رضا کچویی

Fungal diseases, especially superficial and cutaneous forms (dermatophytes) are of great public health importance due to the possibility of transmission from people to others, especially where people live in groups. Lack of knowledge about how to prevent the transmission of these diseases can cause them to be transmitted to numerous people and carry a large financial burden and even significantly reduce the efficiency of people. Therefore, informing different segments of society about the prevalence of fungal diseases, ways and symptoms of its infection, ways to prevent its transmission to others and necessary measures in case of infection, can play a significant role in promoting public health. Dermatophyte infections are one of the most important health problems in the world and affect millions of people worldwide. It is estimated that 10-20% of people will develop these infections in their lifetime. The distribution of these infections varies in different countries in the world and it is more prevalent in developing countries than in industrialized countries. So far, more than 40 species of dermatophytes have been identified, which are classified into three genera: Microsporum, Trichophyton, and Epidermophyton. Dermatophytosis infection has several clinical forms that are classified according to the location of the infection; fungal infection of the feet, arms, legs, trunk, hands, palm and groin area, the scalp and hair the fingernails and toenails, and the nail bed. In addition to dermatophytes, other filamentous fungi can cause superficial and cutaneous infections. Yeasts such as Candida can also cause superficial and cutaneous infections. These diseases always cause problems such as pain, discomfort, disability, time consuming and high treatment costs. To combat dermatophytosis, it is necessary to know the regional dermatophytes and the extent of their spread, and also having instructions for the prevention of fungal diseases that its implementation can be effective in reducing these diseases.

Candida, is a fungal flora in human that can be colonized on the skin and mucosal surfaces. Various factors are involved in causing disease by candida, one of the most important of these factors, are secreted hydrolytic enzymes and aspartyl proteinase enzymes are as a potential factor, but other secretion of hydrolytic enzymes such as phospholipase and lipase are also important pathogen factors. In this study, gene expression of LIP2, LIP3 and LIP5 were studied in isolated C. albicans in 100 samples from patients.
A total of 100 strains of C. albicans was isolated in 2015-2016 years by morphology and molecular methods, from clinical samples. After extracting RNA from them by RNX-Plus kit, expression of LIP2, LIP3 and LIP5genes was evaluated by using RT-PCR test.
From 100 clinical isolates of C. albicans, LIP2 gene expressed 55%, LIP3 gene 41% and LIP5 gene 48%. Among the positive samples, 24 cases (33.80%) were expressed three genes at the same time.
In this study, frequency of lipase genes was different in strains of C.albicans. Lipase gene was not expressed in 30% of cases. It can be concluded that other genes than lipase gene can be involved in the pathogenesis.

The knowledge and recognition of fungal diseases in each area is extremely important. The aim of this study was the survey and molecular identification of Malassezia species which were isolated from pityrosporosis and tinea versicolor diseases in military forces on three Iranian islands of Abu-Musa, Great Tonb and Sirri’s by PCR-sequencing method.
This cross-sectional study was performed during October - September 2011 in Abu-Musa, Great Tonb and Sirri’s islands (Persian Gulf) on military individuals. The samples taken from patients with suspected fungal lesions were studied by mycological (direct, culture) and molecular methods (PCR-sequencing).
In this study, 102 military forces with the mean age of 26 years within the range of 19-53 years old were studied. Among them, 17 patients (16.7 %) were infected with fungal diseases of pityrosporosis (dandruff) (58.8 %), tinea versicolor (35.3 %) and dermatophytosis (5.9 %) respectively. The most frequent species of Malassezia isolated from dandruff and tinea versicolor were M. restricta (80%) and M. globosa (67%), respectively.
In this study, the most frequent Malassezia species isolated from dandruff and tinea versicolor were M. restricta and M. globosa, respectively. In this study, the manual method of DNA extraction from direct sample scrapings was used which has been rarely reported in Iran.

Identification of Nocardia spp. in routine medical laboratories is based on phenotypic methods that is often time- consuming. The objective of this study was to diagnose Nocardia agents by PCR technique in BAL samples of patients with suspected tuberculosis admitted to hospitals in Tehran. In this study, 116 BAL samples of patients admitted in Baqiyatallah, Imam Khomeini and Shariati hospitals were collected within 8 months. Nocardia DNA was extracted by phenol-chloroform protocol. In Duplex PCR, primers NG1 and NG2 were used to amplify a Nocardia genus- specific 598-bp fragment of 16S rRNA gene. BetF and BetR primers were used as housekeeping gene to amplify 204-bp fragment. Gel-purified PCR products were sequenced. Using Duplex PCR, 7 (%6.03) samples were positive for Nocardia spp.. Sequencing results showed that the species identified were N.cyriacigeorgica (6 case) and N. otitidiscaviarum (1 case). In the present study, DNA extracted from Nocardia Spp. in BAL specimens by manual method during minimum time which had not previous record in Iran. The study also was carried out on patients with suspected tuberculosis which restricted work has already been done on them. In this study, N. cyriacigeorgica was dominant species that has been introduced during recent years, which should be considered in clinical laboratories and research centers.

Aspergillus fumigatus is the cause of more than 90% of invasive pulmonary aspergillosis. The aim of this study is identification and detection of A. fumigatus in bronchoalveolar lavage (BAL) samples patients suspected to have tuberculosis by using of Nested-PCR. In this assay، a set of 100 BAL samples which were collected in 9 month، between the years 2012-2013، from patients with suspected tuberculosis in Tehran hospital، were examined by direct، culture and molecular tests. Manual phenol-chloroform method was used in order to extract DNA. Specific primers of Aspergillus genus and beta-actin primers were used for duplex-PCR. A pairs of internal primers were utilized in order to identification of A. fumigatus (Nested-PCR method). Finally، some of the PCR-products were sequenced for confirming results. Overall، 12 samples (12%) were aspergillus-positive by molecular test while 11% and 10% samples were positive by direct and culture test، respectively. The sensitivities and specificities of PCR method compare with direct examination was 100% and 98. 8%، respectively. The study also showed that sensitivities and specificities of PCR method compare with culture was 100% and 97. 7%، respectively. After sequencing PCR product sample، isolated species were recognized as A. fumigatus. This study showed that PCR method in compare with the direct method and the culture has a higher sensitivity in the detection of Aspergillus species، also Present study displayed that can be consider Aspergillus species and A. fumigatus in suspicious to tuberculosis BAL samples

Erythrasma is a chronic superficial infection of the intertriginous areas. Most laboratories use methylene blue stain and 10% KOH smear to identify Corynebacterium minutissimum (C. minutissimum) by direct observation of filamentous bacilli. Occasionally atypical forms can be seen that create problems in diagnosis. This study aims to use the polymerase chain reaction (PCR) method in order to definitively identify C. minutissimum as an agent of erythrasma. This research was performed during 2013 on 100 skin scrapings suspicious for erythrasmawhich were obtained from various medical mycology laboratories in Tehran. Samples were tested by three methods - direct examination، culture and PCR. DNA was extracted by the modified phenol-chloroform method after which PCR was performed using designed primers. We sequenced some of the PCR products. The sensitivity and specificity of the PCR method was compared to the direct and culture examinations. Of the 100 samples، there were 25 positive samples according to PCR analysis، 13 positive by direct examination and 23 that cultured positive. DNA sequencing results showed the presence of C. minutissimum. The PCR method in comparison with direct examination had a sensitivity of 100% and a specificity of 86. 2%. The study also showed that the PCR method in comparison with culture had a sensitivity of 100% and a specificity of 97. 4%. This study showed that the PCR method in comparison with the direct method and culture had a higher sensitivity in the detection of C. minutissimum. The present PCR method confirmed all typical and some of the atypical forms of C. minutissimum which indicated the importance of this method in the definitive diagnosis of erythrasma.

Onychomycosis is caused by dermatophytes, yeasts, and molds. The term onychomycosis refers to nail infections caused by dermatophyte and non-dermatophyte fungi. The aim of this study was diagnosis of onychomycosis agents and also comparison of the accuracy and sensitivity of PCR to conventional methods such as direct examination and culture. Our descriptive-experimental research is based on the usage of standard solid and liquid cultures of Trichophyton, Candida and Aspergillus for molecular testing on clinical specimens from patients. Universal fungal primers (ITS1 & ITS4) and specific Trichophyton rubrum primers (T-rub) were applied in this experiment. For the first time in Iran using an innovative protocol, direct DNA extraction and PCR confirmation of mediums mentioned above was performed in two hours. Furthermore, better sensitivity and precision of PCR rather than routine laboratory results such as direct examination and culture have been proved. Due to the high speed and precision of PCR method, it can be a convenient alternative for conventional methods such as direct examination and culture on clinical samples of onychomycosis.

Fusarium species are prevalent contaminants of foodstuffs and agricultural crops. They produce fumonisins، which are carcinogenic mycotoxins. The present study has evaluated maize and wheat samples from ten provinces in Iran that were contaminated with Fusarium species. Special attention was paid to the ability of the isolates to produce fumonisin B1 (FB1) as a public health hazard. We collected 32 maize and 15 wheat samples from ten provinces that were major cultivation areas. Samples surface disinfected with a 1% sodium hypochlorite solution for 2 minutes. Fusarium species were isolated by the flotation method on malachite green agar. Pure cultures on potato dextrose agar (PDA) were identified using a combination of macroscopic and microscopic morphological criteria. The ability of the isolates to produce FB1 was evaluated by thin layer chromatography (TLC) and the amounts of fumonisin B1 produced were assessed by high performance liquid chromatography (HPLC). A total of 55 Fusarium isolates that belonged to five species were isolated. There were 27 of the 32 maize samples (84. 4%) and 11 of 15 wheat samples (73. 3%) that were contaminated with Fusarium species. Species consisted of F. verticillioides (23 isolates)، F. proliferatum (22 isolates)، F. subglutinans (5 isolates)، F. nygamai (4 isolates) and F. redolens (1 isolate) based on morphological criteria. Twenty-two of the 55 (40%) Fusarium isolates produced FB1 in a total range from 230. 4 to 9565 µg/ml. The highest amounts of FB1 production were related to toxigenic isolates of F. verticillioides and F. proliferatum. Results of the present work indicates a high degree of contamination of maize and wheat with Fusarium strains that belong to the Gibberella fujikuroi species complex وparticularly F. verticillioides and F. proliferatum. This contamination is a potential public health threat due to food spoilage and subsequent production of high levels of carcinogenic FB1.

Approximately three-fourths of women experience an episode of vaginal candidiasis. Candida albicans(C. albicans) is the etiological agent in over 80% of cases. C. albicans has numerous virulence factors such as the agglutinin-like sequence (ALS) genes which code the large glycoprotein family that has a role in the adherence of Candida. The present study aims to evaluate expressions of the ALS 2, 9 genes in C. albicans which have been isolated from vaginal candidiasis.

We collected 150 wet vaginal swabs from patients diagnosed with vaginal candidiasis. Samples were cultured on sabouraud dextrose and CHROMagar for morphological analysis. Then, DNA was extracted by glass bead and lysis buffer. We performed RFLP-PCR to confirm the presence of C. albicans. For investigation of semi-quantitative expression of ALS2 and ALS9 genes, we performed RT-PCR by using specific primers.

From 55 clinical isolates of C. albicans, 36.36% expressed both the ALS2 and ALS9 genes. There were 23 (41.81%) isolates that expressed only the ALS2 gene and 21 (38.1%) expressed the ALS9 gene.

Expressions of the ALS9 (41.8%) and ALS 2 (38.1%) genes in Candida isolates may indicate that these genes play a critical role in adhesion and biofilm formation of vaginal infection. However the presence of both genes in 36.36% of the isolates suggests a positive role for these genes in augmentation of their activities.

Ochratoxin A (OTA) is a mycotoxin that possess a risk to human health due to its nephrotoxic, immunotoxic, mutagenic, teratogenic and carcinogenic properties. This study was undertaken in order to determine the presence and levels of ochratoxin A in different types of bread consumed in Shahrekord city. Eighty six samples of different types of bread purchased in March 2012 from retail bakeshops in Shahrekord city were surveyed for the presence of ochratoxin A (OTA) using ELISA assay. Ochratoxin A was detected in 45 out of the 86 bread samples (52.3%). Levels of OTA in positive samples ranged between 0.19 and 10.37 ng/g and the average contamination of all positive samples was 3.04 ng/g. The highest frequency of positive samples was related to machinery Taftoon (88.8%) and Lavash bread (81.8%). The most contaminated sample (5.39 ng/g) was found in the Iranian Lavash bread. Fifteen of the positive samples exceed the maximum level of 5 ng/g set by European regulations for OTA in cereal and bread. The results of this study indicated that contamination levels of ochratoxin A were high in part of the samples (17.4%). Bread and cereals are considered to be the main and predominant ingredient of Iranian food; therefore, their contamination can have long-term negative impact on people's health.

Dermatophytosis is one of the most common pandemic fungal infections that is a major health problem in cities and villages. This study aims to evaluate PCR sensitivity and accuracy in the detection of nail dermatophytosis compared to conventional direct and culture detection methods، and performs an assessment of Trichophyton rubrum in patients suspected of having nail dermatophytosis. This experiment was a descriptive-experimental study carried out on 71 nail samples obtained from patients with suspected nail dermatophytosis. All clinical samples of nails or chips were divided into three sections and each section underwent direct examination، culture and molecular tests. In the molecular test، we used fungal rRNA universal primers (ITS1 and ITS4) and Trichophyton rubrum-specific primers. In this study، for the first time in Iran and based on a modified protocol، DNA was directly extracted from tissues of infected nails in less than five hours. Additionally a comparison of the results obtained from routine laboratory methods such as direct examination and culture with PCR verified the high sensitivity and accuracy of PCR compared to the other studied methods. PCR، as a rapid، accurate method، can be a good replacement for conventional culture and direct examination.

Mycophenolic acid(MPA), a fungal mycotoxin, is produced by Penicillium brevicompactum and is used for the synthesis of immunosuppressive drugs in pharmaceutical industries. The present study was conducted to evaluate the possibility of mycophenolic acid(MPA) production by standard strains of P. brevicompactum at laboratory level. Three strains of P. brevicompactum were provided from microbial culture collections. To stimulate MPA production, barley was used as culture medium, and dry heat, wet heat, and gamma radiation were used to sterilize the culture medium. Samples were taken from the culture medium at different intervals, and their MPA level was assessed by HPLC method. P. brevicompactum strain which was prepared from Finland(VTT D-061157) was able to produce MPA more than two other strains(from Germany and Iran). The amount of MPA enhanced linearly until day 10, and after that became relatively constant. Gamma radiation was a suitable method to sterilize the substrate, and nylon bags were evaluated as an easy and cheap container for growing the fungus. Production of MPA with simple and cheap culture media to provide primary substance for immunosuppressive drugs such as mycophenolate mofetile and sodium mycophenolate would be possible.

T-2 mycotoxin is the most toxic trichothecene produced by Fusarium species especially Fusarium sporotrichioides. This species is the contaminant of cereal grains that can cause severe diseases among humans and animals and it may lead to death. Tri16 gene encodes an acyltransferase that catalyzes the formation of ester side groups at C-8 in biosynthesis pathway of T-2 toxin. The aim of the present study was to design a molecular method for identification of Fusarium species based on Tri16 gene, capable of producing T-2 toxin. In this experimental study, the DNA of 111 T-2 producing and nonproducing Fusarium strains (including 89 Fusarium isolates and 22 standard Fusarium spp.) were evaluated. After purifying the strains and preparing the cultures in Potato Dextrose Broth (PDB), the DNA was extracted using liquid Nitrogen and lysing buffers and by Phenol, Chloroform and Isoamyl alcohol (PCI) solutions. The gene was detected in all T-2 producing and nonproducing strains using the designed and selected primers based on Tri16 gene and PCR optimization. All T-2 toxin producing Fusarium strains including F usarium sporotrichioides (4 species), F usarium langsethiae (1 species) and F usarium Poae (1 species) contained the Tri16 gene fragment (1380 bp). Important T-2 toxin producing species such as Fusarium sporotrichioides and Fusarium langsethiae could be identified with the designed molecular protocol designed based on Tri16 gene. In addition, weakly T-2 producing species can be identified with this method.

The aim of this research is to explore managerial factors affecting the success of new product development projects in small and medium sized enterprises. To find out these factors, deep interviews are conducted with successful managers in the field of new product development. Afterwards, a survey is conducted to production/sales/marketing/R&D managers and chief executive officers of small and medium sized enterprises to elicit their points of view. This study examines five factors assumed to be critical to influence on the top managers to achieve their success in the new product development. These factors include commitment, entrepreneurial orientation, social capital, international orientation and tendency to future studies of top managers.

Aims. Fusarium species are the most important group of fungi that produced a variety of mycotoxins which can contaminate food. The objective of the present study was presentation and identification of one isolate from Fusarium species belong to section of sporotrichiella, that recently isolated from stored wheat of Tehran province. Methods. This species isolated and purified by freeze blotter and single-spore method, respectively. Isolates cultured onto CLA, SNA, PDA and PSA. All microscopic and macroscopic characters studied. The DNA of this isolate amplified by PCR with F. sporotrichioides and F. langsethiae specific primers. Internal transcribed spacer (ITS) regions of isolate amplified, purified and sequenced using universal primers ITS4 & ITS5 and the fraction of TEF-1α gene also using external and nested primers. Results. On the basis of morphology and ribosomal internal, transcribed spacer (ITS) classified as F. langsethiae but on the basis of partial translation TEF-1α gene grouped with F. sporotrichioides. Conclusion. To our knowledge, this is the first report in Iran and Asia. Therefore, a large-scale monitoring of these fungi and T-2 toxin on wheat in Iran is necessary.

Introduction. Surveillance of natural occurrence of T-2 toxin in cereals intended for human consumption.Materials and Methods. Cereal samples obtained from nine cooking centers of Tehran. The samples were powdered and extracted by methanol-water solution (70:30) and the level of their T-2 toxin assayed using a competitive monoclonal antibody based ELISA.Results. All cereal samples contained T-2 toxin ranging from 7.9 to 65.4 (mean: 17.9±2.1) µg/kg. Wheat samples with mean of 42.4 µg/kg were the most contaminated cereals flowed by barley and rice with 18.3 and 12.5 µg/kg respectively. Conclusion. Although, a minority of cereal samples (13.9 %) was contaminated with levels higher than nation standard of Iran for T-2 toxin, extended contamination with the toxin reveals existence of favorable condition for natural occurrence of mycotoxins in agricultural products, and mentions the risk of chronic effects of the toxin exposure and necessity of foodstuffs mycotoxins assay.

This research was performed to determine the level of the humoral immunity in military persons vaccinated against meningococcal meningitis type A and C in IRAN. Each person was given a subcutaneous injection of 50µg of group A and C polysaccharides vaccine. blood samples were collected from 150 combatants before vaccination as well as at 1,4,7 month after vaccination. Antibody level in the sera was measured by passive hemaglutination. A titer 16 (1-2 µg/ml) in the serum was considered as protective. 3.3% of persons had a protective level of antibody in the circulating before vaccination. One month after vaccination, 89% of persons had protective level of antibody in their sera. The circulating antibody level in only 7.4% of vaccinated did not rise to a protective level. About 3.3% vaccinated did not produce specific antibody at all. The antibody level in vaccinated persons four month after vaccination not only did not rise but also it showed a decrease in circulating antibody level, but the number of combatants with protective levels of antibody did not decrease. The antibody levels in vaccinated combatants seven month after vaccination continued to fall, and in 18% of combatants the antibody levels fell below the protective level. Regarding to a relative low level of circulating antibody after vaccination in immune combatants and a lower rate (82%) of immune individuals (as compared with the results of other investigations who report an immune rate of %90 up to 95%) as well as the continuous fall in circulating antibody after very short rise, the results of present investigation indicate that the polysaccharide vaccine which was used in this study does not provide a high rate of protective immunity. Probably, type of vaccine is important and in other researches used vaccines of another companies.

The most important subcutaneous fungal infections in man are caused by injury due to contaminated thistles and thorns. From an epidemiological point of view , it is important to recognize the fungi as well as their frequency of them in various thistles and thorns

The present research has been conducted on thorns and thistles of 16 regions including cities and villages of Isfahan province. 800 samples have been collected. Specimens were inoculated and incubated at 25◦C in S & SCC medium. In order to isolate and identify the fungi, cultures in specific media, intraperitoneal injections of mice and disc diffusion test have been applied.

Over one year of study, 1676 colonies of actinomycetes and fungi were recognized. The most common fungi were as follows: Alternaria (22/4%),Aspergillus(11/8%) , Cladosporium (10/8%),Esteril mycellium(10/6%) and Penicillium (9/7%). The prevalence was most in Shahreza city(10/2%) , while the least was in Ardestan(3%). The most prevalent yeasts were Candida tropicalis (50%) , Rodotrula rubra (12/5%) , Candida kerusei(11/4%),Trichosporon candida (7/9%) , Unknown yeasts (6/8%), Candida gillermondi (5/7%), Saccharomyces cervisia (3/4%), Geotricum candidum and Trichosporon glabrata (Candida glabrata ) each one (1/1%). The prevalence was most in Khansar city(19/2%). In this study, 4 species similar to Coccidioides immitis, Phialophora verrucosa (4 species) and Exophiala jeanselmei (3 species) were identified.

In this study done for the first time in this area, pathogenic and opportunistic fungi were isolated. Furthermore, Exophiala jeanselmei and Nocardiopsis dassonvillei were isolated for the first time from thorns in the country.

Interoduction. T-2 toxin, an important member of trichothecene mycotoxins family, disturbs ‎structures and functions of many parts of human and animal immune system. Our previous ‎studies showed that peritoneal inoculation of T-2 toxin changes normal proportion of mouse ‎peripheral blood lymphocytes and antioxidants may affect the phenomenon. In the present study, ‎the effects of two well-known antioxidants, Selenium and Vitamin E, on these T-2 toxin induced ‎abnormalities are compared.‎‏ Material and Methods. A single sub lethal dose of T-2 toxin (2 μg/Kg) inoculated to a group ‎of mice and at different times after intoxication the peripheral blood lymphocytes counted by ‎flow cytometry method. Then, 4 separate groups of mice received T-2 toxin and selenium ‎sulphide or vitamin E, and proportion of the lymphocyte subpopulations were determined in their ‎peripheral blood samples. Results. The results showed that T-2 toxin affects normal proportion of all studied lymphocytes. ‎CD19+ and CD8+ lymphocytes decreased and CD4+ cells increased after receiving a single sub ‎lethal dose of the toxin. Peritoneal injection of selenium in both condition, concomitant or before ‎T-2 toxin, prevented CD19+ cells reduction but, vitamin E had no effect on reduction process of ‎this subpopulation. When, selenium inoculated at intoxication time, the increasing of CD4+ ‎population prevented completely but, vitamin E had no efficacy in this condition. In contrast, ‎injection of vitamin E, 24 h before the toxin, significantly affected the CD4+ proportion and ‎prevented the increasing of the subpopulation while, selenium was ineffective in this manner. ‎Receiving of vitamin E in both condition, before and at intoxication, prevented reduction of ‎CD8+ cells while selenium repair the problem only when received concomitant with the toxin.‎ Discussion. It is concluded that to cure T-2 toxin induced abnormalities in lymphocyte ‎subpopulations, efficacy of selenium and vitamin E varies depending on inoculation time of ‎these antioxidants as well as type of disturbed lymphocyte, and it seems that selenium received ‎immediately after intoxication provides more efficacious results while usefulness of vitamin E is ‎more when received prior to intoxication.‎

This study was performed to determine the type and number of the fungus species including saprophyte, opportunistic and pathogen in the salt desert region of Ardestan township in 1375. This descriptive study was conducted by Petri-dish trapping technique. 9 different locations were selected (saltiferous, salt desert forest, agriculture ground, fruit garden, city space, pigeon space, ranches space, poultry space, and canal wells including wild pigeon). From the first 5 locations 75 samples and from the other 4 locations 40 petri-dish were obtained. Totally, 1051 fungus colonies were counted from 115 petir-dishes. The most common fungi were peniccilium spp., cladosporium spp., aspergillus spp., alternaria spp., and yeasts. Number of fungus colonies were as follow: at the evening 45.5%, in the morning 27.7%, and at noon 26.8%. Canal wells including wild pigeons had shown to have the most variable types of fungi, meanwhile, the biggest yeast colony numbers were isolated form this location. Furthermore, 3 fungus species similar to sporothrix schenchii were isolated from this site. The place of sampling and environmental factors may influence the number and type of fungi in the salt desert region. The biggest fungus colonies were isolated form ranches space and the smallest from salt march regions. Meanwhile, some fungi such as peniccilium spp., cladosporium spp., aspergillus spp., alternaria spp are able to grow even in unsuitable environmental conditions.