Molecular Evidence of Emerged Pulmonary Lophomoniasis due to Lophomonas blattarum among Hospitalized Patients in Southwestern Iran: A National Registry-Based Study.

Objectives: Lophomonas protozoan is an emerging pathogen transmitted through arthropods such as cockroaches. Lophomoniasis is still a mysterious disease with many unknown epidemiological aspects. The current study aimed to determine the prevalence of lophomoniasis among patients who were hospitalized in Hajar Hospital, Shahrekord, southwestern Iran, using a conventional PCR technique. Methods: In this retrospective study, 132 frozen bronchoalveolar lavage fluid (BALF) specimens from patients with respiratory disorders hospitalized in Hajar Hospital, Shahrekord district, southwestern Iran, were analyzed during 2020-2021. Samples are referred to the Iranian National Registry Center for Lophomoniasis (INRCL), Mazandaran Province, Northern Iran, for detecting Lophomonas spp. infection by a conventionally small subunit ribosomal RNA (SSU rRNA) PCR test. Results: A total of 132 frozen BALF specimens were examined, 36 (27.3%) tested Lophomonas spp. positive using the conventional PCR technique. Also, based on sequencing data and blast analysis, the presence of L. blattarum species was confirmed. The average age of Lophomonas spp.- positive patients was 67.02 ± 15.14 years. Out of the 36 positive subjects, 63.9% were male and 36.1% female. Male and Lophomonas infection had a significant correlation (p=0.001). Our findings revealed that L. blattarum infected nonsmokers more than smokers (p=0.001). The most common underlying disease was also bronchitis. Conclusion: Our results showed, for the first time, that pulmonary lophomoniasis caused by L. blattarum is a common and emerging disease in the study area, southwestern Iran. Furthermore, our findings support the use of the PCR test to detect Lophomonas infection in archived frozen clinical samples.

In vitro activities of 8 antifungal agents against geophilic dermatophyte isolates.

BACKGROUND: Members of the Nannizzia gypsea complex are globally the most common geophilic dermatophytes which cause infection in animals and human. Although the susceptibility patterns of anthropophilic or zoophilic dermatophyte species to antifungal agents are well documented, the effectiveness of such drugs against geophilic species have rarely been explored. OBJECTIVES: This study was aimed to evaluate the in vitro antifungal activity of common and new antifungals against a set of environmental and clinical geophilic dermatophyte isolates. METHODS: 108 soil and clinical geophilic isolates from two genera Nannizzia (N. fulva n = 59; N. gypsea n = 43) and Arthroderma (A. quadrifidum n = 4; A. gertleri n = 1; A. tuberculatum n = 1) were included in the study. The in vitro antifungal susceptibility patterns of eight common and new antifungals against the isolates were determined according to broth microdilution method and by CLSI M38-A3 (3rd edition) protocol. RESULTS: MIC values across all isolates from five species ranged as: luliconazole: 0.0002-0.002 µg/ml, terbinafine: 0.008-0.125 µg/ml, efinaconazole: 0.008-0.125 µg/ml, ciclopirox olamine: 0.03-0.5 µg/ml, itraconazole: 0.125-1 µg/ml, amorolfine hydrochloride: 0.125-4 µg/ml, griseofulvin: 0.25-2 µg/ml and tavaborole: 1-8 µg/ml, respectively. CONCLUSION: Luliconazole, terbinafine and efinaconazole exhibited the highest in vitro efficacy, regardless of the dermatophyte species. Further surveillance studies are recommended to confirm the implication of such in vitro data for the clinical recovery rate of dermatophytosis with geophilic species following antifungal therapy.

Simultaneous monitoring of SARS-CoV-2, bacteria, and fungi in indoor air of hospital: a study on Hajar Hospital in Shahrekord, Iran.

The novel SARS-CoV-2 outbreak was declared as pandemic by the World Health Organization (WHO) on March 11, 2020. Understanding the airborne route of SARS-CoV-2 transmission is essential for infection prevention and control. In this study, a total of 107 indoor air samples (45 SARS-CoV-2, 62 bacteria, and fungi) were collected from different wards of the Hajar Hospital in Shahrekord, Iran. Simultaneously, bacterial and fungal samples were also collected from the ambient air of hospital yard. Overall, 6 positive air samples were detected in the infectious 1 and infectious 2 wards, intensive care unit (ICU), computed tomography (CT) scan, respiratory patients' clinic, and personal protective equipment (PPE) room. Also, airborne bacteria and fungi were simultaneously detected in the various wards of the hospital with concentrations ranging from 14 to 106 CFU m-3 and 18 to 141 CFU m-3, respectively. The highest mean concentrations of bacteria and fungi were observed in respiratory patients' clinics and ICU wards, respectively. Significant correlation (p < 0.05) was found between airborne bacterial concentration and the presence of SARS-CoV-2, while no significant correlation was found between fungi concentration and the virus presence. This study provided an additional evidence about the presence of SARS-CoV-2 in the indoor air of a hospital that admitted COVID-19 patients. Moreover, it was revealed that the monitoring of microbial quality of indoor air in such hospitals is very important, especially during the COVID-19 pandemic, for controlling the nosocomial infections.

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.

Antifungal Effects of Iranian Propolis Extract and Royal jelly Against Candida albicans In-Vitro.

BACKGROUND: Candida albicans is the most important opportunistic fungal that can establish infection in susceptible individuals. Iranian Propolis and Royal jelly are bee products that are traditionally used against fungal infections. This study was aimed to evaluate the antifungal effects of Iranian Propolis extract and Royal jelly against C. albicans in vitro. METHODS: Antifungal activities of the extracts were performed according to microbroth dilution method in 96-well microdilution plates. The amount of minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) based on counting the number of fungal colonies (CFU) were evaluated for each of Royal jelly and Iranian Propolis extracts against C. albicans compared with the control group. RESULTS: In this study, the MIC, MIC50, and MFC of Royal jelly on C. albicans were, respectively, 80, 103 ± 25, and 160 ± 34 mg/mL and for the Iranian Propolis alcoholic extract were, respectively, 0.030 ± 0.015, 0.0618 ± 0.027, and 0.0833 ± 0.0599 mg/mL. CONCLUSIONS: The results indicate that both Royal jelly and Iranian Propolis alcoholic extract are effective against C. albicans, but the former species has higher antifungal activity. If the clinical trials confirm the results of this study, Iranian propolis, as a new antifungal agent by replacing chemical drugs, can be used to develop antifungal medicinal herbs.

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.

Emergence of Terbinafine Resistant Trichophyton mentagrophytes in Iran, Harboring Mutations in the Squalene Epoxidase (SQLE) Gene.

INTRODUCTION: Trichophyton mentagrophytes and T. interdigitale are important causative agents of superficial mycoses, demonstrating emergent antifungal drug resistance. We studied the antifungal susceptibility profiles in Iranian isolates of these two species. METHODS: A total of 96 T. interdigitale and 45 T. mentagrophytes isolates were subjected to molecular typing by ribosomal ITS region. Antifungal susceptibility profiles for terbinafine, griseofulvin, clotrimazole, efinaconazole, luliconazole, amorolfine and ciclopirox were obtained by CLSI broth microdilution method. The squalene epoxidase (SQLE) gene was subjected to sequencing for mutations, if any, in isolates exhibiting elevated MICs for terbinafine. RESULTS: Luliconazole and efinaconazole showed the lowest MIC values against T. mentagrophytes and T. interdigitale isolates. There were five isolates with terbinafine MICs ≥32 µg/mL in our sample. They belonged to T. mentagrophytes type VIII and harbored two alternative SQLE gene sequence variants, leading to Phe397Leu and Ala448Thr or Leu393Ser and Ala448Thr substitutions in the enzyme. All terbinafine resistant strains could be inhibited by luliconazole and efinaconazole. CONCLUSION: This study documented a step in the global spread of resistance mechanisms in T. mentagrophytes. However, treatment alternatives for resistant isolates were available.

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.

Trichophyton mentagrophytes and T interdigitale genotypes are associated with particular geographic areas and clinical manifestations.

The fungi Trichophyton mentagrophytes and T interdigitale account for significant amount of dermatophytosis cases worldwide. These two dermatophytes form a species complex and have a number of ribosomal internal transcribed spacer (ITS) region genotypes, allowing simultaneous species identification and strain typing. Our aim was to describe the geographic distribution of T mentagrophytes/T interdigitale ITS region genotypes and find an association between the genotypes and clinical presentations of respective infections. We performed rDNA ITS region sequencing in 397 Iranian T mentagrophytes/T interdigitale isolates and analysed all available in GenBank entries with sequences of this kind. For the study, 515 clinical annotations were available. Statistical analysis was performed by chi-squared test and Spearman rank correlation analysis. A total of 971 sequences belonged to genotypes with at least 10 geographic annotations and were classified on the basis of exclusive occurrence in a particular region or high relative contribution to a regional sample. We discerned Asian and Oceanian ("KU496915" Type V, "KT192500" Type VIII, "KU315316"), European ("FM986750" Type III, "MF926358" Type III*, "KT285210" Type VI) and cosmopolitan ("FM986691" Type I, "JX122216" Type II, "KP132819" Type II* and "AF170453" Type XXIV) genotypes. There was statistically significant difference in the ITS genotype distribution between different affected body sites. Trichophyton mentagrophytes "KT192500" Type VIII correlated with tinea cruris, T mentagrophytes "KU496915" Type V correlated with tinea corporis, T interdigitale "JX122216" Type II correlated with tinea pedis and onychomycosis. Trichophyton mentagrophytes and T interdigitale genotypes can be associated with distinct geographic locations and particular clinical presentations.

In Vitro Antifungal Activity of Novel Triazole Efinaconazole and Five Comparators against Dermatophyte Isolates.

The objective of this study was to assess the in vitro activity of the novel triazole antifungal drug, efinaconazole, and five comparators (luliconazole, lanoconazole, terbinafine, itraconazole, and fluconazole) against a large collection of Trichophyton interdigitale and Trichophyton rubrum clinical isolates. The geometric mean MICs were the lowest for luliconazole (0.0005 µg/ml), followed by lanoconazole (0.002 µg/ml), efinaconazole (0.007 µg/ml), terbinafine (0.011 µg/ml), itraconazole (0.095 µg/ml), and fluconazole (12.77 µg/ml). It appears that efinaconazole, lanoconazole, and luliconazole are promising candidates for the treatment of dermatophytosis due to T. interdigitale and T. rubrum.

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.

Mycological aspects of onychomycosis in Khuzestan Province, Iran: A shift from dermatophytes towards yeasts.

BACKGROUND AND PURPOSE: Onychomycosis is fungal infection of the nails with an overall increasing incidence, worldwide. The epidemiological aspects of onychomycosis in Khuzestan, Iran, have not been established. In this study, we aimed to evaluate the clinical and mycological status of fungal nail infection in Khuzestan Province, Iran. MATERIALS AND METHODS: The study population included 433 patients (143 males vs. 290 females). Nail samples underwent primary direct microscopy and culture. The isolated yeasts and dermatophytes were then subjected to additional molecular identification by r-DNA ITS-RFLP. Identification of some non-dermatophytic molds (NDMs) and unknown yeasts was accomplished by ITS and beta-tubulin sequencing. RESULTS: Onychomycosis was confirmed in 154 patients (males: 36.4%; n=56 vs. females: 63.6%; n=98), whose age ranged from 2 to 85 years, with the highest prevalence in the age group of 41-50 years. Infection mostly occurred due to yeasts (57.15%), with Candida albicans as the most frequent (29.35%) species, followed by C. parapsilosis (13.8%) and C. tropicalis (4.5%). Dermatophytes were isolated in 38.35% of the cases; the most common isolates were found to be Trichophyton interdigitale (21.1%), Epidermophyton floccosum (10.5%), T. rubrum (5.25%), and Microsporum canis (1.5%). NDMs were isolated from 4.5% of the cases with Aspergillus spp. as the most common agent. Dermatophytes and NDMs were more frequently seen in toenails, whereas yeasts mostly infected fingernails. Fingernail onychomycosis was significantly more prevalent among females than in males (P<0.05). CONCLUSION: The study highlights that in Khuzestan province, the causative agents of onychomycosis have shifted from dermatophytes to yeasts.