans were randomly divided into two groups by electronic random sampling. In the first group, T. vulgaris essential oil spray and in the second group, 0.2% chlorhexidine (CHX) spray was applied to the appliances. Thereafter, the C. albicans colony count was measured at baseline (T0) and at 2 (T1) and 5 (T2) days after the intervention. Microbial samples were collected by a sterile swab from 3 spots on the internal surface of orthodontic appliances. The Friedman test was used for within-group comparisons and the Mann Whitney test was employed to compare the efficacy of T. vulgaris essential oil with CHX.The results of this study revealed that continuous use of 2%T. vulgaris essential oil or 0.2% CHX significantly decreased the colony count of C. albicans on removable orthodontic appliances; Howevere, no significant difference was noted in the efficacy of 2% T. vulgaris essential oil and 0.2% CHX in decreasing the C. albicans colony count.T. vulgaris essential oil and CHX have favorable antimicrobial activity against C. albicans. Moreover, 2% T. vulgaris essential oil can be considered as an alternative to 0.2% CHX to eliminate fungal contamination of orthodontic appliances.

Background: Vulvovaginal candidiasis (VVC) is a significant health issue due to Candida spp. Although Candida albicans is considered a major causative agent of vaginal candidiasis, non-albicans species have increased during previous decades. Objectives: This research aimed at molecular identification and assessing antifungal susceptibility of VVC isolated Candida spp. Methods: A professional physician examined two hundred and ninety-five suspected females with vaginitis. The specimens were collected by sterile cotton swabs. Swabs were inoculated on Sabouraud dextrose agar plates and then incubated for 48 - 72 hours at 35°C. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect all Candida species. Broth microdilution, according to the M27-A3 and M27-S4 CLSI documents, were employed for determining the antifungal susceptibility tests of caspofungin (CAS), voriconazole (VRC), itraconazole (ITC), fluconazole (FLU), clotrimazole (CLO), ketoconazole (KTO), amphotericin B (AMB), and nystatin (NYS). Results: A total of 295 females suspected of vulvovaginal candidiasis were examined. The culture results were positive in 50.5% (149 of 295) of specimens. According to molecular identification techniques, C. albicans 133/149 (89.2%), C. glabrata 8/149 (5.4%), and C. kefyr 2/149 (1.4%) were the main species. A mixed infection of C. albicans and C. glabrata 6/149 (4 %) was detected. The geometric mean values to all Candida strains were in increasing order as the following: CAS, 0.075 µg/mL; VRC, 0.091 µg/mL; ITC, 0.15 µg/mL; AMB, 0.22 µg/mL; CLO, 0.23 µg/mL; KTO, 0.28 µg/mL; NYS, 0.88 µg/mL; FLU, 1.48 µg/mL. Further, the MIC ranges of all Candida isolates to the tested antifungal agents were in increasing order as follows: CAS: 0.031 - 0.25 µg/mL, KTO and ITC: 0.031 - 2 µg/mL, VRC: 0.031 - 4 µg/mL, CLO and AMB: 0.031 - 8 µg/mL, NYS: 0.06 - 4 µg/mL, and FLU: 0.12 - 128 µg/mL. Conclusions: We reported 1 (7.2 %) C. glabrata isolate resistance to FLU and 2 (14.3%) C. glabrata isolates susceptible-dose-dependent (SDD) to CAS. We also reported 6 (4.5%), 5 (3.8%), and 2 (1.5%) C. albicans resistance to ITC, FLU, and AMB, respectively, but 100% C. albicans susceptible to CAS and VRC.

Background Trichophyton benhamiae is a zoophilic dermatophyte, known as one of the causative agents of dermatophytosis. Objectives The purpose of this study is to explore the genotypes of T. benhamiae strains isolated from geographically different areas of Iran and also to evaluate in vitro antifungal susceptibility profile of these strains against seven antifungal drugs. Methods Twenty-two strains of T. benhamiae and two strains of T. eriotrephon were isolated from patients with distinct types of dermatophytosis. DNA extraction and amplification of rDNA regions using ITS1 and ITS4 primers were conducted on the isolates. The in vitro antifungal susceptibility of posaconazole (PSC), voriconazole (VRC), itraconazole (ITC), ketoconazole (KET), caspofungin (CAS), terbinafine (TRB), and griseofulvin (GRZ) were evaluated according to CLSI M38-A2 protocol. Results The multiple alignment of the ITS-rDNA sequences of T. benhamiae indicated a mean similarity of 99.5%, with 0 to 3 interspecies nucleotide difference. The geometric mean (GM) values of minimum inhibitory concentrations (MICs) and minimum effective concentrations (MECs) across the all isolates were respectively: TRB: 0.025 mg/L, PSC: 0.032 mg/L, ITC: 0.050 mg/L, VRC: 0.059 mg/L with lower values and CAS: 0.31 mg/L, KTZ: 0.56 mg/L and GRZ: 0.76 mg/L with higher values. Conclusion Divers ITS-sequence-types of T. benhamiae was shown in different geographical regions of Iran. The TRB, PSC, and ITC were the most effective drugs against T. benhamiae strains, respectively. Furthermore, in our study, two strains of T. eriotrephon as a scarce dermatophyte species were described.

Objectives: To assess the efficacy of Thymus vulgaris (T. vulgaris) essential oil to decrease the count of Candida albicans (C. albicans) accumulated on removal orthodontic appliances.Methods: In this 2 arm parallel controlled clinical trial, Forty-four patients whose removable orthodontic appliances contaminated with C. albicans were randomly divided into two groups for use of T. vulgaris essential oil and 0.2% Chlorhexidine (CHX) by electronic random sampling. The C. albicans colony count was measured at baseline (T0) and at 2 (T1) and 5 (T2) days after the intervention. Microbial samples were collected by a sterile swab from 3 spots on the internal surface of orthodontic appliances. The Friedman test was used for within-group comparisons while the Mann Whitney test was applied to compare the efficacy of T. vulgaris essential oil and CHX.Results: No significant difference was noted in the efficacy of 2% T. vulgaris essential oil and 0.2% CHX in decreasing the C. albicans colony count on removable orthodontic appliances.Conclusion: T. vulgaris essential oil has favorable antimicrobial activity against C. albicans.

The molecular epidemiology and antifungal susceptibility of Aspergillus nidulans species complex has not been well studied. To evaluate the genetic diversity and antifungal susceptibility patterns of clinical and environmental isolates of A. nidulans complex. Sixty clinical and environmental isolates of Aspergillus section Nidulantes were collected from five countries (Iran, The Netherlands, Spain, Portugal and Greece). The species were molecularly identified by sequencing of β-tubulin gene. The genetic diversity of A nidulans complex isolates (n = 54) was determined with a microsatellite genotyping assay. Antifungal susceptibility profile was determined using EUCAST method. The isolates were classified as A nidulans (46.7%), A spinulosporus (26.6%), A quadrilineatus (10%), A pachycristatus (3.3%), A rugulosus (3.3%), A unguis (5%), A creber, (1.7%), A olivicola (1.7%) and A sydowii (1.7%). Thirty-four sequence types (STs) were identified among the 54 A nidulans complex isolates. A high level of genetic diversity was found among A nidulans sensu stricto strains but low diversity was found among A spinulosporus strains. Amphotericin B showed high MICs to all species. The most active azole was posaconazole (GM = 0.64 mg/L), while itraconazole showed the highest MICs among azoles (GM = 2.95 mg/L). A spinulosporus showed higher MICs than A nidulans sensu stricto for all antifungals except for micafungin and anidulafungin. Interspecies variations may result in differences in antifungal susceptibility patterns and challenge antifungal therapy in infections caused by A nidulans. Differences in the distribution of STs or persistence of multiple STs might be related to the sources of isolation and niche specialisation. This project was supported in part by a research project from the Fondo de Investigación Sanitaria (PI16CIII/00035) and a research fund (No. 2819) from Invasive Fungi Research Center of Mazandaran University of Medical Sciences, Sari, Iran. info:eu-repo/semantics/publishedVersion

Introduction: Usually, for treatment of fungal infections, antifungals such as azoles are used, but one of the biggest problems faced in clinical practice is the emergence of resistance for most of these drugs. Antifungal drugs derived from plants may alleviate this problem. The aims of this study were to analyse the ergosterol and gene expression profiles of ERG genes in Candida albicans treated with carvacrol. Methods: We used carvacrol and conducted a series of follow-up studies to examine the inhibitors of Candida species isolated from immunocompromised patients. Antifungal susceptibility test, time-kill study, ergosterol binding assay and ergosterol content were investigated. Eventually, the expression of ERG3 and ERG11genes was carried out to investigate the inhibitory properties of antifungal activity against Candida albicans using quantitative real time RT-PCR. Results: Carvacrol was able to inhibit Candida species and reduce time-kill kinetic in C. albicans. This phytoconstituent acted by binding to ergosterol in the fungal membrane and caused a reduction of 52% of the ergosterol content compared to the untreated growth control. Finally, carvacrol displayed significant down-regulation of ERG3 and ERG11genes in C. albicans. Conclusion: These results provide proof of concept for the implementation of carvacrol inhibitors of Candida species. In addition, ERG3 and ERG11 genes could be probable target of carvacrol against C. albicans.

Occurrence and diversity of dermatophyte mycoflora in 298 soil samples from Ahvaz, Southwest of Iran was investigated by using the hair-baiting technique. The samples were collected during spring (n = 210) and autumn (n = 88) of 2015, and the fungal isolates were identified based on the macro- and micro-morphology of colonies and with further ITS-rDNA RFLP and sequencing. Totally, 60 soil samples (20.1%) were positive for dermatophyte growth whose pH varied from 7.0 to 7.9. The highest (26.6%) and the lowest (14.3%) recovery rates were from the animal resorts and the streets soils samples, respectively. Seasonally, 16.7% of the spring samples and 28.4% of the autumn samples were positive. Based on molecular identification, three species of two genera were identified viz. M. fulvum (n = 57), M. canis (n = 2) and zoophilic Trichophyton interdigitale (n = 1). As a specific goal in the study, differentiation of the species in Microsporum gypseum complex was established by measuring the mean length and width of macroconidia in some strains of M. gypseum, M. fulvum and M. incurvatum. Mean size for macroconidia length and width in three species showed that M. gypseum and M. incurvatum can morphologically be differentiated from M. fulvum but not from each other. M. fulvum was the most abundant species isolated from the soils of Ahvaz; however, to comprehensively specify the distribution pattern of geophilic dermatophytes in the soils of this city further investigations are needed. Identification based on micro-morphometric is not effective for species distinction in M. gypseum complex, while molecular procedures based on sequencing of certain DNA regions are the most reliable and applicable strategies for this purpose.

Onychomycosis is considered a fungal nail infection caused mainly by dermatophytes, yeasts and non-dermatophyte molds including dematiaceous fungi. Onychomycosis caused by non-dermatophyte molds is a health problem in the medical environment as the patients frequently return to outpatient clinics seeking new therapeutic modalities. Here, we report the first case of onychomycosis caused by a black fungus, Neoscytalidium novaehollandiae, in the right hand finger nail of a 52-year-old Iranian female with no history of immunodeficiency and underlying disease. The pattern of nail involvement was recognized as total dystrophic onychomycosis. Examination of nail scrapings with potassium hydroxide revealed brown, septate and branching sub-hyaline to dark-colored hyphae. The black fungus isolated in culture was identified as Neoscytalidium novaehollandiae by molecular analysis. The patient received oral terbinafine plus ciclopirox nail lacquer twice a week and began responding to the treatment three months after initial antifungal therapy. Additional four weeks’ use of terbinafine plus ciclopirox nail lacquer completely resolved the clinical manifestations of onychomycosis. After four months, both microscopy and culture were negative.

The fungi Trichophyton mentagrophytes and T interdigitale account for significant amount of dermatophytosis cases worldwide. These two dermatophytes form a species complex and have a number of ribosomal internal transcribed spacer (ITS) region genotypes, allowing simultaneous species identification and strain typing. Our aim was to describe the geographic distribution of T mentagrophytes/T interdigitale ITS region genotypes and find an association between the genotypes and clinical presentations of respective infections. We performed rDNA ITS region sequencing in 397 Iranian T mentagrophytes/T interdigitale isolates and analysed all available in GenBank entries with sequences of this kind. For the study, 515 clinical annotations were available. Statistical analysis was performed by chi-squared test and Spearman rank correlation analysis. A total of 971 sequences belonged to genotypes with at least 10 geographic annotations and were classified on the basis of exclusive occurrence in a particular region or high relative contribution to a regional sample. We discerned Asian and Oceanian ("KU496915" Type V, "KT192500" Type VIII, "KU315316"), European ("FM986750" Type III, "MF926358" Type III*, "KT285210" Type VI) and cosmopolitan ("FM986691" Type I, "JX122216" Type II, "KP132819" Type II* and "AF170453" Type XXIV) genotypes. There was statistically significant difference in the ITS genotype distribution between different affected body sites. Trichophyton mentagrophytes "KT192500" Type VIII correlated with tinea cruris, T mentagrophytes "KU496915" Type V correlated with tinea corporis, T interdigitale "JX122216" Type II correlated with tinea pedis and onychomycosis. Trichophyton mentagrophytes and T interdigitale genotypes can be associated with distinct geographic locations and particular clinical presentations.

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.

Background: Vulvovaginal candidiasis (VVC) is a frequent infection in females at the reproductive age. Furthermore, the most common causative agent is Candida albicans yet in the recent years, the incidence of non-albicans species has increased. Objectives: The main aim of this study was the isolation and identification of various Candida species from patients with vulvovaginal candidiasis by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in Yasuj, Iran. Methods: Three hundred and ten suspected women females vaginitis were sampled and examined. Genomic DNA was extracted from fresh colonies by phenol-chloroform glass bead methods. Polymerase chain reaction (PCR) amplification was performed based on the ribosomal DNA internal transcribed spacer (rDNA-ITS), and specific electrophoretic patterns of PCR products after digestion with MspI enzyme used for species identification. Results: The cultures were positive for 160 (51.6%) vaginal samples. Candida albicans (86.8%) was the most common species among the isolates, followed by C. glabrata (3.77%) and C. krusei (3%). Multispecies with two Candida were identified in nine patients. Conclusions: Vulvovaginal candidiasis is more prevalent among females in Yasuj and the predominant agent is C. albicans. In addition, correct identification of Candida species could play an important role in management and treatment of VVC.

Summary Objective Dermatophytes are taxonomically classified in the genera Trichophyton, Microsporum, and Epidermophyton. Pleomorphism, cultural variability, slow growth and sporulation, and the need for additional physiological tests make dermatophytes notoriously difficult to identify. The present study aimed to compare the results of morphological and molecular identification of certain groups of clinical isolates of dermatophytes with a view to evaluating the accuracy of molecular methods. Patients and methods For each sample, the ITS1-5.8S-ITS2 rDNA region was amplified using the primers ITS1 and ITS4. PCR products were subjected to restriction fragment length polymorphism (RFLP) analysis using the enzyme MvaI and isolate identification was performed by comparing the electrophoretic RFLP patterns with reference profiles obtained previously. Finally, paired comparative analyses of molecular and conventional methods were performed. Results While morphology results from routine daily reports of the laboratories indicated that 18 (6.8%) and 136 (52.10%) of the isolates were T. rubrum and T. interdigitale, respectively, PCR-RFLP results suggested that T. rubrum was the most common etiological agent of ringworm accounting for 94 (36.01%), followed by T. interdigitale accounting for 71 (27.20%). Interestingly, 80.8% out of the 94 isolates identified as T. rubrum by molecular testing had been identified by morphological examination as belonging to different species, such as T. interdigitale (75.5%), E. floccosum (2.1%) and M. canis, T. verrucosum, and T. tonsurans (each 1.06%). Ten strains out of 261 (T. interdigitale, n = 8; E. floccosum, n = 2) had been defined as unknown species by morphological tests. Conclusion An unexpected high percent of isolates identified as T. interdigitale by conventional methods were in effect T. rubrum shown by PCR-RFLP, and regarding the necessity of correct identification of dermatophytes recovered from different clinical forms of the infection, we highly recommend ITS-sequencing or ITS-RFLP of the isolates, particularly for epidemiological research studies.

Purpose. We aimed to evaluate the resolving power of the translation elongation factor (TEF)-1α gene for phylogenetic analysis of Aspergillus species. Methodology. Sequences of 526 bp representing the coding region of the TEF-1α gene were used for the assessment of levels of intra- and inter-specific nucleotide polymorphism in 33 species of Aspergillus, including 57 reference, clinical and environmental strains. Results. Analysis of TEF-1α sequences indicated a mean similarity of 92.6 % between the species, with inter-species diversity ranging from 0 to 70 nucleotides. The species with the closest resemblance were A. candidus/A. carneus, and A. flavus/A. oryzae/A. ochraceus, with 100 and 99.8 % identification, respectively. These species are phylogenetically very close and the TEF-1α gene appears not to have sufficient discriminatory power to differentiate them. Meanwhile, intra-species differences were found within strains of A. clavatus, A. clavatonanicus, A. candidus, A. fumigatus, A. terreus, A. alliaceus, A. flavus, Eurotium amstelodami and E. chevalieri. The tree topology with strongly supported clades (≥70 % bootstrap values) was almost compatible with the phylogeny inferred from analysis of the DNA sequences of the beta tubulin gene (BT2). However, the backbone of the tree exhibited low bootstrap values, and inter-species correlations were not obvious in some clades; for example, tree topologies based on BT2 and TEF-1α genes were incompatible for some species, such as A. deflectus, A. janus and A. penicillioides. Conclusion. The gene was not phylogenetically more informative than other known molecular markers. It will be necessary to test other genes or larger genomic regions to better understand the taxonomy of this important group of fungi.

PubMedID: 26767834 The incidence of dermatophytosis due to Trichophyton simii is generally considered to be limited to endemic areas, particularly one area of India. However, the high similarity between the morphological features of atypical T. simii isolates and those of other dermatophytes such as Trichophyton interdigitale and Arthroderma benhamiae may lead to misidentification of the cause of dermatophytosis in many instances. We investigated a rare case of tinea corporis in a 9-month-old female with extensive erythematous lesions. Morphological features of the recovered isolate from the culture resulted in the identification of Trichophyton interdigitale. For accurate identification, the internal transcribed spacer regions (ITS1 and ITS2) of the ribosomal DNA (rDNA) gene were sequenced and the isolate was ultimately identified as T. simii. In conclusion, T. simii, which has been formerly known to be restricted to specific endemic regions, appears to be not infrequent in non-endemic areas but instead simply less well-known and consequently underestimated. To determine its actual prevalence of infection, the application of DNA-based molecular methodologies is required. © 2016, Springer Science+Business Media Dordrecht.

This work aimed to identify the species distribution of common clinical and environmental isolates of black Aspergilli based on simple restriction fragment length polymorphism (RFLP) analysis of the β-tubulin gene. A total of 149 clinical and environmental strains of black Aspergilli were collected and subjected to preliminary morphological examination. Total genomic DNAs were extracted, and PCR was performed to amplify part of the β-tubulin gene. At first, 52 randomly selected samples were species-delineated by sequence analysis. In order to distinguish the most common species, PCR amplicons of 117 black Aspergillus strains were identified by simple PCR-RFLP analysis using the enzyme TasI. Among 52 sequenced isolates, 28 were Aspergillus tubingensis, 21 Aspergillus niger, and the three remaining isolates included Aspergillus uvarum, Aspergillus awamori, and Aspergillus acidus. All 100 environmental and 17 BAL samples subjected to TasI-RFLP analysis of the β-tubulin gene, fell into two groups, consisting of about 59% (n=69) A. tubingensis and 41% (n=48) A. niger. Therefore, the method successfully and rapidly distinguished A. tubingensis and A. niger as the most common species among the clinical and environmental isolates. Although tardy, the Ehrlich test was also able to differentiate A. tubingensis and A. niger according to the yellow color reaction specific to A. niger. A. tubingensis and A. niger are the most common black Aspergillus in both clinical and environmental isolates in Iran. PCR-RFLP using TasI digestion of β-tubulin DNA enables rapid screening for these common species.

Onychomycosis is a common nail infection caused by dermatophytes, non-dermatophyte molds (NDM), and yeasts. Aspergillus species are emerging as increasing causes of toenail onychomycosis. The purpose of this study was species delineation of Aspergillus spp. isolated from patients with onychomycosis.During a period of one year (2012-2013), nail samples were collected from patients clinically suspected of onychomycosis and subjected to microscopic examination and culture. Species identification was performed based on macro- and micro-morphology of colonies. For precise species identification, PCR-amplification and sequencing of the beta-tubulin gene followed by BLAST queries were performed where required.A total of 463/2,292 (20.2%) tested nails were diagnosed with onychomycosis. Among the positive specimens, 154 cases (33.2%) were identified as saprophytic NDM onychomycosis, 135 (29.2%) of which were attributable to Aspergillus. Aspergillus species isolated from the infected nails included Aspergillus flavus (77.3%, n=119), Aspergillus niger (n=4), Aspergillus tubingensis (n=4), Aspergillus terreus (n=3), Aspergillus sydowii (n=2), Aspergillus spp. (n=2), and Aspergillus candidus (n=1). Among the patients diagnosed with onychomycosis due to Aspergillus (average patient age, 47.4 years), 40 had fingernail and 95 toenail involvement. The large toenails were most commonly affected.This study identified a markedly high occurrence of A. flavus, and this fungus appears to be an emerging cause of saprophytic onychomycosis in Iran. The study moreover highlights the necessity of differentiating between dermatophytic and non-dermatophytic nail infections for informed decisions on appropriate therapy.