Low Level of Antifungal Resistance in Iranian Isolates of Candida glabrata Recovered from Blood Samples in a Multicenter Study from 2015 to 2018 and Potential Prognostic Values of Genotyping and Sequencing of PDR1.

Establishing an effective empirical antifungal therapy requires that national surveillance studies be conducted. Herein, we report the clinical outcome of infections with and the microbiological features of Iranian isolates of Candidaglabrata derived from patients suffering from candidemia. C. glabrata isolates were retrospectively collected from four major cities in Iran; identified by a 21-plex PCR, matrix-assisted laser desorption ionization-time of flight mass spectrometry, and large subunit of ribosomal DNA sequencing; and genotyped by amplified fragment length polymorphism (AFLP). Mutations in PDR1, ERG11, and hot spot 1 (HS1) of FKS1 and FKS2 were investigated, and antifungal susceptibility testing (AFST) was performed (by the CLSI M27-A3 and M27-S4 methods). Seventy isolates of C. glabrata were collected from 65 patients with a median age of 58 years. Fluconazole was the most widely used (29.23%) and least effective antifungal agent. The overall crude mortality rate was 35.4%. Only one strain was resistant to fluconazole, and 57.7% and 37.5% of the isolates were non-wild type (non-WT) for susceptibility to caspofungin and voriconazole, respectively. All isolates showed the WT phenotype for amphotericin B, posaconazole, and itraconazole. HS1 of FKS1 and FKS2 did not harbor any mutations, while numerous missense mutations were observed in PDR1 and ERG11 AFLP clustered our isolates into nine genotypes; among them, genotypes 1 and 2 were significantly associated with a higher mortality rate (P = 0.034 and P = 0.022, α < 0.05). Moreover, 83.3% of patients infected with strains harboring a single new mutation in PDR1, T745A, died despite treatment with fluconazole or caspofungin. Overall, Iranian isolates of C. glabrata were susceptible to the major antifungal drugs. Application of genotyping techniques and sequencing of a specific gene (PDR1) might have prognostic implications.

Successful treatment of pulmonary aspergillosis due to Aspergillus fumigatus in a child affected by systemic lupus erythematosus: A case report from Northeastern Iran.

A6-year-old girl affected to systemic lupus erythematosus with symptoms of fever, weakness, and lethargy, cough, chest pain, and abnormalchest x-ray. The isolated Aspergillus fumigatus was identified using partial calmodulin gene sequencing. Gradual improvement was observed onday 19 of treatment with amphotericinB (50 mg /day).

Comparison of the efficacy of indocyanine green-mediated photodynamic therapy and nystatin therapy in treatment of denture stomatitis.

BACKGROUND: Candida species have an influence in the pathogenesis of denture stomatitis. The current study aimed to investigate the efficacy of indocyanine green (ICG)-mediated photodynamic therapy (PDT) in combination with nystatin mouthwash (PDT + nystatin) for the treatment of denture stomatitis in comparison with routine antifungal therapy with nystatin alone. METHODS: In this double-blind randomized clinical trial, 66 patients were randomly assigned into PDT + nystatin (n = 33) and nystatin (n = 33) groups, both groups were treated 3-times a day (15 days) with nystatin mouthwash, and PDT was performed twice once a week for the PDT + nystatin group. Briefly, ICG was applied on the palatal lesion and laser irradiation was performed using a diode laser (810 nm, 56 J/cm2). Nystatin group was also treated with sham laser in order to eliminate the possible psychological effects. The clinical and mycological evaluations were carried out at the baseline, during treatment, and the end of follow-up. Patients who completed the treatment and follow-up were eligible for statistical analysis (each group 28 cases). RESULTS: Patient treatment with nystatin or PDT + nystatin reduced the lesions extension. Candida species were isolated from all patients and the number of Candida CFU in both groups showed a significant reduction at each post-treatment visit; however, the mean reduction achieved in the PDT + nystatin group was significantly higher than nystatin alone. CONCLUSIONS: ICG-mediated PDT in combination with nystatin mouthwash can improve the clinical feature of denture stomatitis with no adverse effects; therefore, it could be used as an alternative to the currently available antifungal therapy using nystatin alone.

Evaluation of biofilm formation in the homozygous and heterozygous strains of vaginal Candida albicans isolates.

BACKGROUND AND PURPOSE: Candida albicans is one of the most opportunistic yeasts around the world. This species has two heterozygous and homozygous strains at hyphal wall protein 1 (hwp1) gene locus. A simple method for the discrimination of these two strains is the amplification of HWP1 gene. Regarding this, the aim of this study was to discriminate C. albicans heterozygous and homozygous strains via the amplification of hwp1 gene and evaluation of biofilm formation between the strains. MATERIALS AND METHODS: A total of 60 homozygous (n=30) and heterozygous (n=30) strains were discriminated among 126 C. albicans vaginal isolates by the amplification of HWP1 gene, using specific primers. The evaluation of biofilm formation was accomplished using the visual method. RESULTS: According to the results, the homozygous and heterozygous strains produced one and two DNA fragments, respectively. The frequency of homozygous strains among the C. albicans vaginal isolates was 76.2%. Biofilm formation activity in the heterozygous strains was more than that in the homozygous strains. However, statistical analysis showed no significant difference between the strains in terms of biofilm formation. CONCLUSION: As the findings indicated, the frequency of the heterozygous strains in C. albicans was lower than that of the homozygous strains. Both of the strains could form biofilm in the different ranges of severity. High activity of biofilm formation in heterozygous strains may set the ground for its pathogenicity.

IL-25 and IL-33 Serum Levels are not Associated with the Type of Allergen Causing Allergic Rhinitis.

BACKGROUND: Fungal aeroallergens might sensitize the airway which in turn produces a specific cytokine profile. OBJECTIVE: To evaluate the IL-25 and IL-33 profile in patients with fungal allergic rhinitis. METHODS: The present study examined patients who were evaluated due to allergic rhinitis (AR) at Emam Reza Hospital of Shiraz, Iran. The allergic patients were categorized based on the skin prick test. Blood samples were collected and allergen-specific IgE and cytokine profiles were analyzed. RESULTS: 184 patients were enrolled in the study and in 35 of whom fungal rhinitis was confirmed. The levels of specific IgE in patients with fungal allergy were statistically significant compared to those in the control group (p<0.000). However, there were no significant differences in IL-25 and IL-33 levels between fungal and none-fungal AR patients. CONCLUSION: Chronic fungal challenge might regulate innate system cytokines in severe persistent AR.

Investigation of cgrA and cyp51A gene alternations in Aspergillus fumigatus strains exposed to kombucha fermented tea.

BACKGROUND AND PURPOSE: Aspergillus fumigatus is one of the most common opportunistic fungus, which causes infection in immunocompromised and neutropenic patients. The current guidelines recommend voriconazole as the initial therapeutic and prophylactic agent for almost all cases, especially in patients with organ transplants, which leads to increased medication resistance in A. fumigatus. The aim of the present study was to evaluate the antifungal activity and effect of kombucha as a natural compound on A. fumigatus growth, as well as on the expression of cgrA and cyp51A genes. MATERIALS AND METHODS: A panel of 15 A. fumigatus strains with two quality controls of CM237 and CM2627 as susceptible and resistant strains were obtained from Tehran Medical Mycology Laboratory, Tehran,Iran(TMML).Antifungal susceptibility testing assay was performed according to the Clinical and Laboratory Standards Institute (CLSI) M38-A2 document. Moreover, the mycelial dry weight of the fungus was calculated before and after being treated with kombucha. In addition, the quantitative changes in the expression of cgrA and cyp51A genes were analyzed by real-time polymerase chain reaction (real-time PCR) technique. RESULTS: In the present study, the minimum inhibitory concentration ranges of kombucha were measured at 6,170 and 12,300 µg/mL for ten A. fumigatus azole-susceptible strains and 24,700 µg/mL for five A. fumigatus resistant strains. Moreover, changes in mycelial dry weight under kombucha treatment conditions underwent a significant reduction (P≤0.05). A coordinate down-regulation of expression in cgrA and cyp51A genes was observed in all azole-susceptible and -resistant A. fumigatus strains, after treating the fungus with different concentrations of kombucha (P≤0.05). CONCLUSION: According to the obtained results, kombucha as a natural antioxidant , can exert inhibitory effects against the growth and expression of some genes in A. fumigatusstrains.

Molecular Identification, Genotypic Diversity, Antifungal Susceptibility, and Clinical Outcomes of Infections Caused by Clinically Underrated Yeasts, Candida orthopsilosis, and Candida metapsilosis: An Iranian Multicenter Study (2014-2019).

Despite the increasing occurrence of Candida orthopsilosis and Candida metapsilosis in clinical settings, little is known about their microbiological and clinical properties. Herein, we conducted a national retrospective study (2014-2019) from multiple centers in Iran. Among the 1,770 Candida isolates collected, we identified 600 Candida parapsilosis species complex isolates. Isolate identification was performed by 9-plex PCR, matrix-assisted laser desorption-time of flight mass spectrometry (MALDI-TOF MS), and rDNA sequencing, and antifungal susceptibility testing (AFST) followed CLSI M27-A3/S4; genotyping was performed by amplified fragment length polymorphism (AFLP) analysis; and clinical information was mined. Thirty-one isolates of C. orthopsilosis from various clinical sources, one mixed sample (blood) concurrently containing C. orthopsilosis and C. parapsilosis and one isolate of C. metapsilosis from a nail sample were identified. Although both 9-plex PCR and MALDI-TOF successfully identified all isolates, only 9-plex PCR could identify the agents in a mixed sample. For the C. orthopsilosis isolates, resistance (non-wild type) was noted only for itraconazole (n = 4; 12.5%). Anidulafungin and fluconazole showed the highest and voriconazole had the lowest geometric mean values. AFLP analysis showed three main and four minor genotypes. Interestingly, 90% of nail isolates clustered with 80% of the blood isolates within two clusters, and four blood isolates recovered from four patients admitted to a hospital clustered into two genotypes and showed a high degree of similarity (>99.2%), which suggests that C. orthopsilosis disseminates horizontally. Supported by our data and published case studies, C. orthopsilosis and C. metapsilosis can be linked to challenging clinical failures, and successful outcomes are not always mirrored by in vitro susceptibility. Accordingly, conducting nationwide studies may provide more comprehensive data, which is required for a better prognosis and clinical management of patients.