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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.

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

Background: Dermatophytosis is one of the most common fungal infections in humans. Antifungals such as fluconazole are effectively used for treating dermatophytosis; however, drug resistance was observed in many cases. Therefore, a newer treatment strategy is essential. Methods: This study (Conducted in the Laboratory of the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran in 2018) evaluated the antifungal susceptibility of nano fluconazole compared to conventional fluconazole on dermatophyte isolates using CLSI M38-A2guidelines. Dermatophyte species isolated from clinical cases of dermatophytosis were identified using PCR sequencing techniques. Zeta potential and size of the nano particles containing fluconazole were measured; scanning electron microscope (SEM) was used to determine nano particle structure. Results: The size of liposomal fluconazole obtained was 88.9 12.14 nm with –20.12 3.8 mV for zeta potential. The encapsulation rate for fluconazole was 75.1 4.2%. MIC50 for the three tested species was 32, 16, and 8 μg/ml for Trichophyton interdigitale, T. rubrum, and Epidermophyton floccosum isolates, respectively. The corresponding values for nano fluconazole were 8 μg/ml for the three tested species. Conclusion: MIC value for nano-fluconazole was lower than conventional fluconazole in all dermatophytes species tested; therefore, nano-fluconazole could inhibit the growth of dermatophytes better than fluconazole at a lower concentration of the drug.

Aflatoxin and ochratoxin are among the most resistant and severe toxins produced by Aspergillus spp. The aim of this study was an evaluation of eliminatory effects of cold argon plasma jet on aflatoxin B1. Sampling of wheat, corn, oatmeal, flour and rice products from northern cities of Iran was carried out. The fungal species were cultured on CHAPK medium and next to sabouraud dextrose broth + malt extract (SB + ME) and also SB + yeast extract (SB + YE) media to obtain aflatoxin B1. The ELISA test was conducted to measure the aflatoxin level. The mean initial concentration of the aflatoxin from charts analysis in the SB +ME medium was 16.106 μg / kg (10.4, 8.32 and 7.55 μg / kg at 30, 60 and 360 sec, respectively), and in the SB +YE medium from 23.699 μg / kg (12.82, 9.93 and 9.54 μg / kg, respectively), at the same time. Furthermore, the mean concentrations of aflatoxin B1 by tables analysis in the SB+ ME medium changed from 20.02 μg / kg to 6.87, 10.46 and 7.55 μg / kg, at 30, 60 and 360 sec, respectively, and in the medium SB +YE from 28.85 μg / kg reached 6.56.6, 8.43 and 7.02 μg / kg, respectively, at the same time. In this study, the change or decrease in the concentration of aflatoxin B1, from Lin/Log and Log/Lin analyses of charts was significant (p

Introduction: Vulvovaginal Candidiasis (VVC) is one of the most common genital tract infections among women, especially in diabetic patients. The increasing prevalence of recurrent infections caused by drug-resistant non-albicans species necessitates further studies on diabetic patients and the identification of causative agents by reliable molecular techniques. The obtained results can assist in adopting proper treatment procedures and prevention of recurrent vulvovaginitis (RVVC). Methods: In a cross-sectional study, 150 vaginal discharge samples were collected from diabetic women suspected of candidiasis referring to health centers in Tehran province. Following the culture of samples on SDA, CHROMagar Candida and PCR-RFLP were used for presumptive and definitive identification of Candida species, respectively. Results: Out of 115 positive patients, 105 showed infection with one species, and 10 had a mixed infection with two species. The frequency of Candida glabrata isolated from non-mixed and mixed infections in RVVC group was higher than Candida albicans (27.8% vs. 9.6%), which contradicted the results of the VVC group (6.1% vs. 24.3%). In the RVVC group, therefore, the patients were more infected with non-albicans species than C. albicans (47.8% vs. 9.6%), while in the VVC group the non-albicans were of lower frequency (18.3% vs. 24.3%). Conclusion: Our findings showed a statistically significant correlation (P

Fungal polysaccharides are well-known for the medicinal properties such as antitumor and immunomodulating effects. Hence, this study evaluated antitumor effects of polysaccharide extracted from Fusarium sp. isolated from soil samples of Karaj district, Alborz, Iran along with its taxonomic study. The filamentous fungus strain FK1 was isolated from the soil sample of Karaj, Iran. The strain was identified based on cultural, morphological and 18 S rRNA gene parameters as Fusarium. Further, the strain Fusarium was cultured in fermented broth of modified (PDB) for 10 days at 25 °C. The polysaccharide of strain FK1 was extracted from the mycelium free supernatant by boiling water method and evaluated for antitoxicity effect on two human cancer cell lines: HeLa cell line and Lymphoblastoid cell line (LCL) by MTT method. Findings revealed that water-extracted from mycelia polysaccharide of strain FK1 had the highest cytotoxicity effect against LCL which is the cause of B lymphocyte cancer, at 50 μg/ml concentration dose (114 ± 1.63) followed by 100 μg/ml (105 ± 0.57) and 10 μg/ml (104 ± 0.57), while it did not have a considerable effect on HeLa cell line. Fusarium could be alternative sources as an antitumor component. Keywords: Polysaccharides, Anti-tumor, Lymphoblastoid cell line, HeLa cell line, Fusarium

The presence of subtilisin genes (SUBs) coding for serine proteases with activities keratinolytic on dermatophyte species gives them the ability to adhere and degrade keratin during infection process. The aim of this study was to determine the presence of SUBs in Trichophyton rubrum and Microsporum canis from dermatophyte infections. In the current study, dermatophytes were isolated from human and animal with dermatophytosis. Isolates were identified by direct microscopic and laboratory examination; then, the isolates were examined for the presence of the SUB gene family by PCR with specific primers. This study showed the most frequency of SUB gene families (11/17). At least one virulence gene was observed for 94% (16/17) of the isolates. Statistical analysis revealed significant associations between SUB presence and isolates, origin of isolates (animal or human infection) and infection type (skin, hair, and nail) (P

Background: Inflammation or infection of the ear that refers to as otitis is a common ear disorder. The otitis externa infection involves outer ear and ear canal. Streptococcus pyogenes, Staphylococcus aureus, and Pseudomonas aeruginosa can be pointed out as the common causative agents of this infection. Due to the detrimental effects of chemical drugs on humans, utilizing the new formulation of herbal essential oils, such as nano-encapsulation is considered owing to the more efficient and reduction of adverse effects resulting from the direct application of essential oils. In this in vitro study, the antimicrobial effects of 4 nano-essences were compared to pure essential oils against four major microorganisms involved in otitis externa (namely, S. pyogenes, S. aureus and P. aeruginosa). Methods: To evaluate the antibacterial effects of nano-essences, minimum inhibitory concentration (MIC) quantitative determination and minimum bactericidal concentration (MBC) were first carried out, and then qualitative disk diffusion tests were conducted. Results: The MIC quantitative test results demonstrated that among the evaluated essential oils, garlic nanoessence had the most antimicrobial effect at the lowest concentration. Besides, the results showed that nanoessences had lower effect compared to pure essential oils. Diffusion disc test results revealed that nano-essences could not be released from paper discs diffusion in solid media. Conclusions: Results suggested great antibacterial effects of nano-essences of garlic, thyme, peppermint, and chamomile on 3 strains of bacteria involved in otitis externa and it can be promised to produce new drugs, with lower side effects in eliminating these pathogens.

Background and aims Despite the use of antifungal drugs, the visceral candidiasis is associated with a high mortality rate. The aims of this study were an evaluation of intrinsic and acquired immune cells infiltration in kidney and spleen of the mice infected with systemic candidiasis and treated with chloroform fraction of Zataria Multiflora Boiss. Materials and methods Candida albicans (C. albicans) ATCC10231 clinical standard strain was isolated. C. albicans LD50 was determined. The laboratory animal (BALB/C mouse) infection with the visceral candidiasis was performed. The kidney and spleen tissues were stained with PAS and prepared for confirmation under the microscope. The Zataria Multiflora Boiss (Shiraz thyme) was prepared and the effects on the infected group were assessed. The kidney and spleen mononuclear cells (MNCs) were prepared and the flow cytometry technique was performed for the assessment of Th1, Th17, and Treg cells. Results The LD50 and LD totals were 1.5 × 108 and 2 × 108 Yeast/0.1 ml, respectively. In mice which had a drug intervention, including chloroform fraction of Zataria Multiflora Boiss, thymol, carvacrol or fluconazole, fungal purification was greater in the spleen than in the kidney. Among those mice without medication intervention, fungal clearance was higher in the kidney. The highest percentage of TH1 cells was in group 1 and then group 4 and in groups 2 and 3 respectively. Moreover, there was a significant difference between groups 4 and 5 and also 6 and 7. The percentage of TH1 cells in the spleen MNCs was higher than that of the kidney cells, which is the difference between the groups except for group 7. The percentage of TH17 cells in the kidney and spleen of all drug-receiving groups exhibited a significant increase compared to groups 6 and 7. The percentage of Treg cells in the kidney and the spleen only in the extract-receiving group had a significant decrease compared to the non-drug receiving group and the other groups receiving group depicted no significant difference in the percentage of Treg cells. Conclusion In addition to the direct effect on the fungus proven in vitro, the extract exhibits immunosuppressive effects, and thus can degrade the fungus through this way. The results demonstrated that the fraction of Zataria Multiflora Boiss can be considered as a powerful alternative to C. albicans therapy along with other therapies.

In Saccharomyces cerevisiae, los1 encodes a nuclear tRNA exporter. Despite the non-essentiality, the deletion of los1 has been shown to extend replicative life span in yeast. Here, we characterized AfuXpot, the los1 homologue in human pathogen Aspergillus fumigatus and found that it is continuously expressed during fungal growth. Microscopic examination of an AfuXpot-GFP-expressing transformant confirmed the nuclear localization of the fusion protein. The targeted gene deletion affirmed the non-essential role of AfuXpot in hyphal growth and sporulation. However, the growth of the deletion mutant was affected by amino acid, but not glucose, deprivation. The susceptibility of the deletant strain to protein and DNA/RNA synthesis inhibitors was also altered. Using bioinformatics tools, some transcription factor binding sites were predicted in AfuXpot promoter. Expression analyses of potential AfuXpot-interacting genes showed a marked down-regulation of sfp1 and mtr10 homologues in ΔAfuXpot strain. Our data demonstrates some conserved aspects of AfuXpot as a tRNA exporter in A. fumigatus.

Chitosan (CS) is a polycation with a pka of about 6.3 with a charge density dependent on pH and the %DA-value which can interact with polyanions to form complex and gels. Nanoparticles (CNPs) in addition can increase the antifungal potential of bioactive compounds like essential oils by increasing cellular interactions between them and the fungal as a result of the very small size that enhances cellular uptake. In this study, was set to investigate the encapsulation of the Gymnema sylvestre essential oils (G.EOs) using Chitosan and Myrestic acid made Nanogel in order to enhance its antifungal activity and stability to the oil against C. albicans strain (ATCC 10231). To procedure this, the self-assembled process of Chitosan and Myrestic acid Nanogel (CS-MA) through the 1- ethyl - 3- (3 dimethyl aminoprophyl) carbodiimide (EDC) was designed. Its physicochemical properties were determined by Fourier Transforms Infrared spectroscopy (FT-IR), X-ray diffraction (XRD), and microscopic methods by Atomic force microscopy (AFM), Transmission Electron Microscopy (TEM), and Scanning Electron Microscopy (SEM). Minimum inhibitory concentration (MIC) at 18.7 to 37.5 µg/ml and 2.3 to 4.6 µg/ml and minimal fungicidal concentration (MFC) at 75 µg/ml and 5.38 µg/ml using by broth micro dilution (BMD) method for G. sylvestre oils (G.EOs) and oil-loaded Nanogels (G.OLNPs) were measurement. The susceptibility of C. albicans biofilm to fractions was examined by 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[8phenylamino) carbonyl] - 2H - tetrazolium hydroxide (XTT) quantitatively.The concentration of G.OLNPs required to inhibit 50 % biofilm formation was 4.68 µg/ml, while that to remove 90 % biofilm growth was 18.07 µg/ml. In addition, it was observed that cell uptake of G.OLNPs was much higher compared with free G.EOs. Reverse transcription polymerase chain reaction (RT-PCR) analysis was performed to determine the effect of sub-MIC concentrations of G.EOs and G.OLNPs on expression of the biofilm–related gene ALS1 / ALS3, and indicated the G.OLNPs down-regulated the expression of hypha-specific gene ALS3. Furthermore, the data strongly suggested that G.OLNPs more effective suppressed C. albicans planktonic cells and reduction biofilm biomass.

Crocus sativus L. (saffron) is a valuable plant which is native to Iran. Saffron is the dried stigmata of the flowering part of the plant that is usually contaminated with different bacteria and fungi through production process. Antimicrobial properties of silver nanoparticles are well recognized. To survey the effects of nanosilver packaging on microbiological status of spiked, saffron samples over a six month period were chosen. Saffron samples from five regions of Khorasan province were purchased and de novo frequencies of microbial contaminants were determined using standard procedures. Totally 35g of saffron was spiked with known numbers of four bacterial and two fungal species and packaged into one gram packets. The packaging materials consisted of polyethylene polymers containing 0, 400, 800, 1200 or 4000ppm nanosilver (as Ag). Total and differential numbers of spiked microorganisms in the packaged saffrons were enumerated at initial and at six time points of seven, 14, 28, 64, 90 and 180days. Baird-Parker agar (BP agar), Kenner Fecal (KF), Salmonella–Shigella agar (SS agar), Violet Red Bile Glucose Agar (VRBGA), and Sabouraud Dextrose agar (SD agar) media were used for enumeration of the six spiked microorganisms including Staphylococcus aureus, Enterococcus faecalis, Salmonella Enteritidis, Enterobacter species and Escherichia coli, Fusarium oxysporum and Aspergillus flavus, respectively. Direct antibacterial activity of the composites was also determined. De novo frequencies of microorganisms in five saffron samples were at acceptable levels with dominance of fungi species. Nanosilver embedded packages accelerated the reduction in live microbial numbers in saffron samples and the efficacy was the best in packages containing 4000ppm nanosilver particles. Nanosilver packaging can significantly reduce microbial burden of saffron.

Over the last decade, communication industries have witnessed a tremendous expansion, while, the biological effects of electromagnetic waves have not been fully elucidated. Current study aimed at evaluating the mutagenic effect of long-term exposure to 900-MHz radiation on alpha-Int1 gene sequences of Candida albicans. A standard 900MHz radiation generator was used for radiation. 10ml volumes from a stock suspension of C. albicans were transferred into 10 polystyrene tubes. Five tubes were exposed at 4°C to a fixed magnitude of radiation with different time periods of 10, 70, 210, 350 and 490h. The other 5 tubes were kept far enough from radiation. The samples underwent genomic DNA extraction. PCR amplification of alpha-Int1 gene sequence was done using one set of primers. PCR products were resolved using agarose gel electrophoresis and the nucleotide sequences were determined. All samples showed a clear electrophoretic band around 441bp and further sequencing revealed the amplified DNA segments are related to alpha-Int1 gene of the yeast. No mutations in the gene were seen in radiation exposed samples. Long-term exposure of the yeast to mobile phone radiation under the above mentioned conditions had no mutagenic effect on alpha-Int1 gene sequence.

Background & aim Chromoblastomycosis is a chronic, progressive cutaneous and subcutaneous fungal infection clinically manifested by nodular, crusted and verrucose lesions appearing as a result of tissue proliferation around the site of traumatic inoculation of the exosaprophytic pigmented fungi. Up to now, little studies available documenting the presence of these organisms from environmental sources in Iran. Therefore, this study was aimed to investigate the presence of black fungi in the nature of Iran. Methods and materials A total of 400 environmental samples including: air, soil, wood, plant matters, grain and organic debris were obtained from different areas (public places) belonging to Babol city of the Mazandaran province in north of Iran during one year. The grown dark colonies were then isolated and exposed to DNA extraction. All culture positive isolates were identified to the species level by DNA sequencing of ITS1–5.8S–ITS2 rDNA region. Results Based on morphological appearances, 79 strains of dematiaceous fungi including 13 genera and 23 species were isolated from 400 included samples. Cladosporium herbarum (17/79) was the most predominantly identified species, afterward Alternaria spp., Curvularia spp. and Ulocladium spp. in descending frequency isolated as well. Summer season was documented as the time of highest fungal isolation rate. Conclusion Although, all the environmental black fungi isolated strains from current study are regarded as nonpathogenic. However, the rise in the reports of chromoblastomycosis cases in both healthy and immunocompromised hosts, caused by these fungi that gain entrance through the skin by traumatic implantation of natural sources, all over the world is alarming.

Background and objectives: Aflatoxins (AFs) are produced by fungi, which may remain in the cow’s milk even after pasteurization. Aflatoxin M1 (AFM1) is specifically of great medical importance, as it is certainly carcinogenic for human. Several strategies have been suggested for its reduction, including the use of probiotics, especially Lactobacilli or lactic acid bacteria (LAB). As this method has not been confirmed as a routine treatment, yet, in this study, we aimed to evaluate the effect of three LABs on reduction of AFM1 in traditional milk and cheese. Materials and methods: In this study, 45 milk samples and 40 cheese samples were purchased from marketplaces of Shiraz city during February 2018-June 2018. Of 50 LABs purchased, the results of antifungal property, and resistance to bile salts, resulted in 5 strains. These 5 strains were tested for mean after addition of 5 ppm AFM1, compared to natamycin. The strains with reduction in AFM1 level were sequenced and registered in NCBI database. Results: The results showed reduced AFM1 level in three LAB strains, sequenced as Lactobacillus fermentum CECT562 (T), Lactobacillus brevis ATCC14869 (T), Enterococcus faecium LMG 11423(T) to 0.05, 0.03, and 0.03, respectively. Conclusion: The three LABs selected in the present study have significant effect on reducing AFM1 level in the traditional milk and cheese.

Objective The effect of candidemia on immunologic parameters in breast tumor bearing patients is not well studied. Here, we hypothesised that candidemia in the tumor background may change the outcome of immunologic parameters and tumor condition. Method Mice were divided into four groups, including normal, tumor, Candida infected (only Candidiasis) and tumor/Candidiasis groups. Tumor changes were recorded daily after tumor transplantation and induction of candidemia. Splenocytes of mice were harvested, cultured, and stimulated with PHA; afterwards, IL-4, IL-10, IFN-γ, TNF-α and TGF-β cytokines were assessed using ELISA kits. We also evaluated the population of CD4+CD25+Foxp3+ regulatory T cells in the tumor infiltrated and splenocytes. Results The results showed that infection with C. albicans decreased the IFN-γ/IL-4 ratio in tumor/candidiasis and candidiasis groups versus their non-infected controls. IL-10, TGF-β and TNF-α levels increased in the candidiasis group. In addition, Candidemia led to an increase in the Treg population in tumor microenvironment and splenocytes of experimental groups compared with non-infected controls. Finally, candidemia increased tumor growth of tumor/Candidiasis group compared with the tumor group. Conclusion It seems that systemic infection with C. albicans could not only induce regulatory T cells but also result in dysregulation of cytokine network and thereby facilitate tumor growth.

Background: Increasing azole resistance among Aspergillus strains was observed over the last decade. For this reason, viable alternatives for the current medicines are required. Objectives: The current study aimed at producing fluconazole-loaded liposomal nanoparticles and comparing in vitro antifungal activity of fluconazole and nano-fluconazole on Aspergillus flavus and A. fumigatus species isolated from patients and poultry. Methods: Fifty isolates of A. flavus and A. fumigatus were collected from visceral and superficial fungal lesions of humans as well as pulmonary fungal infection of poultry. Liposomal nanoparticles were prepared by thin-film hydration method, using soybean lecithin, cholesterol, and fluconazole in a ratio of 10:1:1. The nanoparticles were analyzed in terms of size, zeta potential, and morphology. Antifungal susceptibility testing was carried out to examine and evaluate the minimum inhibitory concentration (MIC) of fluconazole and nano-fluconazole against Aspergillus species by the standard method of broth microdilution as described in CLSI (the clinical and laboratory standards institute) -M38A2. Results: The particle size of liposomes containing fluconazole was 88.9 ± 12.1 nm and its zeta potential was -20.12 ± 1.88 mv. Nano-liposomes containing fluconazole significantly reduced the MIC against A. flavus (P = 0.0005) and A. fumigatus (P < 0.0001). Conclusions: The results of the current in vitro study showed that nano-fluconazole has better antifungal effects than the common form of drug on A. flavus and A. fumigatus species. The current study showed the antifungal activity of nano-drugs.

Background: Due to long-term treatment of dermatophytic lesions, such as tinea unguium and using antifungal drugs, which cause side effects, it is essential to investigate new antifungal drugs with greater absorption and fewer side effects, such as nano-drugs. Trichophyton rubrum and Microsporum canis are important dermatophyte ‎species. To investigate new strategy treatment, researchers have tried to find a new drug with extensive therapeutic effects and short-duration treatment. In this context, nano-drugs have gained great interest in improving the efficacy of antifungal properties compared to commercial drugs. Objectives: The aim of this study was to compare the effects of free terbinafine and nano-liposomal terbinafine on growth of clinical isolates of T. rubrum and M. canis. Methods: In this study, 120 isolates of T. rubrum and M. canis were separated from dermatophytosis lesions. Diagnosis of the strains was based on the morphological structure and molecular identification. Nano-liposomal terbinafine was prepared and analyzed. Antifungal susceptibility testing was performed by broth micro dilution CLSI M 38-A method to compare inhibitory growth effects of nano-liposomal characteristics of terbinafine and free terbinafine. Results: The results showed that the Minimum Inhibitory Concentration (MIC) values of terbinafine against T. rubrum and M. canis strains were 0.0625 to 1 μ/mL and 0.0313 to 0.5 μ/mL, respectively. Also, the MIC values of nano-liposomal terbinafine against T. rubrum and M. canis were 0.0156 to 0.25 μ/mL and 0.0078 to 0.125 μ/mL, respectively. Conclusions: Comparison of the antifungal effects of nano-liposomal terbinafine and free terbinafine against T. rubrum and M. canis isolates showed that the nanopartilces of terbinafine had a greater antifungal activity against the studied isolates and can be used as an alternative agent for dermatophytosis treatments.

Background and purpose Soil is the main habitat of saprophytic and pathogenic fungi. Mucoromycotina constitutes a large group of soil fungi, with certain opportunistic members causing systemic infections in immunocompromised hosts. The majority of human and animal infections are caused by the members of the genera Rhizopus, Mucor, Rhizomucor, Lichtheimia (Absidia), Cunninghamella, and Mortierella. Accordingly, in the present study, we aimed to isolate and identify the main genera of the order Mucorales, using molecular assays and morphological features. Materials and methods In total, 340 soil samples were collected from seven public parks throughout the city and sidewalk gardens in 14 municipal districts in Isfahan, Iran. All the samples were cultured on the appropriate media, incubated at 27°C for 2- 4 days, and examined daily for visible fungal growth. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was applied and macroscopic, microscopic, and physiological characteristics were assessed to identify fungal colonies. Results 400 pure colonies, belonging to the orders Mucorales and Mortierellales, including the genera Lichtheimia, Rhizopus, Rhizomucor, Mucor, Cunninghamella, and Mortierella, were identified. The genus Rhizopus (35.5%) was the most frequent isolate, followed by Mucor (32.25%) and Rhizomucor (27.5%). Conclusion The results emphasize the importance of opportunistic fungi in public areas and indicate the risk of exposure for immunocompromised individuals.

Background: Saccharomyces boulardii, a non-pathogenic yeast, is effective on prevention and treatment of intestinal infections caused by bacterial pathogens and also affects the virulence factors of Candida albicans. SAP2, an important potential virulence factor of C. albicans is a suitable target for a new research. Objectives: The current study aimed at investigating the effect of S. boulardii extract on the expression of SAP2 gene and antifungal susceptibility of C. albicans isolates. Methods: The current study investigated SAP2 relative expression and antifungal susceptibility patterns of C. albicans species isolated from oral lesions of HIV+ patients and the standard strain ATCC 10231 after exposure to S. boulardii extract. Susceptibility of C. albicans isolates to antifungal drugs was assessed according to those proposed in the clinical and laboratory standards institute (CLSI) M44-A2 reference method. The real-time polymerase chain reaction (PCR) was performed to evaluate SAP2 gene expression. Results: Saccharomyces boulardii extract 0.48 mg/mL changed the antifungal susceptibility pattern of C. albicans isolates to ketoconazole and itraconazole. Before exposure, 53.3% of the isolates were sensitive to ketoconazole, while it increased to 73.3% after exposure to S. boulardii extract. The sensitivity to itraconazole increased from 6.7% to 26.7% after exposure to S. boulardii extract. Although susceptibility of C. albicans isolates to other antibiotics (fluconazole, clotrimazole, nystatin, and amphotericin-B) did not change after exposure to S. boulardii extract, there was a significant difference in the average inhibitory zones of the isolates after exposure to S. boulardii extract. Conclusions: The level of SAP2 expression significantly reduced in the isolates treated with 0.48 mg/mL S. boulardii extract. Comparison of SAP2 gene expression between ketoconazole-resistant and -intermediate isolates, and the ketoconazole-sensitive ones demonstrated the relative higher gene expression in resistant and intermediate isolates. Saccharomyces boulardii extract can be considered as a suitable probiotic candidate to prevent and treat C. albicans infections.

With the increase of patients suffering from immune deficiency infections also increased pulmonary fungi even in people with defective immune system can cause fatal and lethal candidiasis. The timely diagnosis of pulmonary candidiasis is one of the problems that has been detected. Polymerase chain reaction (PCR) test and Loop mediated isothermal amplification (LAMP) method optimized on the basis of α INT1 gene and then sensitivity and specificity were evaluated. LAMP is a novel nucleic acid amplification technique with high specificity and sensitivity which has been done under isothermal condition. Samples were the bronchoalveolar lavage suspected of tuberculosis (TB) reviews for TB disease negative have been reported. DNA extraction carried out by standard phenol/chloroform method on samples and PCR test and LAMP was done. PCR and LAMP testing was performed on samples and products of 441 bp were amplified and observed with agarose gel electrophoresis. At the end of the LAMP reaction, SYBR Green was used for identifying negative and positive results. Among the 60 quantities sera, only 7 cases were PCR positive but 8 cases were LAMP positive. In comparison, between LAMP and PCR, the LAMP technique in spite of its simplicity, high sensitivity and specificity, could be an appropriate replacement for PCR.

This study was set to investigate the encapsulation of the Thyme essential oils using chitosan and benzoic acid-made nanogel in order to enhance its antifungal properties and half-life. To achieve this, the self-assembled polymer of chitosan and benzoic acid nanogel (CS-BA) was synthesized, its size and shape were confirmed by spectrometric (FTIR) and microscopic methods (TEM and SEM) and was then used in encapsulating the essence. Under sealed condition, the minimum inhibitory concentration of the CS-BA encapsulated essential oils was recorded at 300 mg/l while the free Thyme extract could only completely prevent the growth of Aspergillus flavus at an elevated concentration of 400 mg/l. Under non-sealed condition, higher concentration of encapsulated Thyme essential oils (500 mg/l) was required to cause complete fungi inhibition and free oils failed to lead to full inhibition even at concentrations as high as 1000 mg/l. In vivo analysis also revealed significant anti-fugal properties of the encapsulated oils at concentrations above 700 mg/l. Overall, due to the volatility and instability of free essential oils, CS-BA nanogel encapsulation was found to have significantly increased half-life and the anti-fungal properties of Thyme essential oils.

Fungi, in particular some of the Penicillium and Aspergillus species, are the main cause of dairy products contamination, they threaten the general health of community and food safety due to the production of mycotoxins. The purpose of this study was to isolate and identify saprophytic fungi contaminating various dairy products in Shiraz in comparison to the National Iranian Standard (2406). In this study, 85 samples (40 cheeses, and 45 milk samples) were collected from Shiraz city of Iran and analyzed for the presence of mold and yeast. After morphological investigation, molecular tests was done by PCR method. The results were compared to the National Iranian Standard and were analyzed by SPSS version 20 and chi-square statistical analysis with about 95% accuracy and significant level (p-value

Aspergillus flavus is a human pathogen, allergen and mycotoxin producer. Plant oils such as peppermint oils are known to possess considerable antifungal properties are increasingly considered as natural agents for food preservation and as alternatives for toxic synthetic fungicides. This study was set to investigate the encapsulation of Mentha piperita essential oils in chitosan–cinnamic acid nanogel in order to enhance antimicrobial activity and stability of the oils against A. flavus. The results obtained showed that the extract possessed remarkable antifungal properties against A. flavus and that due to the volatility and instability of the oils against environmental factors, its encapsulation considerably improved its performance. The minimum inhibitory concentration of free and encapsulated M. piperita essential oils against A. flavus under sealed condition were at 2100 and 500 ppm, respectively. Moreover, the encapsulated oils performed better (800 ppm), when tested under non-sealed condition while the free oils failed to caused complete inhibition within the concentration range tested (up to 3000 ppm). These findings revealed the promising role of CS–Ci nanogel as a carrier for essential oils in order to enhance their antimicrobial properties.

Background and Aim: Soil structure is mainly composed of sand, silt, c lay, and organic materials. Organisms can live in the soil. The large number of stray cats in the cities of Iran is a major environmental and hea lth problem. Toxoplasma oocysts are shed with the feces of cats, so soil is known as a potential source of transmission of toxoplasmosis. The aim of this stud y was to determine the soil contamination with Toxoplasma gondii oocyst in the public parks of the Arak city. Methods: Sixty soil samples were collected from 15 main park s of Arak city. Four soil samples from the children's playground, a potting place, around trash bins, and around toilets were taken. Oocyst was iso lated from soil by floatation in saturated sucrose. Floating debris was tested by tw o methods: staining by the modified Ziehl-Neelsen technique and PCR. The target of PCR was the 122 bp fragment of the B1 gene. Results: From 60 soil samples of public parks of Arak city, 8 samples (13%) were suspected to Toxoplasma oocyst contamination in staining smears. Only 3 samples (5%) of 60 samples were positive in PCR. The result s showed that the staining method is not a good method to detect oocysts in th e soil because the diversity of oocyst in soil is very high and similar in appearan ce. Conclusion: This study showed soils of public parks in the Arak city were contaminated to oocyst of Toxoplasma. Also molecular method for the detection of parasites in the soil was more suitable than sta ining method.

The accumulation of dry waste containing synthetic polymers due to their resistance to microorganisms and other environmental factors has posed some serious problems to the environment in recent years. On the other hand, plastics constitute the foundations of economy as they are widely used in agriculture, constructions, packaging, health care and also medicine. The aim of this research was to investigate the role of different isolated fungi in the degradation of polycarbonate polymers. For this purpose, sampling was done using the garden soil and waste leachate from Isfahan Waste Management Organization. Samples were enriched in the liquid mineral salt medium supplemented with polycarbonate and then were transferred to the same medium solidified with agar to isolate and identify different fungi. Finally, their biodegradation activity was investigated with the help of clear-zone and atomic force microscopic (AFM) techniques, and also lipase and amylase production was tested. Among 15 isolated genera of mold fungi, Fusarium,Ulocladium, Chrysosporium and Penicillium showed biodegradation activity. According to the diameter of clear zone around the fungal colonies and also AFM results, the highest rate of degradation was related to Fusarium. Lipase activity of all isolated fungi was positive, but amylase activity of Ulocladium was negative. It can be concluded that some fungal strains such as Fusarium can be used for the biodegradation of plastic materials as it leads to a very eco-friendly biodegradation process.

Background The Mucorales are an important opportunistic fungi that can cause mucormycosis in immunocompromised patients. The fast and precise diagnosis of mucormycosis is very important because, if the diagnosis is not made early enough, dissemination often occurs. It is now well established that molecular methods such as polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) are feasible and reliable tools for the early and accurate diagnosis of mucormycosis agents. Objectives The present study was conducted to evaluate the validity of PCR-RFLP for the identification of Mucorales and some important Mucor and Lichtheimia species in pure cultures of Zygomycetes. Materials and Methods Specific sense and anti-sense primers were used to amplify the Mucorales, Mucor, and Lichtheimia DNA. The PCR products were digested by AfIII, XmnI, and AcII restriction enzymes, and the resultant restriction pattern was analyzed. Results On the basis of the molecular and morphological data, we identified Mucor plumbeus (10.83%), M. circinelloides (9.17%), Lichtheimia corymbifera (9.17%), M. racemosus (5.83%), M. ramosissimus (3.33%), and L. blakesleeana (0.83%). Conclusions It seems that PCR-RFLP is a suitable technique for the identification of Mucorales at the species level.

Background: The presence of subtilisin genes (SUBs) coding for serine proteases in Microsporum canis DNA, contribute to the adherence of fungi to keratinized tissues. The aim of this study was identifed the presence/absence of subtilisin gene family in M.canis isolated from human and animal source. Materials and Methods: This cross-sectional study was performed on 24 samples of patients and pets with dermatophytosis from September to November 2017. Genomic DNA, pertaining to all pure colonies in Sabouraud agar with cycloheximide and chloramphenicol (SCC) and Sabouraud Dextrose agar(SDA), extracted, using rapid method without liquid nitrogen. M. canis were detected by molecular test. SUB1, SUB2 and SUB3 gene of M. canis amplified. The relative frequency of the genomic sequences also calculated. Results: M. canis DNA were extracted from cats (n=6), dog (n=1) and human (n=3) with tinea corporis and showed a high percentage for SUB2 (90%; 9/10) and the absence of SUB genes in a M. canis isolated from dog with develop dermatophytosis. The same results were observed in the frequency of the SUBs presence from the colonies grown on SCC and SDA. Conclusion: The presence of SUBs in M. canis isolates with human and animal source will be the basal to understand zoonotic infections . The absence SUBs in clinical isolate, indicates that they are indispensable for initiation of the infection.

Mobile phones communicate with base stations using 900 MHz microwaves. The current study was aimed to survey the effects of long-term 900 MHz microwave exposure of mice on experimentally induced cutaneous candidiasis. Forty inbred, male, BALB/c mice were randomly divided into four groups. Cutaneous lesions with Candida albicans were experimentally induced on the lateral-back skin of the 20 mice. One group of the diseased mice were exposed (6 h per day and 7 d per week) to 900 MHz microwave radiation, while the other groups were not exposed. Two unexposed control groups were also included. The skin lesions were regularly monitored and the live candida cell density was enumerated using the colony-forming unit (CFU) assay. The process was repeated after a one week resting interval. One week later, all mice were challenged through intra tail veins using LD90 dose of C. albicans. Mortality of the mice was recorded and the candida load of the kidney homogenates from died animals was counted. 900 MHz microwave exposed mice had 1.5 day and 3.7 day delays on wound healing in stages two. Live Candida inoculated Wave exposed (LCW) mice also showed higher yeast loads in skin lesions at days 5, 7 and 9 post inoculation. Survival analysis of live candida challenged mice showed the radiation exposed group is prone to death induced by systemic infection and candida enumeration from the kidney homogenates showed radiation exposed animals have had significantly higher yeast load in the tissue. In collection, long-term 900 MHz radiation exposure of mice led to longevity of skin wounds and susceptibility of the animals to systemic challenge and higher incidences of microorganisms in internal tissues.

Background: Sinusitis is one of the most prevalent infectious diseases among those who visit doctor, which requires antibiotic intervention. The most common types of bacteria causing this disease are Streptococcus pneumonia, Streptococcus pyogenes, Staphylococcus aureus and Pseudomonas aeroginosa. Nonetheless due to dangerous side effects of chemical drugs on body and increased drug resistance of bacteria, new alternative therapeutic method, such as nano-encapsulating herbal essences is considered to augment effectiveness of treatment as well as bacterial sensitivity. Objectives: In this study, the antimicrobial effects of four encapsulated essential oils including Lavandula angustifolia Mill, Cinnamomum verum J. S. Persl, Artemisia sieberi Besser and Myrtus communis L. were evaluated on microorganisms of S. pneumonia , S. pyogenes, S. aureus and P. aeroginosa, which are among the most common factors for sinusitis. Materials and Methods: First, the Minimum Inhibitory Concentration and Minimum Bactericidal Concentration quantitative tests followed by qualitative test (disk diffusion) were performed to examine drug sensitivity. Results: The quantitative result in examined encapsulated essential oils was 0.097 µg/mL for S. pneumonia with Lavander nano-essence, 0.097 µg/mL, for S. pyogenes with Artemisia and Lavander nano-essences, 1.562 µg/mL for S. aureus with Artemisia nano-essence and 3.215 µg/mL for P. aeroginosa with Artemisia and Lavander nano-essences. Moreover, the results of the disk diffusion test showed that nano-essences did not have the ability to be released from paper disks and diffuse in solid environment. Conclusions: The results show proper effects of encapsulated essential oils on the four leading causes of sinusitis microorganisms (P ≥ 0.01) and also the findings indicate that the essential oils can be used to make more effective drugs with herbal resources and fewer side effects for destroying these microorganisms.

Background: Using garlic is widespread in Iran and other countries as a medicine and a natural spice. Garlic is a potential inhibitor for food pathogens. Foods contaminated with pathogens pose a potential danger to the consumer’s health. The use of garlic can increase the shelf life and decrease the possibilities of food poisoning and spoilage in processed foods. Objectives: The aim of this study was to investigate the antibacterial effect of garlic aqueous extract on growth of Staphylococcus aureus bacteria. Materials and Methods: In this study, the garlic aqueous extract was prepared under sterile conditions and was added in 1, 2, and 3 mL to 100g hamburger samples. A group of samples was prepared to be used as treatment sample, while a group was stored at 4°C and -18°C. The samples were kept in refrigerator for one and two weeks and they were frozen for one, two and three months and then subjected to microbial tests. Results: Statistical evaluation of the first and second week samples indicated a significant growth decreased by all the 1, 2, and 3-mL extracts. In treatment of one, two and three-month samples, the growth of S. aureus was significantly decreased by the 2 and 3-mL extracts. The 1-mL extract was effective in decreasing the growth, and a significant difference was observed in treatments with 2 and 3-mL extracts. However, there was no significant difference between the two and three-month samples, though they were significantly different from the one-month samples. After evaluations, treatment with the 2-mL extract was found to be the best one. Conclusions: Garlic aqueous extract has antibacterial properties against S. aureus present in hamburger. Moreover, garlic aqueous extract can be used not only as a flavor but also as a natural additive for hamburger. In addition, garlic has antibacterial properties against other Gram-positive and Gram-negative bacteria, which must be investigated in further studies.

The present study describes the isolation of microorganisms capable of producing alpha-pinene from beta-pinene. 24 (15 fungi, 9 bacteria) microorganisms were isolated from galbanum, gum and soil, were screened for their ability to transform beta-pinene to alpha-pinene. Biotransformation products were extracted with n-hexan and analyzed by gas chromatography. One microorganism (bacterial strain) were found. The biotransformation medium involved, phosphate buffer pH 6, 100 microL beta-pinene, 1 g biomass of microorganism, 37 degrees C, 150 rpm and 22 h. The experiments were performed in conical flasks. The optimum cell growth were obtained when 30 g L(-1) glycerin applied. The optimum conversion beta-pinene to alpha-pinene was obtained when 20 g L(-1) glycerin applied as carbon source for bacterial strain.

The risk of infection in burnt patients is high and at the present the antibiotic resistance of bacteria isolated from these patients has increased. Therefore, the aim of this study was detection of antibiotic resistance pattern of Escherichia coli, Staphylococcus aereus and Pseudomonas aeruginosa isolated from burnt patients to some current antibiotics. The results showed that all of isolates were resistance to tested antibiotics, therefore the control and preventing of infection in hospital burn units is necessary.