Aspergillus hubkae, a Novel Species Isolated from a Patient with Probable Invasive Pulmonary Aspergillosis.

A 50-year-old man, previously diagnosed with pulmonary tuberculosis and lung cavities, presented with symptoms including fever, shortness of breath, and cough. A pulmonary CT scan revealed multiple cavities, consolidation and tree-in-bud in the upper lungs. Further investigation through direct examination of bronchoalveolar lavage fluid showed septate hyphae with dichotomous acute branching. Subsequent isolation and morphological analysis identified the fungus as belonging to Aspergillus section Nigri. The patient was diagnosed with probable invasive pulmonary aspergillosis and successfully treated with a three-month oral voriconazole therapy. Phylogenetic analysis based on partial ß-tubulin, calmodulin and RNA polymerase second largest subunit sequences revealed that the isolate represents a putative new species related to Aspergillus brasiliensis, and is named Aspergillus hubkae here. Antifungal susceptibility testing demonstrated that the isolate is resistant to itraconazole but susceptible to voriconazole. This phenotypic and genetic characterization of A. hubkae, along with the associated case report, will serve as a valuable resource for future diagnoses of infections caused by this species. It will also contribute to more precise and effective patient management strategies in similar clinical scenarios.

Port implantation-related bloodstream infection caused by Wickerhamomyces myanmarensis: A case report.

Background and Purpose: Wickerhamomyces myanmarensis is a new opportunistic yeast previously named Pichai myanmarensis, which belongs to the order Saccharomycetales. Since its discovery, one environmental isolate of W. myanmarensis has been reported from Myanmar, and one clinical sample from Iran. Case Report: We report a case of bloodstream infection related to an implantable venous access port. W. myanmarensis was isolated from patient's blood after chemotherapy, which was meant to control and heal T-cell lymphoblastic lymphoma. Broth dilution minimum inhibitory concentrations were performed according to the CLSI M27-A3 document. The patient recovered with intravenous voriconazole and was discharged with the recommended prescription of oral voriconazole as a maintenance drug. Conclusion: So far, only one case of W. myanmarensis fungemia has been reported in the world in 2019. This is the second case of bloodstream infection with this yeast from a patient undergoing chemotherapy in Iran.

Iranian National Survey on Tinea Capitis: Antifungal Susceptibility Profile, Epidemiological Characteristics, and Report of Two Strains with a Novel Mutation in SQLE Gene with Homology Modeling.

BACKGROUND: The data on the epidemiological and antifungal susceptibility profile of tinea capitis (TC) in Iran has not been updated in recent decades. This report presents the Iranian epidemiological and drug susceptibility data regarding the distribution of dermatophytes species isolated by six national mycology centers for a period of one year (2020-2021). MATERIAL AND METHODS: A total of 2100 clinical samples from individuals suspeted to TC were subjected to mycological analysis of direct microscopy and culture. For definite species identification, the culture isolates were additionally subjected to PCR-RFLP and PCR-sequencing of the ITS ribosomal DNA (ITS-rDNA) region. Antifungal susceptibility profiles for eight common antifungal drugs were determined by CLSI M38-A3 guidelines. The SQLE gene was partially amplified and sequenced in two terbinafine-resistant and two susceptible T. mentagrophytes isolates to elucidate probable substitutions involved in resistance. RESULTS: TC (n = 94) was diagnosed in 75 children (79.8%) and 19 adults (20.2%) by direct microscopy and culture. Frequency of TC was significantly more among males (66 males = 70.2% vs 28 females = 29.8%). The prevalent age group affected was 5-9 years (39.36%). Thirty-two (34.04%) T. mentagrophytes, 27 (28.7%) T. tonsurans, 14 (14.9%) M. canis, 13 (13.8%) T. violaceum, 5 (5.32%) T. indotineae, 2 (2.1%) T. benhamiae, and 1 (1.1%) T. schoenleinii were identified as the causative agents. MIC values of isolates showed susceptibility to all antifungal agents, except for fluconazole and griseofulvin with GM MIC of 11.91 µg/ml and 2.01 µg/ml, respectively. Terbinafine exhibited more activity against isolates, with GM MIC 0.084 µg/ml followed by ketoconazole (0.100 µg/ml), econazole (0.107 µg/ml), itraconazole (0.133 µg/ml), butenafine (0.142 µg/ml), and miconazole (0.325 µg/ml). Two resistant T. mentagrophytes isolates harbored missense mutations in SQLE gene, corresponding to amino acid substitution F397L. Remarkably, one unique mutation, C1255T, in the SQLE sequence of two terbinafine-susceptible T. mentagrophytes strains leading to a change of leucine at the 419th position to phenylalanine (L419F) was detected. CONCLUSIONS: T. mentagrophytes, T. tonsurans, and M. canis remained the main agents of TC in Iran, however less known species such as T. indotinea and T. benhamiae are emerging as new ones. Terbinafine could still be the appropriate choice for the treatment of diverse forms of TC.

Time and cost-efficient identification of Trichophyton indotineae.

BACKGROUND: During the past 5 years, an outbreak of recalcitrant dermatophytosis due to a novel Trichophyton species generally resistant to terbinafine, T. indotineae, has spread out from South Asia to many countries around the World. These isolates cannot be reliably differentiated from other Trichophyton spp. on the basis of morphological traits and the current laboratory diagnostics relies on sequencing of ribosomal DNA ITS region. OBJECTIVES: In this study, we aimed to introduce two inexpensive and rapid PCR-based assays for differentiation between T. indotineae and other dermatophytes. METHODS: The first introduced assay is based on PCR-restriction fragment length polymorphism (PCR-RFLP) analysis, involving the amplification of TOP2 sequences and the digestion of PCR products by Cfr13I restriction enzyme. The second assay is proposed as conventional endpoint species-specific PCR amplification of the C120-287 intergenic locus. To validate the assays, a total of 191 Trichophyton spp. and 2 Microsporum canis strains with known ITS region sequences were used. From the T. mentagrophytes / T. interdigitale species complex (TMTISC), strains with 18 different ITS genotypes were tested. The sample of TMTISC isolates included 41 T. indotineae strains. RESULTS: TOP2 PCR-RFLP and T. indotineae-specific PCR were positive with testing on DNA of all 41 T. indotineae isolates and two strains of T. mentagrophytes belonging to ITS Types XIII and XVI, but negative with other species and other TMTISC ITS genotypes (n = 152). Therefore, the specificity of both new assays was 99%. CONCLUSION: The two developed diagnostic assays provide accurate and cost-effective means of identifying cultured T. indotineae isolates.

Identification and genetic diversity of Alternaria species recovered from the air of Ahvaz city, the Southwestern part of Iran.

The Alternaria genus has pathogenic, endophytic, and saprobic characteristics. Alternaria genus causes respiratory diseases, fungal allergenicity and the production of mycotoxin in food. Ahvaz city is one of the areas where the presence of dust and high humidity cause the growth and spread of fungal species in the air. Identification of Alternaria species is difficult based on morphology solely. For the first time in Ahvaz, the classification of this fungus was performed using ITS region, alta1 gene, and morphology. For the identification of Alternaria isolates in the Ahvaz city air using morphological and molecular characteristics, potato dextrose agar (PDA) media were used to culture 40 Alternaria isolates recovered from the Ahvaz city air. Afterward, the appearance of the colonies was examined. The DNAs of the isolates were extracted and amplified using the specific primers of the ITS and, Alt a1 regions. The amplified DNA products were sequenced. Then, they were compared with the sequences in the NCBI GeneBank. Based on the morphological results, the isolates included four different species and A. alternata had the highest frequency. Alt a1 gene was present in all the isolates of Alternaria species recovered in our research. Finally, identifying the varieties of Alternaria species based on morphological characteristics as well as ITS or Alt a1 regions is useful but difficult.

Efficiency studies of modified IFAS-OSA system upgraded by an anoxic sludge holding tank.

An upgraded integrated fixed-film activated sludge-oxic settling anoxic (IFAS-OSA) system is a new technology for reducing nutrients and excess sludge. The results showed that the average TN removal efficiency of the IFAS-OSA system was gradually increased up to 7.5%, while the PO4-3-P removal efficiency increased up-to 27%, compared with that of the IFAS system. The COD removal efficiency of the IFAS-OSA system was slightly increased up-to 5.4% and TSS removal efficiency increased up to 10.5% compared with the control system. Biomass yield coefficient (Yobs) in the IFAS and IFAS-OSA systems were 0.44 and 0.24 (gr MLSS/ gr COD). Hence, sludge production decreased by 45%. The average SVI was decreased by 48% in IFAS-OSA system compared with IFAS. This study demonstrated the better performance of the IFAS-OSA system compared to that of the IFAS system.

Genotypic diversity of Iranian Cryptococcus neoformans using multilocus sequence typing (MLST) and susceptibility to antifungals.

Cryptococcus species is an opportunistic yeast pathogen and classified into different molecular types according to typing techniques including multilocus sequence typing (MLST). The study aimed to investigate the genotypes of environmental Cryptococcus isolates using MLST and the relationship between the in vitro antifungal susceptibility and sequence types of isolates. Genotyping Cryptococcus isolates was performed by the MLST method at seven nuclear loci. Antifungal susceptibility was determined by using CLSI broth micro-dilution method for amphotericin B, fluconazole, itraconazole, voriconazole, flucytosine, and luliconazole. Seven sequence types (ST) were detected using MLST analysis, with the most frequent (50%) ST77, followed by ST4 (16.7%) among 30 C. neoformans isolates. All antifungals demonstrated excellent activity against isolates, except for itraconazole and amphotericin B that were non-wild type against 53.3% and 10% of isolates, respectively. Although seven sequence types belonging to C. neoformans isolates were detected, ST77 was the main sequence type in Ahvaz. Also, non-wild type isolates were only found against itraconazole and amphotericin B.

In vitro antifungal susceptibilities of six antifungal drugs against clinical Candida glabrata isolates according to EUCAST.

BACKGROUND AND PURPOSE: Candida glabrata is the second cause of candidiasis. The mortality rate of C. glabrata infections is about 40%; accordingly, it may be life threatening, especially in immunocompromised hosts. Regarding this, the current study was conducted to evaluate the regional patterns of the antifungal susceptibility of clinical C. glabrataisolated from the patients referring to the health centers located in Ahvaz, Iran. MATERIALS AND METHODS: In this study, a total of 30 clinical strains of C. glabrata isolates were recovered from different body sites (i.e., vagina, mouth, and urine). Phenotypic characteristics and molecular methods were used to identify the isolates. The minimum inhibitory concentration (MIC) was determined according to the European Committee on Antimicrobial Susceptibility Testing. RESULTS: Our findings demonstrated that 20%, 80%, and 6.7% of the isolates were resistant to amphotericin B, terbinafine, and posaconazole, respectively, while all the isolates were found to be fluconazole susceptible dose dependent and susceptible to voriconazole and caspofungin. CONCLUSION: Our study suggested that voriconazole had high potency against C. glabrata isolates. Consequently, this antifungal agent can be an alternative drug in the treatment of resistant patients. These results can be helpful for the successful treatment of patients in different regions.

Antifungal susceptibility profiles of otomycosis etiological agents in Ahvaz, Iran.

BACKGROUND AND PURPOSE: Otomycosis is a secondary ear fungal infection among predisposed individuals in humid conditions. Aspergillus species are the most common etiologic agents of this infection. Several ototopical antifungals are currently used for the treatment of this disease; however, recurrence and treatment failure are usually observed in some cases. Regarding this, the present study was conducted to investigate the antifungal activity of caspofungin, azoles, and terbinafine against the isolated agents of otomycosis. MATERIALS AND METHODS: This study was conducted on the specimens collected from 90 patients with otomycosis. The samples were cultured on Sabouraud dextrose agar and identified based on morphological characteristics, physiological tests, and microscopic features. Furthermore, the microdilution method was used for antifungal susceptibility testing according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. Finally, the minimum inhibitory concentration (MIC) and minimum effective concentration (MEC) ranges, MIC/MEC50, MIC/MEC90, and geometric mean (GM) MIC/MEC were calculated for the isolates. RESULTS: According to the results, 77 patients with otomycosis were positive for different Aspergillus (88.3%) and Candida (11.7%) species. Aspergillus niger complex (n=36) was found to be the most common agent, followed by A. flavus, A. terreus, and A. nidulans complexes. Furthermore, epidemiological cutoff values (ECVs) were lower than those presented by the CLSI for itraconazole and caspofungin in 98.5% and 42.6% of Aspergillus species, respectively. Terbinafine exhibited a great activity against Aspergillus species, while fluconazole revealed a low activity against both Aspergillus species. Based on the results, 77.8% of Candida species were resistant to caspofungin; however, miconazole and econazole had low MIC ranges. CONCLUSION: Aspergillus niger and A. flavus complexes were identified as the most common agents accounting for 85.7% of the isolates. In addition, terbinafine was identified as the best antifungal for both Aspergillus and Candida species. Moreover, tested azoles had relatively low MICs, whereas most of the isolates had the MIC values beyond the caspofungin ECVs.

Microsatellite Typing and Antifungal Susceptibility of Candida glabrata Strains Isolated From Patients With Candida Vaginitis.

Vulvovaginal candidiasis (VVC) is a yeast infection with a global reach and millions of dollars are spent annually for its diagnosis and treatment. Recently, Candida glabrata with different degrees of antifungal resistance has been considered as the second most common cause of vaginal infections. The aim of the present study is to determine the antifungal susceptibility and molecular epidemiology profiles of C. glabrata isolates from patients with VVC. Sixty-one C. glabrata isolates were examined for antifungal susceptibility using the EUCAST broth microdilution method. Moreover, microsatellite length polymorphism (MLP) was used for typing the C. glabrata isolates using six microsatellite markers. Overall, 13, 3.3, and 0% of the isolates were non-wild types to itraconazole, posaconazole, and voriconazole, respectively. Sixty (98.4%) isolates were an intermediate phenotype to fluconazole and only one isolate was fluconazole resistant. Microsatellite length polymorphism with a discriminatory power of 0.964 identified 35 distinct types and 24 singleton genotypes. The assessment of the population genetic structure revealed that the non-wild-type population had a moderate genetic differentiation compared to the wild type population (FST = 0.1457). It was also found that the most common genotypes were G27 (eight strains), G12 (six strains), and G4 (five strains). We found that eight strains were resistant/a non-wild phenotype to itraconazole. Five out of eight (62.5%) resistant/non-wild phenotype strains correlated to a predominant genotype (GT27) and the rest belonged to GT11 (12.5%), GT29 (12.5%), and GT28 (12.5%). The current study is the first molecular epidemiology study in the southwest of Iran and demonstrates the antifungal susceptibility profiles of C. glabrata in it. This study shows a wide range of the genetic diversity of C. glabrata (35 different genotypes) from VVC in the southwest of Iran. The majority of the non-wild isolates had a dominant genotype or genotypes related to this dominant genotype (clonal cluster one).

Relationship between environmental Fungi and changes in lung function indices of new referral allergic patients in Ahvaz city under normal and dust conditions.

BACKGROUND: Aim of this study was to determine whether any specific fungal spores could be responsible for changes observed in lung function indices. MATERIALS AND METHODS: 1042 new allergic patients were selected from July 2017 to May 2018 in Ahvaz City, Iran. Fungal samples were collected in normal and dusty condition within 5 and 2 min, respectively. Sampling was repeated once every 6 days and also in the dusty days. RESULTS: Average numbers of fungi colony were 639.86 and 836.44 CFU m-3 under normal and dusty conditions, respectively. Most common fungi in Ahwaz City air were Cladosporium sp., Penicillium sp., Aspergillus Niger, Aspergillus Flavus and Alternaria sp.. Highest fungal mean concentrations, 392 and 480 CFU m-3, were related to Cladosporium sp. under normal and dust conditions, respectively. Average total numbers of colony fungal were 614, 483, 1082, 424 CFU m-3 and 856, 701, 1418, 418 CFU m-3 during the spring, summer, autumn, and winter under normal and dusty conditions, respectively. Patients were evaluated by measured lung function parameters of FEV1 (L), FEV1 (%pred), FVC (L), FVC (%pred), and FEV1/FVC ratio with mean values of 1.85, 58.32, 2.63, 68.18, and 69.43, respectively. CONCLUSION: Increases in mean total spores of fungi in spring were accompanied by decreases in FEV1/FVC ratio. Enhanced spores of Cladosporium sp. in spring led to reduced FEV1/FVC ratio. Increase the spores of Curvularia sp. in summer decreased by FEF25-75%. The augmented spores of Drechslera sp. in summer were associated with declined FEV1 and FEV1/FVC ratio. Enhanced fungal spores of Rhizopus sp. in spring resulted in lowered FEV1, FEV1/FVC ratio, and FEF25-75%.

Antifungal Susceptibility Patterns of Candida Species Recovered from Endotracheal Tube in an Intensive Care Unit.

Aims. Biofilms formed by Candida species which associated with drastically enhanced resistance against most antimicrobial agents. The aim of this study was to identify and determine the antifungal susceptibility pattern of Candida species isolated from endotracheal tubes from ICU patients. Methods. One hundred forty ICU patients with tracheal tubes who were intubated and mechanically ventilated were surveyed for endotracheal tube biofilms. Samples were processed for quantitative microbial culture. Yeast isolates were identified to the species level based on morphological characteristics and their identity was confirmed by PCR-RFLP. Antifungal susceptibility testing was determined according to CLSI document (M27-A3). Results. Ninety-five strains of Candida were obtained from endotracheal tubes of which C. albicans (n = 34; 35.7%) was the most frequently isolated species followed by other species which included C. glabrata (n = 24; 25.2%), C. parapsilosis (n = 16; 16.8%), C. tropicalis (n = 12; 12.6%), and C. krusei (n = 9; 9.4%). The resulting MIC90 for all Candida species were in increasing order as follows: caspofungin (0.5 µg/mL); amphotericin B (2 µg/mL); voriconazole (8.8 µg/mL); itraconazole (16 µg/mL); and fluconazole (64 µg/mL). Conclusion. Candida species recovered from endotracheal tube are the most susceptible to caspofungin.

Biofilm Formation and Susceptibility to Amphotericin B and Fluconazole in Candida albicans.

BACKGROUND: The ability of Candida albicans to form biofilms and adhere to host tissues and biomaterial surfaces is an important factor in its pathogenesis. One of the main characteristics of biofilms is their resistance to broad-spectrum anti-microbial drugs. OBJECTIVES: In the present study the formation of biofilm by C. albicans from different sources was evaluated. In addition, the minimum biofilm inhibitory concentration (MBIC) for two antifungals was evaluated. MATERIALS AND METHODS: In total, 120 isolates of C. albicans from different sources (patients with vaginitis, patients with candiduria, bucal cavity and environmental surfaces) were collected. Biofilm formation was determined by the 96-well micro-titeration plate method. MBIC testing was also performed, using the calorimetric indicator resazurin for amphotericin B and fluconazole. RESULTS: The results indicated that 100% of C. albicans isolates from different sources had the ability to form biofilms in vitro. Amongst these isolates, 83.3% of isolates had the maximum potential (4+) to form biofilms, while only one (0.9%) of isolates had the minimum ability (1+) to form biofilms. Our results showed that 65.0% of the tested isolates are sensitive to amphotericin B at amounts lower than 10 µg/mL, while only 26.7% are sensitive to fluconazole (had MBIC < 10 µg/mL). CONCLUSIONS: Although biofilm formation was detected in all tested isolates, there were differences in the ability to form biofilms between isolates from different sources. In addition, there were differences in the MBIC against the two examined antifungals, amphotericin B and fluconazole.