shirin farahyar

The presence of fungi in the respiratory tract as mycobiome, particularly Candida species (spp.), remains a serious problem due to increasing numbers of immunocompromised pa-tients. The confirmed reliable existence of these pathogens due to frequent colonization is essential. This investigation aimed to recognize Candida spp. among isolates from bron-choalveolar lavage of immunocompromised and critically ill patients and to evaluate their susceptibility to antimycotic drugs.

Bronchoalveolar lavage fluid was collected from 161 hospitalized patients presenting with suspected respiratory fungal infection /colonization. The specimens were examined by standard molecular and mycological assays. Candida spp. were recognized with sequence assessment of the D1-D2 section of the large subunit ribosomal DNA. The susceptibility of Candida isolates to common antimycotic drugs was distinguished by standard broth micro-dilution.

Seventy-one clinical isolates of Candida spp. were recognized. Candida albicans was the most frequent, followed by C. glabrata, C. krusei (Pichia kudriavzevii), C. dubliniensis, C. parapsilosis, and C. tropicalis. We found 5.1% of C. albicans isolates and 8% of C. glabrata isolates to show resistance to fluconazole. The whole of the Candida spp. were sensitive to amphotericin B and caspofungin.

This study demonstrated that C. albicans and C. glabrata are the most common isolates of bronchoalveolar lavage fluid in patients, and the drug susceptibility screening confirmed that amphotericin B and caspofungin are effective against Candida spp. but some C. glabrata and C. albicans isolates showed resistance to fluconazole.

Regarding the wide-spectrum antimicrobial effects of curcumin and silver, this study aimed to evaluate the antifungal activity of green-synthesized curcumin-coated silver nanoparticles (Cur-Ag NPs) against a set of Candida and Aspergillus species.

Cur-Ag NPs were synthesized by mixing 200 µL of curcumin solution (40 mM) and 15 mL of deionized water. The mixture was stirred for 3-5 min, followed by the addition of 2.5 mL of silver nitrate solution (2.5 mM). The resulting solution was incubated for 3 days. Antifungal susceptibility of 30 fungal isolates of Aspergillus and Candida to fluconazole and itraconazole, as well as the activity of Cur-Ag NPs against the isolates, were determined, both alone and in combination, using broth microdilution according to the Clinical and Laboratory Standards Institute guidelines.

Cur-Ag NPs demonstrated promising antifungal activity, particularly against Candida species. The geometric mean value of the minimum inhibitory concentration of Cur-Ag NPs was significantly lower than that of fluconazole for all the studied fungi. Similarly, it was lower than those of itraconazole in C. albicans and A. fumigatus. The minimum fungicidal concentrations of Cur-Ag NPs were markedly better than those of fluconazole but still inferior to those of itraconazole.

Cur-Ag NPs demonstrated indisputable antifungal activity and great potential that can be harnessed to combat fungal infections, particularly those caused by azole-resistant strains of Aspergillus and Candida.

Different studies have shown that despite the expanding number of antifungal drugs, the death rate caused by Candida species has increased during the recent decades due to drug resistance occurrence. The present study aims to identify molecular structure and evaluate drug susceptibility in Candida species isolated from bronchoalveolar lavage fluid in patients diagnosed with hematological malignancies. In this cross-sectional study, 54 clinical specimens were taken from the bronchoalveolar lavage of patients. The suspected colonies were investigated by microscopic examination and subsequent passages were evaluated according to standard operating procedures and specification of the type of colony color prescribed by CHROMagar to isolate the yeast. The sequencing method (ITS1, ITS4) was used to approve Candida species. Finally, susceptibility test was carried out according to M27S-3 and M38-A2 micro-dilution methods. Among 54 samples investigated with culture and PCR methods, 33 Candida species were identified in patients with hematological malignancies. Candida albicans (75.7%) was the most common fungal isolate. Results of drug susceptibility tests showed that the isolated C. albicans (n = 2), C. glabrata (n = 1), and C. tropicalis (n = 1) from patients with hematological malignancies were resistant to fluconazole. The present study showed that the prevalence of C. albicans was higher than other fungal species among patients with hematological malignancies. Candida species are more susceptible to voriconazole, amphotericin B and Caspofungin. Therefore, identification of candida species along with their antifungal susceptibility pattern can help clinicians to better treat patients.

 Vulvovaginal candidiasis (VVC) is an ordinary infection caused by Candida species. Meanwhile, a shift towards non-albicans Candida (NAC) species has been detected in VVC patients.

 This study aimed at molecular identification of Candida isolates, causing VVC.

 Vaginal secretion samples of 320 non-pregnant vaginitis patients at Shahid Akbar-Abadi Obstetrics and Gynecology Hospital in Tehran (Iran) were collected. Samples were evaluated using mycological and molecular approaches. Vaginitis isolates were analyzed with the PCR using NL1 and NL4 primers, and the D1/D2 region of the large-subunit rRNA gene was amplified and sequenced.

 In total, 100 Candida isolates were identified from VVC and recurrent vulvovaginal candidiasis (RVVC). Candida albicans was the most frequent (51%), followed by C. glabrata (36%), C. krusei (Pichia kudriavzevii) (8%), and C. kefyr (Kluyveromyces marxianus) (5%). 51 and 49% of isolates had C. albicans and NAC, respectively.

 Candida albicans and C. glabrata were the most common agents of vulvovaginal candidiasis. NAC spp. (49%) was found as an important agent associated with VVC.

With regard to the increasing number of antifungal-resistant dermatophytes, the requirement for precise identification of causative agents of infections and antifungal susceptibility test is vital. Antifungal susceptibility testing of dermatophytes plays a pivotal role in managing dermatophytosis. The current study aimed at determining antifungal susceptibility profile of 161 important dermatophyte species isolated from Iranian patients.
The current descriptive, cross sectional study was conducted on 508 clinically suspected samples of dermatophytosis collected and identified by conventional methods. All dermatophyte isolates were identified using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The susceptibility of dermatophyte strains to two routine antidermatophyte agents (terbinafine and griseofulvin) was evaluated using micro-dilution method according to CLSI (the clinical and laboratory standards institute) M38-A2 guidelines. Trichophyton rubrum PTCC 5143 and Candida krusei ATCC 6258 were used as quality controls.
Among 161 dermatophyte isolates, T. interdigitale was reported as the most frequent species isolated from patients using PCR-RFLP and Microsporum ferruginum was the least isolated species. The minimum inhibitory concentration (MIC) values of griseofulvin and terbinafine were ranged 0.0312 - 8 and 0.008 - 4 µg/mL, respectively. The most susceptible and resistant species to griseofulvin were T. interdigitale (MIC = 0.0312 µg/mL) and T. interdigitale/T. rubrum (MIC = 8 µg/mL), respectively. The results indicated that T. verrucosum (MIC = 0.008 µg/mL) was the most susceptible species to terbinafine, whereas T. interdigitale and T. rubrum were the most resistant species to it (MIC = 4 µg/mL).
The obtained results assist clinicians to monitor the trend and be able to choose effective medications to treat patients with dermatophytosis, especially in countries such as Iran, where dermatophytosis is still a public health problem.

2-phenylethanol is a colorless and aromatic compound with antimicrobial effects which is used extensively in perfumes and cosmetics, as well as in the food industry. Chronic vulvovaginal candidiasis is a vulvovaginal inflammation which is caused by Candida spp. Resistance to clotrimazole which is one of the most common drugs in the treatment of this disease was reported in many patients. In order to improve the treatment, the effect of 2-phenyl ethanol was investigated in combination with clotrimazole on Candida species isolated from chronic vulvovaginal candidiasis.
This interventional study was performed in Iran University of Medical Sciences from February, 2016 until December, 2016 on Candida species isolated from women with chronic candidial vulvovaginitis who had been referred to Lolagar Hospital of Tehran. All specimens were examined by direct microscopy, culturing on Candida CHROMagar medium (to primary identification), sabouraud dextrose agar medium) to preservation the isolates) and determining the internal transcribed spacer (ITS) sequence (in order to final determination of Candida species). Then clotrimazole and 2-phenyl ethanol alone and in combination, was examined on isolated species, according to Clinical and Laboratory Standards Institute (CLSI) M27-A3 protocol (micro-broth dilution method). Finally, findings were analyzed.
From 40 detected strains of Candida species in this study, 95% were Candida albicans and 5% were Candida africana. The mean minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of clotrimazole were 24.73±28.87 µg/ml and 30.18±33.004 µg/ml, respectively and the mean MIC and MFC of 2-phenylethanol were 2580±932.38 µg/ml and 3200±1403.29 µg/ml, respectively. The MIC50 and MIC90 of clotrimazole were 16 and 64 µg/ml, respectively. The MIC50 and MIC90 of 2-phenylethanol were both 3200 µg/ml. Most of the isolates were resistant to clotrimazole (82.5%). In combination test, the mean MIC of 2-phenylethanol and clotrimazole alone were 3200±0 µg/ml and 56±40.16 µg/ml, respectively. The fractional inhibitory concentration index (FICI) range was 0.14-0.37. Also, there was a significant difference between clotrimazole MIC values alone and in combination (P= 0.021).
The synergistic effect was observed in combination of clotrimazole and 2-phenylethanol.

Vulvovaginal candidiasis (VVC) is an important problem due to Candida spp. The aim of this study was molecular identification, phylogenetic analysis, and evaluation of antifungal susceptibility of non-albicans Candida isolates from VVC.
Vaginal secretion samples were collected from 550 vaginitis patients at Sayyad Shirazi Medical and Educational Center of Gorgan (Golestan Province, Iran) from May to October 2015. Samples were analyzed using conventional mycological and molecular approaches. Clinical isolates were analyzed with specific PCR using CGL primers, and the internal transcribed spacer region and the D1-D2 domain of the large-subunit rRNA gene were amplified and sequenced. Susceptibility to amphotericin B, fluconazole, itraconazole, and clotrimazole was determined by the guidelines of the Clinical and Laboratory Standard Institute.
In total, 35 non-albicans Candida isolates were identified from VVC patients. The isolates included 27 strains of Candida glabrata (77.1%), 5 Candida krusei (Pichia kudriavzevii) (14.3%), 2 Candida kefyr (Kluyveromyces marxianus) (5.7%), and 1 Candida lusitaniae (Clavispora lusitaniae) (2.9%). The fungicides itraconazole and amphotericin B were effective against all species. One isolate of C. glabrata showed resistance to fluconazole and clotrimazole, and 26 isolates of C. glabrata indicated dose-dependent susceptibility to fluconazole. C. lusitaniae was susceptible in a dose-dependent manner to fluconazole and resistant to clotrimazole.
Non-albicans Candida spp. are common agents of vulvovaginitis, and C. glabrata is the most common species in the tested patients.

Background and Candida albicans (C. albicans) is an opportunistic fungus that can colonize women’s mucosal epithelial cell surfaces, causing vulvovaginitis in specific circumstances. The major genes contributing to drug resistance in C. albicans are the candida drug resistance (CDR) and multi drug resistance (MDR) genes. The purpose of this study was to evaluate the CDR-2 and MDR-1 gene expression patterns in C. albicans strains isolated from patients with recurrent vulvovaginal candidiasis.
In this study, 40 isolates of fluconazole-resistant C. albicans were cultured on Sabouraud dextrose agar. These isolates were collected from women with vulvovaginitis who were referred to a clinic in Tehran, Iran, and transferred to a mycology laboratory. Then, RNA was extracted from the isolates using phenolchloroform and glass beads, and the complementary DNA (cDNA) was synthetized. To detect the semi-quantitative expression of CDR-2 and MDR-1 genes, the reverse transcriptase-PCR (RT-PCR) technique was performed using specific primers.
Our findings indicated that of the 40 C. albicans isolates, 35 (87.5%) strains were positive for mRNA of the CDR-2 gene, 32 (80%) strains expressed mRNA of the MDR-1 gene, and 30 (75%) strains were confirmed to express mRNA of both the CDR-2 and MDR-1 genes simultaneously using the RT-PCR assay.
According to the obtained results, the expression rates of CDR-2 and MDR-1 genes were high in fluconazole-resistant C. albicans isolates, which can cause treatments to fail and result in chronic infections.Inhibiting these important genes using novel or natural agents can help with the treatment of chronic and recurrent vaginitis.

Background and Allergy is an undesired immune response to non-pathogenic agents. However, some opportunistic microorganisms such as fungi can also cause allergy. Among those fungi, hyphae form of Aspergillus strains including A. fumigatus, A. flavus, and A. niger could be mentioned. In this study, we aimed to separate allergic proteins from Aspergillus strains and determine their identity.
Standard species of Aspergillus strains were cultivated in optimized conditions and the mycelium was separated by centrifugation. The fungal cells were lysed through physical methods such as freezethawing and grinding to prepare a suitable protein extract. The protein concentration was measured by Bradford method and the electrophoretic pattern of the extract was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins were fractionated by ammonium sulfate precipitation and anion exchange chromatography using fast protein liquid chromatography (FPLC) system. The IgE immunoreactivity of the sensitized patients and controls was studied using the fractionated proteins by enzyme-linked immunosorbent assay (ELISA). Following SDS-PAGE, proteins were electrotransferred onto polyvinylidene difluoride (PVDF) membranes and the strips were blotted with allergic patient's and control's sera. The immunoreactive bands were excised from colloidal coomassie-stained SDS-PAGE gels and studied by mass spectroscopy methods.
Among the studied species, A. fumigatus showed stronger IgE reactivity and more IgE reactive protein bands than others did. The proteins with higher molecular weights showed stronger immunoreactivity in Western blotting. Receiver operating characteristic curve analysis demonstrated a correlation between the results of the applied ELISA methods. One of the most prominent IgE reactive proteins was confirmed to be 45 kDa mycelia catalase.
Our findings confirmed that high molecular weight proteins might play a major role in allergy and IgE reactivity to Aspergillus species. Moreover, the results showed that precipitation and chromatographic methods are applicable for fractionation of fungal proteins such as mycelial catalase.

Considering the increasing drug resistance to Aspergillus fumigatus, search for genes involved in its pathogenicity and identifying alternative antifungal drugs is of utmost importance. The aim of this study was to determine the effects of diclofenac sodium on the growth and sidB gene expression in Aspergillus fumigatus.
In this study, Aspergillus fumigatus was cultured and a fungal suspension prepared, followed by determining the minimum inhibitory concentration of diclofenac sodium at a concentration of 25-1000 µg/ml. Then, RNA was extracted from the suspension at concentrations of 500,700 and 900 µg/ml of the drug. Finally, the extent of expression of the gene was determined by measuring different levels of mRNA-sidB by Real-time PCR.
With increasing concentrations of diclofenac sodium, mycelium production decreased. Concentrations higher than 500 µg/ml had considerable inhibitory effects on the growth of Aspergillus fumigatus.
Findings of this study indicate that diclofenac sodium can cause a sharp reduction in the growth rate of Aspergillus fumigatus. Accordingly, it can be considered as one of the effective pharmacological agents for inhibiting the growth of this fungus.

Background and The presence of Candida yeasts in urine, known as candiduria, is an indicator of infection or colonization of the urinary tract by Candida species. This condition in diabetic patients can be hazardous due to diminished immune system response. The objective of this study was to investigate the incidence of candiduria in diabetic patients and to identify its causative agents. Furthermore, the demographic and laboratory (HbA1c, urine glucose and pH, urine culture colony count, and fasting blood sugar) data and their possible associations with candiduria were investigated.
This cross-sectional, descriptive study was performed on 305 diabetic patients referred to the diabetes research center, Hamedan, Iran, during April 2015 to September 2015. Urine and blood specimens were collected and urine analysis, urine culture, FBS, and HbA1c tests were performed. Positive cases were subjected to colony count and the causative agents were subsequently identified through the routine identification tests, as well as colony color in CHROMagar Candida medium, and the assimilation patterns in API 20C auxanographic method.
Among the 305 cases, 38 (12.5%) were positive for candiduria. Causative agents were identified as Candida glabrata (n=19, 50%), C. albicans (n=12, 31.6%), C. krusei (n=4, 10.5%), C. tropicalis (n=2, 5.3%), and C. kefyr (n=1, 2.6%). According to the results of the statistical analyses, there were significant association between candiduria and female gender, high FBS and urine glucose, uncontrolled diabetes (HbA1c ≥8), and acidic urine pH (P Considering the high incidence rate of candiduria in diabetic patients, control of diabetes, predisposing factors, and causal relationships between diabetes and candiduria should be highlighted.

Acquired azole resistance in opportunistic fungi causes severe clinical problems in immunosuppressed individuals. This study investigated the molecular mechanisms of azole resistance in clinical isolates of Candida glabrata. Six unmatched strains were obtained from an epidemiological survey of candidiasis in immunocompromised hosts that included azole and amphotericin B susceptible and azole resistant clinical isolates. Candida glabrata CBS 138 was used as reference strain. Antifungal susceptibility testing of clinical isolates was evaluated using Clinical and Laboratory Standards Institute (CLSI) methods. Complementary DNA-amplified fragment length polymorphism (cDNA-AFLP) technology, semi-quantitative RT-PCR, and sequencing were employed for identification of potential genes involved in azole resistance. Candida glabrata Candida drug resistance 1 (CgCDR1) and Candida glabrata Candida drug resistance 2 (CgCDR2) genes, which encode for multidrug transporters, were found to be upregulated in azole-resistant isolates (≥2-fold). Fatty acid activator 1 (FAA1) gene, belonging to Acyl-CoA synthetases, showed expression in resistant isolates ≥2-fold that of the susceptible isolates and the reference strain. This study revealed overexpression of the CgCDR1, CgCDR2, and FAA1 genes affecting biological pathways, small hydrophobic compounds transport, and lipid metabolism in the resistant clinical C.glabrata isolates.

Cutaneous fungal infections are common infections, which involve keratinized tissue (skin, hair, and nail). Dermatophytes are important and abundant cutaneous fungal agents. The aim of this study is to assess the epidemiology of dermatophytes and the prevalence of various type of tinea in Tehran province for better identification and treatment of patients.
Five hundred and eight patients suspected to have dermatophyte infections, after clinical examination, were referred to mycology laboratory of Razi Hospital in Tehran city for definite diagnosis, and then using direct examination and slide culture technique, and finally molecular examinations, the causative fungus of infection was diagnosed. Data were analyzed by chi-square statistical test at the significance level of (p£0.05).
Of 161 obtained positive samples, 73 (45.3%) patients were female and 88 cases (54.7%) were male. The mean age of patients was 42 years. The most and least common types of tinea were tinea pedis with 71 cases (44.1%) and tinea faciei with one case (0.6%), respectively. Trichophyton mentagrophytes was the the most isolated species and the least was Microsporum ferrugineum.
According to the results of this study, dermatophyte infections in different parts of the body, especially foot and groin, are considered as an important health problem in Tehran city. It seems that design and implementation of training programs with the purpose of primary and secondary prevention, especially for age group over 20 years, are necessary for reduction of cases of dermatophytosis.

Voriconazole Resistance (VRC-R) in Aspergillus flavus isolates impacts the management of aspergillosis, since azoles are the first choice for prophylaxis and therapy. However, to the best of our knowledge, the mechanisms underlying voriconazole resistance are poorly understood.. The present study was designed to evaluate mRNA expression levels of cyp51A and mdr1 genes in voriconazole resistant A. flavus by a Real-Time Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) technique.. Five A. flavus isolates with resistance to VRC were examined by a RT-PCR approach.. Four out of five isolates revealed cyp51A and mdr1 mRNA overexpression. Interestingly, the isolate, which was negative for cyp51A and mdr1 mRNA expression showed a high voriconazole Minimum Inhibitory Concentration (MIC). Furthermore, a computational-based analysis predicted that voriconazole resistance could be mediated through cooperation with a network protein interaction.. Our experimental and in silico findings may provide new insight in the complex molecular pathways of drug resistance and also could assist design an efficient therapeutic strategy for aspergillosis treatment..

The production and development of an effective fungicidal drug requires the identification of an essential fungal protein as a drug target. Aconitase (ACO) is a mitochondrial protein that plays a vital role in tricarboxylic acid (TCA) cycle and thus production of energy within the cell.. The current study aimed to sequence Candida krusei ACO gene and determine any amino acid residue differences between human and fungal aconitases to obtain selective inhibition.. Candida krusei (ATCC: 6258) aconitase gene was determined by 5’Rapid Amplification of cDNA Ends (RACE) method and degenerate Polymerase Chain Reaction (PCR) and analyzed using bioinformatics softwares.. One thousand-four hundred-nineteen nucleotide of C. krusei aconitase gene were clarified and submitted in Genbank as a partial sequence and then taxonomic location of C. krusei was determined by nucleotide and amino acid sequences of this gene. The comparison of nucleotide and amino acid sequences of Candida species ACO genes showed that C. krusei possessed characteristic sequences. No significant differences were observed between C. krusei and human aconitases within the active site amino acid residues.. Results of the current study indicated that aconitase was not a suitable target to design new anti-fungal drugs that selectively block this enzyme.

Yeasts are one of the most common causes of onychomycosis. In countries like Iran, Saudi Arabia, Italia and Spain, yeasts are reported as the most frequent causes of onychomycosis. Inhibition of the immune system such as chemotherapy, radiation therapy, the use of broad spectrum antibodies, aggressive treatment with corticosteroids, HIV and diabetes melitus predispose the body with these fungal infections. Onychomycosis due to yeast has a higher prevalence in finger nails and its incidence in women is two or three times more than men. In some occupations such as nurses, dish washers, confectionery and housewives it can be more observed. Among the yeasts, Candida albicans is the most common agent onychomycosis. This study has been designed for investigation of prevalence of onychomycosis due to yeast in patients who referred to Razi hospital.

This was a cross sectional study and 700 dystrophic nail samples were examined by both direct examination and culturing. In direct examination KOH (hydroxide potassium) 20% and for culturing saboroud dexterose agar (S) media were used. For identification yeasts complementary examination were done such as: Reynold braude test, API test and culturing on candida chrom agar media and corn meal agar media were used. For investigation of relevance between variables, Chi-square test and Fisher exact test were used.

Of 700 dystrophic nail samples (15.71%), 110 samples were yeast positive by both direct examination and culturing. Thirty one patients were males and 79 patients were females and in both sexes those most infected were between 50-59 years of age (27.3%). Eighty patients had fungal infection of finger nails and 16 patients had fungal infection on toe nails; 14 patients had both infections on finger and toe nails. The most frequent detected yeast species was Candida albicans (42.7%) which was followed by Candida parapsilosis (20.9%), Candida tropicalis (14.5%), Candida krusei (12.7%), Candida glabrata (3.6)%, Candida gillermondi (2.7%), and Candida lousitani, famatata, rodotroula (each 0.9%). The most common clinical type noted was distal subungual onychomycosis in 50% of cases. In this study 52 patients (48.1%) had diabetes which was the most common disease between patients with onychomycosis due to yeasts.

Diagnosis of onychomycosis due to yeast is very important because it shows the person’s immune response. Identification of pathogenic yeast species in terms of epidemiology and selecting appropriate and effective treatment is important.

Background and Dermatophytes are a group of keratophilic fungi some of which produce arthroconidia under invivo conditions and these seem to have an important role in pathogenecity. Arthroconidia formation is a characteristic of dermatophyte infection of skin hair and nail. The present study is intended to study the effects of environmental factors and conventional antifungal drugs on the production of this pathogenic agent in some dermatophytes. Methods and materials: This is a deh1ive analytical study involving the research population of patients with dermatophytosis admitted to Razi hospital in Tehran Iran during 2006-2007. Fifty patients were selected through convenient sampling and were include in the study after direct microscopic examination confirmed the disease. In the present study the environmental factors including the media (SDA SDA+NaCl 1% 3% and 5% Trichophyton agar no.1 and SDB) temperature PH CO2 and the conventional antifungal drugs (Geriseofulvin Clotrimazole Itraconazole Terbinafin and Betametasone) were observed for their effects on arthroconidia production in Trichophyton Verucosum Trichophyton riolaseum Epidermophyton floccosum Trichophyton rubrum and Trichophyton Mentagrophytes. The obtained data were analyzed using Chi- Square and student t-test. The highest production rate of arthroconidia occurred in SDA with PH (7.5) CO2 pressure 10% and temperature of 37ºC after 10 days. No growth was observed at the temperature of 42ºC and in a media of NaCl 3% or higher. Geriseofulvin Clotrimazole and Betametazone stimulated arthroconidia production but Itraconazole and particularly Terbinafin stopped and controlled its production. The results of this study emphasized the importance of arthroconidia production and its being influenced by environmental factors such as PH CO2 pressure and media in dermatophytes.