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Objective: Recent studies determined that the amoeboid form of Blastocystis acts as a factor in stimulating the host’s immune responses and ultimately results in urticaria and other skin disorders. The present study was conducted in order to determine the prevalence of Blastocystis in people referred to Bushehr city health centers and the relationship of this parasite with urticaria. Methods: Fecal samples were collected from 180 males and females referred to Bushehr health centers and a questionnaire containing demographic information was completed for each person. Samples were examined by preparing direct smear (wet mount) and then formalin-detergent sedimentation techniques. Data were analyzed using SPSS 22.0 software and chi-square test. Results: The results showed that 11.1% of cases infected with Blastocystis and 55% of patients with Blastocystis had various gastrointestinal symptoms. Statistical analysis showed that there was no significant relationship between infection with some demographic factors such as sex, age, literacy level and residence, but this was significant with some clinical symptoms such as itching and urticaria. Conclusion: Despite the existence of conflicting information and many ambiguities about the Blastocystis, this emerging pathogen is very important in terms of causing allergic and skin disorders in sufferers, therefore, it is necessary that patients with urticaria be evaluated for Blastocystis along with other diagnostic procedures and physicians should request a test before any medical intervention. Thus, diagnosis and treatment of these people can play an important role in improving the health of society.

Background and Purpose: Candida auris is a multidrug-resistant yeast that rapidly spreads, making it the leading Candidate for the next pandemic. One main leading cause of emerging resistant C. auris isolates is nonsynonymous mutations. This study aimed to detect the Y132F mutation, one of the most important azole resistance-associated mutations in the ERG-11 gene of C. auris, by developing a reliable high-resolution melt (HRM)-based method.Materials and Methods: Five C. auris isolates from Iran, plus three control isolates from other Clades were used in the study. The antifungal susceptibility testing through micro broth dilution was performed to recheck their susceptibility to three azole antifungals,including fluconazole, itraconazole, and voriconazole. Moreover, the polymerase chain reaction (PCR) sequencing of the ERG-11 gene was performed. Following the bioinformatic analysis and HRM-specific primer design, an HRM-based assay was developed and evaluated to detect ERG-11 mutations.Results: The minimum inhibitory concentrations of fluconazole among Iranian C. auris isolates ranged from 8 to 64 μg/mL. The PCR-sequencing of the ERG-11 gene and bioinformatic analyses revealed the mutation of Y132F, a substitution consequence of A to T on codon 395 in one fluconazole-resistant isolate (IFRC4050). The developed HRM assay successfully differentiated the targeted single nucleotide polymorphism between mutant and wild types (Melting temperature [Tm]: 81.79 ℃ - cycle threshold [CT]: 20.06 for suspected isolate). For both mutant and non-mutant isolates, the mean Tm range was 81.79-82.39 °C and the mean CT value was 20.06-22.93. These results were completely in accordance with the findings of DNA sequencing.Conclusion: The fast-track HRM-based method successfully detected one of the most common mechanisms of resistance in the ERG-11 gene of C. auris within 3 h. Finally, the development of more panels of HRM assays for the detection of all azole resistance mutations in C. auris ERG-11 is recommended to expand the scope of the field and facilitate the elaboration of rapid and accurate methods of antifungal resistance assessment.

Candida auris is a newly emerging multidrug-resistant fungal pathogen considered to be a serious global health threat. Due to diagnostic challenges, there is no precise estimate for the prevalence rate of this pathogen in Iran. Since 2019, only six culture-proven C. auris cases have been reported from Iran, of which, five belonged to clade V and one to clade I. Herein, we report a case of otomycosis due to C. auris from 2017 in a 78-year-old man with diabetes mellitus type II without an epidemiological link to other cases or travel history. Short tandem repeat genotyping and whole genome sequencing (WGS) analysis revealed that this isolate belonged to clade I of C. auris (South Asian Clade). The WGS single nucleotide polymorphism calling demonstrated that the C. auris isolate from 2017 is not related to a previously reported clade I isolate from Iran. The presence of this retrospectively recognized clade I isolate also suggests an early introduction from other regions or an autochthonous presence. Although the majority of reported C. auris isolates worldwide are resistant to fluconazole and, to a lesser extent, to echinocandins and amphotericin B, the reported clade I isolate from Iran was susceptible to all antifungal drugs.

Background and Purpose: The frequency and genetic diversity of black fungi in environmental and clinical settings have not been fully studied in Iran. This study aimed to identify and evaluate intra- and inter-species DNA sequence variation and also understand the phylogenetic relationships of melanized fungi and relatives isolated from different geographical regions of Iran.Materials and Methods: In total, 111 clinical and environmental strains of dematiaceous fungi were isolated, and their internal transcribed spacer ribosomal DNA(rDNA) regions were sequenced and analyzed.Results: An inter-species nucleotide sequence diversity rate of 1 to 464 nucleotides was observed between the species. Intra-species differences were found in the strains of Alternaria alternata, Cladosporium cladosporioides, Alternaria tenuissima, Curvularia spicifera, Aureobasidium pullulans, Curvularia hawaiiensis, Neoscytalidium dimidiatum,Alternaria terricola, Alternaria chlamydospora, Didymella glomerata, and Drechslera dematioidea by 0–59, 0–22, 0–4, 0–4, 0–3, 0–2, 0–2, 0–2, 0–2, 0–1, and 0–1 nt, respectively.Conclusion: The internal transcribed spacer rDNA is useful for the discrimination of several taxa of dematiaceous fungi. However, a better understanding of the taxonomy of species of Alternaria requires a larger rDNA region or a library of other gene sequences.

Background and Purpose: The main environmental saprobes, such as Penicillium, play an essential role in natural ecosystems as economically, ecologically, and medically important microorganisms. Biodiversity of this genus has not been described in Bushehr city, Iran. The present study is based on air biodiversity of Penicillium species on culture dependent approach and culture-independent technique using partial b-tubulin sequences.Materials and Methods: By using active sampling with a high volume air sampler, a total of 157 Penicillium isolates were selected and screened for phenotypic characters. For the purposes of the study, 46 strains representative of 11 morphological species wereselected and identified by molecular analysis.Results: Based on the findings, P. crustosum (18 isolates, 39.1%) and P. chrysogenum (15 isolates, 32.6%) were the most common isolated species, followed by P. brevicompactum, P. rubens, P. citrinum, P. italicum (each 2 isolates, 4.3%), P. olsonii, P. expansum, P. griseofulvum, P. palitans, and P. polonicum (each 1 isolate, 2.1%).Except for P. chrysogenum and P. expansum with floccose colony texture, the rest of the isolated species had velutinous texture.Conclusion: This is the first report in southern Iran to identify a large number of Penicillium strains isolated from air samples, showing P. crustosum and P. chrysogenum as the most common isolated species.

Background and Purpose: The taxonomy of Candida is controversial and has undergone changes due to the investigation of the novel species. Candida africana and Candida dubliniensis are new members of C. albicans complex that are currently gaining both clinical and epidemiologic significance. This study reports the prevalence of C. africana among the strains isolated from patients, by using HWP1 gene size polymorphism.Materials and Methods: A total of 235 yeasts confirmed as C. albicans complex based on chromogenic media and ITS-sequencing isolated from various clinical forms of invasive and non-invasive candidiasis mainly candidemia, were re-identified based on HWP1 gene polymorphisms. The Hwp1-PCR amplicons were re-confirmed by sequencing and BLAST analysis.Results: Based on the HWP1 gene size polymorphism, 223 strains were identified as C. albicans (94.89%) from which 7 isolates produced two DNA fragments (850 and 941 bp). C. dubliniensis (n=4, 1.7%), C. africana (n=1, 0.42%) and mix of C. albicans and C. africana (n=7, 2.97%) were also identified. Conclusion: Although C. albicans remains the most common Candida species, C. dubliniensis and C. africana are rarely found among the patient isolates. Due to limited information on the molecular epidemiology of this novel yeast, more studies using molecular methods are recommended.

Diabetes mellitus patients are prone to cutaneous and subcutaneous fungal infections due to pathogenic fungi, including dermatophytes, Mucorales, Candida, Aspergillus, and Fusarium species. Here, we report a case of A. flavus mycetoma confirmed by isolation and molecular identification. The case was a 38-year-old male farmer with a seven-year history of type 2 diabetes mellitus, living in Khuzestan, southwest of Iran. The patient presented with a right foot swelling associated with a nodule and multiple discharging sinuses following trauma sustained on the foot while working barefoot on the rice farm, a year ago. The nodule appeared at the site of the trauma two months after the injury. The initial diagnosis was based on direct microscopic examination of lesions scraping using 20% potassium hydroxide and radiology. Molecular analysis confirmed the isolates to be A. flavus. In vitro susceptibility of the isolate to voriconazole, posaconazole, caspofungin, itraconazole, and amphotericin B was determined. Treatment with voriconazole (200 mg twice daily) stopped the purulent discharge, reduced the swelling, and improved the clinical condition within two months. The study emphasizes the importance of wearing footwear to prevent skin trauma as the main risk factor of patient involvement.

Background and Purpose: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is widely used to discriminate among pathogenic microorganisms in clinical laboratories. The aim of this study was to assess the utility of MALDI-TOF MS in the routine identification of clinical dermatophyte isolates obtained from various geographical regions of Iran. Materials and Methods: A total of 94 isolates, including Trichophyton interdigitale (n=44), T. rubrum (n=40), T. tonsurans (n=4), Microsporum canis (n=4), and Epidermophyton floccosum (n=1), were analyzed in this study. The identity of each isolate was determined by polymerase chani reaction amplification and sequencing of the internal transcribed spacer (ITS) region of nuclear-encoded ribosomal DNA and also MALDI-TOF MS. The obtained data by molecular approach were compared with MALDI-TOF MS. Results: The MALDI-TOF MS led to the identification of 44 (47%) isolates at the species level by generating the spectral score values of ≥ 2.0. However, there was not sufficient agreement between the results of the molecular-based ITS identification methods and MALDI-TOF MS in the species identification of 16 (17%) isolates. The Bruker Daltonics database was also not able to identify protein spectra related to 12 isolates (13%), including T. interdigitale (n=5), T. rubrum (n=4), M. canis (n=2), and T. tonsurans (n=1). Conclusion: According to the results, the utility of MALDI-TOF MS as a routine diagnostic tool for the accurate and reliable identification of dermatophytes can be justified whenever the protein spectra of a large set of worldwide clinical isolates are included in the commercial libraries. In addition, MALDI-TOF MS can be alternatively used to construct an in-house reference database.

Abstract Background and Purpose: Candida parapsilosis is a common cause of candidemia in children and onco-hematologic patients as well as in patients with septic arthritis, peritonitis, vaginitis, and nail and skin infections. Here, we evaluated intra- and inter-species beta tubulin DNA sequence variation within the C. parapsilosis complex with a view to establishing its utility in the identification and phylogenetic analysis of the species. Materials and Methods: The novel primers successfully amplified the target for all 23 strains included in the study. Results and Conclusion: Multiple alignment of 623–629 bp-long nucleotide (nt) sequences reflecting the beta tubulin gene indicated an inter-species divergence ranging from 0 to 68 nt in C. parapsilosis, C. orthopsilosis and C. metapsilosis with a mean similarity of 84.7% among the species. Meanwhile, intra-species differences of 0–20 and 0–6 nt were found within the strains of C. parapsilosis and C. orthopsilosis, respectively. The phylogenetic tree topology was characterized by a clade made up by C. parapsilosis and C. orthopsilosis, while C. metapsilosis formed a related but separate lineage. Our data provide the basis for further discoveries of the relationship between the species belonging to the C. parapsilosis complex and the value of beta tubulin DNA sequence data in the identification and taxonomy of C. parapsilosis and other pathogenic yeasts.

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.

Cutaneous mucormycosis is a rare manifestation of an aggressive fungal infection. Early diagnosis and treatment are vitally important in improving outcome. We report an unusual case presenting with progressive necrotizing fasciitis due to mucormycosis following trauma and dressing by man-made herbal agents.

We study 2-step general Fibonacci sequences in the generalized quaternion groups Q4n. In cases where the sequences are proved to be simply periodic, we obtain the periods of 2-step general Fibonacci sequences.