Sheep serve as a reservoir of Trichophyton mentagrophytes genotype V infection.

Human infections by Trichophyton mentagrophytes occur mainly due to contact with diseased animals. In Iran, T. mentagrophytes genotype V is the most prevalent variant of the fungus. We aimed to determine the animal reservoir of T. mentagrophytes genotype V infection. The study was done on a total of 577 dermatophyte strains obtained from animals with signs of dermatophytosis and human patients. The list of extensively sampled animals included sheep, cows, cats and dogs. For human cases, epidemiological data were collected. All dermatophyte isolates from animals along with 70 human isolates morphologically similar to T. verrucosum and T. mentagrophytes genotype V were identified by rDNA internal transcribed spacer region restriction fragment length polymorphism analysis and DNA sequencing. A total of 334 animal dermatophyte strains were identified as Microsporum canis, T. mentagrophytes genotype V, T. verrucosum, Nannizzia gypsea, T. mentagrophytes genotype II*, T. mentagrophytes genotype VII, T. quinckeanum, and N. fulva. All clinical isolates identified as T. mentagrophytes genotype V originated from skin and scalp infections. Almost all veterinary isolates of T. mentagrophytes genotype V were cultured from sheep, but epidemiological data on animal-to-human transmission of T. mentagrophytes genotype V infection were limited and we found evidence in favor of interhuman transmission. In Iran, sheep maintain T. mentagrophytes genotype V population and therefore serve as animal reservoir of respective infections. The role of sheep as the source of human dermatophytosis due to T. mentagrophytes genotype V isolates is yet to be proven.

In this study, we sampled a variety of animals to determine a reservoir of Trichophyton mentagrophytes genotype V infection. With the use of molecular identification techniques, we show that this infection reservoir is represented by sheep.

Rhizopus microsporus as Causative Agent of Mucormycosis in COVID-19 Patient.

BACKGROUND: The pandemic of coronavirus disease (COVID-19) emerged as a fatal infection, especially in immunocompromised patients. Currently, this infection is managed with systemic corticosteroids. Co-infection of CO-VID-19 with opportunistic fungi is increasingly recognized. METHODS: We describe a case of rhino-cerebral mucormycosis 12 days following severe COVID-19 in a diabetic patient. RESULTS: He received 50 mg amphotericin B and surgical debridement. The patient's symptoms improved following medical and surgical intervention. CONCLUSIONS: Mucormycosis is an uncommon but serious infection that complicates the course of severe COVID-19. Subjects with diabetes mellitus and multiple risk factors may be at a higher risk for developing mucormycosis.

Phenotype, genotype, and mating type determination in oral Candida albicans isolates from pediatric patients with neutropenia.

BACKGROUND: The most frequent species of Candida to infect and colonize patients with neutropenia is still Candida albicans. This study aimed to provide detailed information on the phenotype, genotype, and mating type of oral C. albicans isolated from neutropenic pediatric patients, and to investigate how these characteristics are related. METHODS: Two hundred fifty-four oral samples from patients under 18 years old with neutropenia and malignancies were collected from January to October 2021. Samples were cultured on CHROMagar Candida. Isolates of C. albicans were identified with the germ tube test, chlamydospore production on cornmeal agar, and PCR-RFLP. Genotyping of C. albicans isolates was carried out by amplifying the 25S rDNA gene with specific CAINT-L and CA-INT-R primers. MTLa1 and MTLα1 primers were used to identify each mating type. Yeast peptone dextrose supplemented with phloxine B was used to identify different phenotypes. RESULTS: Ninety-two (36%) patients were positive for C. albicans. The mean age of patients was 7.85. Fifty-three (58.9%) isolates demonstrated type A, 15 (16.7%) type B, 15 (16.7%) types D/E, and 7 (7.7%) type C. Three isolates each (3.3%) were homozygous for MTLa or homozygous for MTLα. All of the MTL-homozygous isolates were genotype A. There was a significant correlation between patients' underlying disease and genotype (p = 0.036). There was a significant correlation between mating type and genotype (p = 0.000). CONCLUSION: Most of the isolates exhibited a white phenotype, noted in the literature as the most virulent. Moreover, heterozygous strains were frequent and may play a role in Candida colonization.

Evaluation of catalase activity of clinical and environmental isolates of Aspergillus species.

Background and Objectives: Catalases are a good scavenger of H2O2 which degrades hydrogen peroxide into water and oxygen. They are considered as a virulence factor that are present in both spores and hypha of fungi. There is limited data regarding catalase activity in Aspergillus species. This study aimed to assess the mycelial catalase activity of clinical and environmental isolates of Aspergillus niger, A. tubingensis, A. flavus, A. luchuensis, A. piperis and A. terreus. Materials and Methods: Briefly, clinical and environmental Aspergillus species were used in the current study. Catalase activity was assessed for both groups of isolates including 13 A. flavus (12 clinical, 1 environmental), 13 A. terreus (8 clinical, 5 environmental), 26 A. tubingensis (13 clinical, 13 environmental), and 44 A. niger (25 environmental, 19 clinical) species. Fungal balls of mycelia were separated from the liquid culture and were crushed using homogenizer. The supernatants were collected and used for a catalase activity assay. Results: Totally, in our study 98 Aspergillus including 45 environmental and 53 clinical isolates were assessed for catalase activity. High catalase activity was detected among environmental Aspergillus species (Mean= 1.62 mU/ml) and the mean of mycelial catalase activity among clinical A. terreus isolates was higher than environmental strains. Conclusion: In summary, mycelial catalase activity varied among species and environmental isolates demonstrated higher catalase activity. Totally a significant difference was found between clinical and environmental Aspergillus isolates.

ABC typing and extracellular enzyme production of Candida albicans isolated from Candida vulvovaginitis.

BACKGROUND: Candida albicans is the most common and virulent genus Candida. Detection of virulence factors in this species plays an important role in the better understanding of pathogenesis and antifungal treatment. Molecular typing investigations are important in the epidemiological interpretation of infection. This study aimed to evaluate extracellular enzyme activity and genotyping of C. albicans species isolated from vulvovaginal samples. METHODS: One hundred and three vaginal C. albicans isolates were tested for esterase, phospholipase, proteinase, and hemolysin activities by specific media. Besides, the DNA of C. albicans isolates was extracted and amplified for ABC genotyping. RESULTS: The highest enzyme production of C. albicans isolates was for proteinase (97.1%) and esterase (95.2%), whereas 59.2% of C. albicans isolates were negative for hemolysin secretion. Genotype C (83.5%) was the most frequent genotype followed by genotype B (12.6%) and genotype A (3.9%). CONCLUSION: It is concluded that genotype C was the predominant genotype in all examined vulvovaginal C. albicans isolates. Also, there was a significant difference between enzyme production in each genotype (except for proteinase).

Pediatric candiduria, epidemiology, genotype distribution and virulence factors of Candida albicans.

The presence of Candida species in urine may be due to colonization of this species in the bladder, urinary catheter, and perineum. Candida albicans has been the most commonly isolated from urine samples in patients with candiduria. Several virulence factors include adhesion to host cells, secreted extracellular enzymes, phenotype switching, and biofilm formation are contributing to the pathogenicity of C. albicans. ABC genotyping is the method based on the determination of 25s rDNA and C. albicans is divided into four genotypes include A, B, C, and E. We aimed to identify Candida species from pediatrics and evaluate extracellular enzyme activities, phenotype switching, biofilm formation, and genotyping in isolates. Urine samples collected, cultured, and yielded yeasts were identified. Phenotype switching, biofilm formation, enzymatic patterns, and genotyping of 50 isolates of C. albicans were evaluated. The Genotyping pattern was compared with extracellular enzymes, biofilm formation, and phenotype switching pattern. 16.2% of urine cultures were positive for the different Candida species. The most common species was C. albicans, followed by C. glabrata. Out of 50 isolates of C. albicans, 72% and 28% isolates were recognized as genotypes A and C. All isolates were produced extracellular enzymes and biofilm formation. In conclusion, candiduria with high colony counts is still a challenge in Iranian pediatrics. Genotype A was the predominant genotype among C. albicans strains. There is a statistical difference between esterase and genotypes of C and A C. albicans.

Discovery of New Trichophyton Members, T. persicum and T. spiraliforme spp. nov., as a Cause of Highly Inflammatory Tinea Cases in Iran and Czechia.

Pathogens from the Trichophyton benhamiae complex are one of the most important causes of animal mycoses with significant zoonotic potential. In light of the recently revised taxonomy of this complex, we retrospectively identified 38 Trichophyton isolates that could not be resolved into any of the existing species. These strains were isolated from Iranian and Czech patients during molecular epidemiological surveys on dermatophytosis and were predominantly associated with highly inflammatory tinea corporis cases, suggesting possible zoonotic etiology. Subsequent phylogenetic (4 markers), population genetic (10 markers), and phenotypic analyses supported recognition of two novel species. The first species, Trichophyton persicum sp. nov., was identified in 36 cases of human dermatophytosis and one case of feline dermatophytosis, mainly in Southern and Western Iran. The second species, Trichophyton spiraliforme sp. nov., is only known from a single case of tinea corporis in a Czech patient who probably contracted the infection from a dog. Although the zoonotic sources of infections summarized in this study are very likely, little is known about the host spectrum of these pathogens. Awareness of these new pathogens among clinicians should refine our knowledge about their poorly explored geographic distribution. IMPORTANCE In this study, we describe two novel agents of dermatophytosis and summarize the clinical manifestation of infections. These new pathogens were discovered thanks to long-term molecular epidemiological studies conducted in Czechia and Iran. Zoonotic origins of the human infections are highly probable, but the animal hosts of these pathogens are poorly known. Further research is needed to refine our knowledge about these new dermatophytes.

Genotyping, antifungal susceptibility, enzymatic activity, and phenotypic variation in Candida albicans from esophageal candidiasis.

BACKGROUND: Esophageal candidiasis is the most frequent form of esophagitis. The pathogenicity of Candida spp. is related to a combination of microbial factors, hydrolytic enzyme secretion and phenotypic switching. This study was designed to investigate esophageal candidiasis, antifungal activity, enzymatic activity patterns, phenotyping, and genotyping profiles of Candida albicans species. METHODS: Nine hundred thirty-three visited patients were evaluated, and esophageal biopsies from patients were included in this study during 2019-2020. Direct smear, Gram staining, and culture on CHROMagar were performed for each sample. Isolated species were identified with conventional procedures and PCR-RFLP. Susceptibility to antifungals was determined according to CLSI guidelines. ABC typing, phenotype switching, hemolysin, proteinase, phospholipase, and esterase activity were also determined with the appropriate protocols. RESULTS: Twenty-three (2.5%) patients (mean age 55.2 years) were diagnosed with esophageal candidiasis. The species isolated were 19(82.6%) C. albicans, 3(13.1%) C. glabrata, and 1(4.3%) C. tropicalis. Genotype A (57.9%) was the predominant type in C. albicans isolates. 50% of C. albicans isolates exhibited a white phenotype. A high level of phospholipase (47.4%), hemolysin (68.4%), and proteinase activity (36.8%) was observed in the C. albicans isolates. Only three C. glabrata isolates displayed non-wild type susceptibility to voriconazole and itraconazole. CONCLUSION: This study shows that C. albicans are still the most frequent isolates from patients with esophageal candidiasis. The predominance of genotype A, the white phenotype, and strong hemolysin activity may indicate a high prevalence of pathogenicity in these isolates. Sensitivity to antifungal drugs was greatest for amphotericin and fluconazole.

Molecular identification, biofilm formation and antifungal susceptibility of Rhodotorula spp.

Luliconazole is an imidazole antifungal agent used in topical form for the treatment of onychomycosis and dermatophytosis. In vitro activity of luliconazole against dermatophytes, Candida, black fungi, Fusarium and Aspergillus species have been investigated. Rhodotorula spp. are environmental yeasts and emerged as opportunistic pathogens among immunocompromised patients. Rhodotorula's human infections are usually resistant to treatment with antifungal drugs especially triazoles and echinocandins. The present study aimed at the molecular detection of environmental isolates of Rhodotorula spp. Then, antifungal efficacy of luliconazole was evaluated against isolates and compared to other routine systemic antifungals including; caspofungin, posaconazole, fluconazole, itraconazole, amphotericin B, and voriconazole. The biofilm production of Rhodotorula isolates was also evaluated. In this study, 39 isolates of Rhodotorula spp. were isolated from the environment, detected using molecular methods, and tested against luliconazole. Then, the anti-fungal activity of luliconazole compared with several routine antifungals. Also, biofilm formation by using a crystal violet staining assay was performed. Our finding showed that luliconazole has a very high minimum inhibitory concentration (MIC) value (1-8 µg/ml) against Rhodotorula spp. Besides, 100% of Rhodotorula strains were resistant to caspofungin, followed by fluconazole 94.7% and voriconazole 74.4%. Amphotericin B was demonstrated excellent in vitro activity against this genus. Our result indicated that 59% of Rhodotorula spp. were in the mid-range of biofilm production. Our results indicated that luliconazole does not effective against the genus Rhodotorula. Furthermore, amphotericin B is the best drug against this genus in comparison to caspofungin and other azole drugs.

Luliconazole, a highly effective imidazole, against Fusarium species complexes.

Luliconazole is a new antifungal that was primarily used for the treatment of dermatophytosis. However, some studies have shown that it has excellent efficacy against Aspergillus and Candida species in vitro. The present study aimed to evaluate of luliconazole activity against some Fusarium species complex isolates. In this study, 47 isolates of Fusarium were tested against several antifungals including luliconazole. All species were identified using morphology features, and PCR sequencing and antifungal susceptibility were performed according to CLSIM38 A3 guideline. Our results revealed that luliconazole has a very low minimum inhibitory concentration value (0.0078-1 µg/ml) in comparison with other tested antifungals. Amphotericin B had a poor effect with a high MIC90 (64 µg/ml), followed by terbinafine (32 µg/ml), posaconazole (16 µg/ml), caspofungin (16 µg/ml), voriconazole (4 µg/ml), and itraconazole (4 µg/ml). Overall, our findings indicated that luliconazole has great activity against environmental and clinical Fusarium species complexes in comparison to tested antifungals.

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

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

Clinical and microbial epidemiology of otomycosis in the city of Yasuj, southwest Iran, revealing Aspergillus tubingensis as the dominant causative agent.

PURPOSE: Otomycosis is a mycotic infection of the external auditory canal and can be caused by a wide range of fungal species. In this study, we aimed to identify fungal isolates from patients suspected of otomycosis. METHODOLOGY: External ear canal samples were taken from patients referred to the outpatient department of Shahid-Mofatteh Clinic in the city of Yasuj, Iran, and examined by direct microscopy and culture. DNA of the isolated fungi was tested by internal transcribed spacer PCR restriction fragment length polymorphism analysis for identification of yeasts and ß-tubulin sequencing for identification of Aspergillus species. RESULTS: Among 275 patients suspected of otomycosis, 144 cases (83 female and 61 male) were confirmed with otomycosis. For 89% (n=128) of positive cultures, microscopy was also positive, while there were no cases with a microscopy-positive and culture-negative result. The predominant predisposing factor was self-cleaning of the external ear using unhygienic tools, and the main risk occupation was 'housewife'. The most common isolated fungi were typically Aspergillus (n=120), including 73 isolates of Aspergillus section Nigri, 43 of section Flavi, 3 of section Terrei and 1 of section Fumigati. After sequencing, 44 out of 73 strains primarily identified as Aspergillus niger turned out to be Aspergillus tubingensis. Thirty-five isolates were identified as Candida, including Candida parapsilosis (n=22), Candida albicans (n=12) and Candida tropicalis (n=1). CONCLUSION: Aspergillus tubingensis was the most common species involved in otomycosis. This work corroborates the difficulty of precise identification of species within the black Aspergilli by morphological characteristics.

Candiduria; a review article with specific data from Iran.

Although, several categories of nosocomial infections are presented during the recent years, urinary tract infections (UTIs) considered as one of the most important systemic infections. The presence of Candida species in the urinary tract system (Candiduria) is seen in only 10-15% of the cases with UTI, however candiduria has been considered as more problematic infection for patients, laboratory workers and physicians. Due to increasing numbers of several predisposing factors, such as antibacterial agents, urinary tract instrumentation, diabetes mellitus, invasive therapies, and prolonged hospital stay, candiduria develops among the hospitalized patients, especially hospitalized in intensive care units (ICUs) and neonatal intensive care units (NICUs). According to the epidemiological studies, Candida albicans is the most common isolated species from candiduric patients. However, during the recent years, due to increasing resistance to antifungal drugs, non-albicans Candida species including, C. glabrata, C. krusei, C. parapsilosis and C. tropicalis have been also implicated. We found that the mean prevalence of candiduria among Iranian patients was lower (16.5%) than worldwide ratio and also males were more frequently affected than females (M:F, 1.2:1). Similar to other countries, C. albicans was most common infectious agent followed by non-albicans Candida species including, C. glabrata, C. tropicalis and C. krusei.

The Frequency, Antifungal Susceptibility and Enzymatic Profiles of Candida Species Isolated from Neutropenic Patients.

BACKGROUND: Neutropenia, as a predisposing factor for invasive candidiasis, is defined as a reduction in neutrophil count to less than 1500/mm3. It is a common condition in patients with hematological malignancy and cytostatic chemotherapy. Extensive chemotherapy and prophylaxis with antifungals have increased the resistance of Candida isolates to antifungal drugs. Although, Candida albicans is the most common causative agent among neutropenic patients, there is an increasing rate of non-albicans species. Extracellular enzymes activity pattern and antifungal agent sensitivity profiles are two important factors for spreading resistant strains. OBJECTIVES: The aim of the present study was to identify the Candida strains isolated from hospitalized neutropenic patients. The patterns of antifungal susceptibility of the causative agents to antifungals and the extracellular enzymes activity of the isolates were also evaluated. PATIENTS AND METHODS: In the present study, 243 urine and 243 oral swab samples were collected from neutropenic patients and inoculated on CHROMagar Candida. In addition, 100 blood samples were also inoculated in biphasic Brain Heart Infusion medium. Several yeast isolates were isolated from samples and identified by classical and molecular techniques. The profiles of extracellular enzymes and the susceptibility of recovered agents to amphotericin B, fluconazole and caspofungin were also evaluated. RESULTS: A total of 110 yeast strains isolated from urine and oral cavities were identified as C. albicans (51.8%), C. krusei (25.5%), C. glabrata (6.4%) and other yeasts (16.3%). No yeast species was isolated from blood samples. Our result showed that in 90% of the isolates, the range of secretion of extracellular enzymes was medium (2+) and high (3+), however only a few isolates were negative for this characteristic. All isolates were sensitive to caspofungin and fluconazole, whereas 54.7% of isolates were resistant to amphotericin B. CONCLUSIONS: We found a marked increase in the incidence of non-albicans species (48.2%) among neutropenic patients. Only a few strains failed to produce extracellular enzymes. Finally, in addition to fluconazole, caspofungin can be considered as the first line treatment against Candida species among neutropenic patients.

Otomycosis in iran: a review.

Fungal infection of the external auditory canal (otitis externa and otomycosis) is a chronic, acute, or subacute superficial mycotic infection that rarely involves middle ear. Otomycosis (swimmer's ear) is usually unilateral infection and affects more females than males. The infection is usually symptomatic and main symptoms are pruritus, otalgia, aural fullness, hearing impairment, otorrhea, and tinnitus. Fungal species such as yeasts, molds, dermatophytes, and Malassezia species are agents for otitis externa. Among molds, Aspergillus niger was described as the most common agent in the literature. Candida albicans was more prevalent than other yeast species. Otomycosis has a worldwide distribution, but the prevalence of infection is related to the geographical location, areas with tropical and subtropical climate showing higher prevalence rates. Otomycosis is a secondary infection and is more prevalent among swimmers. As a result, a higher incidence is reported in summer season, when more people interested in swimming. Incidence of otomycosis in our review ranged from 5.7 to 81 %, with a mean value of 51.3 %. Our results showed that 78.59 % of otomycosis agents were Aspergillus, 16.76 % were Candida species, and the rest (4.65 %) were other saprophytic fungi. Among Iranian patients, incidence of infection was highest in summer, followed by autumn, winter, and spring. In Iran, otomycosis was most prevalent at the age of 20-40 years and the lowest prevalence was associated with being <10 years old. The sex ratio of otomycosis in our study was (M/F) 1:1.53.

Carriage rate of group B streptococci in pregnant women in three teaching hospitals in Shiraz, Iran.

OBJECTIVE: The main goal of this study was to assess differences in the vaginal flora of pregnant women and provide a detailed evaluation of vaginal swabs for the presence of group B streptococcus (GBS) and other organisms in three teaching hospitals in Shiraz, southwest of Iran. SUBJECTS AND METHODS: In a cross-sectional study from April 2006 to March 2007, 310 women with vaginal delivery at three obstetric wards of teaching hospitals in Shiraz were studied. Swab specimens from the vagina and rectum were collected and were examined bacteriologically. All of the newborns were followed up for 3 months. RESULTS: Of 310 pregnant women, 43 (13.8%) were colonized with GBS. There were no differences between GBS carriers and GBS-negative women in risk factors like preterm rupture of membranes (p = 0.77) and preterm labor (p = 0.53). One neonate was hospitalized due to meningitis and GBS was detected in CSF culture and Gram staining. CONCLUSION: Our data showed that the prevalence of GBS carriage rate in pregnant women in Shiraz, Iran was high. The poor correlation between risk factors and GBS carriage requires further investigation in larger groups.