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Background: Despite the unprecedented surge in the incidence of mucormycosis in the COVID-19 era, the antifungal susceptibility patterns (ASPs) of COVID-19 associated mucormycosis (CAM) isolates have not been investigated so far and it is unclear if the high mortality rate associated with CAM is driven by decreased susceptibility of Mucorales to antifungal drugs. Objectives: To describe the clinical, mycological, outcome and in vitro ASPs of CAM cases and their etiologies from Iran. Patients/Methods: A prospective study from January 2020 to January 2022 at a referral tertiary hospital in Tehran, Iran was conducted for screening mucormycosis through histopathology and mycological methods. The identity of Mucorales isolates was revealed with ITS-panfungal PCR& sequencing and MALDI-TOF. The AS for amphotericin B, itraconazole, voriconazole, posaconazole and caspofungin was cleared according to the EUCAST antifungal susceptibility testing protocol. Result: A total of 150 individuals were diagnosed with CAM. Males constituted 60.7% of the population. The mean age was 54.9 years. Diabetes was the leading risk factor (74.7%). The median interval between diagnosis of COVID-19 and CAM was 31 days. The recovery rate of culture was as low as 41.3% with Rhizopus arrhizus being identified as the dominant (60; 96.7%) agent. Amphotericin B (MIC50= 0.5µg/ml) demonstrated the highest potency against Mucorales. Conclusion: Majority of the cases had either diabetes, history of corticosteroid therapy or simultaneously both conditions. Accordingly, close monitoring of blood glucose should be considered. The indications for corticosteroids therapy are recommended to be optimized. Also, an anti Mucorales prophylaxis may be necessitated to be administrated in high risk individuals. Although amphotericin B was the most active agent, a higher rate of resistance to this antifungal was noted here in comparison with earlier studies on mucormycetes from non-CAM cases.

Aim: The aim of this study was to determine whether photodynamic therapy of resistant onychomycosis with Ag@ZnO nanoparticles can promote the treatment procedure and extirpates the recurrence of fungal infection. Methods: Ag@ZnO nanoparticles (NPs) under UVB-radiation were applied to treat T. rubrum and T. mentagrophytes in vitro through photodynamic therapy. In vivo therapeutic efficacy, biocompatibility and biodistribution of Ag@ZnO NPs were studied. Results: 40 μg/ml of UVB-activated Ag@ZnO NPs showed 100% antifungal activity against dermatophytosis in vitro and in vivo followed by complete growth prevention by degeneration of spores and mycelium after 180 days, while posed biocompatibility. Conclusion: This study showed the superiority of photodynamic therapy with Ag@ZnO NPs followed by proper regeneration of the skin with Zinc ion of the shell.

The Article Abstract is not available.

A fatal case of COVID-19-associated mucormycosis (CAM) affected a 40-year-old woman who was initially admitted to our hospital due to a SARS-CoV-2 infection. Her clinical condition worsened, and she finally died because of respiratory failure, hemodynamic instability, and mucormycosis with invasion into the orbit and probably the brain. According to DNA sequence analysis of the fungus isolated from the patient, Apophysomyces variabilis was involved. This is the first published case of CAM and the third case of mucormycosis due to this mold.

Background and purpose: Candidemia is a major cause of morbidity and mortality among patients receiving immunosuppressive therapy and those hospitalized with serious underlying diseases. Here, we investigated the epidemiological, clinical, and mycological features of candidemia in Tehran, Iran. Materials and methods: A prospective observational study of all patients diagnosed with candidemia was performed at two referral teaching hospitals in Tehran, Iran, from February to December 2018. Demographic characteristics, underlying diseases, risk factors, clinical symptoms, and laboratory analyses of candidemic patients with positive culture were mined. Candida isolates were molecularly identified by sequencing of the internal transcribed spacer region (ITS1-5.8S-ITS2). The antifungal susceptibility testing for fluconazole, itraconazole, voriconazole, posaconazole, amphotericin B, caspofungin, micafungin, and anidulafungin against the isolates was performed using CLSI broth microdilution reference method (M27-A3). Results: A total of 89 episodes were identified, with an incidence of 2.1 episodes/1000 admissions. The common underling disease were malignancy (46%), renal failure/dialysis (44%), and hypertension (40%). The overall crude mortality was 47%. C. albicans (44%) was the most frequent causative agent, followed by C. glabrata (21%), C. parapsilosis complex (15%), C. tropicalis (11%), and C. lusitaniae (3.5%). All the isolates were susceptible to amphotericin B. The activity of all four azoles was low against non-albicans Candida species, especially C. tropicalis. Conclusion: The increase in non-albicans Candida species with reduced susceptibility to antifungal drugs might be alarming in high-risk patients. Therefore, accurate knowledge of predisposing factors and epidemiological patterns in candidemia are effective steps for managing and decreasing the mortality rate in candidemia. This study has been funded and supported by Tehran University of Medical Sciences, Tehran, Iran (Grant no. 99-2-99-48944). Sí

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM Objectives Non-albicans Candida species and other rare yeasts have emerged as major opportunistic pathogens in fungal infections. Identification of opportunistic yeasts in developing countries is mainly performed by phenotypic assay, which is time-consuming and prone to errors. The aim of the present study was to evaluate PCR-RFLP as a routinely used identification technique for the most clinically important Candida species in Iran and make a comparison with a novel multiplex PCR, called 21-plex PCR. Methods A total of 173 yeast isolates from clinical sources were selected and identified with sequence analysis of the D1/D2 domains of rDNA (LSU rDNA) sequencing as the gold standard method. Results The results were compared with those obtained by PCR-RFLP using MspI restriction enzyme and the 21-plex PCR. PCR-RFLP correctly identified 93.4% of common pathogenic Candida species (C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, and P. kudriavsevii (=C. krusei)) and was able to identify 45.5% of isolates of the uncommon yeast species compared to the D1/D2 rDNA sequencing. Compared with PCR RFLP, all common Candida species and 72.7% of uncommon yeast species were correctly identified by the 21-plex PCR. Conclusion The application of the 21-plex PCR assay as a non-sequence-based molecular method for the identification of common and rare yeasts can reduce turnaround time and costs for the identification of clinically important yeasts and can be applied in resource-limited settings.

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM Objectives Pseudallescheria boydii keratitis is rare but important type of fungal keratitis because of the inherently resistance of the organism to many existing antifungal agents. We present the clinical characteristics, risk factors, treatment, and prognosis of patients with P. boydii keratitis, and also present the antifungal sensitivities of the isolated strain. Methods Slit-lamp and confocal microscopy were used for clinical examinations. Fungal isolates were identified based on morphological characteristics and DNA sequence of the internal transcribed spacer region (ITS). In vitro antifungal susceptibility testing for fungal isolates was performed according to the Clinical and Laboratory Standards Institute (CLSI). Results Pseudallescheria boydii was identified in four patients. All patients had a history of ocular trauma. In clinical examination hypopyon was seen in three patients. The main antifungal medications were oral and topical voriconazole. After treatment the visual activity of all patients improved in 2-3 weeks. Conclusion These patients bold the importance of determining causative organism of fungal keratitis and their antibiotic susceptibility. Culture findings are limited in identifying organisms. Sequencing of polymerase chain reaction-amplified DNA is good for accurate and rapid identification of species that can be helpful for optimizing treatment.

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM Objective Fungal respiratory infections are being recognized with increasing frequency in parallel with an expanding population of immunocompromised patients. In most cases, colonization is the first step in the progression to pulmonary fungal infection. This study was designed to evaluate the distribution of fungal elements in the respiratory tract of symptomatic patients hospitalized in pulmonary units. Methods This descriptive cross-sectional study was carried out over a period of 2 years, from October 2017 to October 2019 in Guilan province, located in Iran's northern region. In the current study, bronchoalveolar lavage or sputum specimens were collected. All samples were analyzed by direct microscopy using KOH 10% and culture. Fungal identification was accomplished by internal transcribed spacer (ITS) and beta-tubulin sequencing. Also, in patients suspected of invasive pulmonary aspergillosis, BAL specimens were tested for galactomannan (GM) antigen. Results A total of 384 lung specimens [192 bronchoalveolar lavage (BAL) and 192 sputum samples] were obtained from symptomatic patients hospitalized in pulmonary units. Of these, 137 (35.67%) were positive in direct examination and culture. Among the 137 positive cases, most isolates were from male patients 86 (62.77%) and most of them were between 46 and 72 years. Candida albicans (37.22%) and C. tropicalis (21.89%) represent the two most commonly isolated species in the current study. Cough (94.16%), dyspnea (81.02%), purulent sputum (62.04%), and weight loss (56.2%) were the predominant symptoms and tuberculosis (24.81%), chemotherapy (21.89%), and diabetes mellitus (19.70%) were the predominant underlying conditions. Also, 5 cases of invasive pulmonary aspergillosis and 1 case of mucormycosis were diagnosed. Conclusion Candida albicans was the most common fungal species isolated from symptomatic patients hospitalized in pulmonary units. Tuberculosis, chemotherapy and diabetes mellitus were important underlying conditions for pulmonary fungal colonization and/or infection.

Background and Objectives: Monitoring of contagious diseases is important to advance our knowledge of their epidemi- ology and to enable more impressive investigation and prevention efforts. This study aimed to examine antifungal drug sus- ceptibility and molecular analysis of clinical isolates of Trichophyton rubrum and Trichophyton mentagrophytes in humans and cattle. Materials and Methods: A total of 400 patients and 500 cattle were evaluated in this study. Dermatophytosis was confirmed in cases by direct microscopy and culture methods. Antifungal drug susceptibility profiles, MIC , and MIC of isolates were 50 90 determined using the broth microdilution method. Multiplex-PCR, RAPD PCR, and sequencing methods were used for the genetic analysis of virulence genes and the ITS1 and ITS2 regions, respectively. Results: A total of 175 patients and 120 cattle were diagnosed with dermatophytosis. Dermatophytes showed a remarkable rate (30%) of terbinafine resistance. T. mentagrophytes showed lower susceptibility than T. rubrum (MIC=16 μg/mL). Strains harboring Mep1, Mep2, and Mep4 genes had the highest frequency among all genotypes. A RAPD-PCR dendrogram divided T. mentagrophytes and T. rubrum strains into three and six groups, respectively. Conclusion: A notable rate of resistance to terbinafine in isolated dermatophytes was reported in this study. Examination of RAPD-PCR results showed that T. rubrum strains had higher genetic diversity than T. mentagrophytes. Genetic monitoring of dermatophytes must be considered an important factor in providing fungal infection prevention and treatment approaches.

Background: One of the most prevalent infections in hospitalized patients is candiduria. As the prevalence of this infection is increasing, new epidemiologic and therapeutic data can be used as a guide for the management of patients. Objectives: This research aimed to determine the epidemiological and antifungal susceptibility profile of candiduria. Methods: A total of 104 patients admitted to the nephrology and ICU wards of Bu Ali and Labbafinezhad hospitals in Tehran, Iran, were studied in this cross-sectional investigation. Urine samples were examined using direct smear, culture, and PCR-sequencing techniques. The culture plates were subjected to colony count. The clinical and laboratory standards institute (CLSI) document M27 4th ed was used to assess susceptibility to amphotericin B, itraconazole, caspofungin, and fluconazole. Results: Out of 104 patients, 26 (25%) were diagnosed with candiduria. Most patients were between the ages of 64 - 79 years (n = 9, 34.61%) and female (n = 17, 23.94%). Stroke and urinary catheterization were the most common underlying diseases. Candida glabrata (n = 10, 38.64%) was the most common cause of candiduria. Caspofungin and amphotericin B were the most effective antifungal medicines. Conclusions: Candida glabrata has been identified as the most common cause of candiduria. Due to the increasing antifungal resistance in this species, proper treatment of patients is a crucial concern. Caspofungin exhibited potent antifungal activity against all tested isolates. Still, regardless of its favorable in vitro activity, due to its poor glomerular filtration or tubular secretion in vivo, it has sub-therapeutic antifungal concentrations in the urine.

Background Aspergillus fumigatus is the most common species causing invasive aspergillosis (IA), a life-threatening infection with more than 80% mortality. Interactions between A. fumigatus and human blood platelets lead to intravascular thrombosis and localized infarcts. To better understand A. fumigatus pathogenesis, we aimed to analyze the genetic basis of interactions between the pathogen and blood platelets. Methods A bioinformatic pipeline on microarray gene expression dataset, including analysis of differentially expressed genes (DEGs) using Limma R package and their molecular function, as well as biological pathways identification, was conducted to find the effective genes involved in IA. In the wet phase, the gene expression patterns following fungal exposure to blood platelets at 15, 30, 60, and 180 min were evaluated by quantitative reverse transcriptase-PCR analysis. Results Three genes encoding aspartic endopeptidases including (Pep1), (Asp f 13), and (β-glucanase) were the standing candidates. The invasion-promoting fungal proteinase-encoding genes were down-regulated after 30 min of hyphal incubation with blood platelets, and then up-regulated at 60 and 180 min, although only Pep1 was greater than the control at the 60and 180 min time points. Also, the same genes were downregulated in more the clinical isolates relative to the standard strain CBS 144.89. Conclusion Our findings delineate the possible induction of fungal-encoded proteinases by blood platelets. This provides a new research line into A. fumigatus' molecular pathogenesis. Such insight into IA pathogenesis might also guide researchers toward novel platelet-based therapies that involve molecular interventions, especially in IA patients.

Introduction: Colletotrichum species are well-known plant pathogens, which have been increasingly reported as the cause of keratitis or subcutaneous lesions in humans. In this study we reported a rare case of fungal keratitis from Iran and reviewed the literature. Case Presentation: A 69-year-old man whose right eye was injured by herbal material was examined by slit-lamp biomicroscopy and mycology investigation of corneal scrapings was done. The grown filamentous fungal was identified as Colletotrichum gloeosporioides based on morphological characteristics and DNA sequence of the internal transcribed spacer region. The isolated strain was sensitive to amphotericin B, caspofungin, anidolafungin, micafungin, voriconazole, and relatively resistant to fluconazole, and itraconazole. Patient was successfully treated with voriconazole. Conclusions: This report highlights that the early and accurate identification and therapy can helpful to management keratitis caused by C. gloeosporioides.

Acute respiratory distress syndrome is a common complication of severe viral pneumonia, such as influenza and COVID‐19, that requires critical care including ventilatory support, use of corticosteroids and other adjunctive therapies to arrest the attendant massive airways inflammation. Although recommended for the treatment of viral pneumonia, steroid therapy appears to be a double‐edged sword, predisposing patients to secondary bacterial and invasive fungal infections (IFIs) whereby impacting morbidity and mortality. Mucormycosis is a fungal emergency with a highly aggressive tendency for contiguous spread, associated with a poor prognosis if not promptly diagnosed and managed. Classically, uncontrolled diabetes mellitus (DM) and other immunosuppressive conditions including corticosteroid therapy are known risk factors for mucormycosis. Upon the background lung pathology, immune dysfunction and corticosteroid therapy, patients with severe viral pneumonia are likely to develop IFIs like aspergillosis and mucormycosis. Notably, the combination of steroid therapy and DM can augment immunosuppression and hyperglycaemia, increasing the risk of mucormycosis in a susceptible individual. Here, we report a case of sinonasal mucormycosis in a 44‐year‐old woman with hyperglycaemia secondary to poorly controlled diabetes following dexamethasone therapy on a background of influenza pneumonia and review 15 available literatures on reported cases of influenza and COVID‐19 associated mucormycosis.

Background and Objectives: Dermatophytosis induced by Trichophyton mentagrophytes is a major human and animal fun- gal contamination. Antifungals like terbinafine and fluconazole are widely used to treat dermatophytosis; nevertheless, the prevalence of drug resistance has increased. Hence, novel curative strategies are needed. In the present study, we compared the efficacies of conventional and nanoform of antifungals agents in guinea pig model of dermatophytosis. Materials and Methods: Guinea pigs (n=36) were injected (the posterior dorsal portion) with Trichophyton mentagrophytes conidia. The guinea pigs were divided into 6 groups (positive control, negative control, fluconazole 0.5% treated group, na- no-fluconazole treated group, terbinafine 1% treated group, and nano-terbinafine treated group), then were scored both clin- ically (redness and lesion intensity) and mycologically (microscopy and culture) until day 40 of inoculation. The treatment started 5 days after the inoculation and continued until day 40 of inoculation. Results: Assessment of the mean score of clinical lesions in groups treated with nano-drug forms of fluconazole and terbin- afine on the first day of treatment showed a score of 3 (significant redness with large scaling) and for the conventional form of terbinafine and fluconazole had a score of 4 (ulcer and scar). The decrease in lesion score in nano-drug treated groups was observed between days 15 and 20 and continued until day 40. On day 40, all groups had zero scores except the positive control group. Conclusion: This study indicated that nano-drugs are more suitable for the treatment of dermatophytosis and could be con- sidered as future alternatives for the treatment of dermatophytosis.

Background: Candida species are normal vaginal flora in healthy women, which can cause vulvovaginal candidiasis (VVC). The formation of biofilm is a cause of drug resistance in Candida species of vaginal origin. We aimed to specify Candida species cause VVC, detect their biofilm-forming ability, and antifungal susceptibility pattern. Methods: Overall 150 vaginal samples were collected from suspected cases of referring to Bahar Hospital of Shahroud, Iran between Jan 2018 and Jan 2019. Samples were cultured on Sabouraud dextrose agar (SDA), Chrome gar Candida and Corn meal agar (CMA). PCR-RFLP was performed to confirm the identification. Biofilm formation of the identified species was measured by the Crystal Violet method. The susceptibility to fluconazole, clotrimazole, and miconazole was determined based on the CLSI document M27-A3. Results: Of 50 women (33.3%) were suffering from VVC. C.albicans was the predominant species isolated in this study (n=39, 78%) followed by C. glabratia (n=11, 22%). In addition, in 25 (50%) of positive samples, biofilm formation was determined. The mean MIC of fluconazole and clotrimazole for C. albicans was 5.02 μg /mL and 3.92 μg /mL, respectively. Furthermore, the mean MIC related to these drugs for C. glabrata was 12.45 μg / mL and 4.1μg / mL, respectively. The mean diameter of miconazole inhibition zone for C. albicans and C. glabra isolates was 25.13 mm and 24.5mm, respectively and all of them were susceptible to this drug. Conclusion: C.albicans was the predominant Candida species isolated from patients with VVC and also was the predominant biofilm producer species.

The transition of Candida albicans from yeast to hyphae cause the attachment to epithelial cells, forming biofilm and invasion. Therefore, the effect of Pediococcus acidilactici on morphogenesis and pathogenesis of C. albicans were examined. Inhibitory activity of P. acidilactici on Candida species growth was investigated and inhibition of C. albicans biofilm formation was measured by XTT method. Also, expression of seven genes as: Agglutinin-like protein 1 (ALS1,3), hyphal cell-wall protein (HWP1), secreted aspartyl proteinase (SAP4,6), Enhanced filamentous growth 1 (EFG1) and Enhanced activated protein 1 (EAP1) were studied by RT-PCR with different concentrations of P. acidilactici. The experimental activity of this probiotic bacterium was evaluated in an animal model by culture and histopathological methods. P. acidilactici inhibited the growth of Candida species at concentrations of 8-512 µg/mL (approximately 8 × 106 to 5 × 108 CFU/ml). This probiotic bacterium inhibited the germ tube and biofilm formation in a dose-dependent manner. RT-PCR analysis showed a reduction in genes expression. The P. acidilactici reduced the CFUs in mice receiving this probiotic bacterium. Histopathological analyses showed that Candida colonization was diminished in mice following the administration of probiotic. Since the wide range of antifungal activity of this bacterium bacterium, it can be used to manage oral candidiasis.

Introduction. Given the limited number of candidaemia studies in Iran, the profile of yeast species causing bloodstream infections (BSIs), especially in adults, remains limited. Although biochemical assays are widely used in developing countries, they produce erroneous results, especially for rare yeast species.Aim. We aimed to assess the profile of yeast species causing BSIs and to compare the accuracy of the Vitek 2 system and 21-plex PCR.Methodology. Yeast blood isolates were retrospectively collected from patients recruited from two tertiary care training hospitals in Tehran from 2015 to 2017. Relevant clinical data were mined. Identification was performed by automated Vitek 2, 21-plex PCR and sequencing of the internal transcribed spacer region (ITS1-5.8S-ITS2).Results. In total, 137 yeast isolates were recovered from 107 patients. The overall all-cause 30-day mortality rate was 47.7 %. Fluconazole was the most widely used systemic antifungal. Candida albicans (58/137, 42.3 %), Candida glabrata (30/137, 21.9 %), Candida parapsilosis sensu stricto (23/137, 16.8 %), Candida tropicalis (10/137, 7.3 %) and Pichia kudriavzevii (Candida krusei) (4/137, 2.9 %) constituted almost 90 % of the isolates and 10 % of the species detected were rare yeast species (12/137; 8.7 %). The 21-plex PCR method correctly identified 97.1 % of the isolates, a higher percentage than the Vitek 2 showed (87.6 %).Conclusion. C. albicans was the main cause of yeast-derived fungaemia in this study. Future prospective studies are warranted to closely monitor the epidemiological landscape of yeast species causing BSIs in Iran. The superiority of 21-plex PCR over automated Vitek 2 indicates its potential clinical utility as an alternative identification tool use in developing countries.

Microbial species such as bacteria and fungi can be transported by dust storms over long distances, and may change the mycobiota in downwind. This study aimed to evaluate phenotypes and genotypes of airborne fungi during the Middle Eastern dust (MED) events and normal days in Khorramabad, Iran. The samples were collected regularly every six days at three locations during April 2018–March 2019, with additional samplings during MED days. For phenotypic analyses, the Petri dishes were incubated at 25 °C for 72–120 h. Molecular identification of fungi was carried out using polymerase chain reaction (PCR). The average (±SD) of total fungal concentration was 460.9 (±493.2) CFU/m(3). The fungi with the highest average concentrations included Cladosporium cladosporioides, Penicillium brevicompactum, and Cladosporium iridis, respectively. The average concentration of fungi during dust days (967.65 CFU/m(3)) was 3.6 times higher than those in normal days (267.10 CFU/m(3)). During normal and dust days, 61 and 45 species were detected, respectively. Aspergillus and Cladosporium spp. were relatively more dominant during normal and dust days, respectively. Eight fungal species were only observed during MED days, including Talaromyces albobiverticillius that was detected for the first time in Iran. Despite air temperature, relative humidity and wind speed were associated to the fungal concentrations. Dust events lead to the changes in the air pollutants composition and mycobiota, identification of new fungi, and elevated fungal concentrations that may extremely affect the public health. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s40201-019-00428-0) contains supplementary material, which is available to authorized users.

Fusarium genus is a filamentous ascomycete fungi with the ability to produce toxins. Sampling was carried out between 2015-2016 in three tea courses from Mazandaran and Gilan provinces. At intervals of 3, 7 and 15 days, the plates were examined and the colonies grown in each plate were identified and their macroscopic characteristics were recorded. Finally, 150 colonies were selected. Samples cultured in a plate were cultured in liquid medium sB + ME and sB + yE. The environments were examined for successive days and slides were prepared and stained with lacto phenol. This is to ensure the inactivation of fungi exposed to plasma or the removal of spores from the sample surface. There is a significant relationship between the DON produced in the potato medium containing yeast extract and DON contained in the sample taken from the same medium that was exposed to plasma jet for 60 seconds This feature was statistically significant in comparison between changes in the toxicity of T2 and toxin after 60 seconds of treatment with jet plasma and T2 reducibility severity to DON. Plasma treatment results showed a significant reduction in time-dependent toxin concentrations. These results indicate that the plasma system has a high potential not only to destroy mycotoxins, but also toxins produced from fungi, which can be greatly reduced and can be used effectively in the food industry.

Abstract Candida is a common yeast in opportunistic fungal diseases around the world and is usually colonized on the skin and mucosal membranes. The purpose of this study was to synthesize chitosan–silver–copper nanocomposite and to investigate its antifungal effects on Candida albicans. Silver, copper and chitosan nanoparticles were synthesized individually. Then, copper–silver–chitosan nanocomposite was synthesized. These nanoparticles are approved by transmission electron microscope, and nanocomposite structure was also confirmed by scanning electron microscope. Then, the minimum inhibitory concentrations and minimum fungicidal of these nanostructures were examined on C. albicans. The results of this study indicate that the properties and effects of the investigated nanocomposite are comparable to amphotericin B as standard material. The results show that this effect was higher for copper–silver–chitosan nanocomposite than for other nanoparticles studied. Antifungal effect of copper nanoparticles and chitosan nanoparticles was not established separately, but it was found that their composition had antifungal effects that were effective. The combination of nanoparticles of chitosan with silver has been shown to have some antifungal effects. The most antifungal effect for the nanoparticles studied is related to copper–silver–chitosan nanocomposite and, which has had a significant effect on the growth of C. albicans in the laboratory environment compared to other nanoparticles. Graphic abstract

Bacterial or virus co-infections with SARS-CoV-2 have been reported in many studies; however, the knowledge on Aspergillus co-infection among patients with COVID-19 was limited. This study was conducted to identify and isolate fungal agents and to evaluate the prevalence of pulmonary aspergillosis (CAPA) as well as antifungal susceptibility patterns of Aspergillus species in patients with COVID-19 admitted to Shahid Beheshti Hospital, Kashan, Iran.The study involved 119 patients with severe COVID-19 pneumonia referred to the Shahid Beheshti Hospital, Kashan, Iran. A total of 17 Aspergillus spp. that were isolated from COVID-19 patients suspected of CAPA were enrolled in the study. CAPA was defined using ECMM/ISHAM consensus criteria. The PCR amplification of the β-tubulin gene was used to identify the species. The antifungal activities of fluconazole, itraconazole, voriconazole, amphotericin B against Aspergillus spp. were evaluated according to the Clinical and Laboratory Standards Institute manual (M38-A3).From the 119 patients with severe COVID-19 pneumonia, CAPA was confirmed in 17 cases (14.3%). Of these, 12 (70.6%) were males and 5 (29.4%) were females; the mean age at presentation was 73.8 years (range: 45-88 years; median = 77; IQR = 18). Aspergillus fumigatus (9/17; 52.9%), Aspergillus flavus (5/17; 29.4%), Aspergillus oryzae (3/17, 17.6%), were identified as etiologic agents of CAPA, using the molecular techniques. Voriconazole and amphotericin B showed more activity against all isolates. Moreover, the MIC of fluconazole, itraconazole varied with the tested isolates. For 3 clinical isolates of A. fumigatus, 2 isolate of A. flavus and 3 A. oryzae, the MIC of fluconazole and itraconazole were ≥ 16 µg/mL.We observed a high incidence (14.3%) of probable aspergillosis in 119 patients with COVID-19, which might indicate the risk for developing IPA in COVID-19 patients. When comparing patients with and without CAPA regarding baseline characteristics, CAPA patients were older (p =0 .024), had received more frequent systemic corticosteroids (p = 0.024), and had a higher mortality rate (p = 0.018). The outcome of CAPA is usually poor, thus emphasis shall be given to screening and/or prophylaxis in COVID-19 patients with any risk of developing CAPA.

Background and Purpose: The pandemic of COVID-19 has caused a worldwide health crisis. Candidemia is a potentially lethal condition that has not yet been enough discussed in patients with COVID‐ 19. The current study aimed to investigate the prevalence of candidemia among Iranian COVID‐ 19 patients and characterize its causative agents and the antifungal susceptibility pattern. Materials and Methods: The present cross-sectional survey was carried out from March 2020 to March 2021 at Imam Khomeini Hospital, Tehran, Iran. Blood specimens were obtained from patients with confirmed coronavirus infection who also had criteria for candidemia and were examined for any Candida species by conventional and molecular techniques. Susceptibility of isolates to amphotericin B, voriconazole, itraconazole, fluconazole, caspofungin, and 5-flucytosine was tested using the CLSI broth dilution technique. Results: In total, 153 patients with COVID-19 were included and candidemia was confirmed in 12 (7.8 %) of them. The majority of patients were ≥ 50 years of age (n=9) and female (n=8). Moreover, 6 out of the 12 patients were diabetic. The presence of central venous catheters, broad-spectrum antibiotic therapy, ICU admission, andmechanical ventilation was observed in all patients. The C. albicans (n=7, 58.3 %) and C. dubliniensis (n=2, 16.7%) were the most common isolated species. Amphotericin B and 5-flucytosine were the most active drugs. Despite antifungal treatment, 4 out of 12 patients (33.3 %) died. Conclusion: Due to the high mortality, the early diagnosis and proper treatment of candidemia are essential requirements for optimal clinical outcomes in COVID-19 patients.

Background: Tracheoesophageal voice prostheses (TVPs) have been the gold standard in rehabilitation, after laryngectomy, producing faster and premier voicing towards esophageal speech. Fungal colonization shortens the device’s lifetime and leads to prosthesis dysfunction, leakage, and subsequent respiratory infection. Therefore, in the current study, we aimed to investigate the fungal colonization patterns and to propose prophylactic measures that shall increase the longevity of voice prosthesis. Methods: Failed TVPs were removed - due to leakage and/or aspiration - from 66 post laryngectomy patients and examined. They were referred to Amiralam and Rasoul Hospital, the main centers of Ear, Nose, and Throat in Tehran, Iran from April 2018 to January 2020. Fungal colonization patterns were assessed using DNA sequencing techniques. Furthermore, the susceptibility to fluconazole, amphotericin B, nystatin, and white vinegar was evaluated according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. Results: Resident fungal species from the upper airways colonized all the 66 TVPs (100%). Diabetes (31%) and smoking (98%) were the predominant underlying disease and predisposing factors, respectively. Among the 79 fungal agents isolated from the 66 TVPs, Candida glabrata (n=25, 31.7%) was the most common. A significant reduction in minimum inhibitory concentration (MIC) values were observed for white vinegar when used alone (P

Background: With the emergence and spread of coronavirus disease 2019 (COVID-19) globally, health care systems have faced the biggest challenge in recent decades. Objectives: The present study aimed to identify risk factors associated with oropharyngeal candidiasis (OPC) in COVID-19 patients. Methods: The total number of confirmed COVID-19 patients was 218 (105 cases with OPC and 113 controls without OPC). The questionnaire used in this study consisted of demographic data, treatment strategy, clinical and laboratory data, and underlying diseases collected from the onset of clinical OPC until the end of hospitalization. Results: Pseudomembranous candidiasis (77/105, 73.3%) was the most prevalent form of OPC in case patients. The majority of the cases (58.1%) and controls (58.4%) were males. Increasing age (P = 0.03) and hospitalization length (P = 0.016) were significantly associated with OPC in COVID-19 patients. Diabetes (P = 0.003), solid tumor (P = 0.019), and hypertension (P = 0.000) were the most common underlying conditions. The use of dentures (P = 0.003) and poor oral hygiene (P = 0.000) were related to OPC in the case group. Therapy with chloroquine (P = 0.012), IVIG (P = 0.001), diuretics (P = 0.000), and corticosteroid pulse therapy (P = 0.000) were significantly associated with developing OPC in case patients. Conclusions: Old age, hospitalization length, poor oral hygiene, corticosteroids use, diabetes, solid tumor, and hypertension may predispose COVID-19 patients to develop OPC.

The pandemic of COVID-19 has created a global public health crisis. ICU patients with COVID-19 are prone to infections of bacterial and/or fungal origins due to several risk factors. Consequently, the current study was conducted to evaluate the frequency, demographic characteristics, underlying conditions, and etiologic agents of fungal and bacterial co-infections of the respiratory tract among ICU patients with COVID-19 in Iran.From May to October 2020, sputa and endotracheal aspirates were collected from ICU patients hospitalized with COVID-19 who also were suspected of bacterial and/or fungal co-infections according to inclusion criteria. The etiologic agents of bacterial co-infections were identified using the Vitek 2 identification method. For fungal identification, all samples were analyzed by direct microscopy using KOH 10% and culture. Furthermore, all isolates were subjected to sequencing method.A total of 73 lung specimens were obtained from patients who met the inclusion criteria. Of these, in 15 cases (20.54%) fungal and/or bacterial co-infections were confirmed. Males were more infected (73.33%) and all of them were between 49 and 79 years.The frequency of bacterial co-infections of the respiratory tract in ICU patients hospitalized with COVID-19 was relatively high. According to the results, one of the causes of death of these patients could be a secondary infection.

Otomycosis is a fungal infection of the external auditory canal caused mainly by the genus Aspergillus. Aspergillus luchuensis, an industrially important fungus, is a member of Aspergillus section Nigri. In this report, we present a case of otomycosis due to Aspergillus luchuensis in a 43-year-old female patient. We performed a partial PCR-sequencing of β-tubulin and calmodulin genes to identify the isolate to the species level. Further, we determined the in vitro susceptibility of the isolate to nystatin, clotrimazole and itraconazole according to the Clinical & Laboratory Standards Institute (CLSI) M38-A2 protocol. Accordingly, the minimum inhibitory concentrations of clotrimazole, nystatin and itraconazole were 0.25μg/mL, 0.5μg/mL and 1μg/mL, respectively. This is the first report of clinically relevant isolation of Aspergillus luchuensis identified by a molecular technique as a causative agent of otomycosis.

Cerebral mucormycosis (CM) is a life-threatening manifestation of mucormycosis, an angioinvasive fungal infection caused by Mucorales. We sought to systematically review all available case reports to describe epidemiologic features, clinical manifestations, predisposing factors, and diagnostic and treatment strategies of CM. A systematic search was conducted using a combination of the following keywords: "Mucor", "Zygomycetes", "mucormycosis", "cereb*", "brain", "central nervous system", and "intracranial", separately and in combination until December 31

Background: Candida albicans remains the main cause of candidiasis in most clinical settings. Available drugs for candidiasis treatment have many side effects. In this work, novel nitroglycerin derivatives were synthesized and their cytotoxic and antifungal effects evaluated against fluconazole susceptible and resistant clinical C. albicans isolates. Methods: This experimental study was performed in Tehran University of Medical Sciences and Baqiatallah University of Medical Sciences, Tehran, Iran between Feb to Dec 2019. The in vitro activities of two novel nitroglycerin derivatives (1b and 2b) against 25 clinical fluconazole-susceptible and resistant C. albicans isolates and four standard C. albicans strains were determined according to CLSI reference M27-A3 documents. The cytotoxicity of chemical compounds was investigated near the SNL76/7 cells by colorimetric assay. Real-time PCRs were performed to evaluate the alterations in the regulation of ERG11 and CDR1 genes under nitroglycerin derivatives-treated and untreated conditions. Results: The derivatives 1b and 2b exhibited potent antifungal activity against C. albicans isolates; MICs and MFCs varied from 18 μg/ml to 72 μg/ml and 36 μg/ml to 144 μg/ml, respectively. The cell viability evaluation demonstrated that both chemical compounds are safe within 24h. The nitroglycerin derivatives were able to reduce the transcription level of CDR1 and ERG11 genes in all susceptible and resistant C. albicans isolates. Conclusion: Considering the potential and efficacy of these compounds against clinical C. albicans isolates, the complementary in vivo and clinical trials should be investigated.

Glycosylphosphatidylinositols (GPI)-anchored proteins (GpiPs) are related to the cell wall biogenesis, adhesion, interactions, protease activity, mating, etc. These proteins have been identified in many organisms, including fungi such as Neurospora crassa, Candida albicans, Saccharomyces cerevisiae, and Fusarium graminearum. MGL-3153 gene of Malassezia globosa (M. globosa) encodes a protein which is homologous of the M. restricta, M. sympodialis, M. Pachydermatis, and U. maydis GpiPs. Real-time PCR assay showed that the expression of MGL_3153 gene was significantly up-regulated among M. globosa isolated from patients with pityriasis versicolor (PV) compared to a healthy individual, suggesting the contribution of this gene in the virulence of M. globosa. Accordingly, the sequence of this protein was analyzed by bioinformatics tools to evaluate the structure of that. The conservation analysis of MGL-3153 protein showed that the C-terminal region of this protein, which is responsible for GPI-anchor ligation, was highly conserved during evolution while the N-terminal region just conserved in Malassezia species. Moreover, the predicted tertiary structure of this protein by homology modeling showed that this protein almost has alpha helix structure and represented a stable structure during 150 ns of molecular dynamic simulation. Our results revealed that this protein potentially belongs to GPI-anchored proteins and may contribute to the virulence of M. globosa which warrants further investigations in this area.

Background and Objectives: Pneumocystis jirovecii pneumonia (PJP) is a serious infection that usually affects those with a weak immune system. Since the prevalence of this infection in Iran and in the world is not clearly defined, the present study aimed to evaluate the incidence, clinical spectrum, and demographic characteristics of PJP among HIV and non-HIV immunocompromised patients. Materials and Methods: Bronchoalveolar Lavage (BAL) specimens were obtained from 3 groups of immunocompromised patients, including acquired immunodeficiency syndrome (AIDS) patients, diabetic patients, and patients receiving immuno- suppressive therapies. All were hospitalized in pulmonary units. The specimens were examined using microscopic methods (Giemsa and calcofluor white staining) and the nested-PCR technique based on mtLSU-rRNA gene. Results: A total of 120 BAL samples were collected. From 12.5% (5 from 40) of HIV-infected patients, 5% (2 from 40) of patients receiving immunosuppressive therapies, and 2.5% (1 from 40) of diabetic patients Pneumocystis jiroveci was iso- lated. There was not any association between the prevalence of PJP and the patient's gender (p= 0.557) and age (p= 0.681). Fever and dyspnea (n=7, 87.5%), nonproductive cough and abnormal auscultation sound (n=5, 62.5%), and also chills and weight loss (n=2, 25%) were the documented clinical symptoms of PJP. Also, the results showed that none of the samples had positive results for P. jiroveci with microscopic tests while using the nested-PCR method 8 samples had positive results. Conclusion: Since PJP often causes symptoms that are similar to other illnesses, such as the flu or tuberculosis, clinical and laboratory findings should be used simultaneously for making the final decision on drug administration.

BackgroundOnychomycosis is one of the most common and recurrent dermatological diseases worldwide. The antimycotic activity of prescribed medications varies according to the causative agents, and treatment failure rates exceeding 30%. This study aimed to assess the epidemiological profile of onychomycosis in Iran. Also, the susceptibilities to conventional and new antifungals were investigated.MethodsIn this descriptive cross-sectional study, during the period of 18 months starting from September 2019 until March 2020, 594 nail specimens were obtained from patients who presented nail changes compatible with a clinical diagnosis of onychomycosis. The patients were referred from different cities, including Tehran, Kermanshah, Arak, Kashan, Rasht, Qom, Urmia, Zahedan, Hamadan, Zanjan, Borujerd, Bushehr, and Yazd. All the samples were subjected to microscopic examination and fungal culture. Fungi identified were confirmed through the PCR-sequencing method. The susceptibility to itraconazole, fluconazole, terbinafine, griseofulvin, posaconazole, ravuconazole, efinaconazole, luliconazole, and tavaborole was evaluated according to the Clinical and Laboratory Standards Institute (CLSI) guidelines, document M38-A2 for filamentous fungi, and document M27-A3 for yeasts.Results594 patients were included. Of these, in 179 cases (30.1%) (95% CI:0.3 ± 0.037) onychomycosis was confirmed. The majority of patients were ≥ 60 years of age (n=58, 32.6%) and female (n=113, 63.1%). Saprophytic fungi accounted for the vast majority of the nail isolates (n=92, 51.4%) (95% CI:0.051 ± 0.0.073), followed by dermatophytes (n=45, 25.1%) (95% CI:0.25 ± 0.063), and yeasts (n=42, 23.5%) (95% CI:0.23 ± 0.061). Diabetes mellitus (77.3%), hypothyroidism (18.2%), and solid tumors (4.5%) were documented as the most prevalent underlying conditions. Antifungal susceptibility testing was performed against 60 fungal isolates (20 each of Candida species, saprophytic fungi, and dermatophytes). Efinaconazole, ravuconazole, and luliconazole were the most active agents against Candida species. Also, luliconazole, posaconazole, and efinaconazole were most potent against dermatophytes. Luliconazole had the greatest antifungal activity against saprophytic fungi.ConclusionsThe prevalence of onychomycosis in Iranian patients was relatively high. LUL exhibited potent antifungal activity against the three groups of fungi tested, determining its broad-spectrum antimycotic activity and its probable use as the first-line therapy for onychomycosis.