فهرست مطالب somayeh sharifynia

One important complication of the coronavirus disease 2019 (COVID-19) is COVID Associated Mucormycosis (CAM), especially in patients with conditions such as diabetes and in immunosuppressed patients. Systemic acidosis, hyperglycemia, and other biochemical factors such as free iron and β-hydroxybutyrate (BHB) can play a role in this complication.

Rhizopus oryzae was isolated from a patient at Masih Daneshvari Hospital microbiology laboratory and sub-cultured on the Potato Dextrose Agar (PDA) for 48 hours at 37 ◦C. Subsequently, Roswell Park Memorial Institute (RPMI) 1640 Broth medium buffered to pH 7.0 with 3-N-morpholino-propane sulfonic acid. Macrodilution and microdilution methods were performed with 8.4% sodium bicarbonate. After 24 hours of incubation at 35°C, the minimum inhibitory concentration (MIC) and the minimum fungicidal concentrations (MFC) were evaluated.

We found that the minimum inhibitory and fungicidal concentrations are at 1.05 % and 2.1 % respectively. Therefore, the minimum concentration is 2% sodium bicarbonate, which requires achieving the desired environmental pH for fungal inhibition and fungicidal effects.

Regulation of systemic acidosis by sodium bicarbonate could be used to decrease the chance of mucormycosis. In addition, According to our study and some others, an alkaline environment can prevent fungal growth. We found that a minimum concentration of 2% sodium bicarbonate is required to achieve the desired mucosal pH to inhibit the fungus. Therefore, sodium bicarbonate inhalation, as a cost-effective and well-tolerated medicine, is a good candidate for the prevention of mucormycosis. In this regard, extensive clinical and laboratory research is needed to achieve more accurate doses and appropriate administration intervals.

 Nowadays, non-albicans Candida are common in human pathogens, and some of these cases were found in milk. Therefore, as well as the lack of accurate estimates of its global prevalence and severity, the present study aims to assess the demographic features of non-albicans Candida (NAC) spp. and determine the species distribution of NAC. It was also evaluating the in vitro Azole susceptibility of NAC species and identified the erg11 gene and erg11 expression in fluconazole-resistant isolates of NAC spp., in Iran.

 In the present study, non-albicans Candida, including Candida glabrata, Candia krusei, Candida parapsilosis, and Candida tropicalis, were isolated and identified from 14 farms (raw milk) and human patients using culture methods, Real-Time PCR and sequencing. The resistance and susceptibility of the samples to azole were examined and erg11 expression was evaluated by RT-qPCR. The results were analyzed by REST Software to compare the levels of erg11 gene expression involved in drug resistance of NAC.

 74 and 52 NAC strains were isolated in 262 collected milk samples and human samples. Based on ITS sequencing, 0.76% were identified as C. glabrata, 2.29% C. tropicalis, 4.19% C. parapsilosis, and 19.8% C. krusei. The expression of erg11 gene in the NAC was increased in samples isolated from humans compared to samples isolated from livestock (P>0.05), while no significant difference was found in the case of Candida glabrata isolated from both sources (P<0.05). All NAC isolates were sensitive to flucytosine.

 NAC isolates from cows' milk have antifungal resistance genes while they had not taken any antifungal drugs. The resistance gene is transferred from antifungal agents in crop protection medications. Clinical isolates also had increased resistance to antifungal activity. Also, using Azole antibiotics can increase resistance gene level activity. This phenomenon should be considered for treatment program protocols.

Candida albicans complex species are well known as the main cause of candidiasis, particularly among susceptible individuals. In this study, we report the genetic diversity of Candida spp. and the antifungal susceptibility pattern of the cryptic C. albicans complex isolates in Kerman, Iran.

A total of 112 yeast isolates were obtained from different clinical samples, and molecular identification was performed. All C. albicans complex isolates were tested for susceptibility of them to amphotericin B, fluconazole, and itraconazole.

The majority of clinical isolates were C. albicans complex (n=48) followed by C. glabrata complex (n=34), C. parapsilosis complex (n=21), and C. krusei (n=9). Among C. albicans complex, 45 isolates were C. albicans (94%), 2 isolates were C. dubliniensis (4%), and 1 isolate was C. africana (2%). Amphotericin B was the most active antifungal, whereas 8.9% and 6.7% of the isolates were resistant to fluconazole and itraconazole, respectively.

Regarding the high incidence of Candida infections particularly in susceptible populations and the emergence of an infrequent yeast species with elevated MICs, which is indistinguishable with conventional methods, developing accurate molecular methods for laboratory diagnosis should be considered in the clinical setting.

Fusarium species are known to be one of the common causes of keratitis. This study was conducted to identify Fusarium spp. causing keratitis and to investigate their genetic diversity using TEF1 and RPB2 gene sequences.

Twenty-four clinical isolates of Fusarium were isolated from the patient with keratitis. Phylogenetic analysis of two-locus of the 24 clinical isolates and three reference strains was carried out using the maximum parsimony and RAxML methods.

Based on gene sequences of the 24 clinical isolates, 17, 4, and 3 isolates were identified as Fusarium solani species complex (FSSC), Fusarium fujikuroi species complex (FFSC), and Fusarium oxysporum, respectively. FFSC include F. proliferatum (n=1), F. globosum (n=1), F. verticillioides (n=1), and F. brevicatenulatum (n=1), respectively.

Given that sequence of a sole gene can be challenging and on the other hand, due to the high resistance to antifungal drugs, identification of Fusarium species is of substantial significance. In this study, by designing a novel set of primers for the RPB2 area and using TEF1 primer, we were able to differentiate 24 Fusarium spp. isolated from patients with keratitis.

Aspergillus fumigatus as a ubiquitous fungus can be found in the respiratory tract of the asthmatic and healthy people. The inhalation of Aspergillus spores leads to an immune response in individuals with asthma and results in the aggravation of the clinical symptoms. The present study aimed to investigate the prevalence of specific immunoglobulin E and G (IgE and IgG) against A.fumigatus in asthmatic patients. This study was conducted on 200 consecutive patients with moderate to severe asthma referring to Masih Daneshvari hospital Tehran, Iran, from January 2016 to February 2018. Skin prick test (SPT) was performed in all subjects with Aspergillus allergens. Moreover, all patients underwent specific IgE testing for Aspergillus using Hycor method. Enzyme immune assay was applied to measure total IgE and Aspergillus-specific IgG. According to the results, the mean age of the patients was 45.8 years (age range: 18-78 years). The mean levels of total IgE and Aspergillus specific IgE in asthmatic patients were obtained as 316.3 (range: 6-1300 IU/ml) and 1.5 (range: 0.1- 61.3 IU/ml), respectively. Out of 200 patients, 27 (13.5%), 65 (32.5%), 22 (11.0%), and 86 (43.0%) cases had positive Aspergillus SPT, total IgE of > 417 IU/ml, Aspergillus-specific IgE, and IgG, respectively. The level of these variables in patients with severe asthma were 16 (16.5%), 36 (37.1%), 15 (15.5%), and 46 (47.4%), respectively. As the findings indicated, reactivity to Aspergillus is a remarkable phenomenon in asthmatic patients. It is also emphasised that the climatic condition may affect the positive rate of hypersensitivity to Aspergillus.

Rapid and accurate identification and evaluation of antifungal susceptibility pattern of Candida isolates are crucial to determine suitable antifungal drugs for the treatment of patients with vulvovaginitis candidiasis.
Vaginal samples were collected from 150 women with suspicious vaginal candidiasis, and then cultured on Sabouraoud’s Dextrose Agar with chloramphenicol to isolate Candida species. After identification of Candida isolates using polymerase chain reaction?restriction fragment length polymorphism technique, antifungal susceptibility testing of four azolic antifungal drugs was carried out using broth microdilution method according to the CLSI M27?A3.
Candida species were isolated from eighty suspected patients (61.79%). The most common pathogen was Candida albicans (63.75%). Resistance to fluconazole and ketoconazole was observed in 27.5% and 23.75% of Candida isolates, respectively, and only 2% of Candida isolates were resistant to miconazole. Interestingly, resistance to fluconazole in C. albicans was more than other Candida species.
The results indicated that therapy should be selected according to the antifungal susceptibility tests for the prevention of treatment failure and miconazole therapy can be considered as the best therapeutic choice in the
management of vulvovaginitis.

Background and Aspergillosis is one of the most common opportunistic fungal infections in immunocompromised and neutropenic patients. Aspergillus fumigatus (A. fumigatus) is the most common causative agent of this infection. Due to variable CO2 concentrations that pathogens are exposed to during the infection process and to understand the role of CO2, we examined the effects of various CO2 concentrations as one of the environmental factors on morphological changes and induction of antifungal resistance in A. fumigatus.
A. fumigatus strains were cultured and incubated under 1%, 3%, 5%, and 12% CO2 atmospheres, each time for one, two, and four weeks. The control culture was maintained for one week without CO2 atmosphere. Morphological changes were investigated and antifungal susceptibility test was performed according to the recommendations of the Clinical and Laboratory Standards Institute (CLSI) M38-A2 document. The results of different CO2 atmospheres were compared with that of the control sample.
We found that 1%, 3%, 5%, and 12% CO2 atmospheres were associated with morphological colony changes. Macroscopically, the colonies were shallow dark green, smooth, crisp to powdery with reduced growth; microscopic examination revealed the absence of conidiation. The induction of antifungal resistance in the susceptible strains to itraconazole, voriconazole, and amphotericin B increased after exposure to 12% CO2 atmosphere and four weeks of incubation. The MIC values for itraconazole, voriconazole, and amphotericin B were 16 g/ml, 1 g/ml, and 16 g/ml, respectively. These values for the control group were 0.125 g/ml, 0.125 g/ml, and 2 g/ml, respectively.
Exposure to different CO2 atmospheres induced morphological changes in A. fumigatus, it seems to increase the MIC values, as well. In parallel, resistance to both itraconazole and voriconazole was also observed.

Aflatoxins are highly toxic secondary metabolites mainly produced by Aspergillus parasiticus. This species can contaminate a wide range of agricultural commodities, including cereals, peanuts, and crops in the field. In recent years, research on medicinal herbs, such as Pistacia atlantica subsp. kurdica, have led to reduced microbial growth, and these herbs also have a particular effect on the production of aflatoxins as carcinogenic compounds.
In this study, we to examine P. atlantica subsp. kurdica as a natural compound used to inhibit the growth of A. parasiticus and to act as an anti-mycotoxin.
In vitro antifungal susceptibility testing of P. atlantica subsp. kurdica for A. parasiticus was performed according to CLSI document M38-A2. The rate of aflatoxin production was determined using the HPLC technique after exposure to different concentrations (62.5 - 125 mg/mL) of the gum. The changes in expression levels of the aflR gene were analyzed with a quantitative real-time PCR assay.
The results showed that P. atlantica subsp. kurdica can inhibit A. parasiticus growth at a concentration of 125 mg/mL. HPLC results revealed a significant decrease in aflatoxin production with 125 mg/mL of P. atlantica subsp. kurdica, and AFL-B1 production was entirely inhibited. Based on quantitative real-time PCR results, the rate of aflR gene expression was significantly decreased after treatment with P. atlantica subsp. kurdica.
Pistacia atlantica subsp. kurdica has anti-toxic properties in addition to an inhibitory effect on A. parasiticus growth, and is able to decrease aflatoxin production effectively in a dose-dependent manner. Therefore, this herbal extract maybe considered a potential anti-mycotoxin agent in medicine or industrial agriculture.

Candidiasis, the main opportunistic fungal infection has been increased over the past decades. This study aimed to characterize C.albicans species complex (C.albicans, C.dubliniensis, and C.africana) isolated from patients with respiratory infections by molecular tools and in vitro antifungal susceptibilities by using broth microdilution method according to CLSI M27-A3 guidelines. Totally, 121 respiratory samples were collected from patients with respiratory infections. Of these, 83 strains were germ tube positive and green colonies on chromogenic media, so initially identified as C.albicans species complex and subsequently were classified as C.albicans (89.15%), C.dubliniensis (9.63%), and C.africana (1.2%) based on PCR-RFLP and amplification of hwp1 gene. Minimum inhibitory concentration (MICs) results showed that all tested isolates of C.albicans complex were highly susceptible to triazole drugs. However, caspofungin had highest activity against C.albicans, C.dubliniensis, and C.africana. Our findings indicated the variety of antifungal resistance of Candida strains in different areas. These results may increase the knowledge about the local distribution of the mentioned strains as well as their antifungal susceptibility pattern which play an important role in appropriate therapy.

Cryptococcosis is a fungal infection which caused by various Cryptococcus species. The majority of patients are immunodeficienct cases. Cryptococcosis is known as one of the four fatal infectious disease in patients suffering from AIDS. The current study presents a case of cryptococcal meningitis in patient with HIV, and also briefly reviews the cases of cryptococcosis in Iran over the past decades. In February 2017, a patient with fever, chills and headache was admitted to the Dr Masih Daneshvari hospital. The microscopic examination of cerebrospinal fluid (CSF) specimen, showed yeast cells, followed by serological and molecular tests which confirmed the diagnosis. The primary diagnosis was cryptococcal meningitis and patient was treated with amphotericin and fluconazole. Generally in countries with poor epidemiological information on cryptococcosis, which usually have restricted accurate diagnostic facilities in their laboratory centers, the true incidence of the disease cannot be assessed easily. Furthermore, regarding to importance of this disease, especially in immunocompromised individuals; early diagnosis and proper treatment of the patients have crucial roles in mortality rate reduction in infected patients.

As an important health care problem, nosocomial infections are associated with high mortality rate and increasing hospital costs. The pathogenic organisms of these infections usually transmit to patients via hospital personnel and medical devices. Among them, Candida species are the most common fungi isolated from biofilms. Particularly, the biofilms formed by these organisms are identified in immunocompromised patients. The aim of this study was the molecular identification of isolated Candida spp. from endotracheal tubes of ICU patients.Ninety five intubated and mechanically ventilated ICU patients were evaluated for the presence of endotracheal tube biofilms. The endotracheal tubes were cultured on Sabouraud Dextrose agar. Using PCR-EFLP method, the ITS1-5.8S-ITS2 region was amplified and digested by the restriction enzyme MspI.By the evaluation of 51 isolates which were derived from all samples, C. albicans was the most frequently isolated species 20 (39.2%), followed by non albicans spp. of C. glabrata 14 (27.4%), C. tropicalis 8 (15.6%), C. krusei 6 (11.7%), and C. parapsilosis 3 (5.8%).The results show that Candida albicans was the most frequent species isolated from endotracheal tubes. Knowledge about the species identification of Candida biofilms can be helpful for clinicians to manage intensive care unit patients.

Shallots are an important part of the diet of many population and there is long-held belief in their health enhancing properties. At least one episode of vaginal candidiasis is reported in up to 75 percent of women that Candida albicans is responsible for 80 to 92 percent of episodes of vaginal candidiasis in the world. The aim of this study was to determine anticandidal activity of shallot against chronic candidiasis agents. This study is an experimental study. With use of microdilution method, antifungal activities of alcoholic and aqueous extracts of shallot (allium hirtifolium) were tested against 33 candida species in vitro that isolated from patient with chronic candidiasis that referred to Mirza Koochak khan and Lolagar hospitals. The results showed antifungal activity of Allium hirtifolium against all the candida species tested and anticandidal activity of the alcoholic extract was much better than aqueous one. The results indicated that crude juice of shallot has anticandidal activity and might be promising in treatment of candidiasis.