In vitro antifungal susceptibility profile of Iranian Fusarium isolates: Emphasising on the potent inhibitory effect of efinaconazole compared to other drugs.

BACKGROUND: Fusarium species are opportunistic human pathogens that remarkably cause fungal infections ranging from superficial to fatal invasive disseminated infections. Fusarium species are notoriously resistant to the majority of antifungal agents. OBJECTIVES: Therefore, detailed studies regarding in vitro susceptibility are required and may lead to a better prognosis of severe infections. METHODS: We evaluated 25 antifungal drugs in vitro against 282 clinical and environmental Fusarium isolates. RESULTS: Fusarium species demonstrated high MICs/MECs values to the most commonly used antifungal drugs in clinical practice. The geometric mean (GM) MICs for luliconazole (0.004 µg/ml) and lanoconazole (0.012 µg/ml) were the lowest, followed by efinaconazole (0.98 µg/ml) and amphotericin B (1.04 µg/ml). CONCLUSIONS: Efinaconazole, a novel triazole, may be a promising candidate for the treatment of superficial Fusarium infections. Furthermore, the development of systemic formulations of these drugs as well as further in vitro and in vivo investigations could aid in the treatment of systemic fusariosis.

Indoor environment assessment of special wards of educational hospitals for the detection of fungal contamination sources: A multi-center study (2019-2021).

Background and Purpose: The hospital environment was reported as a real habitat for different microorganisms, especially mold fungi. On the other hand, these opportunistic fungi were considered hospital-acquired mold infections in patients with weak immune status. Therefore, this multi-center study aimed to evaluate 23 hospitals in 18 provinces of Iran for fungal contamination sources. Materials and Methods: In total, 43 opened Petri plates and 213 surface samples were collected throughout different wards of 23 hospitals. All collected samples were inoculated into Sabouraud Dextrose Agar containing Chloramphenicol (SC), and the plates were then incubated at 27-30ºC for 7-14 days. Results: A total of 210 fungal colonies from equipment (162, 77.1%) and air (48, 22.9%) were identified. The most predominant isolated genus was Aspergillus (47.5%), followed by Rhizopus (14.2%), Mucor (11.7%), and Cladosporium (9.2%). Aspergillus (39.5%), Cladosporium (16.6%), as well as Penicillium and Sterile hyphae (10.4% each), were the most isolates from the air samples. Moreover, intensive care units (38.5%) and operating rooms (21.9%) had the highest number of isolated fungal colonies. Out of 256 collected samples from equipment and air, 163 (63.7%) were positive for fungal growth. The rate of fungal contamination in instrument and air samples was 128/213 (60.1%) and 35/43 (81.2%), respectively. Among the isolated species of Aspergillus, A. flavus complex (38/96, 39.6%), A. niger complex (31/96, 32.3%), and A. fumigatus complex (15/96, 15.6%) were the commonest species. Conclusion: According to our findings, in addition to air, equipment and instrument should be considered among the significant sources of fungal contamination in the indoor environment of hospitals.

Diversity of Geophilic Dermatophytes Species in the Soils of Iran; The Significant Preponderance of Nannizzia fulva.

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.

Vaginal and oral use of probiotics as adjunctive therapy to fluconazole in patients with vulvovaginal candidiasis: A clinical trial on Iranian women.

Background and Purpose: Vulvovaginal candidiasis is considered the second most prevalent gynecologic infection among women and one of the main reasons for referring to a gynecologist. During recent decades, probiotic usage has been defined as one of the therapeutic regimens for vaginal candidiasis management, but these findings were controversial. The current study was conducted to determine the effect of fluconazole plus vaginal and oral probiotics supplementation on clinical and mycological improvement of vaginal candidiasis concomitant with antifungal susceptibility of Candida species to fluconazole. Materials and Methods: This double-blind, randomized, placebo-controlled trial was conducted on 76 women with vaginal candidiasis admitted to Naghavi and Imam Reza Gynecology Clinics in Kashan, Central Iran, from July 2017 to March 2020. Patients were diagnosed according to vaginal candidiasis symptoms and positive culture for Candida species. The patients were divided into two groups; one of them received fluconazole plus vaginal and oral probiotics, while the other one received fluconazole with placebo. The clinical and mycological findings were recorded before and after the treatment. In vitro, the fluconazole susceptibility test was determined by the microdilution method according to the Clinical and Laboratory Standards Institute (M27-A3) for the baseline Candida isolates. Conclusion: Based on the findings, 35 days after the intervention, a significant reduction was reported in vaginal candidiasis symptoms in the probiotics supplementation group. Although probiotics supplementation therapy was a better mycological cure, compared to the fluconazole with the placebo group, this difference was not significant (68.4% vs. 46.9%, P=0.184). Exclusion of resistant and susceptible dose-dependent strain in the regression model demonstrated a significant reduction in positive culture probiotics in the supplementation group. Results: Oral and vaginal supplementation with probiotics for 4 weeks played a significant role in the elimination of vaginal candidiasis symptoms. Adjustment of clinical and mycological responses with drug resistance patterns of patients could open a promising horizon for probiotics consumption as a complementary treatment.

Microbial contamination of keyboards and electronic equipment of ICU (Intensive Care Units) in Kashan University of medical sciences and health service hospitals.

Microbial contamination of computer keyboards and inanimate surfaces of electronic equipment in ICU (Intensive Care Units) can have a significant role for ICU-acquired colonization and a spectrum of nosocomial infections. The aim of this study was to survey the incidence of bacterial contamination and the distribution of species of computer keyboards and inanimate surfaces of bed side equipment in ICUs in Kashan University of medical sciences and health service hospitals. This descriptive, cross-sectional study was done on 75 computer keyboards and inanimate surfaces electronic equipment in 5 ICUs during 2016-2017. Samples were collected from computer keyboards and electronic equipment with normal saline rinsed swabs. Samples were Cultivated on Blood Agar (BA), and MacConkey Agar (MAC) and growing bacteria were identified based on their morphology and biochemical properties. Seventy six (76%) out of 75 computer Keyboards and electronic equipment were contaminated with bacteria and fungi. The most contamination pertained to gram positive bacteria (70.7%) and the most isolated bacteria were coagulase-negative staphylococci. The highest contamination rates were found on computer keyboards and electronic equipment of which were nurses. •This study demonstrates that monitoring inanimate surfaces and considering these surfaces as source of nosocomial infections is necessary.•In total, Seventy six (76%) out of 75 computer keyboards and electronic equipment in ICUs had positive culture.•It can be concluded that it is necessary for ICUs of Kashan university of medical sciences and Health service hospitals to have practical and regular program to reduce nosocomial infections.

Novel Point Mutations in cyp51A and cyp51B Genes Associated with Itraconazole and Posaconazole Resistance in Aspergillus clavatus Isolates.

Aspergillus clavatus is a common environmental species known to cause occupational allergic disease in grain handlers. We have recently observed azole-resistant isolates of this fungus as a cause of onychomycosis. To further characterize the cause of resistance, the genes encoding 14 α-sterol demethylase enzyme (cyp51A and cyp51B) were characterized and analyzed in 9 ITC-susceptible isolates and 6 isolates with high minimum inhibitory concentrations (MICs) of clinical (nail and sputum) and environmental A. clavatus strains. We found that six isolates with itraconazole MIC >16 mg/L demonstrated nonsynonymous mutations, including V51I, L378P, E483K, and E506G, and synonymous mutations, including F53F, A186A, Q276Q, and H359H. Moreover, P486S was detected in five strains with ITR MIC >16 mg/L. One mutation, F324S, was detected in an isolate with posaconazole MIC >16 mg/L. The effect of E483K and P486S mutations of CYP51A on azole resistance was further investigated using homology modeling and molecular dynamics. We found that E483K and P486S mutations were located near the ligand access channel of CYP51A that could partly lead to narrowing the entry of the ligand access channels. Therefore, we concluded that E483K and P486S mutations may potentially contribute to the limited access of inhibitors to the binding pocket and therefore confer resistance to azole agents.

A Case of Fungus Ball-Type Pansinusitis Due to Fusarium proliferatum.

Incidence of fungal sinusitis due to the genus Fusarium has increased during the last two decades. We report a case of fungus ball sinusitis with multiple sinuses involvement in an Iranian 21-year-old woman. The patient was diagnosed as having a fungus ball-type sinusitis in computed tomography scan. The sinus biopsy revealed fungal structures on histopathological and direct microscopic examinations and a Fusarium species arose in culture. Partial sequencing of the translation elongation factor 1-alpha identified the isolate as F. proliferatum. Removal of all lesions by endoscopic surgery resulted in a favorable outcome. To the best of our knowledge, this is the first case of F. proliferatum-associated fungus ball which involved multi-sinus and highlights the efficiency of molecular methods for discrimination of fungal agents involved.

Fungal rhino sinusitisin in tehran, iran.

BACKGROUND: Fungal rhino sinusitis (FRS) is an important infection of para nasal sinuses, which encompasses two main categories; invasive and noninvasive forms according to histopathological findings. Aspergillus spp are the most common species isolated from noninvasive form, while Mucorales are more frequently isolates from acute infections. METHODS: Four hundred fifty patients suspected to fungal rhino sinusitis were investigated in a cross-sectional prospective study from June 2009 to Sep 2013. All patients under went endoscopic sinus surgery of the middle meatus. Tissue biopsies were investigated for culture, histopathology and molecular examination. RESULTS: Totally, 87 patients were diagnosed with fungal rhinosinusitis. A. flavus was the most common etiological agent of chronic invasive form (CIFRS), allergic fungal rhino sinusitis (AFRS) and fungus ball (FB), while Rhizopus oryze (26.7%) was the most common cause of infection in acute invasive fungal rhino sinusitis (AIFR). However, a few rare species such as Shyzophyllum commune and Fusarium proliferatum were also isolated. CONCLUSION: Diabetes is the most important predisposing factor for patients with acute invasive form of sinusitis and the most involved sinuses were unilateral multiple sinuses and maxillary sinus.

Molecular identification and prevalence of malassezia species in pityriasis versicolor patients from kashan, iran.

BACKGROUND: Malassezia species are lipophilic yeasts found on the skin surface of humans and other warm-blooded vertebrates. It is associated with various human diseases, especially pityriasis versicolor, which is a chronic superficial skin disorder. OBJECTIVES: The aim of the present study was to identify Malassezia species isolated from patients' samples affected by pityriasis versicolor, using molecular methods in Kashan, Iran. PATIENTS AND METHODS: A total of 140 subjects, suspected of having pityriasis versicolor from Kashan, were clinically diagnosed and then confirmed by direct microscopic examination. The scraped skin specimens were inoculated in modified Dixon's medium. DNA was extracted from the colonies and PCR amplification was carried out for the 26s rDNA region. PCR products were used to further restriction fragment length polymorphism by CfoI enzyme. RESULTS: Direct examination was positive in 93.3% of suspected pityriasis versicolor lesions. No statistically significant difference was observed in the frequency of Malassezia species between women and men. The highest prevalence of tinea versicolor was seen in patients 21-30 years-of-age. No difference could be seen in the frequency of Malassezia species depending on the age of the patients. In total, 65% of patients with pityriasis versicolor had hyperhidrosis. The most commonly isolated Malassezia species in the pityriasis versicolor lesions were; Malassezia globosa (66%), M. furfur (26%), M. restricta (3%), M. sympodialis (3%), and M. slooffiae (2%). Malassezia species were mainly isolated from the neck and chest. CONCLUSIONS: This study showed M. globosa to be the most common Malassezia species isolated from Malassezia skin disorders in Kashan, Iran. The PCR-RFLP method was useful in the rapid identification of the Malassezia species. By using these methods, the detection and identification of individual Malassezia species from clinical samples was substantially easier.

A successful treatment of rhinocerebral mucormycosis due to Rhizopus oryzae.

Mucormycosis is an invasive fungal infection caused by filamentous fungi of the Mucoraceae family. The genera most commonly responsible are Mucor or Rhizopus. The disease occurs mostly in association with diabetic ketoacidosis. Mucormycosis has an extremely high death rate even when aggressive surgery is done. Death rates range from 25-85% depending on the body area involved. A case of rhinocerebral mucormycosis in a 65-year-old diabetic male patient typically presenting as headache, especially in parietal and frontal lobes, with nose and left eye discharge. After clinical and laboratory examination, mucormycosis was diagnosed, and Rhizopus oryzae was isolated. Systemic therapy with amphotericin B administered intravenously then replaced by posaconazole by a combination of aggressive surgery. The patient was treated and followed up for one year. We emphasize the importance of early detection and aggressive treatment in the management of this fatal disease.

Candida species in cutaneous candidiasis patients in the Guilan province in Iran; identified by PCR-RFLP method.

Due to the epidemiological alteration in distribution of Candida species as well as significant increasing trend of either intrinsic or acquired in resistance of some of these fungi, the precise identification of Candida species is necessary for effective antifungal therapy and also for prevention of nosocomial infections. PCR-RFLP method is indicated to be a reliable, rapid and simple technique which is able to differentiate the Candida species. In the present study, we applied this method to evaluate the distribution of Candida species in patients affected with cutaneous candidiasis in the Guilan province. 896 clinical cutaneous samples were collected from different parts of skin and nail of suspected patients referred to clinical centers all over the Guilan province during 24 months. Samples were examined directly with 15% KOH and cultured on fungal specific media. Genomic DNA was extracted and the restriction enzyme Msp1 was applied for polymorphism analysis. Totally, 47 yeast strains were successfully isolated from different clinical samples and identified by conventional as well as PCR-RFLP methods. The results indicated that Candida albicans (36.17%) was the most frequent species followed by C. parapsilosis (25.53%), C. tropicalis (19.14%), C. guilliermondii (14.89%), C. famata (2.12%) and C. krusei (2.12%). Female finger nails were the most common location to be affected by Candida species. In conclusion, PCR-RFLP method was successfully used for recognition of clinical Candida species within the Guilan province and obtained results revealed C. albicans as the predominant causative agent of cutaneous candidiasis. However, distribution of other Candida species did not completely consist with the reported distribution of Candida species in other parts of Iran with different climate to the Guilan province.

A case of fungus ball type pansinusitis caused by Schizophillum commune.

Schizophillum commune has been increasingly reported from allergic bronchopulmonary mycosis (ABPM) as well as fungus ball, brain abscess and several cases of maxillary or allergic fungal sinusitis. In the present study, we reported a case of fungus ball type pansinusitis from a 32-year-old woman in Iran. According to computed tomography (CT) scan, fungus ball type pan-sinusitis was likely to be the first diagnosis. Mycological examination revealed hyaline hyphae with small projection and also clamp connection structures on PDA medium. To identify the obtained isolate properly, molecular analysis of the internal transcribed spacer region was performed and indicated that the causing agent of the infection is surely Schizophillum commune. The patient completely recovered after surgical endoscopic operation and consequent post-operation MRI revealed clearance of sinuses.