( ﺃ)

ﻛﺘﺎﺑﭽﻪ ﺧﻼﺻﻪ ﻣﻘﺎﻻﺕ

ﺩﻭﻣﻴﻦ ﻛﻨﮕﺮﻩ ﻗﺎﺭچ ﺷﻨﺎﺳﻲ ﭘﺰﺷﻜﻲ ﺍﻳﺮﺍﻥ

ﺑﻬﻤﻦ 1391 ﺍﻫﻮﺍﺯ- ﺍﻳﺮﺍﻥ

( ﺏ)

ﺭﺋﻴﺲ ﻛﻨﮕﺮﻩ: ﺩﻛﺘﺮ ﻣﺼﻄﻔﻲ ﻓﻘﻬﻲ

ﺩﺑﻴﺮ ﻋﻠﻤﻲ ﻛﻨﮕﺮﻩ: ﺩﻛﺘﺮ ﻋﻠﻲ ﺯﺍﺭﻋﻲ ﻣﺤﻤﻮﺩﺁﺑﺎﺩﻱ

ﺩﺑﻴﺮ ﺍﺟﺮﺍﻳﻲ ﻛﻨﮕﺮﻩ: ﺩﻛﺘﺮ ﺍﺣﻤﺪﺭﺿﺎ ﻣﻬﺘﺪﻱ

( ﺕ)

ﺑﺎ ﻫﻤﻜﺎﺭﻱ:

 ﮔﺮﻭﻩ ﻗﺎﺭچ ﺷﻨﺎﺳﻲ ﭘﺰﺷﻜﻲ ﺩﺍﻧﺸﮕﺎﻩ ﻋﻠﻮﻡ ﭘﺰﺷﻜﻲ ﺟﻨﺪﻱ ﺷﺎﭘﻮﺭ ﺍﻫﻮﺍﺯ  ﻣﺮﻛﺰ ﺗﺤﻘﻴﻘﺎﺕ ﺑﻴﻤﺎﺭﻱ ﻫﺎﻱ ﻋﻔﻮﻧﻲ ﻭ ﮔﺮﻣﺴﻴﺮﻱ ﺟﻨﺪﻱ ﺷﺎﭘﻮﺭ ﺍﻫﻮﺍﺯ  ﺍﻧﺠﻤﻦ ﻗﺎﺭچ ﺷﻨﺎﺳﻲ ﭘﺰﺷﻜﻲ ﺍﻳﺮﺍﻥ

( ﺙ)

ﺍﻋﻀﺎء ﻛﻤﻴﺘﻪ ﻋﻠﻤﻲ: 1) ﺩﻛﺘﺮ ﻣﺴﻌﻮﺩ ﺍﻣﺎﻣﻲ 17) ﺩﻛﺘﺮ ﻣﻬﻨﺎﺭ ﻓﺘﺎﺣﻲ ﻧﻴﺎ 2) ﺩﻛﺘﺮ ﺍﻣﻴﻦ ﺁﻳﺖ ﺍﻟﻬﻲ ﻣﻮﺳﻮﻱ 18) ﺩﻛﺘﺮ ﻋﺒﺪﺍﻟﻤﺠﻴﺪ ﻓﺘﻲ 3) ﺩﻛﺘﺮ ﺣﻤﻴﺪ ﺑﺪﻟﻲ 19) ﺩﻛﺘﺮﻋﺒﺪﺍﻟﺤﺴﻦ ﻛﺎﻇﻤﻲ 4) ﺩﻛﺘﺮ ﭘﺮﻳﺴﺎ ﺑﺪﻳﻌﻲ 20) ﺩﻛﺘﺮ ﭘﺮﻳﻮﺵ ﻛﺮﺩﺑﭽﻪ 5) ﺩﻛﺘﺮ ﻛﻴﻮﺍﻥ ﭘﺎﻙ ﺷﻴﺮ 21) ﺩﻛﺘﺮﺷﻬﻼ ﻣﺤﻤﺪﻱ ﺭﻭﺩﺑﺎﺭﻱ 6) ﺩﻛﺘﺮ ﻋﻠﻴﺮﺿﺎ ﺧﺴﺮﻭﻱ 22) ﺩﻛﺘﺮﺳﻌﻴﺪ ﻣﻬﺪﻭﻱ ﻋﻤﺮﺍﻥ 7) ﺩﻛﺘﺮ ﻣﺼﻄﻔﻲ ﭼﺎﺩﮔﺎﻧﻲ ﭘﻮﺭ 23) ﺩﻛﺘﺮ ﻋﻠﻲ ﻣﻴﻜﺎﻳﻴﻠﻲ 8) ﺩﻛﺘﺮ ﻃﺎﻫﺮﻩ ﺷﻜﻮﻫﻲ 24) ﺩﻛﺘﺮ ﺳﻴﺪ ﺣﺴﻴﻦ ﻣﻴﺮﻫﻨﺪﻱ 9) ﺩﻛﺘﺮ ﻣﻌﺼﻮﻣﻪ ﺷﻤﺲ ﻗﻬﻔﺮﺧﻲ 25) ﺩﻛﺘﺮ ﻋﻠﻲ ﻧﺎﺻﺮﻱ 10) ﺩﻛﺘﺮﻣﻬﺪﻱ ﺭﺯﺍﻗﻲ ﺍﺑﻴﺎﻧﻪ 26) ﺩﻛﺘﺮ ﻣﺤﻤﺪ ﺟﻮﺍﺩ ﻧﺠﻒ ﺯﺍﺩﻩ 11) ﺩﻛﺘﺮﺳﺎﺳﺎﻥ ﺭﺿﺎﻳﻲ 27) ﺩﻛﺘﺮ ﺳﻴﺪﺟﻤﺎﻝ ﻫﺎﺷﻤﻲ 12) ﺩﻛﺘﺮ ﻋﻠﻲ ﺭﺿﺎﻳﻲ ﻣﺘﻪ ﻛﻼﻳﻲ 28) ﺩﻛﺘﺮ ﻣﺤﻤﺪﺗﻘﻲ ﻫﺪﺍﻳﺘﻲ 13) ﺩﻛﺘﺮ ﻋﻠﻲ ﺯﺍﺭﻋﻲ ﻣﺤﻤﻮﺩ ﺁﺑﺎﺩﻱ 29) ﺩﻛﺘﺮ ﻣﺤﻤﺪﺣﺴﻴﻦ ﻳﺎﺩﮔﺎﺭﻱ 14) ﺩﻛﺘﺮ ﻣﺠﻴﺪ ﺯﺭﻳﻦ 30) ﺩﻛﺘﺮ ﺳﻴﺪ ﺍﻣﻴﺮ ﻳﺰﺩﺍﻥ ﭘﺮﺳﺖ

15) ﺩﻛﺘﺮ ﻛﺎﻣﻴﺎﺭ ﺯﻣﺮﺩﻳﺎﻥ 16) ﺩﻛﺘﺮ ﺑﺘﻮﻝ ﺻﺎﺩﻗﻲ ﻧﮋﺍﺩ

( ﺝ)

ﺍﻋﻀﺎء ﻛﻤﻴﺘﻪ ﺍﺟﺮﺍﻳﻲ: 1) ﺩﻛﺘﺮ ﻋﺒﺪﺍﻟﻨﺎﺻﺮ ﻣﻴﺮ ﺷﻜﺎﻙ 2) ﺩﻛﺘﺮ ﻣﻬﻴﻦ ﺩﻳﺎﻧﺖ 3) ﺩﻛﺘﺮ ﻣﻨﺼﻮﺭ ﺍﻣﻴﻦ 4) ﻣﻬﻨﺪﺱ ﻣﺤﻤﺪﺭﺿﺎ ﺍﻓﺸﺎﺭ 5) ﺁﻗﺎﻱ ﻧﻮﺭ ﺍﷲ ﻧﻴﻜﺨﻮﺍﻩ 6) ﺁﻗﺎﻱ ﺣﺴﻴﻦ ﺭﺷﺎﺩﺗﻴﺎﻥ 7) ﺁﻗﺎﻱ ﻋﺒﺪﺍﻟﺮﺣﻴﻢ ﺣﻴﺎﺗﻲ 8) ﺁﻗﺎﻱ ﺣﻤﻴﺪ ﺳﺮﻛﺎﻛﻲ 9) ﺁﻗﺎﻱ ﻋﻴﺴﻲ ﺟﺒﺎﺭﻱ 10) ﺁﻗﺎﻱ ﺟﻤﺸﻴﺪ ﻋﺎﻛﻒ 11) ﺁﻗﺎﻱ ﻣﺤﻤﺪ ﺍﻓﺘﺨﺎﺭ 12) ﺁﻗﺎﻱ ﻣﺤﺴﻦ ﻧﻌﻤﺖ ﺗﺒﺎﺭ 13) ﺁﻗﺎﻱ ﺭﺍﺷﺪ ﺭﺿﺎﻳﻲ 14) ﺁﻗﺎﻱ ﺭﺿﺎ ﻗﺎﺳﻤﻮﻧﺪ 15) ﺧﺎﻧﻢ ﺳﻤﻴﺮﻩ ﺭﺣﻴﻢ 16) ﺧﺎﻧﻢ ﺳﻤﺎء ﻓﺮﺍﻣﺮﺯﻱ 17) ﺧﺎﻧﻢ ﺳﻴﺪﻩ ﻋﺎﻃﻔﻪ ﻣﻮﺳﻮﻱ 18) ﺧﺎﻧﻢ ﺷﻬﺮﺯﺍﺩ ﺣﻴﺪﺭﻱ ﻧﻴﺎ 19) ﺧﺎﻧﻢ ﻓﺎﻃﻤﻪ ﻗﻨﻮﺍﺗﻲ 20) ﺧﺎﻧﻢ ﻣﮋﮔﺎﻥ ﻧﻮﻳﺪ 21) ﺧﺎﻧﻢ ﻣﺮﻳﻢ ﻋﺮﻓﺎﻧﻲ 22) ﺧﺎﻧﻢ ﻧﺪﺍ ﻛﻴﺎﺳﺖ 23) ﺧﺎﻧﻢ ﻣﻬﺴﺎ ﺭﺟﺒﻲ

( ﺏ)

Oral presentation ...... 8 Useful molecular markers for identification of human pathogenic fungi ...... 9 Fungal infection in burns ...... 10 Loop-mediated isothermal amplification and rolling circle amplification, novel molecular techniques for detection and identification of fungal pathogens ...... 11 A focus on new biomarkers of Aspergillus as a diagnostic target in invasive aspergillosis ...... 12 Antifungal activity of nano, micro and macro agents ...... 13 Mycetoma in Iran- a retrospective study of 62 cases ...... 14 Fungal exoenzymes, characteristics and diagnosis ...... 15 Evaluation of Pneumocystis jirovecii in normal population ...... 16 Aspergillus flavus: A universal challenge for public health and agriculture ...... 17 Determining the concentration of ochratoxin a in breast milk of the mothers living in Khorrambid, Fars province, Iran ...... 18 Curcumin regulates aflatoxin biosynthetic gene cluster in Aspergillus parasiticus NRRL 2999 . 19 Aflatoxin detoxification in rice using citric acid ...... 20 Extracellular enzymes production in Rhodotorula species ...... 21 Evaluation of the sensitivity and overexpression of the genes cdr1 and mdr1 to fluconazole in clinical Candida albicans isolates from Iranian oral candidiasis AIDS patients ...... 22 Antifungal bacteria in soil community: Diversity and bioactivity ...... 23 Paradoxical growth effects of the fluconazole, ketoconazole, voriconazole, and itraconazole on clinical isolates of Candida species...... 24 Evaluation of antifungal activity of new derivatives of imidazole using colorimetric method .... 25 In vitro susceptibility of aflatoxigenic and non-aflatoxigenic Aspergillus flavus strains to conventional antifungals...... 26 Clinical and paraclinical aspect of 16 cases of mycetoma during past 3 decades in Mashhad ..... 27 Recovery of Candida and Aspergillus species in Iranian patients suffering from cystic fibrosis . 28 Comparison of Candida colonization in angular cheilitis lesions before and after denture replacement ...... 29 Invasive fungal infection and chronic obstructive pulmonary disease (COPD) ...... 30 Candida species in cutaneous candidiasis patients in the Guilan province in Iran; identified by PCR-RFLP method...... 31 A 5 year epidemiological study of superficial cutaneous in suspicious patients in Tehran (2006- 2010) ...... 32 Identification of Malassezia species isolated from patients with pityriasis versicolor by PCR- RFLP method in Arak...... 33 The new formulation of nanoencapsulated essential oil as green antifungal against Fusarium graminearum ...... 34 Evaluation of the effects of Zataria multiflora, Geranium pellargonium roseum, Myrth and Lemon shell essences on immune system function in experimental animals ...... 35 Assessment of antifungal activities of Urtica dioica against Microsporum canis on the in vitro and in guinea pig model of dermatophytosis...... 36 Expression analysis of hyphae-involved genes in Candida albicans treated by allicin originated from garlic ...... 37 Use of the extract of Myrtus communis to inhabit biofilm formation of Candida tropicalis...... 38 Chemical composition, antifungal and antibiofilm activities of the essential oil of Mentha piperita L...... 39 (1)

RNA-mediated gene silencing in Candida albicans: reduction of fungal pathogenesis by use of RNAi technology ...... 40 The relationship between gene ERG11 and resistance of fluconazole Candida albicans in patients to vaginal candidiasis ...... 41 Application of Real-time PCR, Nested-PCR and mycology assay using BAL samples for detection of Aspergillus flavus and A. fumigatus in hematological malignancies ...... 42 Development of a quantitative TaqMan Real- Time Polymerase Chain Reaction for detection of Pneumocystis jirovecii from bronchoalveolar lavage fluids ...... 43 Fungal genomic projects, metagenomics and gene analyses ...... 44 Molecular identification of Fusarium sibiricum isolated from Iranian grains ...... 46 Development approach of fungal dendritic cell vaccines ...... 47 Evaluation of Candida albicans allergens reactive with specific IgE in atopic dermatitis patients by Immunoblotting ...... 48 Aspergillus monitoring project in an Iranian educational hospital ...... 49 Analysis of airborne allergenic fungi in farms and the rate of their caused allergic diseases in farms’ staffs ...... 50 Propolis efficacy on candidiasis histopathology and on TNF-α, IFN-γ and IL2 cytokines production in old mice with and without systemic candidiasis ...... 51 Study the effects of different types of interferon pretreatments on expression of cathelicidin in murine macrophages following exposure to Candida albicans ...... 52 Poster presentation ...... 53 Activity of Ajowan (Carum copticum) essential oil against Alternaria alternata ...... 54 Identification and antifungal susceptibility profile of Candida species isolated from nosocomial candiduria ...... 55 The effects of Spirulina on nitric oxide in balb/C mice with systemic candidiasis ...... 56 Agar based drug diffusion assay for in vitro susceptibility testing of dermatophytes ...... 57 Comparison between antifungal effect of Alchemilla vulgaris (Paye Shir) with two chemical drugs (ketoconazole, fluconazole) on Candida albicans and Malassezia furfur ...... 58 Evaluation of antifungal activity of essential oil Allim jesdianum on Malassezia furfur and Candida krusei ...... 59 Antifungal activity of silver nanoparticles on Aspergillus niger ...... 60 Antifungal effect of nanosilver on saprophytic and pathogenic fungi ...... 61 Antifungal effect of Cydonia oblonga extracts on Aspergillus niger ...... 62 In vitro antifungal activity of Anthemis gayana flower methanolic and ethanolic extracts on Candida species ...... 63 In vitro anti-Candida activity of Salsola rigida ...... 64 In vitro anti-Candida activity of Prosopis farcta ...... 65 Peganum harmala, a plant with antifungal activity ...... 66 Anticandidal effect of shallot (Allium hirtifolium) against chronic candidiasis ...... 67 Identification and antifungal susceptibilities Candida species isolated from nail infection ...... 68 Antifungal susceptibilities of Candida species causing vulvovaginitis by disk diffusion ...... 69 Susceptibility to fluconazole in Candida species isolated from vulvovaginal candidiasis in Ahvaz ...... 70 Interaction between Candida albicans with different bacteria ...... 71 Investigation the potential of allicin to inhibit the growth of Candida albicans in vitro and in a systemic candidiasis mouse model ...... 72 (2)

Therapeutic effect of Shallomin as an antifungal drug on dermatophytes ...... 73 Azole resistance in Aspergillus fumigatus obtained from hospitals environments ...... 74 Antifungal activity of special propolis extract ...... 75 Antifungal susceptibility profile of Rhodotorula species from Ahvaz ...... 76 Survey the effect of Licorice extract on aflR gene expression and aflatoxin production in Aspergillus parasiticus via Real Time PCR ...... 77 Isolation and identification of soil chitinolytic bacilluses in the Northern Iran and survey of antifungal effects ...... 78 Unsuccessful treatment of co-infection endocarditis due to Candida albicans and C. tropicalis in a drug abusers patient ...... 79 Assessment of antifungal activity of various extracts and fractions of Ephedra major against some dermatophytes moulds and yeasts...... 80 Isolation of anti-dermatophyte compounds from Myrtus communis L. leaves ...... 81 Laboratory study of anticandidal activity of Cinnamon essential oil by microdilution method ... 82 Anti-Candida activity of the hydroalcoholic extracts of Heracleum persicum fruit ...... 83 In vitro effectiveness of Pinus longifolia on Candida species isolated from patients with vulvovaginal candidiasis in Isfahan ...... 84 Antifungal efficacy of Myrtus communis linn. extract against Candida sp. and Aspergillus sp. . 85 Determination of synergistic effect of Myrtus communis essential oil and linalool with itraconazole against azole-resistant Candida species ...... 86 Antifungal and antibacterial activity of Fistulina hepatica (Schaeff.) methanolic extract ...... 87 Antifungal activity of ethanolic extract of leaves "Myrtle" against Microsporum gypseum, M. canis and Trichophyton mentagrophytes ...... 88 Antimicrobial properties of Ixora brachiata Roxb. against bacteria and Candida isolates ...... 89 Study of antifungal activity of essential oils of Iranian endemic medicinal herbs against Candida albicans ...... 90 Different silver nanostructures and growing inhibition of clinical Candida albicans isolates ..... 91 Are some isolates of Candida albicans resistance against silver nanostructures? ...... 92 In vitro inhibitory effect of Echinops cephalotes methanolic extract on Candida spp. isolated from patients with vulvovaginal candidiasis in Isfahan ...... 93 The influence of water ozonation in wheat tempering on reduction of Aspergillus parasiticus growth ...... 94 In vivo antifungal activity of new fluconazole derivatives in systemic murine candidiasis ...... 95 Study on inhibitory activity of propolis alcoholic extract on opportunist fungi of Candida in vitro ...... 96 Effect of fresh and old garlic on germ tube formation in Candida albicans ...... 97 Evaluation of antifungal effect of Cyclamen coum tuber extract against isolated Candida albicans and C. tropicalis ...... 98 In vitro study the effects of tobacco on dermatophytes...... 99 In vitro antifungal susceptibility testing of dermatophytes against eight antifungal drugs ...... 100 Antifungal activity of Scrophularia striata extract on Candida albicans...... 101 In vitro antifungal susceptibility of Candida species isolated from blood cultures of burnedburn patients or burn victims patients in burns unit of Zare Hospital, Sari, Iran ...... 102 The concentration of aflatoxin M1 in the mothers’ milk in Khorrambid city, Fars, Iran ...... 103 The study of temperature on the T-2 toxin in corn...... 104

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Detection and identification of fungal pathogens in clinical samples from patients referred to professor Alborzi mycology research lab, Shiraz, Iran ...... 105 Study the existing fungal flora in the atmosphere of different areas of Kashan in 2001 ...... 106 The study of soil fungi of Khuzestan's oil regions ...... 107 Determine the frequency of blood groups in patients with dermatophytosis in the Glabchy laboratory and private practices Kashan in 2001 ...... 108 Air fungal contamination in Namazi hospital transplantation wards ...... 109 The effects of variable CO2 concentrations on morphological patterns in dermatophyte pathogen Microsporum canis ...... 110 Study on saprophytic fungi in archives of office from Sari city and their role in sick building syndrome ...... 111 Assessment of soil fungi prevalence in different areas of Kashan from 2010 to 2011 ...... 112 Airborne fungal contamination in the operating rooms of Yazd medical university hospitals ... 113 Fungal isolated from a small industry related to car factory in Babol ...... 114 Real Time RT-PCR study of beta-defensin 1 gene expression in murine macrophages pretreated with different types of interferons and treated with Candida albicans and lipopolysaccharide . 115 Identification and detoxification of aflatoxin in bread wastes used in livestock food in Tehran 116 A comparative investigation on total aflatoxin and ochratoxin production spectra in 15 different Asprergillus species from Iranian northern states ...... 117 A comparative investigation on ochratoxin and aflatoxin B production spectra in 15 different species of Aspergillus isolates from Iranian northern states (a case study) ...... 118 A comparative investigation on ochratoxin and aflatoxin B production spectra in culture medium of 15 different Aspergillus species from Iranian northern states ...... 119 Survey the dermatophyte contamination of covered swimming pools in Yazd 2009 ...... 120 Measurement of aflatoxin B1 in flour of Sohan making in Qom ...... 121 Genetic diversity of Aspergillus spp. isolated from ICU, indoor and outdoor environments in hospitals in by PCR and determine drug sensitivity...... 122 An investigation of the ability of Candida albicans in reducing the growth of Aspergillus parasiticus in culture medium and wheat...... 123 Prevalence of fungal elements in Isfahan swimming pools water ...... 124 Qualitative and quantitative study monitoring airborne fungal flora in the kidney transplant unit ...... 125 Toxigenic mycoflora distribution of hazelnut in Isfahan ...... 126 Radionuclide concentration in Agaricus bispora (Agaricaceae); an edible mushroom ...... 127 Characterization and determination of red yeast isolated from leather waste water as potential source of carotenoid ...... 128 Effect of vitamin A, K1 and B1 on Aspergillus parasiticus growth and aflR gene expression .. 129 Hemolytic and esterase activity of Candida species isolated from patients ...... 130 Candida species are transmitted by cell phones in hospitals: A comparison between doctors, nurses, accompanying the patients and patient cell phones contamination ...... 131 Effect of three Iranian honeys on pro-inflammatory cytokines during invasive aspergillosis .... 132 Isolation of keratinophilic fungi and aerobic Actinomycetes from City Park soils in Gorgan, North of Iran ...... 133 The impacts of indoor Aspergillus fungus on human health ...... 134 Evaluation of the keratinase activity in Trichophyton mentagrophytes strains isolates from patients with dermatophytosis...... 135 (4)

Biological control of the Blatella germanica with Beauveria bassiana and Lecanicillium muscarium ...... 136 A study of IgE reactivity to Curvularia lunata antigens in patient with asthma ...... 137 Microbiological contamination in bottled mineral water and assessment of antibacterial sensitivity among the isolates ...... 138 An in vitro study of nematophagous fungi on pathogen larva nematodes ...... 139 Sensitivity and specificity of skin prick test to Aspergillus fumigatus antigens ...... 140 Survey of mycological flora in the student hostel baths in Kurdistan University of medical sciences (2011) ...... 141 A laboratory study on properties of Deoxynivalenol (DON) production in standard culture medium by 24 Aspergillus spp. isolates from Northern Iran ...... 142 A study on properties of Deoxynivalenol (DON) production in culture medium regarding Aspergillus spp. isolates from processing factories of northern Iran ...... 143 Comparison of incubation time and carotenoid production in Rhodotorula slooffiae and R. mucilaginosa isolated from leather tanning wastewater ...... 144 Evaluation of the number of Treg cells in normal mice exposed to AFB1 and treated with aged garlic extract ...... 145 The qualitative and quantitative assessment of airborne fungi in indoor environment of various wards of Imam Reza hospital in Mashhad, 2012 ...... 146 Measurement of Aflatoxin B1 in the flour of Arak traditional bakeries ...... 147 Investigation of contamination of medicinal plants by aflatoxin ...... 148 Effect of silver nano- particles on growth of aflatoxigenic fungus, Aspergillus parasiticus ...... 149 Cellulose production using Trichoderma reesei on solid state fermentation ...... 150 Isolation and molecular identification of keratinophilic fungi from parks of Shiraz city ...... 151 Aflatoxin in raw and roasted with salt nuts (pistachio, peanut and walnut) sold in markets of Tabriz, Iran ...... 152 Mould contamination of untreated and roasted with salt nuts (walnut, peanut and pistachio) sold in markets of Tabriz, Iran ...... 153 Inhibitory effect of vitamin C on Aspergillus parasiticus growth and aflatoxin gene expression ...... 154 Candida africana, a first case from Iran, susceptibility to antifungals, evaluation extracellular enzymes and biofilm formation ...... 155 The inactivation of aflatoxin B1 in food and feed crops by gamma radiation ...... 156 Prevalence of fungal allergens in respiratory allergic patients in Ahvaz ...... 157 Isolation of saprophytic fungi in the soil of Chitgar park, Tehran, Iran ...... 158 A Comparative investigation on DON (Deoxy nivalenol) toxin production patterns of Aspergillus Species biomasses, Northern region of Iran ...... 159 Distribution of outdoor airborne fungi in various regions of Tehran ...... 160 Measurement of aflatoxin B1 in feed for dairy animals in Alborz province ...... 161 Deuterium effect on the growth of the fungus Aspergillus fumigatus and Candida albicans .... 162 Fungal contamination and physicochemical of public swimming pools In Kashan, Iran, 2008- 2009 ...... 163 Subcutaneous phaeohyphomycosis caused by sterile Chaetomium spp. in Iran ...... 164 The high rate of Pneumocystis jirovecii colonization in non-HIV-infected Iranian patients ...... 165 Co- infection of invasive pulmonary Aspergillosis and cutaneous Fusarium infection in a patient suspected to pyoderma gangrenosum: a case report ...... 166 (5)

The first case of onychomycosis due to Exophiala dermatitidis in Iran ...... 167 Phaeohyphomycosis due to dematiaceous fungi; a review of the literatures ...... 168 The frequency of candiduria in hospitalized patients with depressive syndrome ...... 169 Utilization of length polymorphism of ITS1 and ITS2 regions for identification of pathogenic yeast species by normal agarose gel electrophoresis...... 170 Mating type and variety identification of Cryptococcus neoformans isolates ...... 171 Catheter-related candidemia caused by Candida albicans in a coronary artery bypass graft (a case report) ...... 172 A case of fungus ball type pansinusitis caused by Schizophillum commune ...... 173 Dermatophytosis due to Microsporum fulvum, a new case from Iran ...... 174 First case of disseminated phaeohyphomycosis in an immunocompetent individual due to Alternaria malorum ...... 175 Study of the role of Candida species in urinary tract infections with special attention to the predisposing conditions ...... 176 Distribution of Malassezia species in patients with pityriasis versicolor compared to healthy individuals in Yazd, Iran ...... 177 Survey the bacterial, fungal contamination in three dental impression materials ...... 178 Electron microscope studies of actinomycete isolates Kn10 and 115 with antifungal effect against Microsporum gypseum ...... 179 A review on the role of fungi in atopic dermatitis ...... 180 A Survey on oral Candida flora in patients with atopic dermatitis and compared with healthy subjects ...... 181 Gastrointestinal basidiobolomycosis: first culture-proven case in Iran ...... 182 The prevalence of candidiasis in pregnant patients with urinary tract infection referred to Mehrin clinical laboratory ...... 183 Genetic diversity among Iranian Candida albicans isolated from nails and skin ...... 184 Identification of the significant medical dermatophytes from the clinical samples of the skin clinics of the Tonekabon through the Real Time ITS – PCR sequencing method ...... 185 Candida vaginitis due to consumption of azithromycin ...... 186 Actinomycosis in Iran ...... 187 Prevalence of candidiasis in Vali-Asr hospital of Birjand during 1998-2012 ...... 188 Molecular strain-typing of Microsporum canis prevalent in Japan and Iran by randomly amplified polymorphic DNA...... 189 A case of sinusitis caused by Nattrassia mangiferae in Iran ...... 190 Molecular phylogeny of the NADPH oxidase pathway in the filamentous fungus Aspergillus fumigatus, a human pathogen ...... 191 Fungal peritonitis: the disease should be considered in patient with cancer (a case report) ...... 192 Candida species in vulvovaginal candidiasis patients from the town Taibad in Iran; identified by PCR-RFLP method...... 193 Detection of invasive candidiasis in patients with hematological malignancies and bone marrow transplant recipients ...... 194 Detection of Malassezia isolated from patients with pityriasis versicolor and seborrheic dermatitis in Ahvaz using Nested- PCR ...... 195 Identification of Candida species isolated from oral colonization in Iranian HIV-positive patients by PCR-RFLP...... 196

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Study of fungal colonization of burn wound in patients admitted to the burn center of Zare' hospital, Sari, 2011-2012...... 197 Mycotic keratitis among patients referred to mycology laboratory, Emam Reza hospital of Mashhad, Iran ...... 198 Candidiasis prevalence and relationship with clinical and para clinical observations in women supported by selected health centers of Tabriz ...... 199 An evaluation of direct microscopy examination and culture sensitivity in diagnosis of dermatophytosis...... 200 The effects of vaginal candidiasis on newborn ...... 201 Identification of dermatophytes by AP-PCR...... 202 Onychomycosis in Mashhad ...... 203 Study the relationship of Candida species with Lactobacillus species in patients with fungal vaginitis and healthy individuals...... 204 Identification of dermatophytes species by PCR-RFLP during 2011-2012 in Esfahan ...... 205 Molecular identification of Candida species isolated from onychomycosis in Iran ...... 206 Isolation of fungal agents from burn wounds and determining their susceptibility to antifungal drugs ...... 207 Isolation of Malassezia species using modified CHROMagar and confirm by PCR ...... 208 Superficial mycoses in patients who referred to clinic 2 Tabriz University of medical sciences in 2007-2011 ...... 209 β-tubulin restriction mapping for identification of medically relevant dermatophytes...... 210 Use of rolling circle amplification (RCA) to rapidly identify species of Cladophialophora potentially causing human infection ...... 211 Epidemiology of candidiasis in patients referred to the medical mycology laboratory, Afzalipoor faculty of medicine ...... 212 The survey of presence Candida Spp. individuals skin with clinical manifestation of Acne referred to dermatology clinic of Booalh Sina hospital in Sari ...... 213 A survey on fungal burden in the intensive care units environment in Ahvaz University hospitals, 2011 ...... 214 Identification of fungal keratitis agents in the east of Mazandaran area by PCR and determination of their drug sensitivity...... 215 Investigation, isolation and identification of cutaneous mycoses in laboratory animals ...... 216 Frequency of vulvovaginal candidiasis in 19-65 years old women in Lar in 2011 ...... 217 Assessment of oral candidal colonization in diabetic patients and aetiologic factors associated with this colonization...... 218 Palate fistula due to sinonasal mucormycosis, A case report in diabetic patient ...... 219 Molecular determination of dermatophyte species isolated from cutaneous infections ...... 220 Aspergillosis of the paranasal sinuses; report of 7 cases treated by endoscopic sinus surgery (ESS) ...... 221 Epidemiological and microbiological study of fungal disease (inpatient and outpatient) in education hospitals, Kerman, 2012 ...... 222 Evaluation of genetic diversity by use of DNA sequencing and PCR-AFLP with antifungal susceptibility profile for Hortaea werneckii ...... 223 Professional ethics in health system and primary health care ...... 224

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Oral presentation

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Useful molecular markers for identification of human pathogenic fungi

Hossein Mirhendi

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

As the diversity of fungal pathogens that cause human disease continues to expand, we need to expand our ability to detect/identify them in clinical specimens or isolates. Phenotype-based identification of fungi suffers from the characteristics which are influenced by variable natures such as culture conditions and subjective interpretation. Advances in DNA-based techniques have helped us to tracing of evolutionary link between groups at higher taxonomic ranks through phylogenetic investigation and thus improving taxonomy at the level of genera and species, developing diagnostic applications for new defined taxonomic units and improving the epidemiological tools to monitor outbreaks and transmission routes of infection to sub-specific entities. Progresses in advanced molecular methods for diagnosis and epidemiological typing of pathogenic fungi are becoming fundamental for early treatment of patients, controlling fungal clearance, and counteracting resistance to antifungal therapy. Fungal whole genome sequences have provided fantastic resources for developing new phylogenetic markers. In this presentation, recent advances in the DNA-based identification of fungal pathogens were considered. Targets used in molecular diagnostic tests for human pathogenic fungi include both single and multi- copy genes of nuclear and mitochondrial origin such as actin, chitin synthase, cytochrome P-450, lanosterol-14 a-demethylase (L1A1), calmodulin, translation elongation factor 1 alpha, beta- tubulin and different parts of ribosomal DNA gene complex including 18S, D1/D2, 28S, ITS1 and ITS2. The usefulness of the marker for delineating the main pathogenic fungi including the species belong to the genera Aspergillus, Candida, Cryptococcus, Trichosporun, Malassezia, dermatophyte, zygomycetes, Fusarium and dematiaceous fungi is discussed. Keywords: DNA markers, Identification, Pathogenic fungi

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Fungal infection in burns

Tahereh Shokohi

Department of Medical Mycology and Parasitology, Sari Medical School and Invasive Fungi Research Center, Mazandaran University of Medical Sciences Sari, Iran Email: [email protected]

Burns is the most destructive injuries, cause deaths major economic and psychological impact and long-term sequelae. Based on the information that collected from 100 hospitals throughout the Iranian capitals, burns have been considered for 18% of all mortal injury cases and rank second after traffic injuries. Fungal infection now represents a serious problem in burn units and causes a high morbidity and mortality. Blood stream fungal infection often develop in severe burn patient due to humeral and cellular immunodeficiency, loss of skin barrier, gastrointestinal translocation, use of central venous catheter, total parenteral nutrition and invasive monitoring devices, mechanical ventilation. These routes are the common port of entry for fungal pathogens as well. Wound colonization by fungi usually occurs late after gram positive and negative bacteria. The use of broad-spectrum antibiotics alters the intestinal and skin flora and promotes overgrowth of the yeast. Candida spp. are the most common non bacterial colonizers of burned wound and the most common causes of blood stream fungal infection. Candidemia occurs in 1 to 5% of patients with major burns. Candida albicans accounts for the majority of cases with candidemia, but an increasing number of infections due to non-albicans species have been reported. The most common isolated Candida non-albicans are C. glabrata, followed by C. tropicalis, C. parapsilosis, C. krusei, and other Candida spp. The emergence of antifungal resistance within these causative yeasts, especially in Candida non-albicans patients requires an urgent need for laboratory support in the treatment of fungal infections especially in critically ill burn patients. Appropriate systemic antifungal therapy guided by antifungal susceptibility improves the outcome for severely injured burn victims susceptible to fungal infection. This article emphasizes on the importance of early detection fungal infection, identification of causing fungal species and their susceptibility pattern to antifungal drugs. Keywords: Burns, Etiology, Fungi, Infection, Injuries, Candida

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Loop-mediated isothermal amplification and rolling circle amplification, novel molecular techniques for detection and identification of fungal pathogens

Mohammad Javad Najafzadeh

Department of Parasitology and Mycology, Ghaem Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Email: [email protected]

Conventional methods for fungal identification in the clinical laboratory rely on morphological and physiological tests and need several days or weeks and are frequently unspecific. Molecular identification mostly implies sequencing, which is relatively expensive and, time-consuming and impractical for a large number of isolates. In this study we used from Loop-mediated isothermal amplification (LAMP) for rapid detection and identification of Fonsecaea strains (agents of human chromoblastomycosis) and rolling circle amplification (RCA) for rapid detection and identification of Fonsecaea and Exophiala strains. LAMP is a powerful nucleic acid amplification technique that rapidly and accurately amplifies target DNA under isothermal conditions; we used this method for rapid screening of Fonsecaea from other Chaetothyriales (black yeast and relatives). The detection limit was found to be 0.2 fg DNA. The second molecular identification technique that was established is rolling circle amplification (RCA). This technique had primarily been developed for virus diagnostics. Using ITS amplicons as templates, this technique proved to be suitable for the identification of Fonsecaea and Exophiala species. The simplicity, sensitivity, robustness and low costs make RCA an attractive technique for the reliable identification of sibling species and other closely related fungi. LAMP proved to be a fast and sensitive method for direct amplification of fungal DNA from environmental samples, whereas for RCA, ITS amplicons are needed. However, with the LAMP assay, we could not distinguish between the different species of the genus Fonsecaea, because they differ in relatively few nucleotide polymorphisms that were sufficient to allow successful RCA diagnostics. Thus, RCA is more specific than LAMP, but LAMP is more sensitive than RCA. Keywords: LAMP, RCA, Fonsecaea, Exophiala

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A focus on new biomarkers of Aspergillus as a diagnostic target in invasive aspergillosis

Mohammad Taghi Hedayati

Department of Medical Mycology and Parasitology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran; Invasive Fungi Research Center, Mazandaran University of Medical Sciences, Sari, Iran Email: [email protected]

Invasive aspergillosis (IA) is a clinical form due to Aspergillus and a major cause of morbidity and mortality in the severely immunocompromised patients. The diagnosis of IA is still a great challenge in immunocompromised patients, and it is often made late in the course of the infection. The accurate diagnosis of IA can withhold the use of expensive and potentially toxic antifungal drugs. In addition, an earlier diagnosis of IA is very important to amend the patients’ survival. This review is focused on new identified biomarkers of Aspergillus which can be used in diagnosis of IA. The PubMed/MEDLINE database were searched for relevant papers using terms such as invasive aspergillosis, Aspergillus, biomarker, galactomannan, antigen and diagnostic tool to find any biomarkers which they were introduced as a target in IA diagnosis by authors. Traditional diagnostic methods are difficult to interpret, and invasive procedures require to obtain histological specimens which in some cases such as thrombocytopenic conditions endanger patients' life. On the other hand, it is also a time consuming procedure which can delay the proper treatment. But several improvements were observed during the last decade. However, detection of galactomannan as an Aspergillus cell wall antigen in serum or BAL has facilitated the early diagnosis of IA, but it has some limitations. Therefore many researchers have designed to introduce a marker from Aspergillus which can reduce these limitations. Several previous publications have reported a range of antigens from Aspergillus which can be candidate for diagnosis of IA, by detecting specific antibodies or the antigen itself in serum or other body fluids. These candidate antigens such as galactomannan protein (Afmp1p and Afmp2p), gliotoxin and thioredoxin reductase have opened a new window in diagnostic methods which is promising for early and definitive of IA diagnosis. Keywords: Biomarker, Thioredoxin reductase, Afmp, Gliotoxin, Invasive aspergillosis

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Antifungal activity of nano, micro and macro agents

Seyyed Amin Ayatollahi Mousavi

Department of Medical Mycology and Parasitology, Afzalipoor Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran Email: [email protected]

Nowadays, applying standard therapies can be limited because of toxicity, low efficacy rates and drug resistance. Therefore, the search for new antifungal agents has been intensified. The aim of this almost ten years work was to evaluate and compare the antifungal properties of three major groups contain, nano (Ag, Au and Se), herbal extracts (Artemisia absinthium, Eucalyptus spp., Allium cepa, Cinnamomum zelanicum, Curcuma longa, Salvia officinalis, Mentha Piperita, Calendula officinalis and Myrtus communis) and actinomycetes (10 species). The solid and liquid fungal growth was carried out in the presence of different media (S, ME, PD, YE and RPMI) at 25°C and 37°C. Above mentioned plants were prepared and identified by herbarium of Bahonar Kerman University. Extracts from the plants were made by using percolation method with hydroalcolholic solvent followed by evaporation of the organic solvent under vacuum and then to dryness in an Oven 50°C. During a screening, a total of 10 strains were isolated from Kerman soil samples of seven regions. These isolates were also evaluated for their antifungal ability. All ten samples were electron microscopy (TEM and SEM) tested and identified after sequencing. Nano size particles (Ag, Au and Se) were prepared with the average size of 15nm in our lab. The MIC of all agents was tested in micro plates as well. Nano agents were the most active against all kinds of fungi followed by herbal extracts. Against mycelia fungi, nano-Se was found to be the most active as compared to the standard agents. The Streptomyces isolates with most antifungal activity against all kinds of fungi, particularly moulds, were Ks10, Kn10, Ks8, Km2, L1, Ks2, Ks1, D5, Ks4 and Km10. As M. communis had the best antifungal effect among all other herbal extracts that was tested on Guinea pig as well, in vivo. In bioautography method, after separating ethyl acetatic and methanolic extracts which were showed antifungal effect in RF: 0-0. 3, by thin layer chromatography (TLC) method, the TLC papers placed on medium with certain conc. of fungi. The spot of inhibitory zone were shown by using tetrazolium salt. This nearly ten year work demonstrates the antifungal activity of nano, micro (Actinomycetes) and macro (Herbs) agents against most fungi from yeasts (Candida sp.) to mycelial ones (Saprophytic moulds and dermatophytes). Moreover, these agents could have innumerable applications in different areas. Keywords: Nano Ag, Nano Se, Fungi

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Mycetoma in Iran- a retrospective study of 62 cases

Seyed Jamal Hashemi, Mohsen Geramishoar, Ensieh Zibafar, Roshanak Daie-Ghazvini

Department of Medical Mycology, Tehran University of Medical Sciences, Iran Email: [email protected]

We present a retrospective study of 62 cases of mycetoma in Iran that were diagnosed clinically and microbiologically from 1972 to 2005. In 1972 mycetoma has been first report in Iran by medical mycology department of Tehran University. We retrospectively compared the overall prevalence of mycetoma and the prevalence of infective agent in Iran during 33 years. In our study age, sex, job, infective agent and anatomical site of infection have been considered. Retrospective analysis of the records revealed that the ratio of actinomycetoma and eumycetoma was 36:14 that differed significantly (p<0.01), the male to female ratio was 41:21and the peak age for infection was between 40-50 years old but there is not a significant difference between age groups (P>0/05). The single most common site of infection is foot but generally hand and other limbs can be infected and 54. 4% of infected area was in palm and disease can infect other areas with less frequency. The results show that farmers with 45.2% are at greater occupation risk of mycetoma (p<0.01) Keywords: Mycetoma, Actinomycetoma, Eumycetoma, Iran

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Fungal exoenzymes, characteristics and diagnosis

Keyvan Pakshir

Department of Medical Mycology and Parasitology, School of Medicine and Basic Sciences in Infectious Diseases Research Center, Shiraz University of Medical Sciences, Shiraz, Iran Email: [email protected]

The fungi secrets specific enzymes to destroy materials as a part of their life style that play a critical role in their pathogenesis. Invasion by the fungi has been occurred by a combination of mechanical pressure and enzymatic hydrolysis which are important factors for virulence. Some fungi produce proteases and lipases which help them invade the tissues. Candida albicans produces asparatic proteases; lipases and phospholipase which play an important role in their virulence. There are many other exoenzymes such as hemolysin, coagulase, esterase and keratinase that help the fungi for infection. Some of these enzymes are measurable and con be identified by several methods. For example, phospholipase, esterase and hemolytic activity in Candida species con be detected by egg yolk agar, Tween 80 opacity test and blood plate assay, respectively. Fungal exoenzymes give a useful data about fungal pathogenesis and their virulence. Keywords: Fungal exoenzymes, Candida albicans, Phospholipase

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Evaluation of Pneumocystis jirovecii in normal population

Parisa Badiee

Professor Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran Email: [email protected]

Introduction and Objectives: Pneumocystis jirovecii is an important cause of pneumonia in immunocompromised patients. Until recently, the diagnosis of pneumocystosisis has usually been made by microscopic demonstration of the organism by specific staining in bronchoalveolar lavage or trans-bronchial biopsy for the presence of the organism. The PCR assay can detect the low levels of P. jirovecii DNA in respiratory specimens, compared with the routine staining methods, which cannot do so. As this organism exists in the respiratory tracts of the healthy patients, in this study, we try to assess the ability of real-time quantitative PCR method to evaluate P. jirovecii in normal population for distinguishing between active pneumonia and colonization. Materials and Methods: Four hundred and thirty eight bronchoalveolar lavage fluids from normal immunocompotent population without any evidence of clinical or radiological symptoms of lung injury were obtained, when undergoing general surgery and intubation. Both immunofluorescence staining and Real-time PCR were used to evaluate the samples. Results: Of the 438 samples, 7 (1.5%) were positive by microscopic examination and the mean of trophic form count in the whole smears was 2. The lower limitation of diagnosis DNA by PCR was 10 copy numbers/ ml BAL. Quantitative PCR was positive in 12 healthy samples (2.7%), as colonization with CT up to 40 and about 1000-copy number of DNA/ml of BAL. All the colonized cases were asymptomatic, HIV negative, with normal total lymphocyte and CD4+ cell counts. Conclusion: As demonstrated in this study, quantitative PCR assay can differentiate between colonization and infection, which could be particularly helpful for the management of immunocompetent patients. This report provides the first cut off value of P. jirovecii DNA from Iran. Keywords: Pneumocystis jirovecii, Real-time PCR, Immunofluorescence staining

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Aspergillus flavus: A universal challenge for public health and agriculture

Mehdi Razzaghi-Abyaneh, Mojdeh Jamali, Masoomeh Shams-Ghahfarokhi

Department of Mycology, Pasteur Institute of Iran, Tehran 13164, Iran Email: [email protected]

Aspergillus flavus as a human pathogen, allergen and mycotoxin producer is the primary causal agent of hepatocarcinoma in human and a wide range of animal species. The fungus has a world- wide distribution and it is composed of phenotypically and genotypically diverse groups in crops and in the environment. Toxicological data suggest that A. flavus populations largely behave as clones with certain groups scattered in either aflatoxigenic or non-aflatoxigenic strains. Distribution among toxigenic or non-toxigenic is completely depending on biotic and abiotic environmental conditions which affect ecosystems. Niche preference, geographic conditions and agronomic practices all affect the incidence of aflatoxin producing potential of A. flavus populations. Molecular approaches such as random amplified polymorphic DNA (RAPD) provide reliable data for better understanding about genetic diversity and distribution patterns of A. flavus and related aflatoxigenic species. Different genetically divergent groups of A. flavus populations from non-toxigenic to highly toxigenic enable to produce hazardous amounts of AFB1 and CPA may found in soil and air all over the world. These fungi, either toxigenic or not- toxigenic, should be considered as potential threats for public health and agriculture. Keywords: Aspergillus flavus, Genetic diversity, Public health, Aflatoxin, Agriculture

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Determining the concentration of ochratoxin a in breast milk of the mothers living in Khorrambid, Fars province, Iran

Parvin Dehghan, Keyvan Pakshir, Hossein Rafiei, Mostafa Chadeganipour, Mojtaba Akbari

Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran Email: [email protected]

Introduction and Objectives: Mycotoxins, such as Ochratoxins, are toxic secondary metabolites of the fungi which are produced by special species, such as Aspergillus and Penicillium, in suitable conditions. They also play a major role in several human diseases, including different types of toxicities such as nephrotoxicity, suppression of the immune system, and reduction of growth. Thus, the present study aimed to determine the amount of Ochratoxin A in the breast mik samples taken from the mothers living in Khorrambid village, Fars Province, Iran. Materials and Methods: The breast milk samples were taken from the mothers between June and July 2011. The samples were diluted by absolute methanol and after centrifugation; the supernatant was directly used in order to determine the concentration of Ochratoxin A using competitive ELISA. The data were then entered into the SPSS statistical software (v. 16) and analyzed. Results: Among the 87 breast milk samples, 84 ones (96.6%) had Ochratoxin A with the mean concentration of 24.57+13.6 Ng/L ranging from 1.6 to 60 Ng/L. According to the standards of the European Union, 14 breast milk samples (16%) had more than 40 Ng/L contaminations. Conclusion: Existence of Ochratoxin A in the breast milk not only reveals the mother’s consumption of the foodstuffs contaminated with the fungi producing this toxin, but it also shows that both the mother and her infant are contaminated by these toxins which cause irreversible effects on their health as well as the infant’s growth in the long term. Keywords: Mycotoxins, Ochratoxin A, Milk, ELISA

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Curcumin regulates aflatoxin biosynthetic gene cluster in Aspergillus parasiticus NRRL 2999

Zahra Jahanshiri, Masoomeh Shams-Ghahfarokhi, Abdolamir Allameh, Mehdi Razzaghi- Abyaneh

Pasteur Institute of Iran, Tehran 13164, Iran Email: [email protected]

Introduction and Objectives: The effect of curcumin as a natural safe compound with different biological activities was examined on expression of significant genes involved in aflatoxin biosynthetic pathway in Aspergillus parasiticus NRRL 2999. Materials and Methods: A. parasiticus NRRL 2999 was cultured on YES broth in presence of 250 and 1000μg/ml of curcumin in 50 ml Erlenmeyer flasks for 3 days at 28°C. The harvested mycelia mass was flash-frozen in liquid nitrogen and grounded to a fine powder in a porcelain mortar. Total RNA was purified from the homogenized fungal mycelia and First-strand cDNA was prepared according to the protocol. For analysis of the expression of ver-1, nor-1, pksA, omtA and aflR genes in aflatoxin biosynthetic pathway, Real-Time quantitative RT-PCR was carried out using the SYBR Green Master Mix in a final volume of 25 μl for each reaction, and an ABI PRISM 7500 thermal cycler. Results: Analysis of the expression of aflatoxin pathway genes by real time PCR showed that curcumin inhibited the expression of ver-1, nor-1, pksA, omtA and aflR genes at concentrations of 250 and 1000μg/ml. The result showed that curcumin decreased mRNA levels of all tested genes in both concentrations dose-dependently. In 250μg/ml concentration, the expression of ver-1, nor-1, pksA, omtA and aflR was affected by 7.5%, 11.6%, 15.7%, 20.8% and 106.3%, respectively. In concentration of 1000μg/ml, gene expression was reduced by 31.3%, 44.6%, 57.1% 110.9% and 286.7% accordingly. Based on the statistical analyses, reduction in the expression of aflatoxin biosynthesis genes examined in this study was significant only for aflR. Conclusion: Curcumin may be employed successfully as a good candidate in controlling of aflatoxin production in relation to the reduction in the expression of important key genes of aflatoxin biosynthesis pathway in producing fungi. Keywords: Aflatoxin, Aspergillus parasiticus, Curcumin, Aflatoxin inhibitors, Real-Time PCR

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Aflatoxin detoxification in rice using citric acid

Mahin Safara, Farideh Zaini, Seyed Jamal Hashemi, Mahmoud Mahmoudi, Ali Reza Khosravi, Mohammad Hosein Shojai-Aliabadi

Department of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Aflatoxins cause health hazards to human and animals and has also economical problems. Therefore, the detoxification effect of citric acid was investigated in rice as the main food of Iranian people. Materials and Methods: Initially 275 samples of rice were examined for aflatoxins by HPLC. The aflatoxins contaminated samples were later treated by aqueous citric acid and detoxification of aflatoxins were quantified using HPLC. Results: Among the 275 samples analyzed, aflatoxin B1 and aflatoxin B2 were detected in 211 (76.72% of total) samples. Aflatoxin B1 was detected in 203 (73.82% of total) samples with a mean and standard deviation of 2.3±10.21ppb. Aflatoxin B2 together with aflatoxin B1 were detected in only 8 (2.91% of total) samples with a mean and standard deviation of 1.38±2.7ppb of aflatoxin B2 and 2.99±1.56 of aflatoxin B1, respectively. Aflatoxin B1 level in 5 samples (1. 82%) was above the maximum tolerated level of aflatoxin B1 in Iran (5ppb). However considering the Iranian maximum tolerated level for aflatoxins in rice (30ppb), only 3 (1.09%) samples were above the 30ppb and also in regard to the European maximum tolerated level for aflatoxins in rice (4ppb), only 9 (3.27%) samples were considered as higher than 4ppb. Conclusion: The HPLC assay showed that although aflatoxins with a concentration of <30 and <4 ppb in the rice samples were completely degraded, but 97.22% degradation occurred in rice contaminated with 30 and 4ppb when treated with 1N citric acid. These results revealed the efficacy of 1N citric acid in reducing aflatoxins levels in rice. Keywords: Detoxification, Aflatoxins, Aflatoxin B1, Aflatoxin B2, Citric Acid

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Extracellular enzymes production in Rhodotorula species

Zahra Seifi, Sharzad Heydarinia, Ali Zarei Mahmoudabadi

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Rhodotorula species are common airborne contaminant fungi and also considered as normal inhabitants in the skin, lungs, urine and feces in humans. The most common species of Rhodotorula are including; R. mucilaginosa, R. glutinis and R. minuta. Rhodotorula species considered as an important agent for invasive infection among immunocompromised patients. Several studies show that extracellular enzymes such as, phospholipase proteinase and esterase are as pathogenic factors in yeast. The present study, focused on extracellular phospholipase, proteinase, and esterase activities in different isolates of including; R. mucilaginosa, R. glutinis, R. minuta and Rhodotorula species. Materials and Methods: In the present study 68 isolates of different species of Rhodotorula were examined for the production extracellular enzymes. Phospholipase activity assays were performed according to Price et al. using Sabouraud dextrose agar (SDA) supplemented to egg yolk. In addition, proteinase of isolates was evaluated using bovine serum albumin agar medium. Esterase activity was also determined by SDA supplemented to Tween 80. Colony diameter and colony diameter plus precipitation zone were measured for each isolate. Results: The phospholipase activity was detected in 82.4% of Rhodotorula isolates. In the present study phospholipase activity with higher Pz values (<0.70) was seen in 69.1% of isolates. Esterase activity was detected in 29.4% of isolates, whereas 70.6% of them were negative for this enzyme. All isolates of R. mucilaginosa were negative for esterase activity. In the present study all tested isolates were negative for the production of proteinase. Conclusion: Our results show that there are remarkable differences in the enzymatic activity between species of Rhodotorula. In addition, phospholipase activity in Rhodotorula species is remarkable than other extracellular enzymes. Keywords: Rhodotorula, Phospholipase, Proteinase, Esterase

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Evaluation of the sensitivity and overexpression of the genes cdr1 and mdr1 to fluconazole in clinical Candida albicans isolates from Iranian oral candidiasis AIDS patients

Mojtaba Taghizadeh Armaki, Mohammad Hossein Yadegari, Masoumeh Rajabi Bazl, Ehsan Farahbakhsh

Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Email: [email protected]

Introduction and Objectives: Fluconazole-resistant Candida albicans is the cause of oral candidiasis in patients with human immunodeficiency virus infection (HIV). The molecular mechanisms of azole resistance in C. albicans include increased levels of expression efflux transporters MDR1 and CDR1 and alterations in the target enzyme. Materials and Methods: The in vitro activities of fluconazole against 66 isolates of C. albicans were examined. The susceptibility of the antifungal drugs was determined by broth macrodilution and disk diffusion methods according to the Clinical and Laboratory Standards Institute (CLSI). Total RNA was extracted from, drug resistant and sensitive C. albicans. cDNA was synthesized by using the MULV Reverse Transcirptase. Semi quantitative RT-PCR was used to measure expression levels of the genes encoding efflux pumps MDR1 and CDR1 and control gene (ACT1) in 66 isolates of C. albicans. Results: Of the 66 isolates tested with disk diffusion, 45 (68.18%) isolates were susceptible to fluconazole, 5 (7.58%) were susceptible dose-dependent and 16 (24.24%) were resistant to fluconazole. However, in CLSI broth macrodilution test, of the 66 isolates tested, 43 (65.15%) isolates were susceptible, 8 (12.12%) were susceptible dose-dependent and 15 (22.73%) were resistant to fluconazole. Finally, MDR1 and CDR1 genes were overexpressed in 2 (3.03%) and 8 (12.12%) clinical isolates of C. albicans, respectively. Discussion: Nowadays, many advanced and extensive methods are used in reference laboratories to reveal molecular mechanisms beyond drug resistances but Sq-RT-PCR has higher sensitivity relative to Southern blot, Northern blot, general PCR and SDS-PAGE method. Keywords: Candida albicans, AIDS, Resistant genes, Sq-RT-PCR

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Antifungal bacteria in soil community: Diversity and bioactivity

Masoomeh Shams-Ghahfarokhi, Mehdi Razzaghi-Abyaneh

Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran 14115-331, Iran Email: [email protected]

Among the existing biodiversity, bacteria have received major consideration not only for their extremely distribution and population diversity, but also for their capability to produce a wide array of bioactive metabolites with antimicrobial properties. Nowadays, hundreds of chemically diverse antifungal compounds have been isolated from a vast array of bacteria and there are more compounds waiting to be discovered by researchers yet. Since the production of antifungal metabolites in bacteria is quite dependent on the strain and species, ongoing search for finding new bacterial populations to increase the chance of discovering novel antifungals is currently done all over the world. Rapid and accurate screening of a large number of bacteria from soil community could be achieved by visual agar plate assay. Aside from morphology, PCR-based molecular techniques are very suitable for confirmation of identity of isolated bacteria at the genus and species level. Antifungal bacteria to be considered not only as potential candidates for biological control programs, but also for finding rich sources of useful metabolites with potential application in antifungal drug discovery. Here we describe a practical approach for isolation and identification of antifungal bacteria from soil and how we can characterize their bioactive metabolites by novel well-developed techniques Keywords: Bacteria, Antifungal activity, Soil, Aspergillus parasiticus, Bacillus, Pseudomonas

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Paradoxical growth effects of the fluconazole, ketoconazole, voriconazole, and itraconazole on clinical isolates of Candida species

Kamiar Zomorodian, Azadeh Bandegani, Hossein Mirhendi, Keyvan Pakshir

Department of Medical Mycology and Parasitology, School of Medicine, and Basic Sciences in Infectious Diseases Researches Center, Shiraz University of Medical Sciences, Post code 71348- 45794, Shiraz, Iran Email: [email protected]

Introduction and Objectives: In antifungal susceptibility testing of the fungi by broth microdilution methods, a reduced activity of these antifungal agents against certain microorganisms might be seen in high concentrations of these agents which is called as the Eagle effect (paradoxical growth effect). In addition reduced but persistent growth during serial dilution testing might be found which is referred as Trailing growth effect. These effects might affect the results of the test. In the present study, the Eagle and Trailing growth effects were determined during testing of yeasts susceptibility to azoles according to CLSI method. Materials and Methods: A total of 196 Candida isolates were prospectively collected from clinical samples at Shiraz University of Medical Sciences and Tehran University of Medical Sciences. The isolated yeasts were identified by polymerase chain reaction- restriction length polymorphism (PCR-RFLP) as described by Mirhendi et al. Susceptibility to antifungals was tested by the broth microdilution method as recommended by the clinical and laboratory standards institute (CLSI, document M27-A2). The fungal growth in each well was read after 24h and 48h in comparison to the control wells. Results: Eagle effect was found in itraconazole, ketoconazole and voriconazole with frequency of 17% (N=33), 12. 4% (n=24) and 3. 1% (n=6), respectively. The Trilling effect was induced by Voriconazole in 27 isolates (13.8%), by Itraconazole in 26 isolates (13.3%), by fluconazole in 24 isolates (12.2%) and by ketoconazole in 19 isolates (9. 7%). Among the tested Candida, the highest incidence of Trilling effect was found in C. albicans followed by C. dubliniensis and C. tropicalis. Conclusion: Concerning high incidence of Eagle and Trailing growth effects in antifungal susceptibility testing of azoles drugs, the results should be read by experts and the data should be interpreted with caution. Keywords: Candida, Eagle effect, Trailing effect, Broth microdilution

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Evaluation of antifungal activity of new derivatives of imidazole using colorimetric method

Mehraban Falahati, Ensieh Lotfali, Somaye Sharifinia, Azam Fattahi, Sharareh Sayah Alborzi, Mitra Rezaei

Department of Medical Mycology and Parasitology, Tehran University of Medical Science, Tehran, Iran Email: [email protected]

Introduction and Objectives: The incidence of invasive fungal infections has been increasing for two decades. A matter of concern in the treatment of fungal infections is the limited number of efficacious antifungal drugs. Many of available drugs lead to the development of resistance. In order to seek new antifungal agents we assessed the antifungal activity of newly synthesized an imidazole compounds by a colorometric method. Materials and Methods: This was an experimental study that antifungal activity of the new imidazole compounds against Candida albicans, Saccharomyces cerevisiae, Aspergillus niger and Microsporum gypsum was investigated by colorometric method and we compared results with microdilution results. Results: 2-hydroxyphenacyl-azole and 2- hydroxyphenacyl-azolium compounds have been identified as a new class of azole antifungal agents with a good spectrum of activity. The colorimetric method is a simple microtiter method for determining the susceptibility of species of fungal against antifungal agents. Conclusion: In conclusion, most derivatives showed significant in vitro antifungal activities against tested fungi with low MIC values included in the range of 0. 25-32µg/mL is comparable to the reference drug fluconazole. Keywords: Imidazole, Colorometric method, Antifungal activity, Susceptibility tests

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In vitro susceptibility of aflatoxigenic and non-aflatoxigenic Aspergillus flavus strains to conventional antifungals

Azam Fattahi, Farideh Zaini, Parivash Kordbacheh, Seyed Jamal Hashemi, Mahmoud Mahmoudi, Mahin Safara, Roohollah Fateh

Department, of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Iran Email: [email protected]

Introduction and Objectives: Presently appearance of resistance to antifungal agents among Aspergillus species is dramatically increasing. The objective of this study was to look at the in vitro activities of antifungal drugs against Iranian clinical (from nail, bronchoalveolar lavage, paranasal sinus) selected Aspergillus flavus strains. Materials and Methods: The susceptibility of 45 aflatoxigenic and non-aflatoxigenic A. flavus strains were evaluated to six antifungal agents (caspofungin, itraconazole, amphotericin B, ketoconazole, fluconazole, nystatin) using CLSI M38-A2 Broth Microdilution method. Results: The results indicated that 57.1%, 28.6% of aflatoxigenic and 25.8%, 6.5% of non- aflatoxigenic isolates were susceptible to caspofungin, amphotericin B respectively. All isolates but one aflatoxigenic strain were sensitive to ketoconazole. All 45 strains showed to be resistant to nystatin. Also, 64.28%, 92.9% of aflatoxigenic and 64.51%, 100% of non-aflatoxigenic isolates were resistant to fluconazole and itraconazole in ranking order. Conclusion: There was no statistically significant difference between the susceptibilities of aflatoxigenic and non-aflatoxigenic strains of A. flavus to tested antifungal agents. Keywords: Aspergillus flavus, Broth microdilution, Antifungal agents, Susceptibility

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Clinical and paraclinical aspect of 16 cases of mycetoma during past 3 decades in Mashhad

Abdolmajid Fata, Masoud Maleki, Mahmoud Bolursaz, Fariba Berenji, Naser Tayyebi, Sara Fata, Ali Naseri, Mohamad Javad Najafzadeh, Hossein Zarrinfar, Majid Ganjbakhsh

Department of Medical Parasitology and Mycology, Ghaem Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Email: [email protected]

Introduction and Objectives: However mycetoma is the most common disease among sub- cutaneous mycotic infections, but it is a rare sporadic infection. The aim of this study is to investigate the clinical presentation, etiologic agents, epidemiologic factor and the result of treatment among 16 patients admitted at university hospitals of Mashhad. Materials and Methods: During 30 years (1982-2012) a total of 23 patients with deep skin lesions were introduced to Department of Mycology, Ghaem and Emam Reza hospitals, Mashhad Iran. Of those the result of direct examination and culture of 16 patients approved for mycetoma. Demographic clinical and Lab. data of the patients obtained for analysis. Results: Among 23 individual referred to mycology labs, the result of direct examination and culture of 16 patients approved for mycetoma. Of these, 13 individuals (81.25%) were male and 3 (18. 75) were female. Most of them were more than 40 years old. 56% of the patients were farmer. Except one driver who had skin lesions on his hip, in other patients the lesions were located below their knees. Actinomycetoma caused by Nocardia spp., Actinomadura spp. confirmed in 31.2% and 12.5% of the patients, respectively. Eumycetoma due to Fusarium solani, Aspergillus fumigatus and an unknown Dematiaceous fungus confirmed in 3 (18.75%) patients. Most of the Patients received medical treatment, but only two of whom cooperated for follow-up. Surgical treatment and amputation performed for 2 patients. Conclusion: Sporadic cases of Mycetoma still can be observed from many parts of Iran. It should be considered in patients with chronic skin lesions and fistulization. After definite diagnosis, medical treatment could be successful. Keywords: Mycetoma, Actinomycetoma, Eumycetoma, Mashhad

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Recovery of Candida and Aspergillus species in Iranian patients suffering from cystic fibrosis

Melika Laal Karegar, Soheila Khalilzadeh, Somayeh Foladi, Saeed Sadr, Tahereh Shokohi, Hamid Badali

Department of Medical Mycology and Parasitology/Invasive Fungi Research Centre (IFRC), School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran Email: [email protected]

Introduction and Objectives: Cystic fibrosis (CF) is an autosomal recessive disorder due to mutations in the gene cystic fibrosis transmembrane conductance regulator (CFTR) on the chromosome seven. In the respiratory tract, mutations in CFTR result in a defective mucociliary clearance with production of viscous and sticky bronchial mucus that facilitates colonization of the airways by various microorganisms, mainly bacteria, yeast and several filamentous fungal species which contribute to the progressive deterioration of lung function. Therefore the objective of this study was to investigate the frequency of fungi isolated from the respiratory specimens of CF Iranian patients and to determinate the impact of factors including age, gender, inhaled antibiotics and inhaled corticosteroids on fungal colonization. Materials and Methods: 42 patients who have been already diagnosed as CF based on positive sweat chloride tests, characteristic clinical manifestation, or mutations in CFTR protein (29 males and 13 females) enrolled in the Respiratory research centre. Sputum specimens were obtained and direct and culture examinations were performed. Initially, species identification of filamentous and yeast isolates were performed based on conventional mycological morphological, then identification of each isolate was confirmed by sequencing of internal transcribed spacer region and beta Tubulin gene for Candida and Aspergillus species, respectively. Results: A total of 84 respiratory samples were collected with at least two samples per patient. The most predominant recovered fungal genus was Candida (66. 66%) followed by Aspergillus (19.04%). Among Candida species, C. albicans was the most frequently isolated member of the genus. In addition, the most frequently isolated filamentous fungus was A. terreus (37.5%) followed by A. fumigatus and A. oryzae (25%). A higher proportion of patients with persistent Aspergillus and Candida colonization had ABPA than those with no molds. Finally, females were numerically (but not significantly) more often persistently colonized with Aspergillus. Conclusion: An interesting predilection was observed for Aspergillus and Candida species, to preferentially infect patients with CF. However, in the present study, the frequency of Aspergillus species from the sputum secretions of CF patients are low (20.9% of patient), the possibility of fungal colonization and consideration of their clinical significance in these patients is highly recommended. A. terreus was the predominant species which colonized in the majority of CF cases that probably due to resistant to antifungal drug like amphotericin B. Keywords: Candida spp, Aspergillus spp, Cystic fibrosis

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Comparison of Candida colonization in angular cheilitis lesions before and after denture replacement

Abbas Ali Jafari, Mohammad Hossein Lotfi-Kamran, Abbas Falah

Department of Medical Parasitology and Mycology, Medical School, Shahid Sadoughi University of Yazd Medical Sciences, Yazd, Iran Email: [email protected]

Introduction and Objectives: Angular cheilitis is deep fissures with ulcerated appearance, which affected angles of the mouth. Decreasing in vertical dimension of face in denture users is one of the predisposing factors for heavily colonization of Candida in the cheek angles resulted angular cheilitis. Correcting the vertical dimension by a new denture replacement can decrease Candida colonization to prevent or improve the angular cheilitis lesions. The aim of this study was to compare the Candida colonization rate in cheek angles before and after replacing new dentures. Materials and Methods: Twenty eight complete denture users with decreased ridge, and outwear complete dentures, who referred for denture replacement, were randomly selected and their lip angles were cultured before and 3 months after replacing their new denture. The Candida colonization rates before denture replacement and after the vertical dimension correction were compared using Mann-Whitney Wilcoxon statistical tests. Results: All samples were heavily colonized before denture replacement, though 3 months after using new denture only few cultures was positive. A significant statistical difference was observed between Candida colonization before and 3 months after denture replacement (P=0.001). Conclusion: These results suggest that long term using of dentures can cause a wide range of Candida colonization in angular cheilitis lesions resulted angular cheilitis. It can be prevented with changing and replacing a new denture to modify the vertical dimension, and improve the angular cheilitis lesions by decreasing of Candida colonization. Keywords: Candida, Colonization, Angular cheilitis, Denture

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Invasive fungal infection and chronic obstructive pulmonary disease (COPD)

Hamid Badali, Mehrnaz Mohammad Davoudi, Sadegh Khodaveisi

Department of Medical Mycology, School of Medicine, Mazandaran University of Medical Science, Sari, Iran Email: [email protected]

In recent years, the incidence of fungal infection has dramatically increased, especially pulmonary infection by the genus Aspergillus among patients with chronic obstructive pulmonary disease. COPD is a frequent disorder for cigarette smoking which is the most remarkably encountered as a risk factor. Nearly 1%–2% of fatal cases of IPA occur in COPD patients. It has been estimated that COPD will become the 3rd leading cause of death worldwide by 2020. The chronic airflow limitation characteristic of COPD is caused by a mixture of small airway and parenchymal destruction. The latter disorder is also a chronic inflammatory disease which increased the numbers of specific inflammatory cell types in lung airways. These changes in the lungs increase with the disease severity (mild to very sever COPD). Invasive pulmonary aspergillosis (IPA) is a life-threatening pneumonia characterized by lung parenchyma invasion with vasculature erosion and necrosis that is caused by airborne opportunistic fungi belonging to the species Aspergillus. Aspergillus fumigatus is the most common species recovered from patients with IPA. Not only IPA occurs in immunocompromised host, but also healthy individuals like patient with obstructive pulmonary disease and intensive care unit (ICU) patients are as targets. The diagnosis of IPA is hard to establish, because of delay in implementing accurate antifungal therapy and the high mortality rate. Accurate and reliable diagnosis of IPA is adjusted based on tissue biopsy evidencing Aspergillus spp. on microscopic examination or in culture. The results from culture and microscopy of respiratory tract samples have <50% sensitivity and specificity. Other diagnostic approach would be useful such as computed tomography scan, non-culture based methods either in serum or in bronchoalveolar lavage (BAL) for detection of antibody and antigen. Molecular investigations based on PCR have predictive values that need to be further investigated in COPD patients. Antifungal therapy using azole agents like voriconazole is recommended as a first-line treatment of IPA. Keywords: COPD, IPA, PCR, Antifungal therapy

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Candida species in cutaneous candidiasis patients in the Guilan province in Iran; identified by PCR-RFLP method

Ali Akbar Fallahi, Maryam Moazeni, Fatemeh Noorbakhsh, Parivash Korbacheh, Farideh Zaini, Hossein Mirhendi, Hojjat Zeraati, Mehdi Nazeri, Sassan Rezaie

Department of Medical Microbiology and Parasitology and Immunology, Guilan University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Due to the epidemiological alteration in distribution of Candida species as well as significant increasing trend of either intrinsic or acquired in resistance of some of these fungi, the precise identification of Candida species is necessary for effective antifungal therapy and also for prevention of nosocomial infections. PCR-RFLP method is indicated to be a reliable, rapid and simple technique which is able to differentiate the Candida species. In the present study, we applied this method to evaluate the distribution of Candida species in patients affected with cutaneous candidiasis in the Guilan province. Materials and Methods: 896 clinical cutaneous samples were collected from different parts of skin and nail of suspected patients referred to clinical centers all over the Guilan province during 24 months. Samples were examined directly with 15%KOH and cultured on fungal specific media. Genomic DNA was extracted and the restriction enzyme Msp1 was applied for polymorphism analysis. Results: Totally, 47 yeast strains were successfully isolated from different clinical samples and identified by conventional as well as PCR-RFLP methods. The results indicated that Candida albicans (36.17%) was the most frequent species followed by C. parapsilosis (25.53%), C. tropicalis (19.14%), C. guilliermondii (14.89%), C. famata (2.12%) and C. krusei (2.12%). Female finger nails were the most common location to be affected by Candida species. Conclusion: In conclusion, PCR-RFLP method was successfully used for recognition of clinical Candida species within the Guilan province and obtained results revealed C. albicans as the predominant causative agent of cutaneous candidiasis. However, distribution of other Candida species did not completely consist with the reported distribution of Candida species in other parts of Iran with different climate to the Guilan province. Keywords: Candida albicans, PCR-RFLP, Cutaneous candidiasis

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A 5 year epidemiological study of superficial cutaneous in suspicious patients in Tehran (2006-2010)

Seyed Jamal Hashemi, Ayatollah Nasrollahi Omran, Narges Vaseghi

Department of Medical Mycology, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Identification of different species of dermatophytes and cutaneous mycosis agents for finding infection sources and also educating the community in order to familiarize with consequences of contacting infected people or animals will be helpful; and on this basis, the main objective of this study has been determination of distribution and dissemination ways of superficial fungal skin diseases. Materials and Methods: Records of 5500 patients suspected to cutaneous mycosis were evaluated for presence of fungal infection in a 5-year time period from April 2006to March 2010. Skin specimens were collected from patients through skin scraping. Diagnoses were based on direct microscopy examination and culture in accordance with routine laboratory methods of mycology. Results: In total, 2271 cases (41.3%) were inflicted with cutaneous mycosis, of which, dermatophytosis, with 1279 cases (56.31%) was the most prevalent disease of this type. The remaining included 356 cases (15.68%) of tinea versicolor, 283 cases (12.76%) of erythrasma, 243 cases (10.7%) dermal candidiasis and 110 cases (4. 96%) of fungal infections of nails caused from saprophyte moulds. The most common clinical form was tinea cruris with 495 cases (38. 7%). Among dermatophytes isolated from cultures, Trichophyton mentagrophytes was the most prevalent agent with 198 cases (41. 56%). Conclusion: This study shows that dermatophytosis is still an important fungal skin disease among cutaneous mycosis. Keywords: Dermatophytosis, Tinea Versicolor, Candidiasis

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Identification of Malassezia species isolated from patients with pityriasis versicolor by PCR-RFLP method in Arak.

Mojtaba Didehdar, Amir Sayed Ali Mehbod, Zahra Eslami Rad, Mahdi Mosayebi, Behnaz Ghorbanzadeh, Mahmoud Reza Khazaei, Reza Hajihosseini

Molecular and Medicine Research Center, Arak University of Medical Science, Arak, Iran Email: [email protected]

Introduction and Objectives: Malassezia species are lipophilic yeasts and normal flora in human skin. These yeasts are associated with superficial diseases, including pityriasis versicolor, seborrheic dermatitis, atopic dermatitis, folliculitis, dandruff and psoriasis. Identification of Malassezia species by molecular methods is faster and more sensitive than phenotypic methods. In this study, Malassezia species isolated from patients with pityriasis versicolor in Arak; were identified using PCR-RLFP method. Materials and Methods: Skin samples of patients with pityriasis versicolor were cultured on Dixon medium. PCR test was performed directly from Malassezia colonies Targeting D1D2 region of 26SrDNA. Then, the PCR products were examined using enzyme CfoI and create different fragment patterns by RFLP method. Results: From 70 skins sample, 60 samples (85. 7%) were grown. All colonies were positive in PCR test. Malassezia globosa (61. 6%) were the most common species isolated from patients with pityriasis versicolor and the other species were as M. furfur (23. 3%), M. sympodialis (8. 4%), M. restricta (6. 7%), respectively. In one case M. globosa along with M. restricta were isolated. In this study were not isolated M. pachydermatis, M. obtusa, M. slooffiae and other Malassezia species. Conclusion: In our study, similar to other studies that have been done on patients with pityriasis versicolor in Iran, most species isolated were M. globosa and M. fur fur. Keywords: Pityriasis versicolor, Malassezia species, Molecular diagnosis, Arak

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The new formulation of nanoencapsulated essential oil as green antifungal against Fusarium graminearum

Seyed Amir Yazdanparast, Maryam Negahban, Saeed Moharramipour, Seyed Akbar Sadati, Mohamad Pezeshki Modarres

School of Allied Medicine, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Sensu Lato, is a plant pathogen which causes Fusarium Head Blight, a devastating disease on wheat and barley, and the results were compared with effects of Bonmyl. In this research, the essential oil of Artemisia sieberi Besser was encapsulated by insitu polymerization of Oil/Water emulsion in nano scale. Then the antifungal activity of produced nanoencapsuled essential oil (NEO) was examined against Fusarium graminearum. Materials and Methods: Aerial parts of A. sieberi were collected at full-flowering stage in October, 2011. Essential oil was extracted from the plant samples using a Clevenger-type apparatus where the plant material is subjected to hydrodistillation. The nano-encapsulation process was carried out by polymerization technology. Essential oil was used as a core material, and Urea (U) and formaldehyde (F) as shell materials. Antifungal toxicity bioassay was deliberated to assess IP% of NEO. At first, concentrations range of toxicity of nano-encapsulated oil (NEO) and benomyl were determined by using a preliminary experiment and logarithmic distance. Then, the toxicities of the essential oils against the test fungus were tested at 0, 50,80,100, 120, 160, 200, 240, 300, 500,750, 1000, 1500 and 2000 ppm with potato dextrose agar (PDA). In treatment plates, the requi-site amount of NEO was added to PDA to produce a concentration of 2,000 ppm. Results: During screening of NEO and benomyl isolated from different concentration, NEO of A. sieberi possessed absolute toxicity against F. graminearum. The Inhibition Percent (IP) considerably, ranging from 5% to 100% at concentration 50 and 240 ppm respectively. Despite the fact that the benomyl isolated could not completely inhibit the growth of test fungus even at 240 ppm. The NEO at its concentration (160 ppm) showed a 60% inhibition. At the MIC (50 ppm), the oil exhibited its fungistatic nature. The benomyl exhibited a broad spectrum of fungi toxicity by inhibiting completely growth at 1000 ppm. Conclusion: Despite fundamental differences in biology, physiology of tested fungi, the general response of fungi to the toxic effects of nanoencapsulated essential oil were higher than and nanoencapsulated oil in many tests, that shows the potential of this new formulation in different situations. As the new technology of nanoencapsulation of essential oils through the control release of effective components, to overcome the restrictions on plant essential oils usage in agricultural products and farms. Keywords: Nanocapsule, Artemisia sieberi, Anti fungal toxicity, Fusarium graminearum

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Evaluation of the effects of Zataria multiflora, Geranium pellargonium roseum, Myrth and Lemon shell essences on immune system function in experimental animals

Hojjatollah Shokri, Ali Reza Khosravi

Faculty of Veterinary Medicine, University of Mazandaran, Amol, Iran Email: [email protected]

Introduction and Objectives: In this study, effects of some Iranian herbal essences have been evaluated on the function of immune system using experimental animals. Materials and Methods: Fifty rabbits were divided into 10 groups, consist of 5 animals, each group was administrated orally and or subcutaneously by Zataria multiflora, Geranium pellargonium roseum, Myrth and Lemon peel essences and normal saline (control group), 6 times with 6 days of interval. One day before the first administration and 3 days after the last one, rabbits were bleeded from marginal vein. Candida albicans antigens were prepared using standard techniques. Five days after the last injection of the essences, Candida antigens were injected subcutaneously (SC) and intramuscularly (IM) in all animals. Then ELISA test carried out using Candida antigens, sera before and after immunization. Also, phagocytosis and killing assays and lymphocyte transformation test (LTT) were carried out on blood samples. Results: The results indicated that cellular immunity were stimulated against Candida albicans antigens and con-canavalin A (Con-A) mitogen significantly in animals were injected subcutaneously by Z. multiflora and G. pellargonium roseum in comparison with control group, whereas Myrth essence had no considerable effect and Lemon peel essence suppressed the cellular responses. Z. multiflora, Myrth and Lemon peel essences stimulated innate immunity when injected subcutaneously, whereas G. pellargonium roseum essence had no significant effect. Humoral responses to Candida antigens were significantly decreased in animals injected by Lemon peel essence comparing to other essences (P≤ 0. 05). Conclusion: From the present study, it is concluded that Z. multiflora and then Myrth and G. pellargonium roseum essences have immuno-stimulatory effects, therefore it is not recommend for human use. Keywords: Candida albicans, Zataria, Geranium, Myrth, Lemon, Immunomodulatory, Innate immunity

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Assessment of antifungal activities of Urtica dioica against Microsporum canis on the in vitro and in guinea pig model of dermatophytosis

Ali Mikaeili, Isaac Karimi, Masoud Modaresi, Zahra Bagherinasab

Department of Medical Mycology, School of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran Email: [email protected]

Introduction and Objectives: To investigate the antidermatophytic effects of Urtica dioica L. against Microsporum canis in the in vitro and in guinea pig model of dermatophytosis. Materials and Methods: The in vitro antifungal activity of extracts was measured by disc diffusion test and determination of minimum inhibitory concentrations (MIC). The dorsum of animals was infected with Microsporum canis and topically treated as follows: negative control (NC) received no treatment, positive control (PC) received terbinafine 1%, DMSO10% group received DMSO as vehicle, and three other groups: SN10%, SN20% and SN30% which received 10%, 20% and 30% hydroalcoholic extract dissolved in DMSO10%, respectively. Results: Both extracts did not show considerable antifungal activity against M. canis as compared with terbinafine. MICs of aqueous and hydroalcoholic extracts were 30 and 20 mg. ml- 1, respectively. The PC and SN30% groups showed normal hair growth on day 15 while their skin looked healthy on day 30 postinoculation. The lesion severity and clinical scores declined in SN30% and PC groups in comparison to NC group on day 30 postinoculation. On day 30 postinoculation, the clinical efficacy of used formulations increased to 11.76, 23.52, 76.47, 5.88, and 94.11% for SN10%, SN20%, SN30%, DMSO10%, and PC groups, respectively. Conclusion: The hydroalcoholic extract of this plant may possibly be further developed into adjunct drug for topical treatment of tinea corporis. Keywords: Urtica dioica L., Microsporum canis, Kurdish Ethnomedicine, Guinea Pigs

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Expression analysis of hyphae-involved genes in Candida albicans treated by allicin originated from garlic

Alireza Khodavandi, Fahimeh Alizadeh

Department of Paramedical Science, Gachsaran Branch, Islamic Azad University, Gachsaran, Iran Email: dr. [email protected]

Introduction and Objectives: Candida albicans is one of the opportunistic fungi causing candidiasis in humans. It has been demonstrated that one of the main factors for virulence of C. albicans is yeast-to-hyphal-phase dimorphic transition which is controlled by SIR2 and other factors. On the other hand, the ability of Candida to release aspartyl proteinases (SAP) is another strategy for pathogenicity. Allicin, one of the sulfur and volatile compounds from garlic has been shown to possess a broad spectrum of antifungal activity against some pathogenic fungi. In this study, we endeavored to investigate the effect of allicin on the morphology switching phenomenon in C. albicans and to compare the effect of allicin against fluconazole SIR2, SAP1, SAP2, SAP3 and SAP4 gene expression. Materials and Methods: The effect of allicin on growth and hyphae production in C. albicans as compared to fluconazole was investigated using survival time in vitro and microscopic image at time intervals. Time kill study also was performed for both allicin and fluconazole-treated samples in terms of the fungal load reduction. Additionally, the expression of selected genes involved in hyphae formation and development such as SIR2 and SAPs1-4 was evaluated by semi-quantitative RT-PCR and relative real time RT-PCR. Results: Allicin was shown to down-regulate the expression of SIR2 (5. 54 fold), similar to fluconazole (3. 48 fold) at 2× MIC concentrations. Interestingly, allicin had no effect on SAPs1-4 expression, whereas fluconazole was able to suppress SAP4 expression. Conclusion: Our findings showed that allicin was effective in suppressing hyphae development of C. albicans to an extent that is sometimes equal or more than fluconazole. Moreover, allicin and fluconazole seemed to share a common anti-Candida mechanism through inhibition of SIR2 gene, while fluconazole appeared to also exert its fungistatic effect through another pathway that involved SAP4 suppression. Keywords: Allicin, Candida albicans, SIR2 gene, Hyphae

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Use of the extract of Myrtus communis to inhabit biofilm formation of Candida tropicalis

Nahid Shoaie, Parisa Mohammadi, Shahla Roudbar Mohammadi

Department of Biology, Faculty of Science, AL Zahra University, Tehran, Iran Email: [email protected]

Introduction and Objectives: Pathogenic yeasts from the genus Candida can be the reason of serious infections in human especially, in immune compromised patients and are now known as major agents of nosocomial infections. Many such manifestation of infections associate with the formation of Candida biofilms. This structures are extremely resistant to chemical drugs and are cause of many problems in medicine. The aim of this study was to use the extract of Myrtus communis to inhabit biofilm formation by Candida tropicalis (ATCC0750). Materials and Methods: In this study, the extract of M. communis was prepared and then the concentration of extract was measured. The minimum inhibitory concentration (MICs) and minimum fungicidal concentration (MFC) of this natural product was determined according to CLSI by serial microdilution method. After that, biofilm was formed in microtitre plate and antibiofilm activity of this extract was assayed by MTT method. Results: The extract of M. communis has been shown appropriate antifungal and antibiofilm activity against C. tropicalis (ATCC0750). The quantity of MIC50 of this extract was 15/62µg/ml. The result indicated that this extract was capable to inhibit %33.4, %38.9 and 46.7% biofilm formation by this fungal specie in concentrations 1000, 2000 and 4000µg/ml. Conclusion: It can be concluded that extract of M. communis has antifungal and antibiofilm effect and can be used to inhibit biofilm structures. Further investigation must be done to evaluate effect of this extract against other Candida species. Keywords: Myrtus communis, Candida tropicalis, Biofilm

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Chemical composition, antifungal and antibiofilm activities of the essential oil of Mentha piperita L.

Marjan Motamedi, Kamiar Zomorodian, Mohammad Jamal Saharkhiz, Keyvan Pakshir, Ramin Miri, Kimia Hemyari

Department of Medical Mycology and Parasitology, School of Medicine, Shiraz University of Medical Sciences, Postal code 71348-45794, Shiraz, Iran Email: [email protected]

Introduction and Objectives: Medicinal plants have been used for centuries in traditional medicine because of their therapeutic value. Mint species have been exploited by man for more than two thousand years. Peppermint itself has been used for more than 250 years. The goal of this study was to investigate the chemical composition and in vitro antifungal and antibiofilm activities of essential oil of leaves of Mentha piperita collected in the region of Fars province in Iran. Materials and Methods: The variations in quantity and quality of essential oils (EOs) from the aerial parts of cultivated M. piperita were determined. The EO of air-dried sample were obtained by hydro distillation method and analyzed by gas chromatography/mass spectrometry (GC/MS). The antifungal activity of the EO against 25 standard strains of fungi was investigated by broth micro dilution methods as recommended by Clinical Laboratory Standard Institute. A semi quantitative measure of biofilm formation was assayed by using XTT reduction assay. Results: Mnenthol (53.28%) was the major compound of the EO followed by Menthyl acetate (15. 1%) and Menthofuran (11. 18%). The EO inhibited the growth and killed the tested yeasts at concentrations of 0.12–4µL/mL 1-8µL/mL, respectively. No significant differences in inhibitory concentrations were found between azole-resistant and -susceptible strains. In addition, the EO inhibited the growth and killed the standard strain of Cryptococcus neoformans at concentration of 4µL/mL. All of the Aspergillus standard strains were susceptible to M. piperita EO at concentrations of 0. 5-4µL/mL. The EO inhibited the biofilm formation of Candida albicans and C. dubliniensis at concentration up to 2µL/mL. Conclusion: Considering wide range of antifungal activities of the examined EO, it has the potential to be used in the management of fungal infections or in extension of the shelf life of food products. Keywords: Essential oil, Mentha piperita, Candida, Antifungal activity, Biofilm

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RNA-mediated gene silencing in Candida albicans: reduction of fungal pathogenesis by use of RNAi technology

Maryam Moazeni, Mohammad Reza Khorramizadeh, Parivash Kordbacheh, Hojat Zeraati, Fatemeh Noorbakhsh, Laden Teimoori-Toolabi, Sassan Rezaie

Division of Molecular Biology, Department of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: The introduction of RNA silencing machinery in fungi has led to the promising application of RNAi methodology to knock down essential vital factor or virulence factor genes in the microorganisms. Efg1p is required for development of a true hyphal growth form which is known to be essential for interactions with human host cells and for the yeast’s pathogenesis. In addition, it is responsible for positive regulation of the expression of several hyphal-specific genes: SAP5, which encodes secreted aspartic proteinase, and ALS3, which encodes a multi-functional adhesive polypeptide. In this paper, we describe the development of a system for presenting and studying the RNAi function on the EFG1 gene in Candida albicans. Materials and Methods: The 19-nucleotide siRNA was designed on the basis of the cDNA sequence of the EFG1 gene in C. albicans and transfection was performed by use of a modified- PEG/LiAc method. To investigate EFG1 gene silencing in siRNA-treated cells, the yeasts were grown in human serum; to induce germ tubes a solid medium was used with the serum. Quantitative changes in expression of the EFG1 gene as well as two Efg1-associated genes, ALS3 and SAP5, were analyzed by measuring the cognate EFG, SAP3 and ALS3 mRNA levels by use of a quantitative real-time RT-PCR assay. Results: Images taken by fluorescent microscopy method indicated the effectiveness of transfection. Compared with the positive control, true hyphae formation was significantly reduced by siRNA at concentrations of 1 µM, 500 nM, and 100 nM (P\0. 05). According to REST® software data analysis, a considerable decrease in EFG1 gene expression was observed when applying both 500 nM and 1µM of siRNA (P<0. 05). Consequently, the expression of ALS3 and SAP5 were significantly down-regulated both in yeast treated with 500 and 1 µM of siRNA (ρ<0.05). Conclusion: On the basis of the potential of post-transcriptional gene silencing to control the expression of specific genes, these techniques may be regarded as promising means of drug discovery, with applications in biomedicine and functional genomics analysis. Keywords: RNA-mediated gene, Candida albicans, RNAi technology

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The relationship between gene ERG11 and resistance of fluconazole Candida albicans in patients to vaginal candidiasis

Sajad Amani, Ayat Nasrollahi Omran, Ali Nazemi, Parvaneh Afshar, Fatemeh Rahmani, Sahar Yaghobian

Islamic Azad University of Tonekabon Branch, Tonekabon Iran Email: [email protected]

Introduction and Objectives: Drug resistance to azoles in Candida, especially fluconazole is due to different factors, and molecular mechanisms that created these resistance one include excess expression of number of pumping genes towards out of cell or creating point mutation in ERG11 gene in target enzyme or resistance to excess expression. Materials and Methods: 56 prepared samples from infected patients to vulvovaginitis candidiasis have been studied after culturing in medium of Sabouraud’s dextrose agar and CHROMagar. Then based on NSSL protocol, MIC technical was preformed for determining sensitivity of identified yeast. The full duplicated genes ERG11 through Real-time PCR and identify fragments containing the SNP by the High Resolution Melting point difference was the presence SYT_9 color. The different groups based on the melting point of the sequence were determined by PCR-Sequencing. ERG11 gene sequences for each sample were compared with normal sequence aligned with the software and identify the types of mutations in finally done. Results: We have seen in the survey conducted in 8 fluconazole-resistant yeast Candida albicans ERG11 gene changes in amino acid. Conclusion: ERG11 gene expression has seen C. albicans of fluconazole resistance. That may cause mutations in the ERG11 gene of Candida albicans resistant to fluconazole. Keywords: Candida albicans, Fluconazole, ERG11 gene

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Application of Real-time PCR, Nested-PCR and mycology assay using BAL samples for detection of Aspergillus flavus and A. fumigatus in hematological malignancies

Hossein Zarrinfar, Hossein Mirhendi, Abdolmajid Fata, Koichi Makimura, Omolbanin Paknejhad, Sasan Saber, Hossein Khodadadi, Kazuo Satoh, Parivash Kordbacheh

Department of Medical Parasitology and Mycology, Ghaem Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Email: [email protected]

Introduction and Objectives: Invasive aspergillosis (IA) represents a major cause of morbidity and mortality in patients with hematological malignancies. IA is caused by several pathogenic Aspergillus species, mainly A. fumigatus and A. flavus. To evaluate four diagnostic methods: direct microscopy, culture, nested-PCR and real-time PCR for detection of A. fumigatus and A. flavus and to investigate the incidence of pulmonary aspergillosis (PA) on bronchoalveolar lavage (BAL) samples in patients with hematological malignancies in Tehran, Iran. Materials and Methods: During 16 months, 28 BAL samples from patients with hematological malignancies were obtained. Direct wet mounts on %20 KOH and culture on mycological media were performed. The nested-PCR detection of A. fumigatus and A. flavus was based on conserved sequences of internal transcribed spacer 1 (ITS1) ribosomal DNA and the real-time PCR assay targeted β-tubulin gene. Results: Of the 28 samples, 2 (7. 1%) were positive in direct examination having branching septate hyphae, 6 (21. 4%) had positive culture with 5 (83%) A. flavus and 1 (17%) A. fumigatus, 12 (42. 9%) were positive in nested-PCR and 5 (17. 9%) had positive real-time PCR. The incidence of aspergillosis in these patients included probable IPA in 6 (21. 5%), possible IPA in 13 (46. 5%) and not IPA in 9 (32%). A. flavus was the most common cause of PA. Conclusion: Because of high sensitivity of nested-PCR, it may increase the false positive result due to fungal contamination and colonization; therefore, it is not recommend for BAL samples in diagnosis of PA. However, in high risk patients, a positive result of real-time PCR or nested- PCR corresponding with clinical signs or symptoms could be valuable. Keywords: BAL, IPA, Aspergillus flavus, A. fumigates, Hematological malignancies

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Development of a quantitative TaqMan Real- Time Polymerase Chain Reaction for detection of Pneumocystis jirovecii from bronchoalveolar lavage fluids

Hossein Khodadadi, Hossein Mirhendi, Mehdi Mohebali, Koichi Makimura, Hossein Zarrinfar

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of medical sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Pneumocystis jirovecii is an important cause of life-threatening pneumonia in immunocompromised patients. Diagnosis of this fungal infection relies on microscopic examination of respiratory specimens using different staining methods. These methods have low sensitivity and cannot differentiate colonization from infection. Real-time PCR can quantify the DNA burden of the organism and may help to solve this problem. Materials and Methods: A real-time PCR was designed targeting 18s rRNA gene of P. jirovecii for the detection and quantification of P. jirovecii DNA. Tenfold serial dilutions of a standard plasmid containing the target sequence were used for quantification. A total of 458 bronchoalveolar lavage (BAL) samples were examined with this newly designed method. Results: The developed real-time PCR assay was able to detect as low as 10 copies of target DNA of standard plasmid. We consider the threshold cycle 36 (10 copy number) as the cut off point for the detection of P. jirovecii in BAL samples. The assay could detect and quantify specific DNA of P. jirovecii in 47 out of 458 BAL specimens. Conclusion: This novel real-time PCR could detect and quantify the burden of P. jirovecii DNA in respiratory specimens and has a potential to help differentiating between colonization and infection. Keywords: BAL, Pneumocystis jirovecii, Real-time PCR

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Fungal genomic projects, metagenomics and gene analyses

Abdolhassan Kazemi

Department of Medical Parasitology and Mycology, Tabriz University of Medical Sciences, Tabriz, Iran Email: [email protected]

Fungal genomic projects can provide a deep and wide range of knowledge, information and data about biology, physiology, cytology, enzymelogy, biochemistry and fungi for scientists. According to the understanding of fungi genome concepts, the possibility of bioinformatically processed of an exact fungus genome can also provide them with a notable, noteworthy and scientifically predicted data about certain fungous transcriptomic, proteomic, metabolomic, cytomic and system biology. Exactly precise and systematic data in each of the mentioned field (for example proteomic) could be very beneficial to understanding: fungi pathogens, tissue invasion, drug resistance or sensitivity, clinical symptoms process, anti fungal drug design and etc. In addition, the provided detail from fungi genomic projects could be very effective on producing industrial and biotechnological products using fungi as a very important bioreactor. The economical cost of fungal products in biotechnology fields have been estimated more than billions of dollars. Moreover, fungal genes analysis and also genomic meta-analysis are other benefits to fungal genomic projects. The survey of fungal genomic projects (data mining) wing collected genomic data in gene banks and fungal data bases have played an important, comprehensive and invaluable role for the comparison of genomic data with other different microorganism. Comparing various genomic data with each other, could of course be a valuable method for clear understanding of fungal molecular biology and their genes functions. Among 14539 completed Eukaryotic genome projects, 2312 genome projects were about fungi. From the 2312 mentioned fungal genomic projects, 911 were for Ascomycota, 358 for Basidimycota, 5 for Chytridiomycota, 11 for Glomeromycota and 13 about Zygomycota. The shortest fungal genome has been found in Encephalitozoon romaleae (2. 9 Mb, CG= %41. 24) containing 1883 genome with ability for synthesis of 1831 proteins (Bioproject PRJNA 51607). The largest known fungal genome was recognized in Fusarium oxysporum (61. 47 Mb, CG= %48. 23) (Bioproject PRJNA 18813) but proteomic data about the quantity of synthesized proteins by this genome has not yet been released. The difference between the second largest fungal genome with the first is around 20 Mb. By now, the second largest fungal genome has been recognized in Magnoporthe (40. 98 Mb). Magnoporthe genome has 13022 genes with the ability for biosynthesis of 12836 proteins (Bioproject PRJNA 13840). The other largest fungal genome was sequenced in Zymoseptoria tritci (39. 69 Mb) Myceliophthora thermophilia (38. 47 Mb) and Aspergillas oryzae (37. 88 Mb). These genomes have 10964 genes (10963 proteins), 9294 genes (9099 proteins) and 12335 genes (12062 proteins) respectively. Sporisorum reilianam has also presented 23 chromosomes (18. 4 Mb, CG=59. 51%) and then Z. tritci 21 chromosomes (39. 69 Mb, CG=84. 67%), while Schizosacoharomyces pombe only had 3 chromosomes (12. 59 Mb, CG=36. 04%) and two strains of Komagataella pastrosis have 4 chromosomes (slightly more than 9 Mb, CG=41%). Genomic projects, genomic information and also metagenomic analysis/gene analysis are necessary tools for using fungi as bioreactor for producing biotechnological and industrial products (attention to gene promoter structure, designed mutation for protein engineering, codone usage). Also these kinds of analysis using bioinformatical tools can be very useful for (44)

understanding more detail about the natural properties of fungi. Availability of genomic information and detail is of course necessary prior to the molecular study of fungi. Also, metagenomic analysis of fungi genome can create a scientific image about fungi genome/genes structure and arrangement. For example, Z. tritci with 39.6 Mb genome size, has 10964 genes and 10963 synthesed proteins while A. oryzae with 2 Mb lese genome size from Z. tritci genome, has 12335 genes (~1300 more genes) and 12062 syntheses proteins (1100 more proteins). These details could probably show a smaller size of A. oryzae genes, the larger intronic segment and space and also existence of more pseudogenes or processed pseudogenes in the genome of A. oryzea. Keywords: Genomic projects, Genome analysis, Genomic, Metagenome

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Molecular identification of Fusarium sibiricum isolated from Iranian grains

Omid Shirzad, Reza Kachuei, Mohamad Ali Afshari, Sasan Rezaie, Mohamad Hossein Yadegari, Seyed Amir Ghiasian

Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: The type A trichothecenes, in particular T-2 toxin, is the most toxic metabolite produced by Fusarium species. These mycotoxins are certainly produced from Fusarium sporotrichioides and F. langsethiae. In previous studies, isolate of Fusarium genus were isolated from stored grains in Tehran which were produce of T-2 toxin and based on morphology and molecular (primers, ITS and partial EF1-α) characterization have been reported as F. langsethiae. Materials and Methods: In the current study, the available isolate were validated in parallel with the standard strains and available data banks in NCBI as related to F. sibiricum, F. sporotrichioides, F. langsethiae, F. poae and F. graminearum and these were investigated through the Sequencing of IGS rDNA, PHO, RPB2, TUB2, EF-1α and TRI1-TRI16 genes. After manual extraction of the DNA from the Fusarium species under investigation, with the aid of the associated primers (at least 13) the PCR product of each gene in order 2400bp, 800bp, 1900bp, 1400bp, 700bp, 1800bp were extracted from the gel and were sequencing. Results: The sequences were aligned using Mega 5, followed by visual inspection and manual adjustment, and eventually the phylogenetic tree has been mapped out in relation to each gene using the UPGMA method. Investigation results were indicate F. sibiricum which has been introduced and referred in recent times by Mattila et al (2011). Conclusion: According to our knowledge, this is the second report in the literature. The researches show that F. sibiricum in comparison with other generating species, including F. langsethiae and F. sporotrichioides is able to produce high level of T-2 toxin. In such case, it is necessary and mandatory to carry out research in more detail with the objective of identification of T-2 toxigenic Fusarium species in cereal grains. It is also recommended that the growth conditions and producing of T-2 toxin by these species in stored grains are brought to the attention of the relevant authorities. Keywords: Fusarium sibiricum, Iran, Grain, Molecular analysis

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Development approach of fungal dendritic cell vaccines

Ali Reza Khosravi

Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran Email: [email protected]

Dendritic cells are antigen-presenting cells with a unique ability to induce primary immune response. Immature DCs shallow yeast hyphae of Candida albicans and also Aspergillus conidia and induce different Th cell response to the fungus. With respect to different conditions, activated DC result in production of Th1 cytokines and Th1 priming, or Th2 production and Th2 priming. In experimental animal model, injection DC pulsed with Candida blastospore or Aspergillus conidia can induce antifungal protective immunity. DC obtained from spleens or bone marrows transected with yeast or conidia RNA caused express fungal mannoprotein or β- glucan on their surface and undergo functional maturation, as revealed by the unregulated expression of MHC class II antigens, and produce IL-12. In this condition, they are capable of inducing Th1-dependent antifungal resistance in vaccinated animal model. Based on the obtained results, the efficacy of DC pulsed with yeast or conidia RNA as fungal vaccines and point to the potential use of RNA-transfected DC as anti-infective vaccines in some transplant patients could be acceptable.

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Evaluation of Candida albicans allergens reactive with specific IgE in atopic dermatitis patients by Immunoblotting

Ali Naseri, Ali Reza Khosravi, Mohammad Moazzeni, Parvin Mansouri

Department of Medical Mycology and Parasitology, Imam Reza Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Email: [email protected]

Introduction and Objectives: Candida albicans, the major etiologic agent of candidiasis, is a normal inhabitant of the alimentary tract and the mucocutaneous regions. It produces important allergenic components which can induce allergic reactions in sensitized patients. Atopic dermatitis is a chronic, inflammatory and itching disease of the skin with an unknown etiology. It seems that, C. albicans has an important role in pathogenesis of the disease. The goal of this study was to determine the allergenic components of C. albicans that react with IgE antibodies in atopic dermatitis patients. Materials and Methods: In this study C. albicans was cultured massively and then a suitable antigen (cytoplasmic extract) was prepared from it. In the next step, the protein components of crude extract were determined and separated by SDS-PAGE in a discontinuous buffer system using a 5-17.5% gradient gel. Separated proteins were transferred to nitrocellulose sheets and immunoblotting was done with serum samples from 95 patients with atopic dermatitis. The patients were chosen as sequential trial. 70 serum samples from normal none atopic persons were used as negative controls. Results: In SDS-PAGE protein bands with molecular weights between 13-135 KDa were appeared. The specific IgE was detected against 33 of these bands in immunoblotting. 16, 20, 22, 23, 25, 30, 32, 34, 53, 57 and 94 KDa protein bands were the most important allergenic bands in atopic dermatitis patients, among them 25, 34 and 57 KDa bands were the major allergen which reacted with 76. 3, 50, and 42 percent of serum samples respectively. Of 95 sera obtained from patients with atopic dermatitis 38 cases (40%) showed positive results, but all of the healthy controls were negative. There were significant differences between patients and negative controls (p<0. 0001). Conclusion: C. albicans produces several allergenic components that can induce allergic reactions and may be pathogenetically important in patients with atopic dermatitis. Keywords: Candida albicans, Atopic dermatitis, IgE, Immunoblotting

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Aspergillus monitoring project in an Iranian educational hospital

Kambiz Diba, Khadijeh Makhdoomi, Mohammad Hossein Rahimirad, Davood Jabbari

Department of Medical Mycology and Parasitology, School of Medicine, Urmia University of Medical Sciences Email: [email protected]

Introduction and Objectives: The main object was monitoring of Aspergillus infections and epidemiological approaches in the greatest university hospital of Urmia, Iran. The subject of our study included bronchial fluid and sputum were collected from the hospitalized patients with acute respiratory symptoms. Materials and Methods: All clinical specimens were transported to the medical mycology lab, UMSU, for the diagnostic purposes and fungal identifications. For environmental detection of Aspergillus isolates, some specimens were collected from air and environment surfaces. A morphological study was firstly performed including; growth characteristics and microscopic features of Aspergillus species on mycological media. For the confirmation of Aspergillus isolates which similarly found in clinical and environmental sources, PCR-RFLP using a novel restriction enzyme Mwo I and an additional molecular technique of RAPD were carried out. Results: A total of 102 fungal isolates, including Candida species 82 (80%), Aspergillus spp. 20 (19.6%) and the other fungi 2 (0.4%). Among the clinical isolates of Aspergilli; A. flavus (8.4%), A. fumigatus (4.2%) and A. niger (2.8%) were identified, as well as environmental Aspergillus isolates A. flavus (19.4%), A. niger (6.5%) and A. fumigatus (6.5%). Conclusion: Comparing the clinical and environmental findings, 2 of 11 clinical Aspergillus isolates were matched with environmental isolates in RAPD method and the other environmental sources were not confirmed. Keywords: Aspergillus, Epidemiology, Hospital

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Analysis of airborne allergenic fungi in farms and the rate of their caused allergic diseases in farms’ staffs

Hatef Ajoudanifar, Alireza Khosravi, Mohammad Taghi Hedayati, Mansour Bayat, Hojjatollah Shokri

Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran Email: [email protected]

Introduction and Objectives: The spores of various species of fungi such as Cladophialophora, Aspergillus and Alternaria are distributed in air and are considered as important factor in allergenic diseases (airborne fungi). Therefore, long term exposure to spores of some fungi can cause different diseases such as asthma and broncopolmolary Aspergillus. Materials and Methods: In this study, air sampling was done from indoor and outdoor of 20 cattle and, 25 dwelling farms and 10 public places. After 5-7 days from incubation, the resulting colonies were counted and differentially diagnosed. Blood sampling and IgE assessment were done after giving registration forms from all the staffs who worked 6 hours per day at least in one year. Results: In all cultured plates the maximum numbers of counted colonies were specified to Cladophialophora, Aspergillus and Alternaria. Six of staffs showed allergic symptoms. IgE assessment determined that 5 and 3 of these staffs had specific IgE against Cladophialophora and Aspergillus, respectively. The assessed specific IgEs against Cladophialophora and Aspergillus were significantly correlated to each other (p < 0. 05). Discussion: In this study the most prevalent fungi which potentially are allergenic, in indoor and outdoor air of farms were determined. The results show that it is necessary to reduce the amounts of allergenic fungi in indoor air of farms by suitable air conditioning and decreasing of damp. Furthermore, based on the specific IgE analyses in staffs’ blood serum, essential strategies should be considered in order to decrease the time of staffs’ exposure with indoor air dusts of farms. Keywords: Specific IgE, Airborne fungi Cladophialophora, Aspergillus, Alternaria

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Propolis efficacy on candidiasis histopathology and on TNF-α, IFN-γ and IL2 cytokines production in old mice with and without systemic candidiasis

Mahnaz Fatahinia, Ali Reza Khosravi, Hojjatollah Shokri

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Propolis is a resin component collected by honey bees from different part of plants. The immunomodulatory and antimicrobial activities of the propolis have been approved by investigators. The purpose of this study was to evaluate the effects of Iranian propolis on stimulation of IL-2, TNF-α and IFN-γ secretions in aged mice with systemic candidiasis. Materials and Methods: In this study, mice were enrolled to prophylaxis with propolis (100 mg/kg. bw) for 2 days and intraveneously (IV) injected with 2×105 Candida albicans blastospores to induce systemic candidiasis. The aged and young control mice were received phosphate buffered saline (PBS) in rout of oral and IV rout. Five days after injection, the levels of IL-2, TNF-α and IFN-γ were measured by ELISA method. Production of the above cytokines was assessed in spleen cells, with presence or absence concanavalin A (0.5μg/ml) in culture medium. Results: The results demonstrated that the level of IFN-γ, as a key cytokine in innate immunity against systemic candidiasis, was significantly enhanced in infected/ propolis group (P< 0. 001) when compared to infected group. In addition, propolis could decrease the IL-2 and TNF-α when compared to aged control mice, in which approved the anti-inflammatory effects of this agent. According to the obtained results, there were significant differences among various study groups, stimulated and non-stimulated media (P<0.05) that sometimes was followed by cytokine’s pattern in mice sera. Conclusion: Based on histopathological findings, the lesions were categorized in degrees 2 and 3 for the kidneys in infected / propolis group and infected group, respectively. Keywords: Propolis, Candidiasis, IL-2, TNF-α, IFN-γ

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Study the effects of different types of interferon pretreatments on expression of cathelicidin in murine macrophages following exposure to Candida albicans

Kamiar Zomorodian, Elnaz Heshmat Azad, Zahra Rezaei, Bahador Shahriari, Farideh Jalali- Mashayekhi, Keyvan Pakshir

Department of Medical Mycology and Parasitology, School of Medicine and Basic Sciences, Infectious Diseases Research Center, Shiraz University of Medical Sciences. Email: [email protected]

Introduction and Objectives: Antimicrobial peptides (AMPs) are part of the natural defense of the most living organisms. The cellular expression of these peptides could be induced by cytokines and microbial products. Gamma-interferon (IFN) immunotherapy has been used successfully as an adjunctive therapy of invasive fungal infection particularly in CGD patients. The present study was designed to explore the effects of different types of interferon (alpha, beta, and gamma) pretreatments on gene expression of cathelicidin in murine macrophages following exposure to C. albicans yeasts and lipopolysaccharide (LPS) by Real time-RT PCR method. Materials and Methods: Mouse macrophages (J774 cell line) were cultured in RPMI-1640 with GlutaMAXTM-I medium. When cells in 24-cell culture plates reached to confluence of 80%, the plate divided into 6 columns (groups) including 1-negative control (medium), 2-Interferon (medium + IFN), 3-Yeasts (medium + C. albicans), 4-Positive control (medium + LPS), 5-Test-I (medium + IFN + C. albicans) and 6-Test-II (medium + IFN + LPS). RNAs were extracted by Trizole method and first strand cDNA was synthesized by reverse transcriptase. Quantitative Real-time PCR (qRT-PCR) was performed using ABI 7500 real time PCR System. Results: Neither IFNs pretreatment nor C. albicans exposure upregulates cathelicidin gene expression. However, LPS treatment induced expression of cathelicidin about 80-folds and this induction enhanced significantly to more than 110-folds following INFs pretreatment. Conclusion: As C. albicans is a part of mucosal flora, it has some mechanisms for mutual interaction with immune system. One of these mechanisms might be inhibition of AMPs upregulation. However, further studies are needed to clarify such interactions. Moreover, there is a possible clinical use of IFNs in treatment of Gram-negative infections. Keywords: Interferons, Cathelicidin, Real Time-RT PCR

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Poster presentation

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Activity of Ajowan (Carum copticum) essential oil against Alternaria alternata

Reza Mohammad Salehi, Ali Reza Khosravi, Ramak Yahya -Raeeyat

Molecular Microbiology Research Center, Shahed University, Tehran, Iran Email: [email protected]

Introduction and Objectives: Alternaria alternata is a globally prevalent species of mould that can be isolated from plants, soil, food and air. It can cause diseases in humans and plants and by producing toxin, plays a crucial role in the development of asthma, allergic sinusitis and rhinitis. The spores of this mould are among the most common aerobic potential sources of allergy. Alternaria is known as a novel striker pathogen, especially in immunocompromised patients. This species of fungus is also considered a cause of phaeohyphomycosis, keratitis, onychomycosis, infectious sinusitis and hypersensitivity pneumonia. Materials and Methods: In this study, the antifungal effect of the essential oil of Thrachyspermum copticum (Ajowan) was investigated on 6 A. alternata isolates and the standard strain ATCC6663. The essential oil was extracted from 3 ml per 100 g Ajowan. Gas chromatography (GC) and gas chromatography mass spectrometry (GC-MS) were employed to determine chemical composition of essential oil. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of essential oil for A. alternata isolates were determined by the broth micro dilution and macro dilution methods. Briefly, Ajowan essential oil concentrations (2%) diluted with DMSO was incorporated in RPMI 1640. Results: MIC level was measured as 0. 09mg/ml in 71.4% of the cases and 0. 19mg/ml in the remaining 28.6%. MFC level was 0.19mg/ml in 57.1% of the cases, 0.78mg/ml in 28.6%, and 0.39mg/ml in 14.3% of the cases. Gas chromatography (GC) and gas chromatography mass spectrometry (GC-MS) were employed to determine the chemical composition of essential oil obtained from dry fruits of Carum copticum. Thymol (46. 19%), g-terpinene (18. 26%) and P- cymene (25. 53%) were found to be the major constituents of the oil. Conclusion: The results are similar to previous research on the effects of antifungal Ajowan. Potent antifungal effect of the essential oil is used due to its high thymol content. Keywords: Antifungal, Ajowan, Alternaria, Essential oil

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Identification and antifungal susceptibility profile of Candida species isolated from nosocomial candiduria

Farzad Katiraee, Jila Targhibi, Sahar Shadi

Department of Pathobiology, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran Email: [email protected]

Introduction and Objectives: Candiduria is the most common clinical of urinary tract infection caused by Candida species. Candiduria was known as nosocomial fungal infection in hospitalized patients. Also, today’s resistant to antifungal especially between Candida species is a complication for patients with fungal infection. The aim of the study was to investigate the distribution of Candida species isolated from urine specimens of hospitalized patients in some private Hospital, Tehran, Iran, as well as their susceptibilities to antifungal agents. Materials and Methods: During 2008, fifty Candida isolates recovered from urine samples of hospitalized patients and were identified by conventional and a PCR- RFLP method. In the present study, five antifungal drugs (fluconazole, amphotericine B, ketoconazole, caspofungin and nystatin) were used for susceptibility test based on disk diffusion standard method. Results: The most common species recovered was C. albicans 40% of all yeasts, followed by C. glabrata (28%), C. tropicalis (12%), C. krusei (6%), C. kefyr (6%), C. famata (4%), and C. dubliniensis (4%). Based on disc diffusion test, 25% and 32% of C. albicans and C. glabrata isolates were resistance to fluconazole, respectively. A total of 16 (32%) other Candida species, 10% was resistance to fluconazole. In the present study, except one C. albicans and C. krusei all tested isolates were sensitive amphotericine B. Resistant against ketoconazole (12. 5% and28. 4%) were observed among C. albicans and C. glabrata isolates, respectively. On the other hand, all isolates were sensitive to caspofungin and nystatin. Conclusion: It is concluded that caspofungin is the best choice for treatment of Candiduria in hospitalized patients. In contrast, resistance against azole antifungal showed that these drugs aren’t first choice for treatment. In the next step, other suitable antifungal drugs were amphotericine B and nystatin, Keywords: Nosocomial candiduria, Antifungal susceptibilities, Candida species

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The effects of Spirulina on nitric oxide in balb/C mice with systemic candidiasis

Minoo Soltani, Ali Reza Khosravi, Farzad Asadi, Ramak Yahyaraeyat

Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran Email: [email protected]

Introduction and Objectives: Spirulina (Spirulina platensis) is a dietary supplement that is interesting for its immune-enhancing properties. In the present study the effects of Spirulina on nitric oxide (NO) production in peritoneal macrophages of mice with systemic candidiasis were investigated. Materials and Methods: Mices were divided into four groups (G1-G4), 5 mices per each group, animals received a dose of 800 mg/kg of S. platensis for four days and then were inoculated intravenously with 1×106 Candida albicans. After 24 hours, they were euthanized and their intraperitoneal lavage macrophages were collected. Peritoneal macrophages were stimulated by Spirulina (10µg/ml), LPS (2µg/ml), LPS+Spirulina (2µg/ml, 10µg/ml) in vitro. Dulbecco`s modified Eagle medium (DMEM) alone was used for control group. Macrophages were cultivated 24 hours, then the levels of nitrite in culture supernatants were measured by OD reading at 540 nm. Results: The results revealed that, there was significant difference in NO production among the understudy groups (p<0. 05) as well as culture media (DMEM, LPS, Spi, LPS+Spi) (p<0. 001). The NO production synergistically induced when the cells were treated by LPS + Spirulina (9. 6±0. 38) comparing to control culture (7. 39±0. 3). Conclusion: It seems that using of Spirulina+LPS has more acceptable effect on NO production than using Spirulina alone. Keywords: Spirulina, Nitric oxide, Systemic candidiasis

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Agar based drug diffusion assay for in vitro susceptibility testing of dermatophytes

Azar Sabokbar, Seyedeh Shaghayegh Mirabdollahelahi, Majid Riazipour, Masoumeh Alipour

Department of Microbiology, Karaj Branch, Islamic Azad University, Karaj, Iran Email: [email protected]

Introduction and Objectives: A standardized reference method for dermatophyte in vitro susceptibility testing is lacking. In the last two decades a plethora of studies have been done to establish an optimal antifungal susceptibility testing of dermatophytes. However, the earlier studies suffered from drawbacks of being expensive, exposed to contamination, requires long- term laboratory efforts. Materials and Methods: In this study, plates were incubated at 30ºC allowing a reading of the minimal inhibitory concentration (MIC) after 7 and 14 days. Compared to fluconazole, terbinafine possessed the highest antifungal activity against all of the dermatophytes. Results: Our results show that the method is simple, reproducible and cheap and could be used in routine laboratories to determine the antifungal susceptibility of dermatophytes. The objective of the present study was to describe a newly developed agar-based method employing drug diffusion assay to evaluate antifungal activities of terbinafine and fluconazole against 35 clinical dermatophytosis patients in Tehran, Iran. Conclusion: Consequently, it could be concluded that terbinafine revealed to be potentially very good as antifungal agent than other member of azole drug groups (especially fluconazole), so this antifungal agent probably appears to be suitable alternative for the treatment of dermatophytosis. Since agar based drug diffusion is a new method in susceptibility testing; further studies must be conducted in vitro and in vivo as a result of insufficient data in the field. Keywords: Dermatophytes, Terbinafine, Fluconazole

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Comparison between antifungal effect of Alchemilla vulgaris (Paye Shir) with two chemical drugs (ketoconazole, fluconazole) on Candida albicans and Malassezia furfur

Zahra Keshavarzi, Zahra Hesami

Pharmacology Department, Shiraz University, International Division, Shiraz, Iran Email: [email protected]

Introduction and Objectives: Cutaneous fungal agents are a special group of fungus that can attack to epidermal layer of skin, hair in human and animal and can create super facial infection. Although a lot of antifungal are available these days, but still in this course progress is needed. Because of problems that create with synthetic drugs like side effect, creating rigidity and low effect drugs, coming back to nature and special medicinal plants is necessary. Materials and Methods: In this study, standard strain of Candida albicans ACTT10231 and Malassezia furfur were cultured in Sabouraud’s dextrose agar, after that various amount of ethanolic extraction of Alchemilla vulgaris (Paye Shir) were added to these cultures. Results: In this study, MIC of C. albicans and M. furfur for A. vulgaris was 2.5µg/ml for Candida and 5µg/ml for M. furfur. About fluconazole drug, it was 3.6µg/ml for Candida and 2µg/ml for M. furfur and about ketoconazole for both of them it was 2.3µg/ml Conclusion: The result showed that ethanolic extraction of A. vulgaris has antifungal activity. Although, the antifungal activity of A. vulgaris is less than the two chemical drugs, we can use it in treatment of fungal infection. Keywords: Dermatophytes, Alchemilla vulgaris, Fluconazole

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Evaluation of antifungal activity of essential oil Allim jesdianum on Malassezia furfur and Candida krusei

Maryam Shirani, Mahboobeh Madani, Ali Reza Khosravi, Seyed Reza Hoseinidoost

Yong researchers Club, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran Email: [email protected]

Introduction and Objectives: Allium genus some valuable species such as onion and garlic for human being. Allium jesdianum Bois species is a bulbar plant with 2-3 leaves which grows in Zard Kooh Bakhtyary region. The aim of this study was to evaluate the antifungal effect of A. jesdianum Boiss extract on Candida krusei and Malassezia furfur. Materials and Methods: This study evaluated the antifungal activity of essential oil A. jesdianum by means of Inhibitory zone diameter and Minimum Inhibitory Concentration (MIC), using Macrodilution broth and Disc diffusion methods. The MIC of essential oil A. jesdianum was measured by counting the number of colony in agar medium. Results: The minimum inhibitory concentration of ethanol extract on C. krusei is 0.78 but ethanol extract on M. furfur and water extract on C. krusei and Malassezia furfur have no effect. Inhibitory zone diameters in ethanol extract was 30 millimeter for C. Krusei but ethanol extract on M. furfur and water extract on C. krusei and M. furfur no effect, respectively. Conclusion: The results show that ethanol extract A. jesdianum has suitable antifungal effects against standard strains C. krusei. Keywords: Antifungal activity, Allium jesdianum, Candida krusei, Malassezia furfur

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Antifungal activity of silver nanoparticles on Aspergillus niger

Hossein Hamzehei, Hamed Alizadeh, Mohsen Ajalli

Research laboratory, Zanjan University of Medical Science, Zanjan, Iran Email: [email protected]

Introduction and Objectives: Invasive fungal disease represents a major threat to life in immunocompromised patients and is now one of the most common causes of infection in this group. There is a limited range of antifungal agents available to treat disease caused by Aspergillus including the polyenes, flucytosine, azoles and more recently the echinocandins. Therefore, it is necessary to find new ways of treatment for Aspergillus niger. The aim of this study was to investigate the antifungal effects of silver nanoparticles against A. niger. Materials and Methods: Minimum inhibitory concentrations (MIC) of silver nanoparticles against A. niger determined by agar dilution method and minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) data obtained by the broth micro dilution method. Results: The results showed that the silver nanoparticles have fungicidal effect on A. niger in vitro. Conclusion: Silver nanoparticles can be used for disinfection A. niger with fungicidal effect. Keywords: Antifungal Effect, Aspergillus niger, Silver Nanoparticle

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Antifungal effect of nanosilver on saprophytic and pathogenic fungi

Zahra Salehi, Zahra Seifi, Maryam Azish, Ali Zarei Mahmoudabadi

Department of Medical Mycology, School of Medicine, and Infectious Diseases and Tropical Research Centre, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Silver has natural antibacterial and antifungal properties and it has been used for the treatment of several diseases for over 100 years. Many studies have shown the antimicrobial effects of nanosilver, but the antifungal effect of nanosilver is mostly unknown. In the present study, antifungal activities of nanosilver on saprophytic moulds and clinical isolates of Candida species were investigated. Materials and Methods: A total of 85 isolates of saprophytic and pathogenic fungi was used in the present study. Saprophytic moulds were obtained from the culture contaminations in medical mycology laboratory. Clinical isolates of Candida spp. were obtained from the vaginitis infection. Anti-fungal activity of nanosilver was investigated by agar well diffusion method at different concentration. After incubation, 24h for yeasts and 2-4 days for saprophytic fungi bioactivities were determined by measuring the minimal inhibitory concentration. Results: In the present study 21.2% of tested organisms were resistant to the pure concentration of nanosilver followed by; 34.1% to 0.5mg/ml, 13.1% to 2mg/ml, 11.8% to 1mg/ml and 8.2% to 0.125mg/ml. In addition our study showed that moulds were more susceptible to nanosilver at low concentrations than yeasts. Conclusion: The results showed nanosilver have considerable antifungal activity, against saprophytic fungi and pathogenic yeasts. Keywords: Nanosilver, Saprophytic fungi, Pathogenic fungi

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Antifungal effect of Cydonia oblonga extracts on Aspergillus niger

Hamed Alizadeh, Mohsen Ajalli, Hossein Hamzehe

Young Researchers Club, Zanjan Branch, Islamic Azad University, Zanjan, Iran Email: [email protected]

Introduction and Objectives: Aspergillosis causes patient afflictions that are classically defined as invasive, saprophytic, or allergic. Invasive diseases caused by Aspergillus species include infections of the lower respiratory tract, sinuses, and skin as portals of entry. The CNS, cardiovascular system, and other tissues may be infected as a result of hematogenous dissemination or direct extension from contiguous foci of infection. Saprophytic involvement includes Aspergillus otomycosis and pulmonary aspergilloma. Allergic conditions encompass allergic Aspergillus sinusitis and allergic bronchopulmonary aspergillosis. The aim of this study was to evaluate the antifungal effects of Cydonia oblonga on Aspergillus niger. Materials and Methods: The ethanolic and acetonic extracts of C. oblonga leaves prepared. Minimum inhibitory concentrations (MICs) of extracts against A. niger done by agar dilution method and minimum inhibitory concentration (MIC) and minimum cidal concentration (MCCs) data obtained by the broth micro dilution method. Results: The results showed that the C. oblonga extracts can inhibit the growth of A. niger. In addition, ethanolic extract of C. oblonga was more effective than acetonic extracts. Conclusion: Cydonia oblonga extracts can be effective in controlling of Aspergillus niger. Keywords: Antifungal Effect, Aspergillus niger, Cydonia oblonga

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In vitro antifungal activity of Anthemis gayana flower methanolic and ethanolic extracts on Candida species

Zahra Rezvani, Leila Amjad

Young Researchers Club, Falavarjan Branch, Islamic Azad University, Isfahan, Iran Email: [email protected]

Introduction and Objectives: Candida albicans is the most important factor of etiologic candidiasis that it includes 60-75% species separated from candidiasis. Its clinical growth is acute, sub-acute or chronic and unique infection of mouth, throat, skin, and so on and it may be observed as poisonous sepsis, endocarditic and meningitis. So, the aim of this study was In vitro studying of antifungal effects of the flower methanolic and ethanolic extracts of Anthemis gayana. Materials and Methods: In this study, methanolic and ethanolic extracts were obtained by Soxhlet apparatus for 6h and were tested against 10 fungi strains: (SC-2- ; SC-2- ; SC-2- ; SC-2- ; SC-2- ; C. 589; C. 590; C. 592; C. 593; C. 596). Several concentrations of flowers methanolic and ethanolic extracts (50,100,200,400 mg/ml) were prepared by using DMSO solvent and then antimicrobial activity was tested by using agar well diffusion and dilution test. Data were analyzed using AVONA test in the p-value <0/001. Results: The results showed that, flower methanol extracts have more inhibitory activity against SC-2-A6, C. 592, C. 590, C. 589, on the other hand, MIC and MFC were reported for SC-2-A6, C. 592, C. 590, C. 589, 9. 37 and 18. 75 mg/ml and for SC-2-A12, SC-2-A16, SC-2-A17, SC-2-A3, C. 596, C. 593 were reported 18.75 and 37.5 mg/ml, thus, flower ethanol extracts have more inhibitory activity against SC-2-A17, SC-2-A3, SC-2-A12, SC-2A6, C. 596, C. 593, C. 592, C. 590, C. 589 on the other hand, MIC and MFC were reported 9.37 and 18.75mg/ml and for SC-2- A16 were reported 18.75 and 37.5 mg/ml. Conclusion: The results demonstrated that the flowers methanolic and ethanolic extracts of A. gayana have excellent antifungal activity. Hence, this plant may be used further as medicinal plant against Candida spp. Therefore, we will be able to perform researches with extraction of this plant effective compound for the treatment of infectious disease. Keywords: Anthemis gayana, Methanolic extract, Ethanolic extract, Antifungal

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In vitro anti-Candida activity of Salsola rigida

Zahra Seifi, Fatemeh Seifi, Mohammad Javad Mahdavi, Raheleh Seifi, Ali Zarei Mahmoudabadi

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Background and Objectives: The last four decades have witnessed a dramatic increase in the incidence of candidemia and invasive candidiasis, originating in tertiary care centers and now observed in virtually all type hospitals. The spread of resistance to current antifungal drugs and reduced number of drugs available makes it necessary to discover new antifungal drugs and compounds. Traditional medicine is a potential source of new antifungal. Salsola rigida is one of the highly qualitative native species of arid and semi-arid regions of the country. It belongs to the family of Chenopodiaceae. S. rigida is a traditional herbal medicine that it is useful for diseases such as hypertension and leprosy. The aim of study was to assess in vitro anti-Candida activity of the ethanolic extract of S. rigida. Materials and Methods: The aerial parts of S. rigida were collected and its identification was verified. In this study, the Minimum Inhibitory Concentration (MIC) of the ethanolic extract of S. rigida in the concentrations of 25-400 mgml-1 evaluated against various Candida spp collected of urine and vaginitis (Candida glabrata, C. albicans, C. dubliniensis, C. kefyer, C. tropicalis and C. humicola) using the agar well diffusion method. Results: It was found that the ethanolic extract from the aerial parts of S. rigida was exhibited anti-Candida activity by ranges of inhibition zones 1-2. 4mm and MIC by ranges of 100-400 mg ml−1. Conclusion: S. rigida that showed the anti-Candida activity is the worth of further investigations in order to identify the active compounds and their clinical applications for the treatment of candidiasis. Keywords: Salsola rigida, Anti-Candida activity, Ethanolic extract

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In vitro anti-Candida activity of Prosopis farcta

Zahra Seifi, Fatemeh Seifi, Ali Zarei Mahmoudabadi, Mohammad Javad Mahdavi, Raheleh Seifi

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Background and Objectives: The dramatic rise of resistance to current antifungal drugs and reduced number of drugs available and the important this problem in the treatment of infectious diseases in immunocompromised, AIDS and cancer patients makes it necessity to discover new antifungal drugs and compounds. Recently it has been considered as a serious and lethal agent in people with immune suppression. Traditional medicine is a potential source of new antifungal. Recently, biologically active compounds and extracts separated from this plant species served in herbal medicine has increased the attention of researchers. Prosopis farcta is one of the highly qualitative native species of arid and semi-arid regions of Iran that it is native to south-eastern the country. It belongs to the family of Leguminosae. Prosopis farcta is a traditional herbal medicine that it is useful for treatment of stomach ulcers, abortion, dysentery, rheumatism, inflammation of the larynx, cardiac pain and shortness of breath are also investigating other anti- diabetic properties and benefits antispasmodic, analgesic and anti-inflammatory. The aim of study was to assess in vitro anti-Candida activity of the ethanolic extract of Prosopis farcta. Materials and Methods: The fruit of Prosopis farcta was collected and its identification was verified. In this study the Minimum Inhibitory Concentration (MIC) of the ethanolic extract of Prosopis farcta in the concentrations of 25-400 mgml-1 evaluated against various Candida spp collected from patients with vulvovaginal candidiasis (C. glabrata, C. albicans, C. kefyer, C. dubliniensis and Candida species) using the agar well diffusion method. Results: It was found that the ethanolic extract from the fruit of Prosopis farcta was exhibited anti-Candida activity by ranges of inhibition zones 11-17 mm and MIC by ranges of 200-400 mg ml−1. Conclusion: Prosopis farcta that showed the anti-Candida activity is the worth of further investigations in order to identify the active compounds and their clinical applications for the treatment of candidiasis. Keywords: Prosopis farcta, Anti-Candida activity, Ethanolic extract

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Peganum harmala, a plant with antifungal activity

Mehraban Falahati, Roohollah Fateh, Ali Kanani, Jalal Kiani, Fereshteh Zarei

Department of Medical Mycology, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Peganum harmala L. (Zygophylaceae) has been used in a variety of practical applications in medical science. The aim of this study was to determine antifungal activity of seed extract of P. harmala against reference fungal strains. Materials and Methods: Alkaloid extract of P. harmala was tested for in vitro antifungal activities against Aspergillus niger, Microsporum gypseum, Candida albicans and Saccharomyces cerevisiae. Minimal inhibitory concentration (MIC) was determined using broth microdilution method. The effects of extract were compared with those of miconazole. Results: P. harmala alkaloid extract showed antifungal activity against all the fungi species tested with MIC values ranging from 1 to 3.2 mg/ml. Conclusion: The results indicate that alkaloid extract of P. harmala has antifungal activity and might be promising, at least, in the treatment of fungal-associated diseases from mentioned fungi. Keywords: Peganum harmala, Minimal inhibitory concentration, Aspergillus niger, Microsporum gypseum, Candida albicans, Saccharomyces cerevisiae

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Anticandidal effect of shallot (Allium hirtifolium) against chronic candidiasis

Mehraban Falahati, Roohollah Fateh, Somaiieh sharifinia, Ali Kanani, Fereshteh Zarei

Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Shallots are an important part of the diet of many populations and there is long-held belief in their health enhancing properties. The aim of this study was to determine anticandidal activity of shallot against chronic candidiasis. Materials and Methods: Alcoholic and aqueous extracts of shallot (Allium hirtifolium) were tested for in vitro antifungal activities against 33 Candida isolates obtained from patient with chronic candidiasis that referred to Mirza kochank khan and Lolagar hospital. Minimal inhibitory concentration (MIC) was determined using broth microdilution method. Results: A. hirtifolium showed antifungal activity against all the Candida species tested and anticandidal activity of the alcoholic extract was much better than aqueous extract. Conclusion: The results indicate that crude juice of A. hirtifolium has anticandidal activity and might be promising in the treatment of candidiasis. Keywords: Shallot, Chronic candidiasis, Minimal inhibitory concentration

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Identification and antifungal susceptibilities Candida species isolated from nail infection

Farzad Katiraee, Adel Ghaderi, Shahla Ghaderi

Department of Pathobiology, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran Email: [email protected]

Introduction and Objectives: Onychomycosis is a fungal persistent infection of the nail bed and plate by dermatophytes, yeasts or mould species. This study presents ethologic agents of Candida onychomycosis and the determination of in vitro susceptibility patterns of isolated Candida species in multiple private laboratories in Tehran. Materials and Methods: Nail samples were taken from 120 patients with suspected fingernails and toenail onychomycosis during 2011. The direct microscopy with KOH and culture examinations of the nail samples was performed to identify the etiologic agent. Candida species identified by conventional methods. To select antifungal agents, the in vitro susceptibility of the isolates was performed by disc diffusion method. Results: Samples from 35 patients (29.1%) were positive for Candida onychomycosis. Candida albicans (54.3%), C. glabrata (11. 4%), C. parapsilosis (11.4), C. tropicalis (8.6%), C. guilliermondii (5. 8%), C. famata (5.8%), and C. Kefyr (2.9%) were the most frequent species isolated. Females affected more frequently than male and in both sexes, the age group of 20- 40 years old were more affected. Fingernails were affected more frequently than toenails (P < 0. 05). The majority of isolates were susceptible to amphotericin B and nystatin (97.1%), itraconazole (88.6%), fluconazole (80%), and ketoconazole (82. 8%), clotrimazole (82.8%). Conclusion: The yeasts of genus Candida are one of the most important causal agents of onychomycosis and susceptibility testing of nail isolates of Candida species could be considered in the successful treatment in case of relapse or chronic disease. Keywords: Candida species, Onychomycosis, Antifungal

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Antifungal susceptibilities of Candida species causing vulvovaginitis by disk diffusion

Ehsanollah Fakhari, Arjang Nakhaoda, Masoud Emami

Azad University of Tehran, Science and Research Branch, Tehran, Iran Email: [email protected]

Introduction and Objectives: The incidence of vulvovaginal candidiasis suggest approximately two-thirds of women experience at least one episode during their lifetime and nearly 50% of women have multiple episodes. The majority of cases of vulvovaginal candidiasis are caused by Candida albicans; however episodes due to non-albicans species of Candida appear to be increasing. Most non-albicans Candida species have higher azole MICs, and infections they cause are often difficult to treat. The purpose of this study was to determine the species distribution and prevalence of antifungal resistance among a collection of yeast isolates from patients with Candidal vulvovaginitis. Materials and Methods: Susceptibility testing was performed on vaginal yeast isolates collected from 71 patients with suspected vulvovaginal candidiasis. The number of different species of Candida determined by differentiation methods including production of germ tube and chlamydoconidia, growing in 45°C and carbon assimilation. Antifungal susceptibility testing by the agar diffusion method (with 25µg fluconazole disk, 10µg itraconazole disk and 10µg Ketoconazole disk) was performed according to the NCCLS M44P guidelines. Results: The yeasts isolated were C. albicans (38), C. glabrata (10), C. parapsilosis (10), C. tropicalis (8), C. krusei (3), and C. guilliermondii (2). Most resistance cases in several species of C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. krusei and C. guilliermondii were ketoconazole, itraconazole, fluconazole and itraconazole (equal), fluconazole and itraconazole respectively. The correlation between 24h and 48h diameter of inhibition for all drugs in all species was good (p<0. 05). Conclusion: Statistical analysis indicated an increased prevalence of vulvovaginal candidiasis caused by non-albicans Candida. High frequency of C. albicans and susceptibility to azoles in this study indicate possibility of continuous use of azoles in the treatment of uncomplicated candidiasis. Considering the high prevalence of this disease and increasing of drug resistance, it is recommended to use disk diffusion method as a screening method in laboratories. Keywords: Disk diffusion, Candida albicans, Azole, Vulvovaginitis

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Susceptibility to fluconazole in Candida species isolated from vulvovaginal candidiasis in Ahvaz

Zahra Seifi, Ali Zarei Mahmoudabadi, Majid Zarrin

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Vulvovaginal candidiasis is considered as a common opportunistic mycosis in women genital system in the world. Disease is usually associated with considerable morbidity, healthcare cost, distress, pain and sexual functioning. Candida albicans is the most agent of Candida vaginitis followed by C. glabrata and C. tropicalis. Fluconazole is a triazole antifungal that are used for the treatment of Candida vaginitis as a single dose. Several reports shows that the incidence resistant to fluconazole rising during two last decade. In the present study, susceptibility Candida vaginitis isolates evaluated against fluconazole. Materials and Methods: Vaginal samples were collected using sterile cotton swabs and inoculated on CHROMagar Candida. Cultured media were incubated at 30-35°C for one week, aerobically. Colonies producing a green coloration were presumptively identified as Candida albicans. In addition, this species was confirmed by germ tube and chlamydoconidia formation. Other species were detected using CHROMagar Candida, ID 32C and API 20C AUX kits. Susceptibility to fluconazole was determined using ATB Fungus 3 kit. Results: In the present study C. albicans (47.2%) was the most common species among the isolates followed by C. glabrata (24.1%), C. dubliniensis (12%) and Candida species (5.6%). In the present study resistant to fluconazole was found in 34.8% of isolates. C. dubliniensis was the most common isolate (6 of 11 isolates) that show resistant to fluconazole. Conclusion: Based on the present study, resistant to fluconazole among vulvovaginal candidiasis agents is considerable. In addition, this resistant was more common against C. dubliniensis than other species of Candida. Keywords: Vulvovaginal Candidiasis, Candida albicans, Fluconazole

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Interaction between Candida albicans with different bacteria

Hassan Ghorbani Choboghlo, Mehdi Taghavi, Donya Nikayin

Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran Email: [email protected]

In nature, most microorganisms exist in polymicrobial communities. Candida albicans, the most common human fungal pathogen, is a prototypical opportunistic organism that lives harmlessly in the human gastrointestinal tract but has the ability to cause life-threatening invasive disease. The present study aimed to improve our understanding of interaction between C. albicans and bacteria. In this regard review in litterateurs showed there is an antagonistic relationship between C. albicans and nonfermenting, gram-negative bacteria, such as Pseudomonas aeruginosa and Acinetobacter baumannii, whose interactions are likely found in the clinical environment, especially in the oral cavity, respiratory tracts of critically ill patients and on wounds of patients with burn injuries. It is also known that bacteria such as Pseudomonas aeruginosa and Escherichia coli can significantly influence and modify candidal growth and biofilm formation. Moreover, it is known that a secondary metabolite of Pseudomonas species, Pyrrolnitrin, may be a key anticandidal factor. On the other hand, the interaction of C. albicans with bacteria might shape microbial virulence and alter filamentation by Candida spp in the intestinal tract. In fact S. Typhimurium is capable of significantly reducing the viability of C. albicans and inhibiting its ability to form biofilm, a key virulence determinant of this pathogen, in co infection. Additionally, it is becoming clear that the C. albicans epithelial cell interaction at mucosal surfaces is critical for commensalism, pathogenicity and host defense. This relationship is complex depending on the bacterial species and its numbers, and may affect the morphogenesis of the yeast. Identifying the molecular mechanisms of this interaction may provide important insights into microbial pathogenesis. Keywords: Candida albicans, Bacteria, Interaction

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Investigation the potential of allicin to inhibit the growth of Candida albicans in vitro and in a systemic candidiasis mouse model

Alireza Khodavandi, Fahimeh Alizadeh

Department of Paramedical Science, Gachsaran Branch, Islamic Azad University, Gachsaran, Iran Email: [email protected]

Introduction and Objectives: Recently, the incidence of systemic candidiasis which is caused by Candida albicans has increased. Usually, for treatment of systemic fungal infections, azoles such as fluconazole are used, but one of the biggest problems faced in clinical practice is the emergence of resistance due to mutation for most of these azole drugs currently used. Therefore the most urgent challenge in pharmaceutical research is the discovery and development of new antifungals from plant and microbial sources. Allicin, one of the sulfur compounds from garlic has been shown to possess antifungal activity. In the present study, the anticandidal activity of pure allicin demonstrates its strong potential activity both in vitro and in vivo. Materials and Methods: The in vitro efficacy of allicin and fluconazole was measured against C. albicans ATCC 14053 and nine clinical isolates using CLSI M27 A2 documents for yeast and MIC50 and MIC90 of allicin and fluconazole were determined. Subsequently, Scanning electron microscopy for morphology observation of Candida after treatment with allicin was done. For the animal model of systemic candidiasis, female Balb-c mice, 4-6 weeks old, were infected intravenously and then treated by allicin and fluconazole in different dosages. Results: Both allicin and fluconazole showed different MICs that ranged from 0. 05-12.5µg/ml and 0.25-16µg/ml, respectively. Time-kill study also showed a significant effect of allicin (p<0.01) against C. albicans comparable to fluconazole. Scanning electron microscopy observation revealed that similar to fluconazole, allicin produced structural destruction to C. albicans cell surface at low MIC and lysis or puncture at high MIC concentrations. Treatment of C. albicans systemically infected Balb-C mice showed that although the allicin treatment (at 5 mg/kg/day) had slightly less efficacy when compared to fluconazole treatment in terms of the fungal load reduction and host survival time; was however, still effective against C. albicans in terms of mean survival time increase from 8.4 to 15.8 days. Conclusion: These results demonstrate the efficacy of anticandidal effects of allicin both in vitro and in an animal model of candidiasis and affirm the potential of allicin as an adjuvant therapy to fluconazole. Keywords: Allicin, Fluconazole, In vitro, In vivo

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Therapeutic effect of Shallomin as an antifungal drug on dermatophytes

Mansour Amin, Raheleh Sheikhi, Alireza Kalamizadeh

Department of Microbiology, School of Medicine, Infection and Tropical Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Dermatophytes are a keratinophilic and keratinolytic of fungi that produce superficial cutaneous infections and are the principle etiologic agents of the dermatomycosis. Although many antifungal agents have been developed during the last decades and have become available for dermatophytosis, they are limited to a relatively few chemical groups. Therefore investigation for novel antifungal agents has always been one of the major preoccupations of medical society. Materials and Methods: Thirty volunteer patients were obtained from clinically suspected superficial cutaneous fungal infections especially dermatophytosis. All collected specimens were analyzed by direct microscopy and culture. These specimens were cultured on Sabouraud’s Dextrose agar with chloramphenicole and cyclohexamide. Cultures were incubated at 25°C for up to 28 days and checked twice weekly for growth. Patients with fungal positive culture were treated by 0.5% Shallomin (antifungal fraction of Allium hirtifolium). Shallomin was scrubbed to the site of infection twice a day for 10 days. After this time, patients were re- examined for fungal cultures. Susceptibility of isolated fungi to Shallomin was determined by modified E-test. Results: In this study, the most frequent dermatophytes isolated from patients were Trichophyton mentagrophytes, T. rubrum, Epidermophyton floccosum, M. gypseum (12,6,5,2 cases respectively). All isolated microorganisms were sensitive to Shallomin. The mean of MIC values of Shallomin ranged from 2.66-11.50µg/ml. T. mentagrophytes (2. 66µg/ml), T. rubrum (5.16µg/ml), E. floccosum (4.40µg/ml) and T. gypseum (11.50µg/ml). Conclusion: The finding of our study has demonstrated that Shallomin is a useful and effective treatment modality with more rapid and less after-use scaring. Based on this Shallomin can be an effective medicine for the treatment of dermatomycosis. Keywords: Allium hirtifolium, Dermatophytes, MIC

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Azole resistance in Aspergillus fumigatus obtained from hospitals environments

Afsane Vaezi, Hamid Badali, Bita Mousavi, Saba Mayahi, Saham Ansari, Bahram Ahmadi

Department of Medical Mycology and Parasitology/Invasive Fungi Research Center (IFRC), School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran Email: [email protected]

Introduction and Objectives: Aspergillus fumigatus is a saprotroph widespread in nature typically found in soil and decaying organic matter causes severe infection like invasive aspergillosis, especially in immunocompromised host. Environmental study of azole-resistant A. fumigates and other Aspergillus species which are surrounded at the high risk host would be essential study to explore the prevalence of azole-resistant Aspergillus. Materials and Methods: In the present study, we obtained all samples from the soil and air in different hospitals of Tehran and Sari, Iran. The obtained soils were plated on Sabouraud’s dextrose agar (SDA) and SDA supplemented with chloramphenicole and itraconazole (4 mg/liter), incubated at 37°C and checked for azole resistance Aspergillus species. Aspergillus species were diagnosed by conventional and molecular tools. Those A. fumigatus species which grew on multidish azole agar were tested for in vitro antifungal susceptibility based on CLSI method M38-A2 method according to the manufactures’ instructions, and then PCR amplification and DNA sequencing of cyp51A gene were performed to identify azole-resistant A. fumigatus isolates. Results: A. fumigatus was isolated in 20 out of 70 samples. The majority of A. fumigatus isolates obtained from the hospital soil samples rather than environment. In vitro susceptibility results have shown that MIC for itraconazole >8 mg/L and for voriconazole 4 mg/L. Results of sequenced cyp51A gene from 7 strains compared with susceptible wild type of A. fumigatus and revealed a single mutation at codon 98, which was substituting the amino acid leucine to histidine (L98H). Therefore, due to multi-azole-resistant A. fumigatus has increased in hospital area where patients are in the high risk. Discussion: Due to multi-azole-resistant A. fumigatus has increased in hospital area where patients are in the high risk. Thus, understanding of the prevalence of azole-resistant organisms which, are in the first line to cause severs disorder with a potential efficacy is significantly recommended. With present results we suggest that in vitro antifungal susceptibility testing of clinical samples is crucially important and have to be done routinely at all hospital labs before start the treatment. Keywords: Aspergillus fumigatus, Soil samples, Azole resistance

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Antifungal activity of special propolis extract

Akram Astani, Jurgen Reichling, Stefan Zimmermann, Karl Heinz Sensch, Paul Schnitzler

Shahid Sadoughi University of Medical Science, Yazd, Iran Email: [email protected]

Introduction and Objectives: Bee propolis is a remarkable natural substance collected and produced by honeybees. Propolis composition is highly variable, depending on the plant species and on the season of collection. A special purified Propolis extract (Gh 2002) preparing in a special procedure was examined to evaluate the antifungal activity against Malassezia fur fur and Candida reference strains and clinical isolates. Materials and Methods: The microdilution method was used to determine minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC). In this study, standard Candida strains included 3 Candida reference strains C. albicans, 90028, C. glabrata, 2950 and C. parapsilosis, 22019; 6 Candida isolates from patients and M. fur fur were checked. Results: Propolis extract had high activity against Candida strains in the range of 0.15-1.2 mg/ml. M. furfur were inhibited by propolis in the range of 4 mg/ml of propolis. Propolis was fungicidal against most Candida strains and M. furfur in the range of 0.6-2.4 and 4mg/ml. Conclusion: It may be concluded that this special propolis extract is suitable for the treatment of fungal isolates from patients Keywords: Propolis Extract, Antifungal Activity, MIC, MFC

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Antifungal susceptibility profile of Rhodotorula species from Ahvaz

Sharzad Hydrinia, Zahra Seifi, Ali Zarei Mahmoudabadi

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Rhodotorula species are common airborne contaminant fungi and also considered as normal inhabitants in the skin, lungs, urine and feces in humans. The most common species of Rhodotorula are; R. mucilaginosa, R. glutinis and R. minuta. Rhodotorula species considered as an important agent for invasive infection among immunocompromised patients. Both amphotericin B and flucytosine have good activity against Rhodotorula in vitro, whereas fluconazole is inactive. There are few studies in the literature on the susceptibility Rhodotorula against antifungal drugs. In the present study sensitivity profile of Rhodotorula species were evaluated against several antifungal agents including; amphotericine B, nystatin, miconazole, clotrimazole, fluconazole and terbinafine. Materials and Methods: In the present study 69 isolates of Rhodotorula including; R. glutinis (59), R. mucilaginosa (5), R. minuta (3) and Rhodotorula species (2) were examined for susceptibility tests against amphotericine B, nystatin, clotrimazole, miconazole, terbinafine, and fluconazole antifungals. In vitro susceptibility testing was performed by the method disc diffusion. The antifungal agents used in the study were as follows: amphotericin B, fluconazole, itraconazole, ketoconazole, nystatin and lamisil. The beginning of 24-hour culture of yeast was prepared a suspension equivalent to a 0.5 McFarland. 100 µl of the suspension was inoculated on Sabouraud dextrose agar medium and this was spread evenly on the surface medium. Discs containing antifungal agents were placed on medium. The inhibition zone was evaluated after 24-48 hours manually. Results: In the present study 65 (94. 2%) of isolates were sensitive to clotrimazole followed by nystatin 49 (71.0%), miconazole 46 (66.7%), amphotericine B 29 (42.0%) and terbinafine 18 (26.1%). In our study, 26 (37.7%) of isolates were resistance to terbinafine whereas resistance to miconazole was only detected in 1 (1.4%) of isolates. All tested isolates were resistance to fluconazole. Conclusion: All the tested antifungal agents, except fluconazole, were effective against the tested strain of Rhodotorula in vitro. Based on this result we can state that all the antifungal agents tested, except fluconazole, are useful medicaments for the treatment of infections by the Rhodotorula genus. Keywords: Rhodotorula, R. mucilaginosa, Susceptibility profile, Antifungals

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Survey the effect of Licorice extract on aflR gene expression and aflatoxin production in Aspergillus parasiticus via Real Time PCR

Rashin Mohseni, Fatemeh Noorbakhsh, Ayatolah Nasrollahi Omran, Maryam Moazeni, Sasan Rezaie

Young Researchers Club, Tonekabon Branch, Islamic Azad University, Tonekabon, Iran Email: [email protected]

Introduction and Objectives: Aflatoxin is a fungal metabolite from Aspergillus species that is toxigenic, mutagenic, carcinogenic, immunosuppressive and even teratogenic in human and various species of animals. So the aflatoxin is important in food industries, animal husbandry and medical area and there are enormous economical detriments due to this toxin. These factors resulted in many studies on the extracts and plant compounds to reduce the growth of aflatoxin producing organisms, inhibit the toxin production and also suppress the major toxin encoded genes like aflR in these organisms. Licorice is an important medicinal plant in our traditional medicine and it posses many antimicrobial activities. There is no report around the licorice effects or its mechanism on the aflatoxin producing Aspergillus species. The present study focuses on the licorice extract inhibitory effects on the aflR gene expression and the growth and survival of Aspergillus parasiticus. Materials and Methods: After the culture of A. parasiticus in the toxin inducer medium, the minimal inhibitory concentration and minimal fungicidal concentrations for licorice extract was measured. The aflatoxin concentration in the control and treated media was determined using HPLC method. Also after harvesting the fungi from toxin inducing medium its mRNA was extracted and the cDNA was synthesized by universal primers. The quantitative change in the aflR expression was analyzed via real time PCR. The statistical analysis was performed by SPSS program. Results: The production of fungal mycelium was decreased by increasing the concentration of licorice extract. The minimum fungicide concentration was 500mg/ml of the extract. The HPLC analysis revealed that the 10mg/ml of licorice extract could inhibit the toxin production by 99. 9%. Conclusion: In this concentration aflR gene expression was suppressed up to 40% that was revealed by quantitative RT-PCR analysis. Keywords: Licorice extract, Aflatoxin, Aspergillus parasiticus, aflR gene

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Isolation and identification of soil chitinolytic bacilluses in the Northern Iran and survey of antifungal effects

Ayat Nasrollahi Omran, Hamid Pordeli, Jalil Ieri, Nargas Vaseghi

Department of Medical Mycology, Faculty of Medical Sciences, Islamic Azad University of Tonekabon Branch, Tonekabon Iran, P. O. Box: Tonekabon 46815-559. Email: [email protected]

Introduction and Objectives: Chitin, a linear polymer of N-acetylglucosamine residues, has been the most abundant polymer in nature after cellulose. In the recent decades, chitinase have received increased attention because of their wide range of applications especially as biocontrol against fungi. Materials and Methods: In this cross-sectional– Analytic study, isolation Bacilluses producing chitinolytic enzymes screening procedure was performed using 40 soil samples collected from various regions of Gorgan (northern Iran). Primary screening of the chitinolytic potential of the isolates was done by observation of clear zone around colonies in colloidal chitin agar medium Identification of selected strains was performed by polyphasic taxonomy. Extraction DNA was used to perform for subtler identification and sequensing. Antifungal potential evaluated with well method against Candida albicans (ATCC 10231), Aspergillus niger (ATCC 2029) A. flavus (IR6) Fusarium oxyporum (PTCC 5115) and Alternaria alternate (PTCC 5224). Results: The results showed that selected strains were belonged to members of Bacillus Genus. 9strain of chitinase positive Bacillus were isolated on choloidal Chitin Agar (CCA) 5 strains showed antifungal effects and R6 had highest effect and R2 and R3 had lowest effect on fungi. The sequential analysis of 16S rRNA gene was also studied. Conclusion: The 16S rRNA gene sequence of these isolation with known Bacteria have 95-97% sequence similarity and with consideration of morphological, physiological and genetically differences with those probably that isolates are unknown and need too identification. Soil Bacteria can have antagonestic effects with Pathogenic fungi in soil specially human and phytopathogenic. Keywords: Bacillus, Chitinase, Soil, Antifungal potential

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Unsuccessful treatment of co-infection endocarditis due to Candida albicans and C. tropicalis in a drug abusers patient

Seyed Jamal Hashemi, Imaan Haghani, Bita Mousavi, Mohammad Ali Boroumand, Maryam Sotodeh Anvari, Keyvan Abbasi, Hamid Badali

Department of Medical Mycology and Parasitology, School of Medicine/Molecular and Cell Biology Research Center, Mazandaran University of Medical Sciences, Sari, Iran Email: [email protected]

Introduction and Objectives: During recent decades, the incidence and the diversity of Candida endocarditis has increased dramatically, especially in patient infected with human immunodeficiency viruses, underlying valvular heart disease, intravenous drug abusers, implantation of prosthetic valves, prolonged use of intravenous catheters, patients using broad spectrum antibiotics, undergoing solid organ transplantation, preexisting bacterial endocarditis or patients with severe underlying diseases. Despite the application of surgery and antifungal therapy, Candida endocarditis is a life-threatening infection with significant morbidity and mortality which have been caused by Candida species. Materials and Methods: We report a 37 year-old drug abuser male referred to Tehran Heart Center presenting with high fewer, chest pain and loss of appetite with cardiac failure. Results: His echocardiography revealed large tricuspid valve replacement. Although, his blood culture for bacterial agents was negative, diagnosis for fungal endocarditis was performed by culturing of respected vegetation which confirmed by growth of mix Candida albicans and Candida tropicalis in blood culture. Histopathology of the tricuspid respected vegetation revealed septate hyphae with neutrophils infiltration and eosinophils with focal granulation tissue formation. These strains were initially identified by culturing on CHROMagar Candida medium, germ tube, chlamydoconidia formation, and identity was confirmed by sequencing of the internal transcribed spacer (ITS rDNA and D1/D2) regions. The patient was treated by ceftriaxone, gentamicin, cefixime and fluconazole. In vitro antifungal susceptibility testing showed that posaconazole, itraconazole and fluconazole had the highest activity against these isolates; however, their clinical effectiveness in the treatment of C. albicans and C. tropicalis infections remains to be evaluated. Conclusion: The patient failed amphotericin B and fluconazole therapy combined with surgical excision despite the drug’s low MICs in vitro. Keywords: Fungal endocarditis, Candida species, Sequencing, Susceptibility

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Assessment of antifungal activity of various extracts and fractions of Ephedra major against some dermatophytes moulds and yeasts.

Ali Mikaeili, Masoud Modaresi, Korosh Mozafari Isaac Karimi

Department of Medical Mycology, School of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran Email: [email protected]

Introduction and Objectives: To investigate the antidermatophytic effects of Ephedra major against some dermatophytes, moulds and yeasts in the in vitro. Materials and Methods: The in vitro antifungal activity of extracts was measured by disc diffusion test and determination of minimum inhibitory concentrations (MIC). Results: Hydrophilic extracts did show considerable antifungal activity against Trichophyton rubrum, T. mentagrophytes and Epidermophyton floccosum as compared with terbinafine. MIC of hydroalcoholic extracts was 20, 30 and 20 mg. ml-1, respectively. Hydrophilic extracts did show considerable antifungal activity against A. fumigatus and Candida albicans as compared with ketoconazole. MICs hydroalcoholic extracts were 20 and 20 mg. ml-1, respectively. Conclusion: After animal model assessment, the hydroalcoholic extract of this plant may possibly be further developed into adjunct drug for topical treatment of dermatophytosis. Keywords: Ephedra major, T. rubrum, T. mentagrophytes, E. floccosum, Candida albicans, Kurdish Ethnomedicine

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Isolation of anti-dermatophyte compounds from Myrtus communis L. leaves

Mitra Mehrabani, Seyyed Amin Ayatollahi Mosavi, Hosein Gholamzadeh

Herbal and Traditional Medicines Research Center, Kerman Medical University, Kerman, Iran Email: [email protected]

Introduction and Objectives: Fungi are very important in production of acute and chronic infections. But there is a few numbers of drugs for treatment of fungi diseases that are fungistatic or produce constant resistance. Therefore wide range of investigations especially on plants should be made to find an effective compound with suitable anti fungal effect. In recent research, the anti fungal effects of ethyl acetate fractions of hydroalcholic extract of leaves of Myrtus communis in 0 to 0.3 RF against some dermatophyte has been proven. In this study, this RF component separation was performed by HPLC and their antifungal effects were studied. Materials and Methods: Methanolic extracts of the dried leaves of the plant were prepared by maceration. After drying the extract, various fractions obtained based on polarity. Ethyl acetate fraction was found in previous research is the most effective antifungal, on the prepared plates was isolated. RF 0 to 0. 3 carved, components were extracted with methanol. The material was injected into the HPLC with preparative column and 5 components that show the best peaks were isolated and collected. These compounds were studied with micro dilution methods for antifungal effect on Microsporum canis, M. gypseum and Trichophyton mentagrophytes and MIC were determined. Results: The study identified two compounds of 5 compounds isolated from ethyl acetate fraction have good antifungal effects on the three strains. Conclusion: According to the studies of basic patterns that can be observed by the UV detector, these compounds can be considered in the class of polyphenolic compounds. Keywords: Antifungal, Myrtus communis L., HPLC, Microdilution

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Laboratory study of anticandidal activity of Cinnamon essential oil by microdilution method

Shrareh SayahAlborzi, Zeinab Sadeghi Ardestani, Seyed Amir Yazdanparast

School of Allied Medical Sciences, Tehran University of Medical Science, Tehran, Iran Email: [email protected]

Introduction and Objectives: Patients with candidiasis could be treated with antifungal drugs. Due to prevalence of fungal infection, high cost of antifungal drug, side effects and drug resistance, It seems that Herbal medicines are suitable for the treatment of fungal infections and many studies have been led to consider essential oils antifungal effects. Materials and Methods: In this study Cinnamon essential oil, on Candida species were investigated experimentally by Micro Dilution Method. Results: Cinnamon essential oil had the most anti- candidal activity. Candida albicans was the most sensitive species. Conclusion: Because of the strong anti- candidal effects of Cinnamon in this study, conducted using in vitro tests, it is proposed to treat candidiasis with Cinnamon experimentally. Keywords: Cinnamon, Anti-candidal, Essential oil, Microdilution method

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Anti-Candida activity of the hydroalcoholic extracts of Heracleum persicum fruit

Mahsa Rajabi, Batool Sadeghi-Nejad, Majid Zarrin, Ali Zarei Mahmoudabadi

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Nowadays, Candida albicans has been resistant to the toxic and expensive commercial drugs of anti-Candida available in the market. Hence, it is necessary to search for new anti-fungal agents. Materials and Methods: In this study the hydroalcoholic extracts of Heracleum persicum fruit was investigated for anti-Candida activity of 46 pathogenic Candida species such as C. albicans, C. glabrata and C. tropicalis by agar well diffusion method. Results: The Minimum inhibitory concentration (MIC) values at 24 and 48 hours were showed 0.625-20µg/µl for C. albicans, 0.625-40µg/µl for C. glabrata, 5.0-20µg/µl for C. tropicalis. Conclusion: The results of this survey confirmed that tested plant extract had potential anti- Candida activity. Hence, necessary to isolate and identify the active compounds of this plant extract in the future. Keywords: Anti-Candida activity, Heracleum persicum, Hydroalcoholic extract, Candida sp

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In vitro effectiveness of Pinus longifolia on Candida species isolated from patients with vulvovaginal candidiasis in Isfahan

Zohreh Norouz, Mahboobeh Madani‚ Leila Amjad

Department of Microbiology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran Email: [email protected]

Introduction and Objectives: Vulvovaginal candidiasis is a serious public health problem in women that caused by Candida species specially Candida albicans. The plant products and their active constituents play an important role in maintaining perfect health. The present investigation was an attempt to evaluate the in vitro effectiveness of female cones of Pinus longifolia extract on the growth of Candida species isolated from patients with vulvovaginal candidiasis in Isfahan. Materials and Methods: Clinical isolates of Candida were evaluated with mycological algorithms, including morphology on corn meal ager and CHROMagar, germ tube production and biochemical methods. Female cones of P. longifolia were extracted by Soxhlet apparatus using ethanol and water as solvents. The anticandidal properties were evaluated using the agar well diffusion method. Determination of minimum inhibitory concentration was performed using broth microdilution assay. Fluconazole was used as positive control. Results: Four species of Candida were isolated from patients and recognized as Candida albicans, C. parapsilosis, C. glabrata and C. tropicalis. Maximum inhibition zone of ethanolic and aqueous extracts of P. longifolia were 16, 14, 15, 12 and 15, 13, 14, 11 mm and the minimum inhibitory concentration were 5,5,10,10 and 5, 5,10, 20 mg/ml for above-mentioned species, respectively. Conclusion: The results of this work showed antifungal activity of ethanolic and aqueous extracts of female cones of P. longifolia on Candida species. Keywords: Candida, Microdilution, Pinus longifolia, Extract

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Antifungal efficacy of Myrtus communis linn. extract against Candida sp. and Aspergillus sp.

Maryam Erfani-Nejad, Batool Sadeghi-Nejad, Sedigheh Nanaeie, Majid Zarrin, Ali Zarei Mahmoudabadi

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Myrtus communis Linn. (Family, Myrtaceae) is an aromatic evergreen perennial shrub or small tree, 1.8-2. 4m in height with small foliage and deep fissured bark. It is native to widespread in the Mediterranean region. Use of herbal medicines in Asia has been used for long time. The aim of this survey was the in vitro antifungal activity of the Ethanolic extracts of Myrtus communis leaves against strains of Candida sp. such as Candida albicans, C. glabrata, C. tropicalis and three species of Aspergillus including Aspergillus niger, A. flavus and A. terreus. Materials and Methods: The ethanolic extract of myrtle leaves was prepared by maceration method and Minimal Inhibitory Concentration (MIC) of M. communis leaves extract was determined by agar-well diffusion assay. Amphotericin B and clotrimazole were used as the positive control in this experiment. Results: The minimal inhibitory concentration (MICs) values of M. communis leaves extract ranged from 0. 625-5.0µg/µl and 5-20µg/µl against tested Candida sp. and Aspergillus sp. respectively. Conclusion: Results revealed that the ethanolic extracts of M. communis have been antifungal potency against both pathogenic tested fungi and it can be used as natural antifungal agent. Keywords: Myrtus communis, Antifungal efficacy, Aspergillus sp., Candida sp.

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Determination of synergistic effect of Myrtus communis essential oil and linalool with itraconazole against azole-resistant Candida species

Kamiar Zomorodian, Mahmoud Moein, Mohammad Javad Rahimi, Maryam Esmaeilbeig, Mohhamad Hadi Amir-Shahpari, Hanieh Bazrafshan, Nilofar Namazi, Kayvan Pakshir

Department of Medical Mycology and Parasitology, School of Medicine and Basic Sciences in Infectious Diseases Research Center, Shiraz University of Medical Sciences, Shiraz, Iran Email: [email protected]

Introduction and Objectives: Candida species are responsible for a variety of infections in humans ranging from simple dermatosis to life threatening candidemia. Emerge of resistance of Candida species to current antifungal drugs including itraconazole (ITC) is a universal crisis. Combination therapy is successfully used previously to overcome this resistance. In the present study the synergistic effects of the essential oil (EO) of Myrtus communis (myrtle) and linalool- one of its major constituents-in combinations with itraconazole against azole-resistant Candida spp. were determined. Methods: The minimum inhibitory concentration (MIC) values of ITC, plant EO and linalool was determined by broth microdilution assay as recommended by the Clinical and Laboratory Standards Institute (CLSI). We evaluated the synergistic effect between the essential oil, linalool and the antifungal compound ITC by checkboard micro titer assay. To evaluate the combined effects of EO/linalool with ITC, fractional inhibitory concentration index were calculated. Results: Treatment of ITC-resistant isolates with the combination of plant EO and ITC resulted in 60% decline of resistance rate to lower than 0. 5µg/ml. Combination therapy of linalool with ITC exhibited a strong synergetic activity and resulted in reduction of the ITC MICs to >0. 125µg/ml in 6 of 10 studied Candida isolates. Conclusion: Considering the strong synergistic activities of ITC with each of EO of M. communis and linalool (FICI≤0. 5), they might have potential to use in antifungal products to enhance the ITC anticandidal activities. Keywords: Synergistic effect, Candida, Itraconazole, Essential oil, Myrtus communis, Linalool

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Antifungal and antibacterial activity of Fistulina hepatica (Schaeff.) methanolic extract

Mitra Noori, Abolfazal Taghavi

Department of Biology, Faculty of Science, Arak University, Arak, 38156-88349, Iran Email: [email protected]

Introduction and Objectives: Fistulina hepatica (edible beefsteak fungus) has been used as a meat substitute in the past, and can still be found in some French markets. It has a sour, slightly acidic taste. Edible mushrooms may have antioxidant properties. In this research, antifungal and antibacterial activities of a wild edible mushroom (F. hepatica) extract were investigated. Materials and Methods: F. hepatica was collected from Mazandaran province. Methanolic extract was prepared from mushroom dried powder and in vitro antifungal and antibacterial activities of the extract were examined against two bacteria strains (Staphylococcus aureus and Escherichia coli) and two mold species (Aspergillus flavus and Rhizopus stolonifer) by paper disc method. Results: F. hepatica methanolic extract showed antibacterial effects on Gram-positive bacterium (S. aureus) and also antifungal effects on A. flavus while it was not effective on Gram-negative bacterium (E. coli) and R. stolonifer. Conclusion: This study showed the antifungal and antibacterial effects of F. hepatica. It could be suitable as an agent for agricultural crops protecting and also in treatment of bacterial infections. Keywords: Fistulina hepatica, Fistulinaceae, Antimicrobial activity

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Antifungal activity of ethanolic extract of leaves "Myrtle" against Microsporum gypseum, M. canis and Trichophyton mentagrophytes

Bahareh Zihayat, Akram Noski, Mitra Mehrabani, Mehdi RezaieFar, Amin Ayatolahi Mousavi

Department of Medical Mycology and Parasitology, Afzalipoor Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran Email: [email protected]

Introduction and Objectives: Although fungi play a great role in acute and chronic infections, but there is few antifungal drugs in markets for treatment. That’s the reason of evaluating anti- fungal effect of a great number of herbs; one of these herbs is Myrtle. In this investigation we studied anti-fungal effect of Myrtle leaf hydro alcoholic extract by the methods bioautography and disc diffusion. Materials and Methods: Methanol extract of dry Myrtle leaf was prepared by percolation. After the extract dried, different fractions were separated. For determination of the anti-fungal effect of fractions a disk diffusion method was used ten microliter of each fraction was putted on sterile paper disks (6 mm diameter) on pre-cultured fungi on Sabouraud dextrose agar plates. Anti- fungal effects of the plant extract were expressed as inhibition zones around fungal colonies. In autobiography method, the fraction with the greatest inhibition zone was separated by TLC method. After putting TLC sheets in medium, inhibition zone was appeared by tetrazoliom and indicated as Rf. Results: In the disk diffusion method, the ethyl acetate extract had the best anti-fungal effect while in autobiography both ethyl acetate and methanol extracts in Rf=0.03 had anti-fungal effects and inhibition zone on three examined fungi. Conclusion: Antifungal activity of Myrtle may be attributed to flavonoid compounds as the main constituents of plant ethyl acetate extract. Keywords: Autobiography, Disk diffusion, Myrtle anti-mycosis effect

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Antimicrobial properties of Ixora brachiata Roxb. against bacteria and Candida isolates

Ameneh Hardani, Batool Sadeghi-Nejad

Department of Public Health, School of Health, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: The antimicrobial effects of root and leaves extracts of Ixora brachiata against a few common clinical isolates of Gram negative and Gram positive pathogenic bacteria including Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Klebsiella sp. and 10 Candida isolates were studied. Materials and Methods: The plant materials were air-dried, ground to fine powder and prepared for bioassay by agar diffusion technique. Results: Antibacterial activity of the ethanolic extracts of I. brachiata root and leaves were detected by the inhibition zones of 10-16 mm and minimum inhibitory concentration (MIC) ranges of 1.25-10 mg ml-1 for I. brachiata root extracts, 11-17 mm and 0. 15-10 mg ml-1 for I. brachiata leaves extracts. The anticandidal effects of the plant revealed the growth inhibition zone of 12-14 mm and MIC values of 5. 0-10 mg ml-1 for both root and leaves extracts of I. brachiata at 24 hours incubation period. Conclusion: These results revealed the potential anticandidial and antibacterial properties of this I. brachiata in treating of microbial infections. Keywords: Antimicrobial effects, Ixora brachiata, Bacteria, Candida

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Study of antifungal activity of essential oils of Iranian endemic medicinal herbs against Candida albicans

Mehdi Tavalla, Rahman Abdizadeh, Behzad Hamedi, Abdollah Ghasemi Pirbalouti

Department of Medical Parasitology, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Candida albicans is a dimorphic fungus that causes broad spectrum of opportunistic fungal infections in immunocompromised individuals due to the extensive utilization of antibiotics and development of anti-fungal drug resistance. Therefore, searching for the new sources of anti-yeast is required. In this regard, plant extracts as well as essential oils and other compounds are of considerable interest because of their anti-fungal activity. This study aimed to investigate the anti -Candidal activity of the essential oils of six plant species, which are of Iranian endemic plants. Materials and Methods: Six Iranian endemic plants including Melisa officinalhs, Satureja hortensis, Thymus caramanicus, Curum carvi, Artemisia aucheri and Chamaemelum noble were collected from mountainous areas of Central Zagross, Chaharmahal va Bakhtiari and Kerman districts. Subsequently, leaves and flowers were dried at room temperature for one week and the essential oils were isolated by water distillation. The oils were analyzed by Gas chromatography (GC) and their activity was evaluated as in vitro with eight different concentrations (4, 7. 8, 15. 6, 31. 25, 62. 5, 125, 250 and 500µg/ml) against C. albicans, strain PTCC5027, by disc diffusion method in Sabouraud dextrose agar medium. Results: All tested essential oils showed antifungal activities with the diameters of the inhibition zone ranging from 8 to 45 mm in concentrations from4µg/ml to 500µg/ml. Melisa officinalhs and Thymus caramanicus oils exhibited significant antifungal activities (8-35mm. MIC ≥4µg/ml) and (8-42mm, MIC≥4µg/ml) respectively. Also the results of GC analysis revealed that Carvacrol (66. 67%), 1,8-Cineole (21. 52%), lavandlyl acetat (27%), cuminaldehyde (62%) and Beta-Caryophyllene (24%) are the main components of T. caramanicus, A. aucheri, C. noble, C. carvi and M. officinalhs oils, respectively. Conclusion: Drug resistance in pathogen fungi as well as undesirable effect of antifungal agent caused important of find new agents, therefore plant-derived because of the components and fractions of them can be useful candidates for this aim. Findings of the present study indicated relatively high anticandida activity of essential oils of traditional medicinal plants from the Iranian flora that this effect is because of high percentage of their main components especially Carvacrol and Beta-Caryophyllene. Keywords: Candida albicans, Antifungal activity, Essential oils, Iranian endemic plants

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Different silver nanostructures and growing inhibition of clinical Candida albicans isolates

Ali Jebali, Farzaneh Haji Esmaeil Hajjar

Pars Hospital Lab, Tehran, Iran Email: [email protected]

Introduction and Objectives: The Silver nanoparticle is effective on different microorganisms and lead to cell apoptosis and death by destruction of cell membrane, proteins and DNAs. Silver ions can be reduced differently and made different nanostructures depend on chemical agents and reaction condition. Nano wires, nanoplates, nanocubes, nano rods, nanodots are examples of silver nanostructures. The amount effectiveness of all nanostructures on Candida albicans is unclear. The aim of this research was the evaluation of different silver nanostructures on growing inhibition of clinical Candida isolates. Materials and Methods: Thirty clinical C. albicans isolated from candidiasis patients at different origin. Colony isolation was carried out on Sabouraud dextrose agar (SC) and SC with cyclohexamide medium and germ tube test was done for each isolate. For assessment of growth inhibition of different silver nanostructures, disk diffusion method was done on Muler Hinton medium. Nanodots, nanocubes and nanowires of silver were synthesized by reduction reaction and confirmed by scanning electron microscopy and 10µL of each nanostructure was added to Whatman paper (1×1 cm) and dried at room temperature. Disk were hold on Muler Hinton medium and incubated at 37°C for 48 hours and finally inhibition zone diameter of each isolate was recorded for three nanostructures. Results: The mean of inhibition zone diameter of silver nanodots. Silver nanocubes and silver nanowires was 19.7mm, 20.8mm and 17.9mm, respectively. This means silver nanocubes are better growing inhibitors than nanodots and nanowires. Conclusion: This research showed different silver nanostructures had been different growing inhibition of clinical C. albicans isolates, and silver nanocubes are the better inhibitors. Keywords: Silver nanostructures, Nanotechnology, Candida albicans

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Are some isolates of Candida albicans resistance against silver nanostructures?

Farzaneh Haji Esmaeil Hajjar, Ali Jebali

Pars Hospital Lab, Tehran, Iran Email: [email protected]

Introduction and Objectives: Pathogenic fungi like Candida albicans are the cause of life- threatening infections in an increasing number of immunocompromised patients. One important issue in administration of antifungal agents is drug resistance that lead to failure of treatment. Different antimicrobial nanomaterials were studied and no drug resistance has been observed. The best antifungal nanoparticles that were evaluated are silver nanoparticles that can destroy fungal cell by binding protein and membrane. The aim of this research was the assessment of resistance against silver nanostructures on clinical Candida isolates. Materials and Methods: Thirty two clinical C. albicans isolated in Pars hospital lab from candidiasis patients. Firstly approved isolates of C. albicans were grown in Sabouraud Dextrose Agar for 2 days at 37οC and secondly, resistance assessment of different silver nanostructures were carried out by disk diffusion method on Muler Hhinton medium. Nanodots, nanocubes and nanowires of silver were chemically synthesized and confirmed by SEM microscope and then each nanostructure was added to Whatman paper (1×1cm) and were hold on medium and incubated at 37οC for 48 hours and finally resistance of each isolate was reported for nanostructures. Results: All isolates were sensitive to silvernanocubes and silvernanowires but two isolates of them (6%) were resistance to silvernanodots Conclusion: This research showed that nanodots, the most used antimicrobial nanoparticle, are not effective on all isolates of C. albicans. Keywords: Silver nanostructures, Resistance, Candida albicans

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In vitro inhibitory effect of Echinops cephalotes methanolic extract on Candida spp. isolated from patients with vulvovaginal candidiasis in Isfahan

Sara Heshmati‚ Mahboobeh Madani‚ Leila Amjad

Department of Microbiology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran Email: [email protected]

Introduction and Objectives: Nearly three-quarters of all women experience at least one episode of vulvovaginal candidiasis during their life time. Therefore, identification of Candida species in the clinical object is important. This study reports the in-vitro inhibitory effect of Echinops cephalotes methanolic extract on Candida spp. isolated from patients with vulvovaginal candidiasis in Isfahan. Materials and Methods: Mycological algorithms, morphology on Corn meal agar and chromogenic agar, germ tube production and biochemical methods were used for identification of clinical isolates. Antifungal activity of the methanolic extract of E. cephalotes was studied against isolated Candida by well diffusion and determination of minimum inhibitory concentration (MIC). Results: Candida spp. isolated from patients with vulvovaginal candidiasis were C. albicans, C. glabrata, C. parapsilosis and C. tropicalis. The tested extract showed significant antifungal activity against Candida spp. isolates. Inhibition zone of growth of methanolic extract of E. cephalotes were 10-36, 16, 19 and 16 mm respectively and the best result showed against C. albicans. Minimum inhibitory concentration (MIC) values varying from <7/8 to 7/8 mg/ml. Conclusion: This research is the first study on antifungal activity of the methanolic extract of E. cephalotes. Hence, E. cephalotes might be a promising material for controlling Candida spp. Keywords: Vulvovaginal candidiasis, Antifungal activity, Extract, Echinops cephalotes, Candida

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The influence of water ozonation in wheat tempering on reduction of Aspergillus parasiticus growth

Matin Mohammadi Kouchesfahani, Sasan Rezaie, Mahmood Alimohammadi, Gholamreza Jahed Khaniki, Ramin Nabizadeh Nodehi, Zeinab Aghamohseni

Department of Environmental Health Engineering, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Food and feed spoilage moulds cause great economic losses worldwide. It is estimated that between 5 and 10% of the world's food production is wasted due to fungal deterioration. Wheat is important cereal in Iran and very sensitive to mould contamination consist of aflatoxigenic moulds. Ozone (O3) is a strong oxidant and disinfection recognized since 1997 as a GRAS substance. Ozone is effective in gaseous state and aqueous solution. It decomposes rapidly to molecular oxygen without leaving a residue. The objective of this study was to evaluate the effectiveness of ozonated water on reduction of Aspergillus parasiticus. Materials and Methods: In order to study the effect of ozonated water on growth of A. parasiticus, 5g of cleaned and pured wheat placed in each falcon tubes and autoclaved at 121°C for 20min. Then each falcon include of wheat inoculated with spore suspension (10 spore/gr wheat) of A. parasiticus and incubated at 25-30°C for 10 days. Contaminated samples incubated at <20, 25,40°C for 24h, after tempering with ozonated water to concentrations of 0, 1, 2, 2.5mg ozone/l. To evaluate of ozone effectiveness on reduction of A. parasiticus, weighing of mould mycelium method was used in petri dishes include of Sabouraud dextrose broth after drying. Results: Results indicated that ozonated water can be inhibited A. parasiticus growth and this reduction can be increased due to increasing of ozone concentration. A statistically reduction in quantity of mould mycelium were obtained after tempering with ozonated water (p<0. 05). Conclusion: The results obtained in the present research conformed the results of other researches in effect of ozonated water on reduction of wheat total bacteria and yeast/mould counts. Therefore, the ozonated water usage in wheat tempering can be caused reduction of A. parasiticus growth. Keywords: Wheat, Ozonation, Tempering, Aspergillus parasiticus

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In vivo antifungal activity of new fluconazole derivatives in systemic murine candidiasis

Keyvan Pakshir, Maryam Jamshidi, Akram Jamshidzadeh, Zahra Rezaei, Zohreh Eymeripour

Basic Sciences in Infectious Diseases Research Center, Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran Email: [email protected]

Introduction and Objectives: Candidiasis is an opportunistic fungal infection mostly caused by Candida albicans. Recently, efforts have focused on the development of new, less toxic and more efficacious antifungal, with novel in mechanism of action. In this study, for the first time in Shiraz, in vivo antifungal activity of five fluconazole derivatives were evaluated against C. albicans in mouse animal model. Materials and Methods: The mice were divided in 19 groups of five and immunocompromised by cyclophosphamide via intra-peritoneal injection during 5 days, prior to infection. The suspension of C. albicans was prepared and injected to each mouse via tail vein. After two hours, each derivative was injected in three doses via peritoneal. After 24 hours, the mice were killed and its kidneys were homogenized and the suspension was cultivated on Sabouraud dextrose agar. After incubation period, the number of colonies was counted and results were expressed as CFU/two kidneys of each mouse. The fluconazole was used as drug control. Results: Among the tested compounds, one derivate "2- (2,4-difluorophenyl)-1- (3-nitro-1H- 1,2,4-triazol-1-yl)-3- (1H- 1,2,4-triazol-1-yl)propan-2-O" exhibited the best antifungal activities while the others showed less activity in comparison with fluconazole. Conclusion: Fluconazole derivatives could nominate as antifungal drugs in treatment of systemic candidiasis but more studies are needed to confirm these results. Keywords: Candida albicans, In vivo, Fluconazole, Antifungal

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Study on inhibitory activity of propolis alcoholic extract on opportunist fungi of Candida in vitro

Sedigheh Saberi, Mohsen Mahmoudi, Zahra Mosavi, Reza Arjmand, Seyed Hossein Hejazi

Department of Mycology and Parasitology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran Email: [email protected]

Introduction and Objectives: Propolis or bee glue is a production of hive with resin nature was used in treatment of infectious diseases from many years ago. At present time it is still widely used in the eastern part of Europe. The use of propolis for its antibacterial, antiviral, and certain other useful pharmacologic effects is increasing. Because of difference between the sorts of propolis from various countries, we decided to investigate the propolis of specific part of Iran. Materials and Methods: In this experiment, first, the propolis extract was prepared as a thick solution and then dissolved in Ethylic alcohol 96% in order to obtain the alcoholic extract for different dilutions. Next, the fungus suspensions were prepared and homogenized using the McFarland tuibidimetric standard of 0. 5. To determine inhibitory and fatality dose of propolis extract, the serial dilution method was employed. The dilutions includied1:20, 1:40, 1:80, 1:160 and 1:320 which were prepared in a Sabouraud dextrose broth medium. Then 1ml of the each fungus suspension with a 0.5 McFarland standard was added to each tube containing a certain dilution of the extract in aseptic conditions. Results: In this study which was carried out using the statistical test of Kruskal Wallis, it was observed that the 1: 20 dilution of the extract had inhibitory and fatality effect on up to 98% of the isolates whereas in the 1: 40 dilution, the growth of 67% the isolates were inhibited and 57% of them were killed. However, in 1: 320 dilution of the extract, neither inhibitory nor killing effect was observed on the zymogenic isolates. Some changes were observed on the morphological features of colonies (depends on the extract dilution) as well as quantitative effects of dilutions of extract. Conclusion: The general conclusion is that the alcoholic extract of propolis on the growth of Candida isolates had prominent antifungal and inhibitory effects (MIC and MFC was 1:40). Keywords: Minimal inhibitory concentration, Minimal fungicidal concentration, Propolis, Candida

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Effect of fresh and old garlic on germ tube formation in Candida albicans

Majid Riazipour, Zahra Mottaghiyan

Department of Parasitology and Mycology, Medicine School, Baqiyatallah University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Several studies have shown garlic antifungal effects, but there is little information about the effect of this medicinal plant on fungal germ tube production. The objective of our study was to investigate the effect of fresh and old garlic on germ tube phenomenon in Candida albicans. Materials and Methods: Bushes of fresh and old garlic extracted by a household juicer. The extract was centrifuged and clear supernatant placed at 40°C to dry. Serial dilutions of the precipitate prepared in fetal calf serum and fresh cells of C. albicans (1×105 cells/ml) added to each tube. After two hours incubation at 37°C, a drop of preparation examined under optical microscope and by viewing at least 200 yeast cells, the percentage of germ tube production was calculated. Results: In concentrations below 3mg/ml, the garlic extracts did not affect germ tube phenomenon, but in higher concentrations there was a significant negative correlation between the extract concentration and percentage of germ tube production by yeast cells of C. albicans. Old garlic completely inhibited germ tube production at 50mg/ml concentration, while fresh garlic inhibited the phenomenon at 100mg/ml. Conclusion: Old garlic shows more potent inhibitory effect on germ tube production. This unexpected result necessitates further studies on relationship between maintenance time and antifungal effects of garlic. Keywords: Candida albicans, Garlic, Germ tube

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Evaluation of antifungal effect of Cyclamen coum tuber extract against isolated Candida albicans and C. tropicalis

Toktam Sajjadi, Azra Saboora, Parisa Mohammadi, Zahra Fallahi

Department of Biology, Faculty of Science, Alzahra University, Vanak, Tehran, Iran Email: [email protected]

Introduction and Objectives: Candida species have emerged as the most common cause of systemic infections in the last 2 decades. Emergence of pathogenic fungi that are resistant to common therapies during current decades has intensified the investigation of novel antifungals with natural origin. In this research, evaluation of antifungal effect of Cyclamen coum tuber extract, as a medicinal plant from Myrsinaceae family, in glycosidic, n-butanolic and aqueous phases, and aglycone, ethyl acetate and aqueous phases, fractions was carried out. Materials and Methods: Antifungal effects were evaluated using microdilution method, preparing serial dilution of drug in RPMI, and MIC and MFC of the extract was determined against standard strains of C. albicans (ATCC 10231), C. tropicalis (ATCC 0750), three clinical isolates of C. albicans and six clinical isolates of C. tropicalis. Ketoconazole was used as a control, after preparation in 512µg/ml and dilution. Results: Glycosidic aqueous phase in concentration of 0/008-0.032 mg/ml inhibited the growth of all species, while aglycone ethyl acetate phase was ineffective toward all of them. Reduction of antifungal effect was observed with increase of the extract concentration of glycosidic n- butanolic and aglycone aqueous phases with MIC 0.0049 and 0.0002-0.0004mg/ml, respectively. One isolate of C. tropicalis was regarded as the most resistant isolate because its growth was inhibited only by glycosidic aqueous phase with concentration 0. 008 mg/ml. Conclusion: For all tested Candida, the estimated MFC was equal to MIC of extract. More investigations should be carried out to find the exactly effective fraction of plant extract as well the mechanisms of the function of this herbal medicine. Keywords: Antifungals, Candida albicans, C. tropicalis, Cyclamen coum, MIC, MFC

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In vitro study the effects of tobacco on dermatophytes

Fariba Berenji, Abdolmajid Fata, Mohammad Rahimizadeh, Giti Haririzade, Majid Ganjbakhsh, Khadijeh Nejatipour

Parasitology and Mycology Lab, Emam Reza Educational, Research and Treatment Center, Mashhad University of Medical Sciences, Mashhad 91379, Iran Email: [email protected]

Introduction and Objectives: Today the cutaneous fungal infections of man include a wide variety of disease in our World. The infections transmitted from animal, man and soil. Because of high cost of chemical drugs and drug resistance, in this investigation we have studied the antifungal effects of methanolic and aqueous extracts of Nicotiana tabacum leaves on Trichophyton mentagrophytes ( PTCC 5054 ), T. verrucosum (PTCC 10694) and T. violaceum (PTCC 10735) Materials and Methods: The culture media was Mycobiotic agar. Miconazole nitrate was used as positive control and methanol was used as negative control. In both cases the result is given on the basis inhabitation of the growth compared with methanolic reference (in the case of methanolic extract) and water reference (in the case of aqueous extract). Results: The results obtained from methanolic and aqueous extracts were compared with different concentration of standard antibiotic. It was observed that both extracts had good effect on the studying fungi. Methanolic extract had better effect than the aqueous one. Among the considered dermatophytes T. violaceum was the most resistant fungus to the standard drug and to the herbal extract. Conclusion: Authors believe that these extract can be useful for treatment of dermatophytosis. Keywords: Dermatophyte, Tobacco, Traditional medicine, Herbal therapy

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In vitro antifungal susceptibility testing of dermatophytes against eight antifungal drugs

Imaan Haghani, Hamid Badali, Mehrnaz Mohammad Davoudi, Melika Laal Kargar

Department of Medical Mycology and Parasitology / Invasive Fungi Research Center, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran Email: [email protected]

Introduction and Objectives: Dermatophytosis caused by the genus of dermatophytes is thought to be one of the most significant public health problems yet not solved. The most frequent dermatophytes such as Trichophyton mentagrophytes and T. rubrum were known to cause multiplicity of cutaneous infection in humans and animals. Despite worrying clinical pictures associated with the genus of dermatophytes, there is little information regarding its susceptibility patterns against currently available antifungal agents, with only a small number of strains having been studied. Therefore, in vitro antifungal susceptibility tests could help to optimize the therapy and to select appropriate antifungal agents. Materials and Methods: A collection of 88 isolates of different species dermatophytes were performed to determine the MIC for AmB, FLU, ITC, VOC, POS, TER and MECs for CAS and ANID based on CLSI M38-A2 guidelines. Inoculum was adjusted spectrophotometrically (530 nm) to optical densities that ranged from 65-75% T in RPMI 1640 MOPS broth with L- glutamine without bicarbonate. Plates were incubated at 35°C for 72 h. Drug and fungus free controls were included. Results: The resulting MIC ranges for all strains were as follows, in increasing order: Terbinafine (0.008-0.031), posaconazole and voriconazole (0.008-2μg/ml); anidulafungin (0.008-8μg/ml); itraconazole and caspofungin (0.031-16μg/ml); amphotericin B (0.125- 16μg/ml); and fluconazole (2-64μg/ml) without any significant differences in the pattern of susceptibility between different species (p>0.05). The results showed that the activity of terbinafine was significantly higher than other tested antifungal agents and fluconazole was the drug that presented the highest MIC values in comparison with the other tested agents, with an MIC of 64μg/ml. Discussion: The present study based on in vitro activity showed that terbinafine followed by posaconazole, itraconazole, anidulafungin and caspofungin might have a potent activity with a best choice of alternative to previous antifungal drugs, for dermatophytosis and confirmed that the resistance of dermatophyte species to fluconazole, and are contradictory to cure rates achieved by patients who have used it to treat dermatophytosis. However, clinical effectiveness of new generations remains to be determined. Keywords: Dermatophytosis, Dermatophytes, Antifungal susceptibility, Terbinafine

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Antifungal activity of Scrophularia striata extract on Candida albicans

Ali Akbar Jafari, Ali Mohammad Latifi, Majid Shohrati, Reza Hajhosseini, Mohammad Eftekhari, Meysam Sarshar

Young Club Researchers, Islamic Azad University, Parand Branch, Tehran, Iran Email: a. [email protected]

Introduction and Objectives: Candida albicans which resides as commensal in the mucocutaneous cavities of skin, vagina and intestine of humans, can cause infections under altered physiological and pathological conditions such as infancy, pregnancy, diabetes, prolonged broad spectrum antibiotic administration, steroidal chemotherapy as well as AIDS. Scrophularia striata is one of the oldest drugs in Iranian traditional medicine. S. striata is used for the treatment of eczema, wounds and has shown various biological activities such as antimicrobial, antitumoral and anti-inflammatory properties but there are a few studies about anti fungal effects of S. striata, so this study was designed for evaluation of the antifungal effect of S. striata in vitro. Materials and Methods: In this study we use culture C. albicans, after preparation of S. striata extract, for comparison of antifungal effect of S. striata and amphotricin B we used of paper disc method. Results: Diameter of inhabitation zones were 22±0.09 in amphotricin B group and almost 33±0.11 in S. striata group. This study showed there is a statistically significant difference between Amphotricin B and S. striata extract (P=0.002). Conclusion: It was found that S. striata extract was in the case of antifungal effect was more effective than control group of fungicide and our results encourage examination of the efficacy of S. striata extract in other forms of systemic and superficial fungal infections and exploration of its broad spectrum effect against other pathogenic manifestations including malignancy. Keywords: Scrophularia striata extract, Antifungal, Candida albicans

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In vitro antifungal susceptibility of Candida species isolated from blood cultures of burnedburn patients or burn victims patients in burns unit of Zare Hospital, Sari, Iran

Nazanin Lotfi, Tahereh Shokohi, Seyed Zahra Nouranibaladezaei, Ayatolah Nasrollahi Omran

Faculty of Medical Sciences, Azad University-Tonekabon Branch, Department of Medical Mycology, Tonkabon, Iran Email: [email protected]

Introduction and Objectives: Burn patients are ideal hosts for opportunistic agents and blood infections. The aim of this study was to identify the Candida species involved in candidemia of burn patients and to assess the isolated species of Candida against four antifungal agents. Materials and Methods: In a cross-sectional study, blood samples of burn patients obtained. The yeast isolated from blood cultures identified to the species level by conventional procedure and API 20C AUX. In vitro antifungal susceptibility of the Candida isolates to Amphotricin B, fluconazole, voriconazole and caspofungin was determined using Etest. Results: During the present 9-month prospective survey (June 2011-March 2012), a total of 405 blood samples obtained from 113 patients. 27 (6. 7%) of the blood cultures, belong to 13 (11.5%) patients were positive for Candida species. The distribution of Candida spp. in 27 positive blood cultures was 12 (44%) C. parapsilosis, 7 (25. 9%) C. tropicalis, 5 (18. 5%) C. guilliermondii and 3 (11. 1%) C. albicans. E test antifungal susceptibility testing for the yeast isolated showed that all of Candida isolates were sensitive to amphotricin B, fluconazole and voriconazole, all ( 100%) of C. parapsilosis and 60% of C. guilliermondii found to be resistant to caspofungin. Conclusion: In the present study, Candida isolates showed the least resistance against voriconazole followed by amphotricin B. Keywords: Candidemia, Burn, Susceptibility test, Amphotricin B, Fluconazole, Voriconazole, Caspofungin

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The concentration of aflatoxin M1 in the mothers’ milk in Khorrambid city, Fars, Iran

Hossein Rafiei, Parvin Dehghan, Keyvan Pakshir, Mostafa Chadeganipour, Mojtaba Akbari

Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran Email: [email protected]

Introduction and Objectives: Aflatoxins are secondary toxic metabolites produced by certain group of Aspergillus species in suitable conditions. These toxins are highly toxic, immunosuppressive, mutagenic, teratogenic and carcinogenic metabolites. The purpose of this study was to detect aflatoxin M1 concentration in mother's milk from rural area of Khorrambid town of Fars Province. Materials and Methods: In this study, 87 milk samples of mothers were collected by cluster sampling methods in the period between June and July 2011 and the amount of aflatoxin M1 was measured by a competitive ELISA method. Results: From 87 mother's milk, 24 (27. 6%) samples were contaminated with aflatoxin M1 with mean concentration of 0. 56±1. 23pg/ml (range 0.13-4.91pg/ml). Conclusion: The amount of aflatoxin M1 in mothers, milk was lower than 50ng/l (Europe Union and Iranian standard). Detection of Aflatoxin M1 in mothers, milk is due to consuming contaminated food. This contamination not only threatens the health of the mothers but also has irreversible effects on the growth and health of their babies. Keywords: Mycotoxins, Aflatoxin M1, Mothers, Milk, ELISA

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The study of temperature on the T-2 toxin in corn

Fatemeh Zaboli, Issa Gholampour Azizi, Shima Khalatbari

Department of Biology, Ayatolah Amoli Branch, Islamic Azad University, Amol, Iran Email: m. [email protected]

Introduction and Objectives: T-2 toxin is one of the most important fungal toxins produced by some species of Fusarium, Trichoderma, Acremonium, Trichothecium and also Stachybotrys, that causes hormonal disorders, acute and chronic toxicity, weak immune system, carcinogenicity and also hepatic/ pelvic complications. The aim of this research is to test the fungal T-2 toxin in 22 samples of raw corn and popping corn by through Competitive ELISA method and also to study the effects of thermal process on it. Materials and Methods: Samples of raw corn samples of raw corn gathered random and the amount of T-2 toxin measured and the amount of T-2 toxin in raw corn was compared. Results: It was found that if standards of many countries to be considered, all of the raw corn samples and 8 of 11 pop corn samples had infection more than authorized amount (100µg/kg) but according to national standard of Iran (25µg/kg) all of the raw corn and pop corn samples were infected so it is showed high levels of T-2 toxin more than authorized amount- and ought not to be used even for stock. But, as we know, the samples investigated in this study were for human consumption. It is found that, heating has an important role in reducing the amount of T-2 toxin, so that The maximum concentration of T-2 toxin in raw corn and popping corn were respectively 296/75 ppb and 205/1 ppb; and their minimum concentration were respectively 103/3 and 56/05µg/kg; and finally their average concentration were respectively 219/7 and 135/85µg/kg and the percent of decrease of T-2 toxin from pop corn to raw corn is 36/95%. Conclusion: Considering the great importance of T-2 toxin in public health, observing sanitary standards in preparing popcorn and also in preserving conditions of raw corn seems essential. Keywords: T-2 toxin, Raw corn, Pop corn

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Detection and identification of fungal pathogens in clinical samples from patients referred to professor Alborzi mycology research lab, Shiraz, Iran

Parisa Badiee, Hadis Jafarian, Zahra Jafarpour

Professor Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran Email: [email protected]

Introduction and Objectives: Over the recent years, deaths due to fungal infections have increased. As yet antifungals are used widely as prophylaxis in immunocompromised patients. This study seeks to evaluate the efficacy of such important drugs. Materials and Methods: Clinical samples from the patients attending mycology division within 1390 were examined for fungal pathogens. KOH smear and culture were carried out on each arriving sample. The samples consisted of fluid collection 48%, cutaneous specimens 15%, sputum 13%, tissue 9%, blood 5%, CSF 4%, wound 3%, and others 3%. Species were determined with microscopic examination and API 20C AUX. Also, their susceptibility to 7 important antifungals were determined using E-TEST strips. Results: In total, 32. 29% of KOH smears and cultures were positive in 223 patients. Among the isolated yeasts there were 60% Candida albicans, 10% C. famata, 7. 5% C. glabrata, 5% C. guilliermondii, 5% Cryptococcus laurentii, 5% Rhodotorula mucilaginosa, and 7.5% others. The important isolated moulds were 40% Aspergillus fumigatus, 27% A. niger, 20% A. terreus, and 13% A. flavus. Their MICs were indicated as follows: MIC90 for moulds: amphotericin B, ketoconazole, caspofungin, itraconazole, posaconazole, voriconazole, and fluconazole were 0.75, 2. 0, 1. 0, 0. 75, 2. 0, 0. 125, 256, and for yeasts: 1. 0, 32, 0. 125, 32, 32, 32, and 256, respectively. Conclusion: In view of the emerging potentially fatal fungal strains that are resistant to many commonly used antifungal agents, the above mentioned data can be used in developing programmes for prevention and control. Keywords: Fungal infections, Aspergillus, Candida, Antifungal

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Study the existing fungal flora in the atmosphere of different areas of Kashan in 2001

Mohamad Ali Asadi, Azam Samei, Afshin Salehi, Fariba Raygan, Mohamad Reza Sharif

Kashan University of Medical Sciences, Kashan, Iran Email: [email protected]

Introduction and Objectives: All kinds of micro-organisms and fungus spore can be found almost everywhere. Kashan is a city near the central desert of Iran. The level of fungus spore in atmosphere is an important factor for public health. Fungus spore is one of the most important contaminators of atmosphere, which can cause different disease in human and animal. This research was carried out to identify the level of fungus spore in the atmosphere of different areas in Kashan. Materials and Methods: Samples were taken from different areas like, agricultural, industrial, animal farms, mountains and some wild areas. Samples were taken in the morning, noon and night. Cultural plates were exposed to atmosphere in a position 1.5m high from ground level for 15mins. Plates were covered with their lids, transmitted to Kashan medical mycology laboratory and kept in room temperature for 3-5 days. After growing fungus and establishing colonies two techniques of teased mount and slide culture, were used to diagnose the fungi species. Results: In this study 1890 colonies were separated from 270 plates and their species were identified as; Aspergillus (22.7%), Penicillium (20.31%), Alternaria (14.86%), Cladosporium (10.95%), Rhizopus (9.36%), Yeast (6.77%), Mucor (4.7%), Fusarium (4.02%), Scopulariopsis (3.22%), Acremonium (1.79%), Curvularia (1.16%) and unknown fungus (0.8%). Conclusion: This study showed that, most of the time fungi spore exists in the atmosphere in different areas. Diversity and level of fungi spore changes in different climates, geographical situations and the time of day. Further studies are needed to cover more geographical areas and epidemiologic research. Keywords: Fungal flora, Atmosphere, Saprophytic fungi

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The study of soil fungi of Khuzestan's oil regions

Vida Dawoodi, Mahboobeh Madani, Arezoo Tahmourespour

Department of Microbiology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran Email: [email protected]

Introduction and Objectives: Crude oil is a naturally occurring complex mixture of hydrocarbon and non-hydrocarbon compounds which possesses a measurable toxicity towards living systems. The dominance of petroleum products in the world economy creates the conditions for distributing large amounts of complex compounds consist of hundreds of different hydrocarbon molecules, and a huge volume of oily sludge, a carcinogenic and a potent immunotoxicant. Fungi are found ubiquitously in the environment. They execute a wide range of important ecological functions especially ones associated with decomposition of organic substrates in both terrestrial as well as marine environments, and occasionally in extreme conditions. The objectives of this study were therefore, to isolate and identity the indigenous fungal flora of petroleum contaminated soils in Khuzestan province. Materials and Methods: The eight oil contaminated soil samples used for the isolation were from eight different sites located at the Khuzestan's oil regions. Enumeration of soil heterotrophic fungi was estimated by soil dilution plate count method. Potato dextrose agar (PDA) medium supplemented with 1% streptomycin was used for isolation and enumeration of total heterotrophic fungi. Results: Total fungal counts ranged from 0.41 to 3333.33×102 cfu/gsoil. A total 60 fungal isolates (13 genera) were isolated from contaminated soils that belong to genera Aspergillus, Penicillium, Fusarium, Acremonium, Candida, Rhodotorula, Mucor, Aureobasidium, Cunninghamella, Rhizopus, Alternaria, Beauveria and Paecilomyces. Conclusion: Significantly total heterotrophic fungi in different sites, depending on the amount of oil contamination were different. The highest fungal populations were belonged to soil sample of Hafgel with 20.51% organic carbon. The genera Aspergillus and Penicillium were the most dominant in this study. According to Chaillan et al. (2004), Aspergillus and Penicillium are the most commonly found fungi in tropical soil. Keywords: Fungi, Soil contaminated, Khuzestan, Crude oil

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Determine the frequency of blood groups in patients with dermatophytosis in the Glabchy laboratory and private practices Kashan in 2001

Seyede Marzie Jabbari Shiade, Rezvan Talaei, Mohamad Ali Asadi, Abbas Droodgar, Mehdi Salehi

Student Research Committee, Kashan University of Medical Sciences, Kashan, Iran Email: [email protected]

Introduction and Objectives: Dermatophytosis is a kind of skin diseases the prevalence of which is high in all over Iran and the world and due to the high social and personal problems, prevention and treatment of this disease is important. Materials and Methods: During this study, from the patients who were suspected to dermatophytosis lesions, were sampled, prepared smear and cultured by an expert mycologist then the blood group of positive samples were determined. Then the questionnaire was filled with the patient assistance. Results: From the 40 patients with dermatophytosis, 19 cases (47. 5%) had O blood group, 13 patients (32. 5%) had A blood group, 5 cases (12. 5%) had B blood group, 3 patients (7. 5%) had AB blood group. Percentage of male patients was 57. 5% and female patients were 42. 5%. Conclusion: Results obtained during this study indicate that blood groups could be a basic factor in the incidence of dermatophytosis therefore this study suggests that more research should be done in future. Keywords: Dermatophytosis, Blood groups

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Air fungal contamination in Namazi hospital transplantation wards

Parisa Badiee, Hadis Jafarian

Professor Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran Email: [email protected]

Introduction and Objectives: Certain fungi are the most dangerous pathogenic agents in transplant patients. Suppression of immune system for the prevention of transplanted organ rejection is necessary to perform transplantation. Air acts as the main source of contamination in transplant wards. Therefore, the refinement of air is one of the most important ways to control infections in transplant wards. The aim of the present study is to investigate air contamination. Materials and Methods: Air samples were taken from bone marrow and solid organ transplant wards and control air samples were taken from hospital environment six times in two seasons using two methods (settle down and suction). Results: The results of the two above mentioned methods were similar in percentage of contamination and etiologic agents. Air fresh samples from Namazi hospital contained 6 fungal species common to ward results. In this study, it was found that the air in organ transplant ward, bone marrow ward and the environment had 98.49%, 92.23% and 100% contamination, respectively. The six common in all wards were Penicillium sp, Cladosporium sp, Aspergillus niger, A. flavus, Rhizopus and Alternaria sp. Other isolated fungi were included Handersonella sp., Paecilomyces sp., Chaetomium sp., Stemphylium sp., A. terreus, Mucor sp., and Candida albicans. Conclusion: Considering that the transplant patients are so sensitive to pathogenic factors and respective costly hospitalization of such patients, a suitable way to reduce the fungal contamination in these wards is suggested. The presence of more fungal species diversities in this wards shows that source of fungal contamination is probably organized from inside of wards. We recommend using efficient air filtration in the transplant wards to prevent the fungal infection. Keywords: Transplant, Air contamination, Saprophytic fungi

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The effects of variable CO2 concentrations on morphological patterns in dermatophyte pathogen Microsporum canis

Sima Darabian, Sasan Rezaie, Parivash Kordbachche, Mohammad Reza Safari

Department of Medical Mycology and Parasitology, School of Medical, Zanjan University of Medical Sciences and Health services, Zanjan, Iran Email: [email protected]

Introduction and Objectives: The dermatophyte pathogen Microsporum canis is a zoophilic dermatophyte which is transferred from dog and cat to human. Some properties of M. canis have been investigated so far; however, no information is available regarding the effects of CO2 on morphological patterns in this fungus. In this study we want to examine the environmental and iatrogenic factors such as CO2 on the growth and morphological aspects of M. canis. Materials and Methods: Strains used in this study were obtained from the patients with tinea capitis and tinea corporis and the spores have been isolated from samples by standard methods. Spores were cultured on Sabouraud dextrose broth (SDB) with pH 5.6 ±0.2 and incubated at 25°C for 2, 4 and 7 days under the 3%, 5% and 10% of CO2 concentrations. Control growth was maintained for 7 days without CO2 concentration. Results: The results of this study clarified the fact that the maximum variations under 3%, 5% and 10% of CO2 concentrations in maximum 7 days incubation period and the variations of morphological showed different variations under different CO2 concentration. Conclusion: Our result showed the additional effect of CO2 concentrations in the production of arthroconidia, chlamydoconidia and microconidia in M. canis comparing with the controls in all conditions during this study. Keywords: Dermatophyte, Microsporum canis, Microconidia, Arthroconidia

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Study on saprophytic fungi in archives of office from Sari city and their role in sick building syndrome

Somayeh Fooladi Rad, Mohammad Taghi Hedayati, Tahereh Shokohi

Department of Medical Mycology and Parasitology/ Invasive Fungi Research Center, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran Email: [email protected]

Introduction and Objectives: Moulds are responsible for diseases in humans through the three mechanisms of infection, allergy, and toxicity. Indoor airborne mould spores cause human disease such as sick building syndrome. Sick building syndrome (SBS) is a term used to describe the reduced comfort and health status associated with an individual's work place or residential building. This syndrome includes symptoms such as headache, nausea, eye, throat and nasal irritation, tiredness and odor. Sick building syndrome is attributed to inadequate ventilation, chemical contaminants from indoor sources, and biological contaminants such as saprophytic molds, VOC and other biological agent. Materials and Methods: Indoor of 20 archives were sampled using a SKC impactor which draws air at 20 lit /min and impacts the sampled material onto Petri dishes containing Malt Extract Agar. Surface samples were also collected by pressing a sterile cotton swab on different areas of archives and maintained into tubes containing PBS+ 0.1% Tween 80 until they were identified in the laboratory. The collected swabs also cultured on MEA. The plates were incubated at 27- 30°C for 7-14 days, after which the resulting colonies were counted. The fungi were subsequently identified by both microscopic and macroscopic observation. For air samples, the counted fungal colonies were converted to CFUs per cubic meter. Results: 44 plates contained MEA were collected from Indoor air and surfaces of archives. In indoor air, Cladosporium (25.1%), Aspergillus spp (22.9%) and Penicillium (22.9%) had the most frequencies. In surface samples Cladosporium (27.9%), Penicillium (20.5%), Aspergillus spp (19.9%) and Stachybotrys (7.9%) were the most common isolated fungi. Cladosporium had the highest total CFU concentration in indoor air of archive (1227/m³). Conclusion: In the present study, saprophytic fungi is characteristically found in indoor air and surfaces from archive. Archive environments with a main source of organic material provide a nourishing media for fungal growth. Handling of contaminated files, paper, the high concentration levels of airborne fungi and VOC in archives of offices might put the workers at risk and cause adverse effects on the health, such as respiratory diseases and sign of SBS. Therefore, the exposure level to fungal spores on archives of offices must be carefully considered. Keywords: Air borne Fungi, Archives, SBS, VOC

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Assessment of soil fungi prevalence in different areas of Kashan from 2010 to 2011

Mohamad Ali Asadi, Abbas Droodgar, Afshin Salehi, Azam Samei

Kashan University of Medical Sciences, Kashan, Iran Email: [email protected]

Introduction and Objectives: Soils of different areas may contain saprophytic, pathogen or opportunist fungi which are mainly considered for their medical applications. Methods and Materials: In this study, five desert and five mountaineer villages were randomly selected and soil sampling was done from their different desert, residential, livestock and agricultural points. Sampling was carried out both in summer and autumn and six samples were prepared from each, i. e. total ten samples. Also, thirty soil samples were gathered from different points of Kashan (six ones from each point). Then all the samples were studied via two experimental floatation and decoy with hair methods used by mycology experts and after creating colonies and identification of the fungi by teased mount or slide culture in floatation method, the following results obtained: Results: Of the total 484 colonies in mountain regions the most prevalent fungi in order of frequency were: Penicillium 192 colonies, Aspergillus. Sp. 143 cases, Alternaria 68 cases, mycelium sterile 28 cases, Rhizopus 23 cases, Chrysosporium 18 cases and Fusarium 12 cases. Also, in floatation method, of the total 315 colonies in desert areas the most prevalent fungi in order of frequency were: different Aspergillus. Sp. 129 cases, yeast cases 87 cases, Penicillium 27 cases, Fusarium 25 cases, Acremonium 19 cases, Mucor 11 cases, mycelium sterile 8 cases, Cladosporium 6 cases and Alternaria 3 cases. Conclusion: Regarding the variation of saprophytic, opportunist and also pathogen fungi such as dermatophytes in different area soils, it is recommended that more researches should be conducted in this field. Keywords: Soil, Saprophytic, Dermatophyte, Kashan

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Airborne fungal contamination in the operating rooms of Yazd medical university hospitals

Abbas Ali Jafari, Farzanah Mirzaeii Department of Medical Parasitology and Mycology, Medical School, Shahid Sadoughi University of Yazd Medical Sciences, Yazd, Iran Email: [email protected]

Introduction and Objectives: Skin is one of the natural defense mechanisms of the human body act as an important barrier to prevent the penetration of pathogen microorganisms. Damaging of the skin in the patient, who is under operation and as a result of the post operating treatment usually using wide spectrum antibiotics, the opportunistic nosocomial agents especially the airborne fungal spores are a treat for patient. The most disinfectant agents that uses for scrubbing of operating rooms are antibacterial. The general objective of the present study was to evaluate the types and frequency of filamentous fungi and yeast, present in the environment of the operating room of three major hospitals in Yazd. Materials and Methods: In the current cross sectional descriptive study we used the plating method for air sampling collection. We used 921 air samples from 7 operating rooms of 3 hospitals and cultured. The fungi, which isolated were diagnosed by microscopic and macroscopic methods and the results were analyzed using SPSS software. Results: Totally, 885 colonies of fungi from 13 types were isolated that Penicillium (22. 4 %), yeast (19. 4 %) and Cladosporium (18. 2 %) and Aspergillus niger (13. 8%) were the most frequently isolated fungi. Operating rooms in hospital 2 showed the most contaminated rooms in this study. There was seen a statistical significant differential between operating room of 3 hospital (F=43.7, Pv=0). The average isolated fungal spores was 2 in hospital 2 surgery operating room known as most contaminated room but in women’s operating room this was 0. 6 CFU in each sample (F=17.6 %, Pv=0). However, on Wednesdays more contamination was observed in comparison with Mondays and Saturdays but there wasn’t statistically significant this difference (F=0.45, P=0.665). Conclusion: It seems that the operating rooms in the studied hospital one highly contaminated with fungal spores and it is necessary to have more attention especially in open heart surgery operating room. Keywords: Operating room, Airborne, Fungi, Contamination

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Fungal isolated from a small industry related to car factory in Babol

Saeid Mahdavi Omran, Seyed Mohammad Bagher Norozian Amiri, Mohammad Reza Yousefi, Seyed Javad Mosavi

Infectious Diseases and Tropical Medicine Research Center, Babol University of Medical Sciences, Babol, Iran Email: [email protected]

Introduction and Objectives: Fungal air contamination can cause some degradation in rainwater quality, if it use from roof for drinking and also for some delicate usage. According to the report around the abnormality of solution for car staining, the present study was carried out for detection of causative agent. Materials and Methods: twenty one samples were taken from ten tanks in a small factory related to car industrial, Babol. Ten milliliter of each sample selected according the standard sampling of water and the solution. After removing of large particles, the samples were precipitated by centrifugation. All precipitated samples were examined microscopically and 50µl of them plated on Sabouraud dextrose agar supplemented with chloramphenicole for growth of fungi, and fungi identified according to macroscopic and microscopic appearances. Results: Thirteen isolates of fungal colonies were isolated from these tanks, which yeast, Penicillium and Cladosporium were the most prevalent fungi. Rinsing tank 2, activation tank and fixation tank were the most contaminated with fungi. Some of the tanks were without any fungi contamination at one or two time sampling. Conclusion: According to the results of present study, the fungal contamination can change the quality of rainwater such as color and suitable usage. Thus we need possess some step such as filtering for removing and deletion of fungi and also bacteria from this water, specially according the regional weather conditions. Keywords: Rainwater, Fungi, Industry

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Real Time RT-PCR study of beta-defensin 1 gene expression in murine macrophages pretreated with different types of interferons and treated with Candida albicans and lipopolysaccharide

Elnaz Heshmat Azad, Kamiar Zomorodian, Zahra Rezaei, Bahador Shahriari, Farideh Jalali- Mashayekhi, Keyvan Pakshir, Marjan Motamedi Department of Medicinal Chemistry, Faculty of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran Email: [email protected]

Introduction and Objectives: Antimicrobial peptides (AMP) are important components of the natural defense of the most living organisms. The cellular expression of these peptides could be stay constant or be induced by cytokines and microbial products. The present study was designed to explore the effects of different types of interferons (IFNs) treatments on gene expression of AMP, mouse beta defensin 1 (mBD-1), in murine macrophages following exposure to Candida albicans yeasts and lipopolysaccharide (LPS) by Real time-RT PCR method. Materials and Methods: After cultivation of murine macrophages, 18 hrs pretreatments with IFNs and 6 hours treatments with C. albicans and LPS were carried out in 6 groups: negative control (cells + medium), positive control (cells + medium + LPS), Interferon (cell + medium + IFN), Yeast (cell + medium + C. albicans), Test1 (cell + medium + IFN + C. albicans) and Test2 (cell + medium + IFN + LPS). Experiments, all were performed triplicate. Then, cellular RNAs were extracted by Trizole method and the first strand complementary DNAs (cDNAs) were synthesized by Reverse Transcription method. After that, the changes in expression of the mBD- 1 gene were studied in comparison with the house-keeping gene, beta-actin by Real Time PCR method and ΔΔCT analysis. Results: Significant changes were limited to Test 2 groups: 1.89 ± 0.02 and 1.51±0.05 for IFN-α + LPS and IFN-γ + LPS respectively. Conclusion: This study showed that the mBD-1 expression was not upregulated by IFNs, C. albicans and LPS treatment alone. But pretreatment with IFNs and treatment with LPS, could raise mBD-1 expression significantly. This effect was higher for IFN-α. Whether, due to low expression of this gene in macrophages, study in other type of cells, like epithelial cells is suggested. Keywords: Beta defensin 1, Interferons, Real Time RT-PCR

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Identification and detoxification of aflatoxin in bread wastes used in livestock food in Tehran

Atena Alirezaei, Seyed Jamal Hashemi, Ayatolah Nasrollahi, Ehsanollah Fakhari

Department of Mycology, Azad University, Science and Research Branch, Tehran, Iran Email: [email protected]

Introduction and Objectives: Mycotoxin is secondary metabolites of Aspergillus flavus and Aspergillus parasiticus that detected of livestock foods. Aflatoxin is leader to most of health problems such as cirrhosis. Aflatoxin M classified as Aflatoxin B is carcinogen for liver and kidney that find in corn, beans, peanuts, rice and bread. In this study we want to identiy and detoxify AFT in the bread that used by livestock₁ in Tehran. ₁ Materials and Methods: During this research, the sampling is done by following 3 steps of 17 stages. The wastes bread 1 step in put and 2 stages output were selected from the crusher and packaging machine. For each input waste bread, 50Kg is given selected to the machine, 100 grams in the beginning and 1000 grams after heating process. Identification was done in all type of Aflatoxin AFT, G , G , B , B using high performance liquid chromatography (HPLC) method. The output waste bread was held detoxification of Aflatoxin using normal 1 citric acid. Results: According to₂ surveys₁ ₁conducted,₂ to the output of waste bread from machine showed significant reduction in comparison with the input of waste bread in terms of Aspergillus flavus and A. Parasiticus contamination. We detected (5.88%) AFB , (29.41%) AFG , (100%) AFB and AFT. The process for exclusion of Aflatoxin was 100% successful on all waste bread samples. ₂ ₁ ₁ Conclusion: Mycotoxins creates a health risk for humans and livestock human exposure to Aflatoxin can result from direct consumption of Aflatoxin contaminated foods, whether processed unprocessed. Keywords: Detoxification, Aflatoxin, Waste bread

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A comparative investigation on total aflatoxin and ochratoxin production spectra in 15 different Asprergillus species from Iranian northern states

Zohre Zohari, Arash Chaichi Nosrati, Leila Modiri, Naser Ghaemi, Soheil Shokri

Department of Microbiology, Faculty of Basic Sciences, Lahijan Branch, Islamic Azad University, Lahijan, Gilan, Iran Email: [email protected]

Introduction and Objectives: Members of the genus Aspergillus have been of interest because of positive impact, toxicity, and pathogenicity. There are multiple factors significantly involved in the development of toxigenic section-species and secondary metabolites biosynthesis. Ochratoxins are the second mycotoxin group in importance after aflatoxins of which are the most studied metabolites due to their occurrence in food and feed and toxicological significance in human and animal diets. Besides environmental factors, biological factors have a noticeable influence on growth and toxins production. Some are depended only on the genetic basis of the fungal strain which vary in their ability to produce as well as the quantity produced. Materials and Methods: In order to explore the biochemical scope of toxin producing Aspergilli, we screened 106 isolates using indirect competitive ELISA detection with a limit of detection: 2.5 ppb. per assay. Results: Fungal species contaminations were highly variable. Some contain both afla- ochratoxins named as section Circumdati, Candidi, Flavi, Fumigati, Nidulantes, Terrei, Wentii, Ornati and Nigri, only or typically contain extremely low levels ranging by firstly 200-300 and secondarily 200-400 ppb. The 31 isolates could be grouped into a large and clearly separated with a small outlying isolates of ranged between 500-700 ppb as high ochre-afla toxin producing isolates belonging to section Flavipedes and Circumdati. The finest group, containing 5 isolates, mainly originating from the same subgenus sources included the bank culture of Iranian northern Aspergilli. We review isolates distribution, biosynthesis, toxicity and other aspects focused on toxins, since is becoming a key in its tremendous intraspecific variation in production due to different species and sections of Aspergilli. As with aflatoxins, this variation is a combination of phenotypic plasticity and genetic variation. it is entirely possible that, ochratoxins will have overshadowed aflatoxins in terms of public interest and debate. Conclusion: Assigning combined toxin producing is very important because it emphasizes that despite sharing the ability to produce both toxins, We suggest that the groups represent the species, belong to the series or subgenus groups of fungi are biosynthetically and ecologically very different produce nobel secondary metabolite profile suggesting that usually occupy same ecological niches. Keywords: Ochratoxins, Aflatoxins, Aspergillus

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A comparative investigation on ochratoxin and aflatoxin B production spectra in 15 different species of Aspergillus isolates from Iranian northern states (a case study)

Sareh Akhavan, Leila Modiri, Arash Chaichi Nosrati, Soheil Shokri

Department of Microbiology, Faculty of Basic Sciences, Lahijan Branch, Islamic Azad University, Lahijan, Gilan, Iran Email: [email protected]

Introduction and Objectives: Mycotoxins are natural contaminants found in different type of foods in which the risk of its exposures continues in the developing countries. The European Union (EU) limits ochratoxin in imported foods, with a maximum of 5lg/kg (=ppb) in feed,3lg/kg in processed foods strongly suggests that OTA given uncertainties and trends, will have overshadowed aflatoxins in terms of public health interest and debate. Materials and Methods: In order to explore the biochemical scope of OTA producing Aspergilla and their metabolic correlations to AFLA-B productivities, we screened 46 isolates. OTA and AFLA-B detection was developed and performed based on indirect competitive ELISA as an integrated detection with a limit of 2.5 ppb in cellular extracts for the production secondary metabolites. Results: Fungal metabolites contaminations were highly variable. Some of species isolates mentioned here rarely contain ochratoxin-aflatoxin named as Section Circumdati, Candidi, Flavi and Nigri isolates, only or typically contain extremely low levels ranging by firstly 300-35 and secondarily 0-500ppb,one or both. The other 10 isolates could be grouped into a large and clearly separated group and a small outlying isolates of ranged between 2000-2500 ppb as high- ochratoxin producing isolates belonging to Circumdati section. The solis group, containing1 isolate, mainly originating from the same subgenus sources included the bank culture of Iranian northern Aspergilla. Conclusion: The major objective of this research is to review isolates distribution, toxicity and regulatory aspects of ochratoxins focused on ochratoxins in all isolates biomasses, since is becoming a key in ochratoxin-Aflatoxin bounded research. There is tremendous intraspecific variation in toxin production due to different species and sections of Aspergilli even though the cellular metabolites to culture media. Keywords: Ochratoxin, Aflatoxin B, ELISA, Aspergillus

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A comparative investigation on ochratoxin and aflatoxin B production spectra in culture medium of 15 different Aspergillus species from Iranian northern states

Hedyeh Jafarzadeh, Arash Chaichi Nosrati, Leila Modiri, Soheil Shokri

Department of Microbiology, Faculty of Basic Sciences, Lahijan Branch, Islamic Azad University, Lahijan, Gilan, Iran Email: [email protected]

Introduction and Objectives: Aflatoxin and Ochratoxin a diverse group of toxic compound contaminants in four sections of Aspergillus and in Penicillium spp, with a curiously distribution also the areas their biosynthesis, toxicity are still not completely elucidated and the regulations be used to control contamination. The risk of mycotoxin exposures continues due to lack of food security and there is a strong need for further proper management practices using biocontrol agents to protect consumer’s health even few of these have practical application. From an economics point of view, it is necessary to salvage mycotoxin-contaminated commodities and to avert health risks associated with the toxins. Ochratoxins-Aflatoxins, are gaining importance so we consider them in the context of which are better understood in a combination of phenotypic plasticity. Many sampling strategies are not designed to detect consistently these few, highly contaminated at a range of daily OTA intake estimated at 0. 7–4.6ng/kg body weight, based on a typical European diet in various foods, in certain places and circumstances. Materials and Methods: In order to explore the biochemical scope of Ochratoxin-Aflatoxin producing Aspergilli, we screened 66 isolates. Toxin detection was developed and performed based on indirect competitive ELISA as an integrated detection with a limit of 2.5 ppb in culture media extracts for the production of released cellular secondary metabolites. Results: Some of species isolates mentioned here rarely contain Ochratoxin named as section Fumigati, Nidulantes, Terrei, Candidi, Flavi, Wentii, Nigri,Ornati isolates, only or typically contain extremely low levels ranging by firstly 300-400 and secondarily 200-300 ppb or both. Conclusion: The 21 isolates could be grouped into a large and a small outlying isolates of ranged between 500-700 ppb as high-Ochratoxin producing isolates belonging to section Flavipedes and Circumdati. The latest group, containing 4 isolates, mainly originating from the same subgenus sources included the bank culture of Iranian Northern Aspergilli. Keywords: Ochratoxin, Aflatoxin B, Aspergillus species

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Survey the dermatophyte contamination of covered swimming pools in Yazd 2009

Abbas Ali Jafari, Mohammad Hassan Ehrampoosh, Mohammad Taghi Ghaneian, Somaya Rahimi

Department of Medical Parasitology and Mycology, Medical School, Shahid Sadoughi University of Yazd Medical Sciences, Yazd, Iran Email: [email protected]

Introduction and Objectives: Swimming public covered pools are known as one of important entertainment centres for most people. Contaminated pools could transmit a number of contagious diseases; particularly fungal disease. This study was conducted in order to determine the dermatophyte contamination and few variables such as the mean value of residual chlorine, pH, turbidity and temperature in the Yazd covered pools in summer 2009. Materials and Methods: In current descriptive study four covered swimming pools were randomly selected for detecting fungal contamination in summer 2009. Samples were collected from 25 different areas of each pool (foot-bath, showers, dressing room and sauna) using carpet collection method. Several physico-chemical parameters of water in swimming pools such as turbidity, pH, temperature and residual chlorine were also determined in this study. Results: The data indicate that 7% of the total samples were contaminated with dermatophytes mostly in dressing rooms, Sauna and foot-bath parts. Surprisingly Trichophyton mentagrophytes were isolated from 6 and Epidermophyton floccosum from one sample. Conclusion: Results demonstrated the dermatophyte contamination in Yazd covered public pools, which can cause cutaneous infections and pose a significant risk to the health of swimmers. It seems that more strict supervision including decontamination of pools, sanitation and controlling the residual chlorine is necessary by the health authorities. Keywords: Swimming pools, Dermatophytes, Contamination

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Measurement of aflatoxin B1 in flour of Sohan making in Qom

Nasrin Bitarafan Qomi, Mojgan Saghazadeh, Majid Riazipour

Department of Microbiology, Islamic Azad University, Qom branch, Qom, Iran Email: [email protected]

Introduction and Objectives: Aflatoxins are toxic secondary metabolites. It is a matter of importance that Aflatoxin B1 is the most toxic and carcinogenic of them. It has mutagenic and carcinogenic properties, so it is a potent hepatocarcinogen agent in mammals. Oil seeds, cereals and dried fruits are susceptible for contamination of Aflatoxin B1 if they confront with adverse condition during storage. The aim of this study is the investigation of contamination of flour which is used in Sohan making in Qom province. Materials and Methods: In this study, a number of 80 samples of wheat flour from 80 different Sohan making stores have been collected randomly. After extraction with methanol, the amount of Aflatoxins were measured using competitive ELISA, also more information about the type of flour, the place of wheat flour producing and collecting, storage duration were collected with the help of special staff in Sohan making stores. Statistical analysis, were performed using SPSS software, curve software, Excel and student T test. Results: All of the samples were contaminated with some level of aflatoxin B1 but level of contamination for B1 in all sample were determined lower than 5 ppb (IRAN) and 2ppb in (EUROPEAN) standard. The average concentrations of total mycotoxins in all samples were determined 0.75μg/kg with 0.37 standard deviation. The highest level of aflatoxin B1 was 1.79ppb and was related to Sohan making shop Number 74 using NOL POUYA flour and the lowest level of aflatoxin B1 was 0. 22ppb and was related to Sohan making shop number 8 using Laleh Isfahan Nol flour. Conclusion: Although aflatoxin B1contamination in flour which used in Sohan making was in Permitted limit, regarding to this fact that even low amount of aflatoxin can jeopardize human health, so control and evaluation of aflatoxin B1 level in Sohan making shop could be necessary. Keywords: Aflatoxin B1, Wheat flour, Sohan

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Genetic diversity of Aspergillus spp. isolated from ICU, indoor and outdoor environments in hospitals in by PCR and determine drug sensitivity

Ghazaleh Mahmmodi, Ayatolah Nasrollahi Omran, Mohammad Taghi Hedayati, Ali Nazemi, Azar Karmi

Department of Microbiology, Islamic Azad University, Tonekabon Branch, Iran Email: [email protected]

Introduction and Objectives: Over the past decade, systemic infections caused by fungal pathogens such as Candida and Aspergillus species have become a prominent cause of disease, especially in severely ill or immunocompromised patients. High genotypic diversity indices have been described among clinical and environmental isolates of Aspergillus spp., usually as the result of studying outbreaks at ICU. Materials and Methods: From May 2012 through October 2012, 100 cases Aspergillus strains (20 cases A. fumigatus, 40 case A. flavus and 40 cases A. niger) were obtained from ICU, indoor and outdoor environments hospitals in Mazandaran province. Stock solutions of itraconazole for MIC were prepared in DMSO. The broth microdilution assay was performed according to the Clinical and Laboratory Standards Institute M38-A reference method. Miniprimer PCR method was also conducted for the differentiation of species. Sequences were compared to those derived from type strains deposited in the Gen Bank data base to identify isolates to the species level. Results: The minimal inhibitory concentration (MIC) ranges for itraconazole against A. fumigates 0.25 -2, A. flavus 0.5 – 0.125, A. niger (0.5-2) were, respectively. Genetic diversity of Aspergillus species may represent a useful tool for illustrating the presence or absence of specific clones in clinical wards. Conclusion: Advances in health care technology and life support systems have increased the risk of nosocomial fungal infection, which are now a major cause of morbidity and mortality in the ICU. Considering the high dispersion capability of Aspergillus spp.in indoor environments, more attention should be given for preventative measures in restricted wards that are mainly used for admission of hematological and transplant-risk patients. Keywords: Genetic diversity, Aspergillus spp., ICU, Hospitals, Drug sensitivity

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An investigation of the ability of Candida albicans in reducing the growth of Aspergillus parasiticus in culture medium and wheat

Zeinab Aghamohseni, Sasan Rezaie, Gholamreza Jahed Khaniki, Mahmood Alimohammadi, Fatemeh Noorbakhsh, Matin Mohammadi Kuchesfahani

Department of Environmental Health Engineering, Faculty of Health, Tehran University of Medical Sciences and Health Services, Tehran, Iran Email: [email protected]

Introduction and Objectives: Great economic losses occurred around the world because of Food spoilage moulds. It is estimated that between 5 to 10% of the world’s food production is wasted due to fungal deterioration. These fungi cause losses in dry matter or quality and some species like Aspergillus parasiticus can produce health-damaging mycotoxins. In this study, the ability of Candida albicans in reducing the growth of A. parasiticus in culture medium and wheat is investigated. Materials and Methods: A. parasiticus and A. parasiticus were cultured into Sabouraud dextrose agar (SDA) and were incubated at 28°C for 3 days for C. albicans while A. parasiticus were incubated at 28°C for 5 days. The inoculum was adjusted to 104 spores/gr for A. parasiticus and 105, 103, 107, spores/g for A. parasiticus by using neubauer lam and added to culture medium SDA and wheat that autoclaved at 121°C for 20 min then incubated for 5 days in 28- 30°C. Fungal growth was determined at the end of the incubation period by measuring weight of Aspergillus mycelium. Results: The results from both culture media and wheat indicated that all three concentrations of Candida used can have growth inhibition of the mycotoxin-producing Aspergillus strain (p<0.05) and the reduction of growth is related to the concentration of Candida respectively. Conclusion: It is shown that C. albicans might be employed as biological tool to lower Aspergillus growth. Keywords: Aspergillus parasiticus, Reduction, Yeast

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Prevalence of fungal elements in Isfahan swimming pools water

Parvin Dehghan, Mehrnoosh Maherolnaghsh, Zahra Musavi, Hossien Saffari, Hossienali Yousefi, Gholamreza Ahmadi, Zahra Jabalameli

Department of Mycology and Parasitology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran Email: [email protected]

Introduction and Objectives: The widespread use of recreational waters and swimming pools in recent years could be considered as an important factor of health risk relating to water and microorganisms entering the pool directly or indirectly through contaminated water, air, soil, dust and individual bathers. Our purpose is to these problems, focusing on fungi. Materials and Methods: 399 liters of water samples (133 samples, 3 liter) were collected aseptically from 29 different swimming pools and also their internal waters were examined. Out of 29 pools, 17 had Jacuzzi. Filtration technique (Millie-pore) was used for both direct examination and cultural method. Results: The ranges of free chlorine residual, pH and temperature of water pools were 0-6. 8, 5.6-7 and 25-31°C, respectively. In direct examination, out of 29 pools, 8 internal water were positive considering fungal elements and the range was from trace to 4+, Also Fungal elements were seen on 14 of 29 (48.2%) of water sediments and just 4 pools (13.7%) showed positive for yeasts by direct smears (ranges 33-400 yeasts/lit). By cultural method all pools except 10 of internal waters were polluted by viable fungal elements. The number of fungal elements in water pools was accounted ranged from 6-820 colonies/lit. Yeasts were grown in more than half of pools (51. 7%) and Candida albicans were isolated just from water of one Jacuzzi. Among the mycelial fungi, Aspergillus species particularly A. flavus was prominent in the swimming pools water of Isfahan. Conclusion: Our results suggest that the safety and cleanness of internal water is very important and the population of all the microorganisms and physico - chemical conditions of water should be monitored, managed and balanced regarding the population of bathers, microorganisms and their interactions in water. Keywords: Prevalence, Swimming pool, Fungus, Isfahan

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Qualitative and quantitative study monitoring airborne fungal flora in the kidney transplant unit

Mohammad Ali Afshari, Majid Riazipour, Reza Kachuei, Mojtaba Teimoori, Behzad Einollahi

Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Solid organ transplant patients are at high risk for opportunistic air-borne fungal infections due to using the potent immunosuppressive agents. The aim of the current study was to quantitative and qualitative evaluations of fungal flora presenting in the air of Kidney transplant unit of Baqiyatallah hospital. Materials and Methods: In this prospective study, air samples from patient room, baths site, ICU and isolated room, corridor site and outside the ward were obtained by settled plate technique using plates containing Sabouraud's dextrose agar medium. In this study, the number of the 36 agar plates containing Sabouraud dextrose agar medium was used. The plates were exposed for 20 min at the height of 100-150 cm above the ground in units of hospital. All samples were immediately incubated at 27±2˚C for 4 weeks . The slide culture method and Lacto-phenol cotton blue were used for definitive identification and staining fungal cultures, respectively. Results: The mean of colony forming units (CFUs) on indoor and outdoor plates was 6.6 ±1.3 and 6 ±1.9 / plate, respectively. Statistical analysis showed that the observed difference is not significant. Also, the results showed the mean of CFUs in the air of patient's rooms (6. 8±1.7), halls (4.5±1.7), bathrooms (6.8±1.5), and ICU rooms (3.2±1.8) have not significant differences. The mean of different fungal genera isolated from indoor and outdoor plates were 1. 9±0.2 and 4±0.5 genera/plate respectively, that indicates significant difference between indoor and outdoor air quality (P<0. 001). Conclusion: Lack of difference between quantity of outdoor and indoor air fungi indicates inefficacy of air control measures and indoor lower genus diversity in comparison to outdoor air shows that there may be conditions that facilitate fungal growth in the environment of kidney transplantation unit. Keywords: Air fungal flora, Opportunistic fungi, Renal transplantation unit, Hospital

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Toxigenic mycoflora distribution of hazelnut in Isfahan

Elham Saffari‚ Mahboobeh Madani‚ Vajihe Karbasizade

Department of Microbiology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran Email: [email protected]

Introduction and Objectives: Nuts can be contaminated by fungi under appropriate conditions, such as temperature and humidity. Some fungal species such as Aspergillus flavus and A. parasiticus can produce aflatoxin. Aflatoxins are genotoxic, carcinogenic and mutagenic. So the prevention of contamination with fungi is necessary. The aim of this research was to determine toxigenic mycoflora distribution of hazelnut in Isfahan. Materials and Methods: One hundred samples of huzelnut were collected randomly from supermarkets in Isfahan. All samples were immersed in sodium hypochlorite solution (5%) for 3 minutes, rinsed in sterile distilled water and dried between sterile papers. Samples were cultured on sabouraud dextrose agar and incubated at 25oC for 4 days. Then, the fungal colonies that developed from the infected grains were isolated, purified and maintained on slant potato dextrose agar (PDA) for identification trials. Isolated fungi were evaluated by mycological methods. Results: In this study, 78% of the samples were contaminated and 9 genuses of fungi, including Aspergillus, Penicillium, Rhizopus, Ulocladium, Alternaria, Drechselera, Trichothecium, Scopulariopsis, and Mucor were identified. Isolated Aspergillus were A. flavus, A. niger, A. fumigatus and A. terreus. The most predominant isolated fungus was Aspergillus niger, followed by Penicillium, Rhizopus and Alternaria respectively. Conclusion: Some of these fungi are toxigenic and threaten heath of humans. So fungal and mycotoxin contamination of hazelnut should be considered. Keywords: Fungal species, Huzelnut, Aflatoxin

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Radionuclide concentration in Agaricus bispora (Agaricaceae); an edible mushroom

Mitra Noori, Reza Poorimani, Sana Rahimi

Department of Biology, Faculty of Science, Arak University, Arak 38156-88349, Iran Email: [email protected]

Introduction and Objectives: Living organisms are ever exposed to nuclear radiations that are radiated from primary radionuclide unstable nucleus. Entrance and distribution of radionuclides in environment is influenced on ecosystem and living things. Fungi have a good potential in radionuclide collection and can absorb some radiant elements of their environment. Therefore, in this study radionuclide activity of a nourished mushroom (Agaricus bispora) was determined. Materials and Methods: An edible mushroom (A. bispora) samples were collected from a Mushroom Cultivation Center in Kermanshah for two cultivation periods. Dried mushroom powder was used and then spectrometry was done using HPGe gamma ray spectrometer with 30% relative efficiency. Obtained spectra were analyzed and according to them samples nuclei activities were determined. Results: Analysis of the results showed special activity means were 16.04 ± 0.08 Bq/kg for 137Cs, 4592 ± 11 Bq/kg for 40K, 81.33 ± 0.01 Bq/kg for 238U and 204.84 ± 0.06 for 232Th in the studied species. Conclusion: This study has shown existing 137Cs, 40K and natural radioisotopes chains in the studied samples and confirms pollution of some regions of our country to artificial radioisotopes. Also studies on using fungi for cleaning radionuclide pollutant is needed and suggested. Keywords: Radionuclide, Edible mushroom, Nuclei activities, HPGe

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Characterization and determination of red yeast isolated from leather waste water as potential source of carotenoid

Farzaneh Sadat Naghavi, Parichehr Hanachi, Mohammad Reza Sodi, Ozra Sabora, Atefeh Ghorbani

Faculty of Basic Science, Biology Department, Alzahra University, Tehran, Iran Email: [email protected]

Introduction and Objectives: Red yeast in the genera Rhodotorula, Rhordosporidium, Sporobolomyces, Sporidium, Sporobolomyces and Phaffia are known as potential natural alternative sources of carotenoids. Rhodotorula, an anamorphic genus of heterobasidiomycetous yeasts is characterized by the absence of ballistoconidia, fermentation ability, and starch-like compounds, although Biology of the species, per se, does not describe it; molecular identification must be carried out. Sequence analysis of the D1/D2 region can be used for the identification of the majority of the species of Basidiomycetous yeasts. The increase of demand for carotenoids obtained from natural sources has promoted major efforts to recognize potential microbial sources. The aims of this study were identification of a strain which had been isolated from leather waste water and investigation of the ability of carotenoid production in the strain. Materials and Methods: Strain was determined morphologically, physiologically and biochemically. Molecular taxonomic analysis was carried out based on the 26S rDNA D1/D2 domain. To isolate the carotenoid pigment, cells were suspended in acetone and broken using a homogenizer, followed by centrifugation. The supernatant was separated. For Analytical method pigments were measured spectrophotometrically at 450 nm using the extinction coefficient 1% E 450=2500. Results: The morphological and biochemical characterization indicated that this strain belonged to the genus Rhodotorula. Molecular analysis based on the 26S rDNA D1/D2 domain sequence represented strain SG006 as a Rhodotorula slooffiae. The sequence was set down in the Gen bank database with Accession number JX997835. Conclusion: Among pigmented yeasts, only some small taxonomic groups have been investigated for the carotenoid pigment content. The type of carotenoid and their relative amount may vary depending on genus of yeast and environmental condition. It is obvious that SG006 has the ability to produce carotenoid, however further study is needed in order to utilize of this strain in food industry, pharmaceuticals and cosmetics. Keywords: Carotenoid, D1/D2 domain, Rhodotorula

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Effect of vitamin A, K1 and B1 on Aspergillus parasiticus growth and aflR gene expression

Ladan Nazemi, Maryam Moazeni, Sassan Rezaie

Division of Molecular Biology, Department of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran E-mail: [email protected]

Background and Objectives: Aflatoxins are teratogenic, mutagenic and hepato carcinogenic fungal secondary metabolites. They are produced by certain species of some fungi such as Aspergillus parasiticus. Recently, the elimination of aflatoxin contamination in food and feed has been considered by scientists worldwide. In this study, the effect of some vitamins such as; vitamin A (as a powerful antioxidant), vitamin K1 and vitamin B1 (thiamin), was studied on A. parasiticus growth and aflatoxin production. Materials and Methods: Standard strain of A. parasiticus ATCC 15517 was applied for performing antifungal susceptibility test due to the vitamin A, K1 and B1. Vitamin K1 had been provided in suspension whereas vitamin A were dissolved in DMSO containing Tween 40 and absolute alcohol and vitamin B1 w dissolved in sterile diluted water, respectively. Antifungal susceptibility test was performed according to CLSI document M27-A3. Vitamin A and B1 were dissolved in their own solvent to get the diluted concentration of 64, 32,16,8 mg/ml and 500, 250, 125, 62.5 mg/ml, respectively. For vitamin K1 different concentration (500, 250, 125, 62.5 mg/ml) were prepared. Quantitative changes in expression of the aflR gene were analyzed by quantifying the aflR mRNA level by use of a quantitative real-time RT-PCR assay. Results: It was revealed that vitamin A has the best inhibitory effect on A. parasiticus growth at 32 mg/ml. However, applying vitamin B1 leads to a relative increase in fungal growth in a dose dependent manner. In case of both vitamin B1 and K1, the minimum inhibitory concentrations were yield at >500 mg /ml. In addition, the level of aflR gene expression was significantly decreased after treating the fungus with 500 mg/ml of both vitamin A and K1.In case of vitamin B1, the level of cognate aflR mRNA was decreased at ≥250 mg/ml. Nevertheless, the expression of aflR gene was up-regulated at the concentration of 125mg/ml and below. Conclusion: Based on the obtained results, vitamin A has a significant inhibitory effect on A. parasiticus growth as well as aflatoxin production. Although vitamin K1 and B1 could not inhibit fungal growth completely, the rate of aflR gene expression was significantly reduced after treating with the two mentioned vitamins. In conclusion, using natural compounds such as vitamins may be regarded as potential anti-toxic agents in food and agriculture-related industries. Keywords: Aflatoxin, Aspergillus parasiticus, Vitamins, Gene expression

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Hemolytic and esterase activity of Candida species isolated from patients

Keyvan Pakshir, Maryam Jafari, Kamiar Zomorodian, Hamideh Taraz, Hooman Ebrahimi, Mohsen Karamitalab

Department of Parasitology and Mycology, School of Medicine Shiraz University of Medical Sciences, Shiraz, Iran Email: [email protected]

Introduction and Objectives: Candidiasis is an opportunistic fungal infection caused by many species of Candida that affects different sites of body. Secretion of exoenzymes plays an important role in virulence and pathogenesis. The aim of this study was to evaluate the esterase and hemolytic activity of Candida species isolated from onychomycosis and oral lichen planus patients. Materials and Methods: A total of 84 Candida spp including 24 C. albicans and 46 C. parapsilosis from onychomycosis and 14 C. albicans from oral lichen planus patients were included in our study. Tween 80 opacity test and blood agar plate assays were used for determining esterase and hemolytic activity, respectively. The isolates were subculture on Sabouraud medium and 10µl from each colony was prepared and cultured on specific medium for evaluation of hemolysin activity. A sterile swab did used for preparation of inocula for esterase activity. Opaque halos around the colonies were graded as positive esterase activity. A ring of lyses around the colonies were considered for hemolysin activity. Results: C. parapsilosis isolates had the least exoenzyme activity among the isolates. Candida albicans isolates from lichen planus showed less exoenzyme activity in comparison to the onychomycosis isolates. All of the C. albicans isolates from onychomycosis patients had beta hemolysin activity but only 56. 52% of C. parapsilosis isolates had beta hemolysin activity. 43. 47% of parapsilosis isolates had esterase activity. Among C. albicans isolates, 87.55% and 78.5% from onychomycosis and oral lichen planus had esterase activity, respectively. Conclusion: Candida isolates reveals different enzymatic patterns in clinical lesions. Keywords: Esterase, Hemolysin, Candida albicans, Onychomycosis, Lichenplanus, C. parapsilosis

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Candida species are transmitted by cell phones in hospitals: A comparison between doctors, nurses, accompanying the patients and patient cell phones contamination

Seyed Reza Aghili, Mitra Shokri, Bahar Salmanian

Department of Medical Mycology and Parasitology, Faculty of Medicine, Mazandaran University of Medical Sciences Sari, Iran Email: [email protected]

Introduction and Objectives: Cell phones are increasingly becoming an important means of communication by medical personnel in the world. Sharing of cell phones between health care workers (HCWs) and patient may directly facilitate the spread of potentially pathogenic fungi to the community. Candida species is the fourth most common source of hospital-acquired infections. This is probably the first study in Iran that attempts to study the Candida contamination of cell phones of HCWs and to compare it with that found on cell phones of non HCWs in hospitals. Materials and Methods: A total of 247 cell phones of all participants from the three educational Hospitals of Mazandaran University of Medical Sciences were screened. A piece of sterile carpet was rolled over all exposed outer surfaces of the cell phones used for at least 1 month. The sterile carpet was removed by tapping and shaking and placed on the surface of Sabouraud’s dextrose agar medium containing chloramphenicole. Plates were incubated at 25-27°C aerobically a period of 2 weeks. Results: 116 (96. 6%) of the cell phones of participants in the special care units (SCU) and 124 (97. 6%) of participants in the non-special care units (NSU) showed positive growth for Candida species. The most common Candida species found on cell phones were Candida krusei, C. albicans and C. glabrata (41.7, 30, and 17%). 68.4% samples in SCU and 57.5% in NSU had higher than +4 scores of Candida growth. However in comparison, doctor’s cell phones in SCU had less contamination than candidal contamination to doctor’s cell phones in NSU. But nurse’s phones in SCU had higher than candidal contamination to nurse’s phones in NSU. Samples that have no growth or less than the scores of +2 were seen in the cell phones of personnel who have jobs that require more frequent wash or disinfect their hands. Conclusion: This study indicates that the carriage of Candida species on the cell phones of populations present in hospitals can happen possibly and act as vehicles for transfer of potential fungal associated with nosocomial infections. More frequent wash or disinfect of hands can reduce cell phone contamination. Keywords: Cell phone, Mobile phone, Candidal contamination, Nosocomial infection

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Effect of three Iranian honeys on pro-inflammatory cytokines during invasive aspergillosis

Donya Nikaein, Ali Reza Khosravi, Hojjatollah Shokri, Ahmad Erfanmanesh, Hassan Ghorbani Choboghlo, Hadi Bagheri

Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran Email: [email protected]

Introduction and Objectives: In recent years, invasive aspergillosis has become an emergence in immunocompromised patients. Because of increased antimicrobial resistance, natural agents with medicinal and immunomodulatory effects has gained more attention. Honey is a natural substance with documented antimicrobial, anti-inflammatory and wound healing effects. In this study the effects of three Iranian honeys on pro-inflammatory cytokines during invasive aspergillosis was investigated. Materials and Methods: Mice were divided into 10 groups (honey alone, honey and infectious, negative and positive controls) each containing 10. Mice were pretreated with honey (1.5g/kg BW/orally) for 5 days. At day 6 Aspergillus fumigatus conidia (5×105/ml) was injected intravenously to infectious groups. Mice were euthanized at day 11, and spleen and peritoneal cell culture was done. Pro-inflammatory cytokine production using ELISA method was performed. Results: Results showed that honey treatment could significantly increase IL6 and IL1β production in infectious mice as well as improve macrophage killing (p<0.05). Also mice treated with honey had a greater survival time than infectious group. Conclusion: Regarding our results it is suggested that honey could boost the immune system and increase survival time in invasive aspergillosis. Keywords: Honey, Aspergillus fumigatus, Invasive aspergillosis, Immunomodulation, Inflammatory cytokines

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Isolation of keratinophilic fungi and aerobic Actinomycetes from City Park soils in Gorgan, North of Iran

Esmat Malek, Mahnaz Moosazadeh, Parastoo Hanafi, Ziba Abbasi Nejat, Abolfazl Amini, Rasool Mohammadi, Frahad Niknejad

Laboratory Science Research Center, Golestan University of Medical Sciences, Gorgan, Iran Email: [email protected]

Introduction and Objectives: Keratinophilic fungi are a group of fungi that colonize various keratinous substrates and degrade them to components of low molecular weight. Dermatophytes are recognized to cause superficial infection of keratinized tissues in humans and animals. The soil is also regarded as the reservoir of most pathogenic microorganisms including non- dermatophytic fungi. This study was conducted to determine the prevalence of keratinophilic fungi and aerobic Actinomycetes in soil of city parks in Gorgan. Materials and Methods: A total of 244 soil samples were collected from 22 different parks in Gorgan, North of Iran. The soil samples were collected from the superficial layer with depth not exceeding 0-10 cm in sterile polyethylene bags. The samples were transferred to the laboratory for processing. We used hair bait technique for isolation keratinophilic fungi. Colony identified by macroscopic and microscopic characterization after slide culturing. Actinomycetes were isolated by antibiotic dilution methods and recognized with physiological tests such as Lysozyme, Casein, Xanthine, Hypoxanthine, Gelatin, Urea Broth, and modified acid-fast stain. Results: 75 isolates of aerobic Actinomycetes were detected that Nocardia brasiliensis was the most strain (12%). Microsporum gypseum was more frequent than other keratinophilic fungi (26. 2%) and Aspergillus spp. was the most species of saprophyte fungi (18. 4%). Conclusion: This study showed that the soil of studied areas was rich of keratinophilic fungi and Actinomycetes, therefore hygiene protocol should be taken to prevent the spread of pathogenic and saprophytes fungi in the environment of susceptible person. Keywords: Keratinophilic fungi, Actinomycetes, Soil, Gorgan

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The impacts of indoor Aspergillus fungus on human health

Bita Mousavi, Mohammad Taghi Hedayati

Department of Medical Mycology, School of Medicine Molecular and Cell Biology Research Center, Mazandaran University of Medical Sciences, Sari, Iran Email: [email protected]

The growth of fungal molds is possible in homes, official buildings, schools, automobiles and other places where water and organic materials are available. So there is a considerable concern regarding the potential health outcomes of exposure to materials existing in the air which have biologic origin. Inhalation exposure to mycotoxins occurs by inhaling airborne particulates containing mycotoxins, including dust and fungal components. Sterigmatocystin which is the most notable Aspergillus versicolor mycotoxin is able to prevent from hair slips in respiratory system. Volatile organic compounds (VOC) may cause headache, lack of concentration, inattentiveness and dizziness. Sensitivity to Aspergillus is related to allergic diseases and Aspergillus might be a significant source of internal allergens. Aspergillus broncho-polmunary allergic (ABPA) and allergic fungal sinusitis (AFS) might also happen. Exposure to fungi occurs both in occupational settings and in domestic environments. So the quality of indoor air is one of the major health concerns. More studies have to be achieved from research to practice about indoor effect of fungi in order to improve the diagnosis of these health outcomes and to prevent further new cases of disorders. Keywords: Aspergillus, Allergen, VOC, Mycotoxin

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Evaluation of the keratinase activity in Trichophyton mentagrophytes strains isolates from patients with dermatophytosis.

Shahnaz Parsa, Masood Emami, Alireza Khosravi, Masoomeh Shams Ghahfarokhi

Islamic Azad University, Science and Research Branch, Tehran, Iran Email: [email protected]

Introduction and Objectives: Keratinase is the most important enzyme produced by dermatophytes. It can degrade the keratin of stratum corneum, nail and hair and use the resulted material. In this survey extracellular activity of Trichophyton mentagrophytes isolated from patients was evaluated. The aim of this study is the comparison of keratinase activity between different strains isolates of T. Mentagrophytes obtained from acute and chronic dermatophytosis Materials and Methods: Specimens were cultured on Sabouraud glucose agar containing cyclohexamide for 21 days at 30ºC, and then the colonies were inoculated on inducer medium with standard and synthesized keratin. Enzyme activity of understudy dermatophytes was assayed by spectrophotometry method. Results: Based on our results there were no significant differences between enzyme activities of dermatophytes in media containing either standard or synthesized keratin P>0. 05, also no correlation was observed between keratinase activity and the morphology of colonies. With respect to the site of infection and severity of lesions, they were not considered as important factors to release keratinase enzyme P>0.05. Conclusion: In order to determine the exact role of enzyme activity in pathogenesis of T. mentagrophytes, this study should be considered by different T. mentagrophytes obtained from various subjects. Keywords: Keratinase enzyme, Dermatophytes, Trichophyton mentagrophytes

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Biological control of the Blatella germanica with Beauveria bassiana and Lecanicillium muscarium

Sadegh Khodaveisi, Behroz Davari, Farzad Ala, Khorosh Javan, Noshin Abdolmalaki

Department of Medical Parasitology and Mycology, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Lecanicillium muscarium and Beauveria bassiana grow naturally in soils throughout the world and acts as a parasite on various arthropod species, causing white muscardine disease; thus belongs to the entomopathogenic fungi. This study evaluates the entomopathogenic fungus L. lecanii and B. bassiana for effectiveness in controlling Blatella germanica. Materials and Methods: Conidial formulations of L. muscarium (PTCC 5184) and B. bassiana (PTCC5197) were prepared in aqueous suspensions with Tween 20. German cockroaches were immersed in a 5 ml oil-based conidial suspension at one of several aqueous conidial suspensions at 1×105, 106, 107 or 108 conidia ml or bait method. The control groups were immersed in water or oil solutions with no conidia. Treatments with aqueous conidial suspensions were conducted with 10 cockroaches per group (1×105, 106, 107, 108 conidia ml or control). Bioassays were repeated twice on different days with different batches of conidia. After treatment, the biological parameters were evaluated. Data were analyzed by Probit program and LC50 and LC90 concentrations were estimated. Results: The results indicate that all of the treatments significantly different from the control (P<0.01). Better results were observed when conidial B. bassiana suspension was used. By increasing the concentration suspension B. bassiana, mortality rate increases and LC50 and LC90 level were 1.5x107 and 6x108 spore/ml, respectively. Conclusion: The lethal concentrations (LC50 and LC90) observed with the aqueous conidial solution of L. muscarium and B. bassiana in this study are potential to control B. germanica, therefore, be seriously considered for the control of B. germanica. The entomopathogen can propound in biological control of the pest and supplementary studies must be continued, because factors responsible for host susceptibility are not known. Keywords: Lecanicillium muscarium, Beauveria bassian, Blatella germanica, Biological control

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A study of IgE reactivity to Curvularia lunata antigens in patient with asthma

Ali Reza Khosravi, Hassan Ghorbani Choboghlo, Mehdi Taghavi, Donya Nikaein

Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran Email: [email protected]

Introduction and Objectives: Fungi are involved in hypersensitive reactions in about 20–30% of the atopic cases worldwide. Also fungal allergens play an important role in the pathogenesis of allergic airway inflammation and asthma. Among asthmatics, mold allergy affects 70% of adults and 45% of children. Curvularia lunata, one worldwide fungus, belonging to class Deuteromycetes is an inhalant allergen implicated in allergic rhinitis, asthma and allergic bronchopulmonary mycoses. Materials and Methods: In order to assess the hypersensivity of asthmatic patients to C. lunata allergens skin prick test in 36 patients with asthma were carried out. Enzyme-linked immunoadsorbent assay (ELISA) test by patient's sera and control group were done, as well. Results: From 36 patients, 17 (47. 2%) cases were positive in SPT. Based on ELISA results, 13 patients (36. 1%) had positive test by anti human IgE conjugate with HRP. There is no significant difference between positive results in two tests. Conclusion: Last decade has seen an unprecedented increase in population suffering from allergy and asthma. Recently recombinant allergens have been derived from various sources such as tree pollen, molds, foods, pet danders, etc. These results (47 %and 36%) approved that asthma patients had allergic reactions to Curvularia. It is suggested to use these fungal allergens in allergic tests. Keywords: IgE, Curvularia lunata, Asthma

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Microbiological contamination in bottled mineral water and assessment of antibacterial sensitivity among the isolates

Keyvan Pakshir, Mahbobeh Pishva, Kamiar Zomorodian, Maryam Jafari, Mohsen Karamitalab, Reza Behjat, Mosa Rahimi Ghiasi

Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran Email: [email protected]

Introduction and Objectives: Many of gastrointestinal diseases are transferable by drinking water. Nowadays, consumption of bottled mineral waters is common worldwide. In spite of quality controls, several studies had documented fungal and bacterial contamination in bottled waters. The aim of this study was evaluation of fungal and bacterial contamination in bottled mineral water and determination of antibacterial resistance among the isolates. Materials and Methods: A total of 120 bottled mineral water of 12 commercial brands in Shiraz city were purchased and assessed for fungal and bacterial contaminations, using membrane filtration technique. The filters were cultured on Sabouraud and brain heart infusion agar. ITS regions of DNA were amplified by specific primers and DNA sequence analysis performed for fungal identification. Identification of bacteria did performed by conventional methods and their sensitivities to eight antibiotics were evaluated by disk diffusion method. Results: Three brands of bottled water were contaminated with Aspergillus, Phoma and Penicillium species and 9 brands were contaminated with bacteria such as Bacillus and Diphtheroid species. Diphtheroid had the most resistance to penicillin and cephalothin and was sensitive to ciprofloxacin and gentamycin. Bacillus had the most resistance to clindamycin and was sensitive to Ciprofloxacin. Conclusion: The bacteria such as Diphtheroid considered as normal flora of human skin and can contaminate the water during preparation and packaging process. Air and soil are the other source for fungal contamination. Keywords: Bacillus, Diphtheroid, Bottled water, Antibiotic, Fungi, Sequencing

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An in vitro study of nematophagous fungi on pathogen larva nematodes

Majid Zarrin, Mahmoud Rahdar, Abbas Gholamian

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Ability of fungi in decreasing of nematodes larvae has been reported in many researches and it has been confirmed that bacteria and nemathodes have the ability to control many parasitic nemathodes. Nemathodes-destroying fungi produce traps along the hyphae. In the present study we used animal-parasitic nemathodes as a model because they are simple for work. This work aimed to confirm the nematophagous effect of some saprophyte fungi on nematodes larve stage that could be a good way to biological control of nemathodes. Materials and Methods: To investigate the effect of fungi on infectious stage of Trichostrongylidae family, the work was performed in 3 stages: Preparation of larvae, preparation of fungi, and evaluation of the effect of fungi on larvae. After preparation of infective Trichostrongylidae larva stage and fungal culture, the larva were added to fungi media. The numbers of live larva were counted on different days. K square statistical test was used for analyzing the data. Results: The most reduction in number of live larva was seen for Alternaria and Acromonium with more than 90% in 14 days after challenging. A. niger, Paecilomyces sp., Fusarium sp. also showed a significant effect on reduction of live larva. There are no significant different in reduction of live larva for Trichotishium sp. and Curvularia sp. when compared with control group (p>0. 05). Conclusion: The present results indicated that fungi can destroy the nematodes parasites in laboratory condition. The method can be used as a novel strategy for control of nematode diseases in plants and animals but it is slowly and need prolonged time when compared with the chemical controls. Keywords: Nematophagous fungi, Nematodes, Saprophytic fungi

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Sensitivity and specificity of skin prick test to Aspergillus fumigatus antigens

Javaher Chabavizadeh, Ali Reza Khosravi

Department of Medical Mycology and Parasitology, School of Medicine, University of Medical Sciences, Isfahan, Iran Email: [email protected]

Introduction and Objectives: Since in public areas dust and pollens, there are a lot of allergic particles including fungi, and among the fungi, Aspergillus fumigatus was reported to be the commonest allergenic fungus, therefore this study was carried out to determine A. fumigatus antigens in predisposed patients by using accurate, easy, early diagnostic specific and sensitive method. Materials and Methods: Present study was done on 105 patients and 76 control groups. A. fumigatus extracts were prepared and after standardization, used as crude antigens for Skin Prick Test (SPT) (in vivo), specific and total IgE by ELISA (in vitro) method were done as well. Results: SPT results showed: 55. 2% (n=58) of patients for A. fumigatus, were positive (P<0. 01). IgE levels raised in a greater number of patients (105 subjects) in comparison to the control group (76 cases). Specific IgE in sera to A. fumigatus antigens Correlation Coefficients were determined with Pearson’s Linear Regression analysis. The sensitivity and specificity for A. fumigatus allergens skin test was 95.9% and 80.4%, respectively. Conclusion: The skin test results strictly, correlated with the presence or absence of A. fumigates allergen specific IgE in serum. Skin Prick Test with 95.9% and 80.4% sensitivity and specificity is recommended as a valid screening test for A. fumigates. Keywords: Aspergillus fumigatus, Total IgE, Specific IgE, ELISA test, Skin Prick Test

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Survey of mycological flora in the student hostel baths in Kurdistan University of medical sciences (2011)

Faeze Mohamadi, Sadegh Khodaveisi, Esmaiil Ghahramani, Pari Abdolahi, Shiva Sori, Somayeh Sharifinia, Marjan Motamedi

Department of Medical Mycology and Parasitology, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Damp environments such as swimming pools and public baths are thought to be one of the major places where fungi are spread. It is well-known that baths are major sources of fungal infections. This study was performed to find the mycological flora in the student hostels in Kurdistan University of medical sciences. Materials and Methods: A total of 256 samples were collected from floor and walls were screened for fungal pathogens. Samples were collected from wet surfaces with a sterile cotton swab and dry surfaces, swabs were moistened with sterile saline. Samples were inoculated on Sabouraud dextrose agar with chloramphenicole and cyclohexamide (SCC). All cultures were incubated at 28°C, and were observed weekly for fungal growth. All fungal isolates were identified using colony morphology and microscopic morphology. Results: A total of 256 samples were positive. The most common fungi recovered were as follows; Cladosporium sp. 65.6%, Exophiala sp. 46%, Rhodotorula sp. 31%. Also Trichophyton mentagrophytes and Microsporum gypseum were isolated in baths samples. Conclusion: To prevent mycotic infections among bathers, adequate preventive measures like use of private or disposable slippers and effective cleaning after bath must be taken. Keywords: Baths, Mycological flora, Student hostel

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A laboratory study on properties of Deoxynivalenol (DON) production in standard culture medium by 24 Aspergillus spp. isolates from Northern Iran

Raheleh Firouzmand, Leila Modiri, Arash Chaichi Nosrati

Division Microbiology, Department of Molecular and Cell Biology, Lahijan branch, Islamic Azad University (IAU), Lahijan, Guilan, Iran Email: [email protected]

Introduction and Objectives: According to the increased fungi contaminations and related damages, microbiologist's incentive in considering the fungal contaminations in human habitats had increased. Some of fungi cause disease through production of toxins in animals and thus in humans. Since these toxins are not easily distinguishable, then it is crucial to study their characteristics. Aspergillus are among the most important toxigenic fungi which are found abundantly in northern Iran habitats that is one of most important habitats of Iran and which is the main source for many feed and food stuffs in the state. Thus, we have decided to study the DON production characteristics in culture medium by northern Iran's Aspergillus isolates collection. Materials and Methods: Samples were collected from Northern Alborz and southern Caspian sea agricultural plants culturing and processing centers. The mould samples were then isolated and identified based on CBS environmental sampling rules and ICPA diagnostics standards then were cultured to stimulate the toxin production till the targeted toxin to be measured at culturing substrate and fungi biomass. Afterward, they were exposed to extraction and then existing DON size was measured by ELISA technique. Results: Amongst studied species, A. melleus (subgenus Circumdati/mean 82.581 ppb) had the highest DON toxin production followed by S. ornata (subgenus Ornati/56. 777 ppb) and A. sojae (subgenus Circumdati/54. 661 ppb). Conclusion: With this idea that it is possible for the DON production genes to be active in Aspergillus isolates it could be noticed in addition to Fusarium ingredients as the second genera of DON production causative agents. Keywords: Growth media, DON, Aspergillus, Species, Iran

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A study on properties of Deoxynivalenol (DON) production in culture medium regarding Aspergillus spp. isolates from processing factories of northern Iran

Raheleh Firouzmand, Leila Modiri, Arash Chaichi Nosrati

Division Microbiology, Department of Molecular and Cell Biology, Lahijan branch, Islamic Azad University (IAU), Lahijan, Guilan, Iran Email: [email protected]

Introduction and Objectives: According to the increased fungi contaminations and related damages, microbiologist's incentive in considering the fungal contaminations in human habitats had increased. Some of fungi cause disease through production of toxins in animals and thus in humans. Since these toxins are not easily distinguishable, then it is crucial to study their characteristics. Aspergillus are among the most important toxigenic fungi which are found abundantly in northern Iran habitats that is one of most important habitats of Iran and which is the main source for many feed and food stuffs in the state thus, we have decided to study the DON production characteristics in culture medium by northern Iran's Aspergillus isolates collection. Materials and Methods: Samples were collected from Northern Alborz and southern Caspian sea agricultural plants culturing and processing centers. The mould samples were then isolated and identified based on CBS environmental sampling rules and ICPA diagnostics standards then were cultured to stimulate the toxin production till the targeted toxin to be measured at culturing substrate and fungi biomass. Afterward, they were exposed to extraction and then existing DON size was measured by ELISA technique. Results: Amongst studied species, A. melleus (subgenus Circumdati/82. 581 ppb) had the highest DON toxin production followed by A. spp. VI (subgenus Unclassifiable/50. 803 ppb) and A. parasiticus (subgenus Circumdati/49. 108 ppb). Conclusion: This report gives value to saying idea about mouldy of solid food by DON. DON presence is a danger in Feed-Food processing; storing or packing that is controllable by regular examination and toxin measuring for DON. Keywords: DON, Aspergillus, Species, Processing factories, Iran

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Comparison of incubation time and carotenoid production in Rhodotorula slooffiae and R. mucilaginosa isolated from leather tanning wastewater

Farzaneh Sadat Naghavi, Parichehr Hanachi, Mohammad Reza Sodi, Ozra Sabora

Faculty of Basic Science, Biology Department, Alzahra University, Tehran, Iran Email: [email protected]

Introduction and Objectives: Carotenoids naturally synthesized by all photosynthetic organisms and fungi such as yeasts can act as an antioxidant that property of which is closely related to their ability to decreased risks for a variety of degenerative diseases. In recent years, the increase of demand for carotenoids obtained from natural sources has promoted major efforts to improve carotenoid production from biological sources and Pigmented yeasts are an interesting subject from this point of view. The aims of this study were to compare between amount of total carotenoid and evaluating the effect of incubation period on pigmented yeasts isolated from leather tanning wastewater. Materials and Methods: Biomass was determined by optical density using a spectrophotometer at 600 nm. To isolate the carotenoid pigment, cells were suspended in acetone and broken using a homogenizer, followed by centrifugation. The supernatant was separated and process was repeated. For Analytical method pigments were measured spectrophotometrically at 450 nm 1% using the extinction coefficient E 450=2500. For studied of effect of incubation time, Samples were held at 30 ̊С in a shaker at 150 rpm for 24h, 48h, 72h, 96h, and 120h. Extraction and quantification of Carotenoids were carried out as outlined. Results: The results showed that the content of total carotenoid in R. slooffiae was the highest when samples incubated for 72 h but R. Mucilaginosa has more potential to produce carotenoid generally. The best incubation periods were 72h and 48h for R. slooffiae and R. Mucilaginosa respectively. Conclusion: Among pigmented yeasts, only some small taxonomic groups have been investigated for the carotenoid pigment content. The type of carotenoid and their relative amount may vary depending on genus of yeast and environmental condition. The cost of carotenoid production and the amount of yield can be changed positively by optimizing its process. Keywords: Carotenoid, Rhodotorula sp., Incubation time

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Evaluation of the number of Treg cells in normal mice exposed to AFB1 and treated with aged garlic extract

Mohaddeseh Larypoor, Mansour Bayat, Zahiur Mohammad Hassan, Abbas Akhavan Sepahy, Masoud Amanlou

Department ofMedical and Veterinary Mycology, Faculty of Veterinary Sciences, Islamic Azad University, Science and Research Branch, Tehran, Iran Email: [email protected]

Introduction and Objectives: Aflatoxin B1 (AFB1) suppresses immune system. To decrease such suppressive effects on the immune system, a wide range of herbal medicine like garlic is utilized. Biological activities of garlic in vitro and in vivo have also been verified. Our previous studies demonstrated that aged garlic (preserved dry garlic bulbs in the freezer for six months in -20 ºC) have increased immunostimulator fractions and reduced immunosuppressor fractions. This study focuses on immunosuppressor activity of AFB1 and immunostimulator activity of aged garlic extract (AGE) through the evaluation of the number of CD4+ CD25+ FoxP3+ T regulator cells (Treg) and cytokine production pattern in Balb/c normal mice. Materials and Methods: In this experimental research, AFB1 was separated from Aspergillus flavus (PTCC 5004) by HPLC, AGE by Mantis method, and DTH was carried out to determine the effectiveness of AGE and AFB1 dosage, which can have an effect on the immune system. Subsequent experiments were carried out on 20 Balb/c mice to estimate the effects of AGE and AFB1 on the number of Treg cell in 4 groups: 10µl/kg/day of AFB1 and AGE diluents were administered for 4 consecutive days to group 1: AFB1, 2: control; 3: AGE + AFB1and 4: AGE via intraperitoneal (IP) route, respectively. Mice were sacrificed and splenocytes harvested and the percentage of splenic Treg cells was measured by flow cytometry analysis. ELISA method was utilized to measure Cytokine production. Results: The findings revealed that AGE increased the level of IFNγ and IL-4 cytokines produced from stimulated splenocyte by specific tumor antigen and decreased the number of Treg in spleen (P<0. 05). AFB1 increased the Treg number in spleen and decreased cytokine production (P <0. 05). In group 2 (control) and 4 (AGE) the Treg rate decreased (P value<0. 05) and in groups 1 and 3 Treg number was increased (P <0. 05) Conclusion: This study indicated that AGE is able to alter the cytokine production in normal mice into a Th1 protective pattern is favorable to immune system in general and anti-tumor immunity in particular. AFB1 is able to alter the cytokine production into a Th2 protective pattern. Therefore, AGE might be used as herbal medicine with few side effects as compared to chemotherapy in treating cancers caused by substances like AFB1. Keywords: Aflatoxin-B1, Garlic, Treg, Immunotherapy

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The qualitative and quantitative assessment of airborne fungi in indoor environment of various wards of Imam Reza hospital in Mashhad, 2012

Hossein Zarrinfar, Nasrin Rostami, Hossein Alidadi, Pegah Salehi

Student Research Committee, School of Health, Mashhad University of Medical Sciences, Mashhad, Iran Email: [email protected]

Introduction and Objectives: The air quality of the hospital wards is important for patients, because they are susceptible to fungal infections that are transmit able through the air. The objective of this study was determination of types and concentration of fungi in air of various wards of Imam Reza hospital in Mashhad in 2012. Materials and Methods: In this cross sectional study, hematology, infection, ENT and NICU wards were selected and samples were collected by open plate technique (on Sabouraud dextrose agar) once a week. Finally, 84 samples were collected from indoor environmental air and immediately transported to laboratory. Then plates were placed in incubator for 7-10 days at 32.5ºC. The cultures were examined microscopic and macroscopic methods. Results: Our results showed that almost all of the parts were polluted by various fungi. The highest indoor fungi contamination was observed in hematology ward. Higher rate of fungi identified was in ENT and infection wards are Penicillium spp, and in hematology and NICU wards is Aspergillus flavus and Chrysosporium spp respectively. Conclusion: According to the results, control of fungal contamination in various wards of the hospitals especially those in which patients with immunocompromised or other predisposing factors are hospitalized, via suitable filtration and ventilation air and proper disinfection of the wards seem is necessary. Keywords: Hospital wards, Airborne fungi, Mashhad

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Measurement of Aflatoxin B1 in the flour of Arak traditional bakeries

Riazipour Majid, Bagheri Mohammad Kamal

Research Center for Applied Microbiology, Baqiyatallah University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Wheat is one of the most consumed cereals in Iran, thus wheat flour contaminated with mycotoxins can endanger the health of a large part of our community. Measurement of Aflatoxin B1 contamination of the flour used to make bread in Arak traditional bakeries was the objective of this study. Materials and Methods: Flour samples were collected from Arak traditional bakeries. The samples were extracted with 80% methanol and the level of aflatoxin B1 in the extracts was measured using a competitive ELISA. Results: All flour samples were contaminated with aflatoxin B1, but none of them exceed Iran nation standard (5µg/kg). Mean of the mycotoxin amount in samples was 0.4 (± 0.06), ranging from 0.3 to 0.6µg/kg. Flour samples used to make bread "Lavash" with average contamination of 0.38 (±0.08) were the highest contaminated flours flowed by the flours of "Sangak" and "Barbari" breads, with an average contamination of 0.36 (±0.02) and 0.35 (±0.05) respectively. But these differences were not significant. Conclusion: Prevalence of contamination with small amounts of aflatoxin B1 indicates presence of favorite conditions for growing fungi in various stages of wheat production to consumption and storage conditions requires improvement. Keywords: Aflatoxin B1, Bakery, Bread, Flour

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Investigation of contamination of medicinal plants by aflatoxin

Arezou Azarm, Hoda Azarm Department of Microbiology, Flavarjan Branch, Islamic Azad University, Flavarjan, Isfahan, Iran Email: [email protected]

Introduction and Objectives: Plants have been used in the prevention, treatment and cure of disorders and diseases since ancient times. Fungi of the genera Aspergillus and Penicillium are known to contain strains that produce mycotoxins. In this study the contamination of some herbal drugs by fungi and aflatoxin that tradititionally used in medicine were evaluated. Materials and Methods: For this purpose 45 different medicinal plants were collected from local market in Isfahan. After initial preparation, samples were inoculated into Sabouraud dextrose agar, MEA using the spread plate method and the incubated at desired temperature. The grown fungi on the media were determined by macroscopically and microscopic methods. Results: In this study about 11% of all medical plants used were contaminated by Aspergillus flavus. Aflatoxin contamination was present only in one sample. Conclusion: Due to the increased use of herbs as alternative medicine, Evaluation of fungal growth and production of mycotoxins are necessary for reducing the health risk to consumers. Keywords: Medicinal plants, Aspergillus flavus, Aflatoxin, Isfahan

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Effect of silver nano- particles on growth of aflatoxigenic fungus, Aspergillus parasiticus

Somayyeh Pourtalebi, Seyyed Amin Ayatollahi Mosavi, Sanaz Hadi Zadeh

Department of Medical Mycology and Parasitology, School of Medicine, Medical University of Kerman, Kerman, Iran Email: [email protected]

Introduction and Objectives: Aflatoxins are poisons which are produced as secondary metabolites by filamentous fungi and because of carcinogenic, mutagenic effect, in human and animals, have great importance. Aspergillus flavus and A. parasiticus are the main aflatoxigenic fungi which can grow on different materials especially corn, pistachio, almond, peanut, and cotton to produce poison. So growth control of these fungi leads to control of aflatoxin production and eliminate harmful effects of them. Materials and Methods: In this study, by using standard A. parasiticus (PTCC5286) and colloid solution of silver nano-particles, minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) by micro-dilution broth, macro-dilution broth and disk diffusion methods, were performed. Moreover, beside negative control, ketoconazole was also used as positive control. Results and Discussion: Colloid solutions of silver nano-particle in 180 ppm concentration prevent growing of A. parasiticus and results of MFC (300 dilution) shows fungistatic characteristic of silver nano-particle. In disk diffusion method, MIC result was similar to micro- dilution and macro -dilution methods which was 3.3mm diameter halo. The results of this research are similar to results of previous studies on silver nano-particle on A. flavus. Conclusion: This research shows colloid solution of silver nano-particle has inhibitory effect on growth of aflatoxigenic fungus: A. parasiticus. So it is effective on decreasing aflatoxin production. Keywords: Silver nano-particle, Aspergillus parasiticus, Aflatoxin, MIC, MFC

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Cellulose production using Trichoderma reesei on solid state fermentation

Elham Ghorbani Totkaboni, Gholam Khayati, Ayatolah Nasrollahi Omran

Department of Biology and Microbiology, Tonkabon Branch, Iran Email: [email protected]

Introduction and Objectives: The fungi are the most important group cellulase producing. The Most fungi producing are Trichoderma and Aspergillus that have been studied expensively. Trichoderma species are opportunist and plant symbiosis. Now, agricultural waste such as rice straw and wheat straw are suitable solid state for growth and cellulose production. The objective of the study was cellulose production under SSF conditions by different agricultural waste using Trichoderma reesei as the microorganism. Materials and Methods: Rice straw and wheat straw were used as solid substrates. The flasks were sterilized, and after cooled to room temperature and inoculated with desired volume of inoculums to obtain 106 spores /g substrate for each flask under aseptic conditions. After extraction, filter paper activity (FP activity) and Carboxymethyl cellulose (CMC) or cellulose gum were measured. Results: Selection of substrate depends upon several factors mainly related with cost and availability and thus may involve the screening of several agro-industrial residues. The results showed that the maximum enzyme activity for FP activity with rice straw and wheat straw were found 808 and 678 U/g, respectively. Carboxymethyl cellulose (CMC) was found 162 and 84 U/g for rice straw and wheat straw respectively. Conclusion: In comparison with two different solid substrates, the rice straw was the best one with 1.2 and 1.9 fold greater enzyme activities. Keywords: Trichoderma reesei, Enzyme, Cellulose, Solid, Fermentation

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Isolation and molecular identification of keratinophilic fungi from parks of Shiraz city

Keyvan Pakshir, Moosa Rahimi Ghiasi, Kamiar Zomorodian

Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran Email: [email protected]

Introduction and Objectives: Keratinophilic fungi are an ecologically important group of fungi found in soil and cab be transferred to human and make cutaneous diseases accidentally, specially the children who play in the parks. The aim of this study was to isolate and identify of keratinophilic fungi from soil of different parks around the Shiraz city. Materials and Methods: A total of one hundred and ninety six soil samples were collected from different parks of Shiraz and hair baiting method was used for fungal isolation. The colonies around the hairs were purified and transferred to new medium. Identification of keratinophilic fungi was performed by morphologic and molecular method, using DNA sequence analysis. ITS regions of DNA were amplified by specific primers and PCR products were sequenced. Results: Four hundred and eleven fungal isolates from 22 genera were isolated and identified as follows: Acremonium, Aspergillus, Alternaria, Bipolaris, Paecilomyces, Penicillium, Fusarium, Chrysosporium, Pseudallescheria, Mucor, Cephalosporium, Phialophora, Rhizopycnis, Microsporum, Chaetomium, Malbranchea, Dothrideamycetes, Tritirachium, Ochroconis, Verticillium, Scopulariopsis, Scedosporium and Bionecteria. Conclusion: In this study, 47% of the places were contaminated with keratinophilic fungi. These areas have potentially high risk for human and animals and could be considered as a source of fungal infections. Keywords: Keratinophilic fungi, Soil, Park, Shiraz, Sequence

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Aflatoxin in raw and roasted with salt nuts (pistachio, peanut and walnut) sold in markets of Tabriz, Iran

Abdolhassan Kazemi, Alireza Ostadrahimi, Fereshteh Ashrafnejad Asl, Reza Mahdavi, Nafiseh Sargheini, Mohammadreza Farshchian, Sepideh Mahluji

Tabriz University of Medical Sciences, Tabriz, Iran Email: [email protected]

Introduction and Objectives: Nuts are one of the main consumed snacks worldwide and also have an important role among Iranian. Natural contamination of nuts with aflatoxin is unavoidable and causes a special challenge to nuts safety and quality. The purpose of this research is to study aflatoxin contamination in commercially available nuts (pistachio, walnut and peanut) in the markets of Tabriz, Iran. Materials and Methods: 62 samples of 50g roasted with salt peanut and pistachio and 109 samples of 50 gr pure pistachio, walnut and peanut were collected from different areas of local markets. After initial preparation, ELISA test was used for aflatoxin measurement. Results: Result has shown that walnut (90%) and pure pistachio (2.3%) were the most and least contaminated samples respectively. Mean of aflatoxin contamination in roasted with salt samples (19.88 ± 19.41) was significantly higher than pure ones (6.51 ± 9.4) (p<0.001). Respectively 58.6%, 48.4% and 7.6% of roasted with salt pistachio, peanuts and walnut samples had aflatoxin contamination more than the Maximum Tolerable Level of Iran (MTL, 15 ppb). Conclusion: It was concluded that aflatoxin content of nuts should be monitored regularly to minimize the risk of aflatoxin hazard, ensure food safety and quality. Results of the current survey could be useful to minimize the aflatoxin contamination and also educate people on the danger of aflatoxin in nuts. Keywords: Aflatoxin, Raw, Roasted, Salt, Nuts

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Mould contamination of untreated and roasted with salt nuts (walnut, peanut and pistachio) sold in markets of Tabriz, Iran

Abdolhassan Kazemi, Alireza Ostadrahimi, Fereshteh Ashrafnejad Asl, Reza Mahdavi, Nafiseh Sargheini, Mohammadreza Farshchian, Sepideh Mahluji

Tabriz University of Medical Sciences, Tabriz, Iran Email: [email protected]

Introduction and Objectives: Nuts are one of the main consumed snacks worldwide and also have an important role among Iranian food habits. Natural contamination of nuts with fungus is unavoidable and causes a special challenge to nuts safety and quality. The purpose of this research is to study fungal contamination in commercially available nuts (pistachio, walnut and peanut) in the markets of Tabriz, Iran. Materials and Methods: 100 samples of 50 gram roasted with salt peanut and pistachio and 300 samples of 50 gram pure pistachio, walnut and peanut were collected from different area of local markets. After initial preparation, the samples were cultured on Sabouraud dextrose agar (SDA). 19 fungal isolators were identified. Results: Results have shown that Aspergillus niger was the predominant mould among the pure (44%) and roasted with salt (14%) nuts (p<0/001). In addition, percentage of mycotoxigenic fungal contamination was 18% in roasted with salt nuts and 11% in pure samples. Conclusion: The overall result of the analyzed samples showed that the rate of fungal contamination in pure samples was higher than roasted with salt ones (p<0. 005). Results of the current survey could be useful to minimize the fungal contamination and also educate people on the danger of mold in nuts. Keywords: Mould, Raw, Roasted, Salt, Nuts

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Inhibitory effect of vitamin C on Aspergillus parasiticus growth and aflatoxin gene expression

Maryam Akbari, Maryam Moazeni, and Sassan Rezaie

Div. of Molecular Biology, Dep. of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Aflatoxins are one of the natural toxic molecules which are produced by a number of species belong to the genus Aspergillus. These toxins which are well known to be mutagenic, carcinogenic, hepatotoxic, and immunosuppressive, are leading cause of some liver cancers and serious gastrointestinal problems. Antimicrobial and antiviral activity of vitamin C has been documented in several documents. However, the antifungal and anti-toxic effect of the mentioned vitamin has not yet been reported. In the present study, the antifungal and anti-toxic effect of vitamin C is investigated against Aspergillus parasiticus as well as aflatoxin gene expression. Materials and Methods: According to CLSI document M27-A3, standard strain of A. parasiticus ATCC 15517 was used for performing antifungal susceptibility test due to the vitamin C. Vitamin C was dissolved in sterile diluted water to get the concentration of 400 mg/ml, and then diluted to the final concentrations of 200 to 12. 5 mg/ml. Quantitative changes in expression of the aflR gene were analyzed by measuring the cognate aflR mRNA level by use of a quantitative real-time RT-PCR assay. Results: Our obtained results demonstrated that vitamin C had inhibitory effect on A. parasiticus growth at 100 mg/ml. In addition, the level of aflR gene expression was significantly decreased after treating the fungus with 100 mg/ml of the mentioned vitamin. Conclusion: We evaluated the antifungal and anti-toxin activity of vitamin C against A. parasiticus. It was revealed that vitamin C has antifungal properties and is significantly able to reduce the expression of the gene aflR which encodes aflatoxin. To sum up, it may be considered as an effective potential anti-mycotoxin agent in medicine or industrial agriculture. Keywords: Aspergillus parasiticus, AflR, Vitamin C, Gene expression

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Candida africana, a first case from Iran, susceptibility to antifungals, evaluation extracellular enzymes and biofilm formation

Zahra Seifi, Ali Zarei Mahmoudabadi

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Candida africana was firstly isolated from patients in Africa and Germany by Tietz et al., as atypical C. albicans. Several researchers have also isolated and described atypical isolates of C. albicans as C. africana that causes of vaginitis. C. africana is most often recovered from vaginal swabs from patients with Candida vaginitis. Researchers have described germ tube positive and chlamydospore-negative for C. africana. C. africana has atypical carbohydrate assimilation profile (unable to assimilate the N-acetylglucosamine, glucosamine, trehalose and, DL-lactate. This report describes the isolation of C. africana from an Iranian healthy woman without vaginitis. In this report, the sensitivity to four antifungals are also studied. To our knowledge, this is the first report of the recovery of C. africana in Iran. Materials and Methods: Vaginal sample was cultured onto CHROMagar Candida and recovered yeast detected using morphological characteristics, growth at different temperatures, and ID32C identification kit (bioMérieux, France). Disk diffusion testing of fluconazole, miconazole, econazole, nystatin, amphotericin B, clotrimazole and Lamisil was used for susceptibility test. Phospholipase activity assay was performed according to Price et al., method. The acid proteinase activity of the isolate was also measured by Ombrella et al., method. The coagulase and hemolytic activity of the isolate was assessed using the rabbit plasma and blood plate. Esterase activity was determined by visual methods presented by Slifkin. Biofilm formation was determined by visual methods using Falcon conical tubes. Results: This isolate has slow grown (especially at 30-37°C) and was unable to produce chlamydoconidia on cornmeal agar. Atypical carbohydrate assimilation profiles (ID 32C profile, 7046340011) also characterize this species of Candida. The production of extracellular proteinase, phospholipase and hemolytic were confirmed in our strain; however it was without coagulase and esterase activities and biofilm formation. Our isolate fully resistant to fluconazole, dose depended to amphotericin B, sensitive to miconazole and highly sensitive to Lamisil. Conclusion: To our knowledge, this is the first isolation of C. africana in Iran. In this study, we presented several data about extracellular enzymes, antifungal susceptibility and biofilm formation in this strain. Keywords: Candida africana, ID 32C, Extracellular enzymes, Biofilm formation

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The inactivation of aflatoxin B1 in food and feed crops by gamma radiation

Seyed Amir Yazdanparast, Mojtaba Barzegar, Farzan Saeedi

Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Aflatoxins were reported to be among the most potent mycotoxins. AFB1 is one of four toxic, secondary metabolites produced by the food spoilage fungi Aspergillus particularly A. flavus and A. parasiticus. The Aflatoxins are highly carcinogenic agents, and AFB1 is listed as a carcinogen of group 1 by the International Agency for Research on Cancer. Food and Agricultural Organization (FAO) estimated that as much as 25% of the world's agricultural commodities are contaminated with mycotoxins, leading to significant economic losses. The purpose of the present study was to assess the effect of gamma radiation on the breakdown of AFB1 in several foods and feed products following inoculation, in the lab, of these products with a strain of A. flavus known to produce Aflatoxins B1. Materials and Methods: Following thorough mixing, each sample was divided into 25g sub- samples, each of which was placed in transparent plastic bags. Samples were irradiated in a gamma chamber which has 6OCo as the radiation source. Doses of radiation applied were 4, 6, and 10 kGy. The experiments were done on three replicates for each radiation dose and for each food and feed sample. AFB1 was extracted from food and feed samples using the method described in by De Gastro and Vargas with some modifications. The sample was evaporated to complete dryness using a stream of nitrogen. The sample was reconstituted in 5ml of chloroform. Aflatoxins recovery using the above method on corn sample spiked with 20ppm of Aflatoxins B1 was 80%. AFB1 was quantified using TLC according to CAMAG method. Concentration of Aflatoxins was estimated from a calibration curve created by measuring the absorbance at 254 nanometer of known concentrations of Aflatoxins standards. Results: It was found that the source of carbohydrates available to A. flavus greatly influences Aflatoxins production. In oil-crop food samples, a negative correlation was observed between percentage of oil content of these crops and percentage of AFB1 degradation. Percentage of oil content in those crops was estimated based on the weight of the solvent extractable oil during Aflatoxins extraction related to the weight of the sample before extraction. Thus, percentages of AFB1 degradation at 10 kGy was 80%, 69%, and 58%, corresponding to percentages of oil content of 16%, 32%, and 38% for corn, pistachios, and peanuts, respectively. Conclusion: From the results presented in the present study, it can be concluded that it is possible to use gamma radiation, within the maximum permitted dose, as a means for breakdown of AFB1 in contaminated food and feed stuff to below the maximum permitted levels. The degree of breakdown of AFB1 in food and feed varies according to the commodity and it may be related to oil content in oil crops. Keywords: Gamma radiation, Aspergillus flavus, Aflatoxins B1, Food feed

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Prevalence of fungal allergens in respiratory allergic patients in Ahvaz

Abdolhussein Shakurnia, Mohammad Ali Assarehzadegan

Immunology Department of Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Fungi are ubiquitous in our environment and they are one of the important causes of allergic diseases. Identification of the most common aeroallergens to which patients are sensitized in a specific area is important in the diagnosis and treatment of allergic rhinitis. The purpose of this study was to determine the prevalence of skin reactivity to common fungal allergens and total IgE in patients with allergic rhinitis in Ahvaz city. Materials and Methods: In this cross sectional study, 285 volunteers with the signs and symptoms of allergic diseases in Ahvaz during 2010 were investigated. All patients were subjected to skin prick test (SPT) with common allergenic extracts. Data were analyzed by SPSS-18 software using Chi square test. Results: Seventy subjects, comprising 23. 7% of the study group had positive skin test to at least one of the fungal allergens. The prevalence rate of sensitivity to fungal allergens was as follow: Cephalosporium 11.5%; Penicillium mix 9.5%; Alternaria mix 8.1%; Aspergillus 5.1% and mold mix 9.8%. Mean total IgE in patients with positive SPT was significantly higher than in patients without any positive skin prick test (251 vs. 125 IU/ml, p = 0.001). There was no statistical difference in the prevalence of sensitization to these allergens between two sexes; whereas, 15-35 age groups had significantly higher rates of allergy to fungal allergens (p=0.047). Conclusion: Fungal sensitization is a relatively common finding among patients with allergic rhinitis. Elimination or reduction of mold exposure in allergic patients is of special consideration and measures to reduce environmental factors which facilitate mold growth and proliferation is very important. Keywords: Allergy, Fungal allergen, Skin prick test

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Isolation of saprophytic fungi in the soil of Chitgar park, Tehran, Iran

Zeinab Sadeghi Ardestani, Shrareh Sayah Alborzi, Mehraban Falahati

Department of Medical Mycology and Parasitology, Tehran University of Medical Science, Tehran, Iran Email: [email protected]

Introduction and Objectives: Soil is well known to support the transient or ongoing existence of saprophytic fungi. Fungi in soil are found in all regions of the world, but its abundance in different regions in terms of Environmental and materials for organism growth is different. Materials and Methods: In this study, One hundred and twenty soil samples were collected from surface 10 and 20 cm depth to determine the prevalence of saprophytic fungi at Chitrgar Park in the west part of Tehran. The soil suspensions were prepared by sterile saline and then cultured on Sabouraud dextrose agar medium containing chloramphenicol, then incubated at 35 for 2-3 weeks. The fungal isolates were identified by colony morphology, slide cultures, and differentiation tests. Results: A total of 220 fungal colonies including 12 genera were isolated as follows: Aspergillus flavus (4.09%), A. niger (5.40%), A. fumigatus (2.27%), Fusarium sp. (10%), Cunninghamella sp. (3.63%), Alternaria sp. (2.72%), Paecilomyces sp. (1.36%), Rhizopus sp. (25%), Mucor sp. (14.09), Penicillium sp. (3.18%), Trichoderma sp. (1.36%), Stemphylium sp. (3.63%), phoma sp. (0.45%) and Monilia sp. (22.7%). Conclusion: It can be concluded that soil of Chitgar Park is rich in saprophytic fungi including Rhizopus sp. and the selection of certain saprophytic isolates could become useful in managing polluted soil of parks. Keywords: Saprophytic fungi, Soil, Chitgar Park

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A Comparative investigation on DON (Deoxy nivalenol) toxin production patterns of Aspergillus Species biomasses, Northern region of Iran

Parvaneh Rahsseparpoor, Leila Modiri, Arash Chaichi Nosrati, Seyed Hamed Shirazi Beheshtiha

Division of Microbiology, Department of Molecular and Cell Biology, Lahijan Branch, Islamic Azad University (IAU), Lahijan, Guilan, Iran Email: [email protected]

Introduction and Objectives: Mycotoxins are important chemotoxicant since they are mainly produced in large feed stock subtracts where fungus producing these toxins find optimum conditions for growth and development and they may become the cause of occurrence of some disease in humans and animals. Thus, their presence in foods and animals feed is a subject of global or national importance. Among them, Aspergillus has particular importance. In present study we will study the production of DON toxin in cell extract of Aspergillus isolated ecogeneous in Northern Iran. Materials and Methods: Firstly, sampling, culture and isolation was performed in Guilan and Mazandaran provinces. After recognition of the species, using ELISA and Ridascreen Kit, we quantitively analyze DON produced by available species. Results: Frequency distribution based on biomasses DON concentration in each interval between 0 to 100 ppb indicates that among all the toxigenic isolates and 83. 2 % have produced the toxin in the range of 0-10 ppb that more than 85 % among all the toxigenous isolates in 0-10 ppb range. In the range lower than acceptable standard rates it was 1-3 ppb and only 2. 5 % of studied fungi samples have toxigenic ability in acceptable range of 3-5 ppb. No one had measurable toxin production in 5-10 ppb range. Conclusion: According to fact that the greatest number of isolated had produced toxin in 0-10 ppb range (in 0-3 ppb range), despite its low production rate, we can concern seriously due to the collective effect of the toxin, as a critical risk factor, is not negligible. Since DON standard rate in foods is 3-5 ppb. Keywords: DON, Asperjillus, Species, Biomass

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Distribution of outdoor airborne fungi in various regions of Tehran

Sanaz Aghaei Gharehbolagh, Narges Aslani, Mehdi Razzaghi-Abyaneh, Masoomeh Shams- Ghahfarokhi

Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran 14115-331, Iran Email: [email protected]

Introduction and Objectives: Airborne fungi are pathogenic for immunocompromised patients and nosocomical infections. The presence of fungal spores in the air, as allergens particles, can be a risk factor especially for susceptible patients. In addition, some of the fungal genera can produce mycotoxin that threat human and animal life. The aim of this study is identification of these fungi to find out their distribution and the presence in the air in Tehran. Materials and Methods: For identification of airborne fungi, we used an open plate method. The PDA or MEA plates were located in 93 different region of Tehran at the height of about one meter above the floor, for 30 min during summer. All cultured plates were incubated at 28°C for 3-10 days. Fungal isolates were identified on the basis of their microscopic and macroscopic morphological criteria according to standard methods. Results: On the basis of our results 6485 colonies, belonged to 26 fungal genera were isolated. Aspergillus (31.3%) was the most frequent genus followed by Cladosporium (22.1%), Penicillium (13.8%), Alternaria (12.2%), Monilia (2.3%), Drechslera (1.6%), Rhizomucor (1.6%), Fusarium (1.4%), Exophiala (0.8%), Paecilomyces (0.8%), Acremonium (0.6%), Ulocladium (0.6%), Trichothecium (0.6%), Mucor (0.4%), Cladophialophora (0.4%), Phialophora (0.4%), Curvularia (0.4%), Helminthosporium (0.4%), Epicoccum (0.2%), Torula (0.2%), Cleistothecium (0.4%), Rhizopus (0.2%), Absidia (0.2%), Phoma (0.2%), Stemphylium (o.2%), Basidiobolus (0.2%), and Unknown (6. 2%). Conclusion: Results of the present study clearly showed that Aspergillus was the most prominent isolated fungal genus followed by Cladosporium, Penicillium and Alternaria. Knowledge of outdoor airborne fungi of Tehran can assist the recognition and management of fungal contamination problems in public health and detect an association between some diseases such as respiratory symptoms and moulds. Keywords: Outdoor air, Distribution, Fungal airborne, Tehran

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Measurement of aflatoxin B1 in feed for dairy animals in Alborz province

Mojgan Saghazadeh, Azar Sabokbar, Ramin Hajikhani, Omid Nilchi Zadeh Rahbar

Microbiology Department, Qom Azad University, Iran Email: [email protected]

Introduction and Objectives: While there is lot of carcinogens and teratogenic agents in daily consumes, but there is still Aflatoxin B1 the most important dairy toxin. However among all Mycotoxins there is Aflatoxin family that has more essence for search and elimination. These Toxins are secondary metabolites of Aspergillus parasiticus and A. flavus as a result of growing on foods Materials and Methods: In this study, we try to detect the presence of AF B1 among 75 cow feed samples in Alborz province by the Enzyme Linked Immuno Assay method. All samples have been selected randomly last of spring 12 and stored for 3 months in dark and dehumidified area before tests. Results: 69 samples exposed only 3.0µg/Kgr AF B1, and only 6 samples exposed 3.9 to 6.1µg/Kgr AF B1 that indicate all in admissible rate of standards. Conclusion: More over all Alfalfa samples have been detected as a high risk feed. Keywords: Aflatoxin B1, Feed, Aspergillus parasiticus, A. flavus

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Deuterium effect on the growth of the fungus Aspergillus fumigatus and Candida albicans

Farzad Doostishoar, Abdolreza Hasanzadeh, Seyed Amin Ayatolahi Mousavi

Department of Medical Mycology and Parasitology, Afzalipoor Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran Email: [email protected]

Introduction and Objectives: Deuterium has different action from its isotopes hydrogen in chemical reactions and biochemical processes. It is not a significant difference in heavier atoms between the behavior of heavier isotope and the lighter One but for very lighter atoms it is significant. According to that, most of the weight of all creatures body is water natural rate can be significant. In this article we want to study the effect of reduced deuterium on the fungus cell. If we saw the dependence of deuterium concentration of environment on the cells growth we can test this in in vivo models too. Materials and Methods: At first we measured deuterium concentration of the distillated water. This analysis was operated by Arak’s heavy water company. Then the deuterium was diluted to ½,¼,1/8 and 1/16 by adding water free of deuterium for making media. In three of samples the deuterium concentration was increased by adding D2O up to 10, 50, 100 times more concentrated. For Candida albicans growth, we used Sabouraud dextrose agar medium and for Aspergillus fumigatus growth we used Sabouraud dextrose agar medium containing chloramphenicol. After culturing the fungal species we put the mediums for each species in the shaker incubator for 10 days in 25 centigrade. In different days and times the plates were studied morphologically and some microscopic characteristics were studied too. The experiments and cultures were repeated 3 times. Results: Statistical analyses by paired-sample T test showed that A. fumigatus growth was decreased in concentration of 72 ppm (half deuterium concentration of negative control) significantly. In deuterium concentration reduction the growth reduce into the negative control significantly. Conclusion: The project results showed that C. albicans was sensitive to reduction and decrease of the deuterium in all concentrations. Keywords: Deuterium, Cancer cell, Growth

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Fungal contamination and physicochemical of public swimming pools In Kashan, Iran, 2008-2009

Sima Rasti, Mohammad Ali Assadi, Lila Iranshahi, Hamid Reza Gilasi

Department of Parasitology and Mycology, Kashan University of Medical Sciences Email: [email protected]

Introduction and Objectives: Swimming pools can transmit contagious diseases such as fungal disease, otitis, and conjunctivitis. The aim of this study was to find out fungal contamination and physicochemical condition of swimming pools. Materials and Methods: In this cross – sectional study, 200 water samples were collected from four swimming pools of Kashan, Iran. Temperature, pH and residual chlorine and turbidity of the pools were examined. Samples were concentrated through membrane filter and sedimentation to test the fungal contamination by culturing in Sabouraud dextrose agar and Sabouraud dextrose agar medium containing chloramphenicol and cycloheximide. The data were analyzed by X2 and Fischer exact tests. Results: The results of the study showed that the mean of physicochemical parameters except in temperature was standard more than 60%. Average temperature was higher than standard. The highest level of chlorine was recorded in summer. Prevalence of Saprophytic and opportunistic fungi was 27%. Twelve species of fungi were isolated; the most common were Aspergillus, Penicillium sp, Rhizopus sp, and Fusarium sp, and the highest fungi pollution was observed in the summer. There was significant association between Fungal contamination with residual chlorine (P= 0. 014). Conclusion: The results showed that the prevalence of opportunistic saprophytes fungal contamination was relatively high. Such condition may be attributed to high temperature and insufficient water treatment. In addition, there is a need to monitor water quality and staff training to increase user's knowledge and awareness of the risks. Keywords: Physicochemical, Fungal contamination, Swimming pools, Kashan

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Subcutaneous phaeohyphomycosis caused by sterile Chaetomium spp. in Iran

Mohammad Javad Najafzadeh, Maryam Saradeghi Keisari, Shirin Farahyar, Majid Ganjbakhsh, Masood Ziaee, Ali Naseri, Fariba Berenji, Sybren G. De Hoog, Abdolmajid Fata

Department of Parasitology and Mycology, Ghaem Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Email: [email protected]

We report a case of a 66-year-old Iranian woman with phaeohyphomycosis, there was a mass (approximately 1×1 cm in size) on the right lateral of neck. She had dysphagia, hoarseness and no pain. She complains about black discharge, irritation, headache, obscurity and periodical vertigo. Histological examination of biopsy fragments from the lesions showed septate branched, dematiaceous, hyphae. Direct microscopy of mass in 20% potassium hydroxide (KOH) and Histological examination showed septate light brown hyphae. Sporulation of the fungus isolated from the patient was unsuccessful in a 1-months culture on SDA, MEA, PDA, water agar with paper sterile filter and plant steam agar. In slide culture we just find sterile mycelia. Based on molecular identification, Chaetomium spp was confirmed by sequencing of the ITS (Internal Transcribed Spacer) domain of the rDNA gene and comparison with sequences held at gene bank and the Centraalbureau voor Schimmelcultures (CBS). ITS sequencing show that the isolate clustered within the ascomycete genus Chaetomium. The sequence did not fully match with any sequences of available ex-type strains of Chaetomium, Thielavia and Papulaspora and hence might belong to an undescribed species. After local excision of the mass and Antifungal therapy by ketoconazole (200 mg twice a day), the lesion healed completely. Keywords: Phaeohyphomycosis, Chaetomium

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The high rate of Pneumocystis jirovecii colonization in non-HIV-infected Iranian patients

Hossein Khodadadi, Hossein Mirhendi, Mehdi Mohebali, Hossein Zarrinfar

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Pneumocystis jirovecii causes Pneumocystis pneumonia (PCP) in immunocompromised patients with a high rate of morbidity and mortality. In addition colonization with this fungus may stimulate pulmonary inflammation or lead to PCP in susceptible patients. The epidemiology of this infection and routs of its transmission has poorly studied in Iran. We examined the prevalence of Pneumocystis colonization in patient with various lung underlying diseases. Materials and Methods: Bronchoalveolar lavage (BAL) fluids of 458 patients with different pulmonary signs and without PCP symptoms collected between August 2010 and January 2012. Patients were divided to four groups (Transplant recipients, malignant patients, immunosuppressed patients and immunocompetent patients). A sensitive nested-PCR method targeting 18s rRNA gene of P. jirovecii was used for investigating Pneumocystis DNA. Results: In 57 (12. 5%) of all samples P. jirovecii specific DNA was detected with nested-PCR. Pneumocystis colonization rate in malignant patients, transplant recipients, immunosuppressed patients and immunocompetent patients was 21.7%, 20.3%, 12.7% and 7.3% respectively. Conclusion: A significant number of patients with pulmonary diseases are colonized by P. jirovecii and this colonization may develop to PCP in these patients or they may transmit the fungus to other susceptible patients. The study showed that malignant and transplant recipient patients had a higher rate of P. jirovecii colonization among Iranian patients. Keywords: Pneumocystis jirovecii, Colonization, Nested-PCR

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Co- infection of invasive pulmonary Aspergillosis and cutaneous Fusarium infection in a patient suspected to pyoderma gangrenosum: a case report

Tahereh Shokohi, Nasrin Amirrajab, Masoud Aliyali, Sabah Mayahi, Narges Najafi, Ruhollah Abdi

Department of Laboratory Sciences, School of Paramedical Sciences and Tropical Medicine Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

A 32-year-old man with previous history suspected to chronic PG was admitted with cough, sputum and dyspenia for past four days. The patient received methylprednisolone pulses one week before admission. His current medication was oral prednisolone and cyclosporine. Large and diffuse deeply necrotic skin lesions were found on upper limbs and over the chest and back. A computed tomography (CT) scan of the lungs, revealed multiple 1-2 cm nodules in various stages with feeding vessels sign and cavitations in some of them. The halo sign (an area of low attenuation surrounding a nodular lesion) and the air- crescent signs were characteristic. The diagnosis of IPA caused by Aspergillus flavus was made on the basis of direct microscopic examination, culture of endobronchial washing and broncho alveolar lavage (BAL) samples and was confirmed by PCR sequencing. The diagnosis of cutaneous fusariosis was made on isolation of Fusarium sp. from skin lesion biopsy in multiple media. This fungus was confirmed by In situ hybridization (ISH) PCR from a histological specimen as well. He underwent systemic antifungal therapy with voriconazole. Unfortunately, the treatment was not successful and he expired after 12 days of hospitalization. This report highlights the rarity of coexistence of IPA and a chronic skin lesion such as fusarial skin infection and physicians should be alert to the possibility of invasive fungal infection in immunosupressed patients. Keywords: Aspergillus, Fusarium, IPA, Pyoderma gangrenosum

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The first case of onychomycosis due to Exophiala dermatitidis in Iran

Mohammad Javad Najafzadeh, Mehraban Falahati, Shirin Farahyar, Farideh Zaini, Zeinab Ghasemi, Ali Rezaei-Matehkolaei, Somayeh Dolatabadi, Maryam Saradeghi Keisari and Jacques Meis

Department of Parasitology and Mycology, Ghaem Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Email: [email protected]

We report a rare case of onychomycosis due to Exophiala (Wangiella) dermatitidis in a 54-year- old Iranian female with a history of no immunodeficiency, without underlying disease presented in 2011 to Razi hospital in Tehran, Iran. The patient was mountaineer, she has blackish pigmentation in toenail and distal area of the nail was empty. Scrapings were collected deeply from hyperkeratotic distal areas. Examination of potassium hydroxide mounts from the samples revealed brown, septate, branching hyphae. The scrapings were cultured on malt extract agar at 25°C. The colonies were initially moist and gray, becoming black or dark-green with dull surface. The fungus was identified as E. dermatitidis by its morphological characteristics and through DNA sequencing of the internal transcribed spacer region of rDNA. In vitro antifungal susceptibility has shown that itraconazole and posaconazole (0. 063μg/ml) had the highest and fluconazole (16μg/ml), caspofungin (2μg/ml) and micafungin (4μg/ml) the lowest activity against E. dermatitidis. Onychomycosis was considered as a fungal nail infection mainly caused by dermatophytes, sometimes yeasts and rarely caused by nondermatophyte molds such as dematiaceous fungi. This is the first report of onychomycosis due to E. dermatitidis in Iran. Keywords: Exophiala dermatitidis, Black yeast, Onychomycosis

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Phaeohyphomycosis due to dematiaceous fungi; a review of the literatures

Mohamad Hossein Afsarian, Tahereh Shokohi, Mehdi Arzanlou, Mehdi Taheri Sarvtin, Hamid Badali

Department of Medical Mycology and Parasitology/ Invasive Fungal Research Center (IFRC), Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran, Departments of Microbiology, Fasa University of Medical Sciences, Fasa, Iran Email: [email protected]

Introduction and Objectives: Melanized or dematiaceous fungi are frequently considered ubiquitous saprobes inhabiting living and dead plant material and soil. The clinical syndromes caused by latter fungi are differentiated based on histological criteria into phaeohyphomycetoma, chromoblastomycosis, and phaeohyphomycosis. Phaeohyphomycosis is a term generally reserved for the remainder of clinical disorder due to black yeast like fungi and relatives which ranging from mild allergic disease to disseminated brain fatal infection. Over 150 species and 70 genera of dematiaceous fungi have been implicated in human and animal disease. Materials and Methods: In the current review article, we searched of Pubmed Medline, Scopous, Google scholar, Elsevier databases, Irandoc, Iranmedex, Magiran, SID, MEDLIB and other sites of relevant articles by Keywords: Phaeohyphomycosis, melanized fungi, black yeast and Antifungal therapy. Results: Melanized fungi are common in the environments which become increasingly recognized as important pathogens, particularly in immunocompromised patients, though individuals with apparently “normal” immune systems have also been reported to have invasive, often fatal infections. Subcutaneous lesions are the most common case reports of infection due to melanized fungi which tends to be more frequent among farmers and gardeners as a result of local trauma and exposure to the potential etiologic agents during their work activities. For example Alternaria sp, is frequently isolated from subcutaneous lesions, Bipolaris and Curvularia from allergic diseases and Cladophialophora and Scedosporium most often from brain abscess and disseminated infection. The clinical manifestations in these infections consist of black grains in phaeohyphomycetoma, sclerotic bodies in chromoblastomycosis and presence of black hyphae in Phaeohyphomycosis. The timely and accurate diagnosis of fungal infections by melanized fungi consists of a multifaceted approach. Standard conventional diagnostic procedures include direct microscopy, histopathological examination, radiographic and computerized tomography (CT) findings, and isolation procedures to recover the fungus and identify the etiologic agent by different criteria. Conclusion: Recent assays have focused on non-culture based methods, in particular PCR and nucleic acid sequencing. Most of the invasive infections due to dematiaceous fungi are treated by triazole antifungal, e. g. voriconazole, itraconazole and posaconazole demonstrated the highest in vitro activity. However combination therapy is crucially recommended. Keywords: Phaeohyphomycosis, Melanized fungi, Black yeast, Antifungal therapy

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The frequency of candiduria in hospitalized patients with depressive syndrome

Mojgan Navid, Ali Zarei Mahmoudabadi, Ahmad Fakry

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Background and objectives: The presence of different species of Candida in urinary system is considered as candiduria. Depressive syndrome is an unpleasant and debilitating disease that causes several depressive disorders in patient behavior. Depressive syndrome increased appetite, weight gain and constant sleeping with the dominant group "atypical" fit. The aim of present study was to identify and compare candiduria in patients with depressive syndrome with healthy individuals. Materials and Methods: In this study on 131 patients admitted to Salamat and Boustan hospitals were sampled. In addition 105 healthy individuals were also sampled as control. Urine samples were collected in sterile urine bottles and transferred medical mycology laboratory, department of medical mycology, Ahvaz. 50µl of uncentrifuged urine samples was inoculated on CHROMagar Candida and incubated at 37°C for one week. All yielded yeasts were colony counted and identified based on morphology on CHROMagar Candida, production of germ tubes on serum, and formation chlamydoconidia on cornmeal agar with 1% Tween 80. Results: Our study shows that 2.3% and 13.3% of patients and control were respectively positive for candiduria. In the present study, 14 (13.3%) urine samples from control group was positive for Candida species, whereas only 2.3% of cases were positive. In both groups, females were affected by yeasts. The most common isolated yeasts were C. albicans and C. glabrata. Conclusion: Candiduria was most prevalent in control group than patients, treated for depressive syndrome by several drugs. Keywords: Candiduria, Candida albicans, Depressive syndrome

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Utilization of length polymorphism of ITS1 and ITS2 regions for identification of pathogenic yeast species by normal agarose gel electrophoresis

Ladan Karimi, Hossein Mirhendi, Hossein Khodadadi, Nilofar Jalalizand, Maiken Cavling Arendrup

Department of Biology, Faculty of Basic Sciences, Tehran Payame Noor University, Tehran, Iran Email: [email protected]

Introduction and Objectives: Yeast species play an important role in the morbidity and mortality of immunocompromised patients. In the present study, we used fragment size polymorphism (FSP) of internal transcribed spacer (ITS) of rDNA for identification of most important clinically yeast species among clinical isolates. Materials and Methods: The ITS1 and ITS2 regions of 214 yeast strain including 30 standard strains and 184 clinical isolates were amplified by PCR with two primer pairs (ITS1-ITS2) and (ITS3-ITS4) separately. PCR products of two regions were mixed equally and their electrophoretic patterns on agarose gel were analyzed according to the size of ITS regions that extrapolated from GenBank database. Results: The sizes of each ITS1 and ITS2 regions were enough different for each pathogenic yeast species so that identification of common clinical isolates was possible by specific pattern of normal agarose gel electrophoresis. We differentiated nearly all (about 35) pathogenic yeast species by this method, avoiding use of expensive tools such as sequencing or capillary electrophoresis. The patterns achieved for clinical isolates were in accordance with predicted patterns of sequence analysis. The most common etiologic yeast species such as C. albicans, C. tropicalis, C. glabrata, C. kefyr, C. krusei, C. lusitaniae, and C. guilliermondii were easily identified with FSP methods. Conclusion: We think that the method is an easy to perform molecular approach for identifying a wide range of yeasts species. This method was applicable for diagnostic or epidemiological purposes in any reference laboratory. Keywords: Identification, ITS1, ITS2, PCR, Pathogenic Yeasts

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Mating type and variety identification of Cryptococcus neoformans isolates

Setareh Agha Kuchak Afshari, Tahereh Shokohi, Seyed Reza Aghili, Hamid Badali

Department of Medical Mycology and Parasitology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran Email: [email protected]

Introduction and Objectives: Cryptococcus neoformans is the basidiomycetous yeast-like that can cause cryptococcosis. The major variety of clinical and environmental is C. neoformans var. grubii (Serotype A) with worldwide distribution and responsible for 95% of all C. neoformans infections. This study of Iranian Cryptococcus isolates was performed as an effort to determine the characteristics environmental Cryptococcus strain, including the varieties and mating types within the northern region of country. Materials and Methods: For 20 samples of C. neoformans isolated from pigeon excreta in Mazandaran province we used MATα and MATa specific primers for determining mating type. The two primers NL-1 and NL-4 were used to amplify the D1/D2 regions of large subunit (26 S) rDNA and for amplification of the IGS1 regions the two primers IGSF and IGSR were used. The sequence analysis of the D1/D2 and intergenic spacer regions were done. Results: All the samples were α mating type. The sequencing findings were compared with reference data obtained from GenBank database using BLAST and all DNA sequences were submitted to GenBank. Sequence analysis of the D1/D2 and IGS1 regions indicates only two isolates were C. neoformans var neoformans and 18 isolates were C. neoformans var. grubii. Conclusion: C. neoformans var. grubii is the most common variety in environmental sample in Mazandaran. Such a study would offer insight into the importance of this strain in relation to infection. However, we do not know whether this strain is the most common variety in clinical sample or not. Therefore further clinical and environmental investigations are warranted to determine the extent of C. neoformans variety in climatically different regions of Iran. Keywords: Cryptococcus neoformans, Molecular characterization, Mating type, Iran

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Catheter-related candidemia caused by Candida albicans in a coronary artery bypass graft (a case report)

Seyed Reza Aghili, Mohammad Ali Boroumand, Tahereh Shokohi, Shirin Sadat Hashemi Fesharaki, Bahar Salmanian

Faculty of Medicine, Mazandaran University of Medical Sciences Sari, Iran Email: [email protected]

A forty six-year-old male with 15 pack-month smoking, dyspepsia, unstable angina and resting retrosternal chest pain of 30 minutes’ duration that did not respond to sublingual nitroglycerin referred to the Heart Center Hospital (HCH), Tehran, Iran, in July 2012, for further evaluation. After angiography patient was found have atherosclerosis with three coronary vessels disease. An urgent coronary artery bypass grafting (CABG) was done for him with grafting three vessels without any complication intraoperative. After induction of anesthesia and tracheal intubation, a three-lumen central venous catheter was placed through the right sub-clavian vein. On the 12th postoperative day, the patient was suffering from an infection in the sternum area without fever. Staphylococcus aureus was isolated from culture of the discharge. Therefore, the antibacterial treatment was prescribed clindamycin and ciprofloxacin. Within 3 weeks of antibiotic therapy, the patient had no fever. Patient was receiving antibiotics, and removed the central venous catheter but he still had a fever. The catheter tips rolled on BHI agar, blood agar and MacConkey agar plates, then suspension of internal material of tube cultured in the biphasic Brian Hart (BBHI) media bottle. Growth of yeast and bacteria was obtained on BHI agar and BBHI after 72 h of incubation at 37°C. The man was started on fluconazole infusion in 5% dextrose. After 4 days, he became afebrile. Subsequent blood cultures after 7 days of starting fluconazole was reported sterile. Scraping of skin era in catheter penetration place in the patient was cultured in Sabouraud dextrose agar (SDA) containing the antibiotic chloramphenicol and incubated at temperature 37° C. Yeast colonies were grown for 48 hours. For further identification of yeast species, the isolates were sent to Medical Mycology Department, Medicine Faculty, Mazandaran University of Medical Sciences, Sari, Iran. Based on morphological characteristics of colonies on SDA and CHROM agar, germ tube test positive, and urease test positive, both of isolated yeast, Candida albicans was distinguished. The patients who usually have≥1 intravascular catheter, candidemia is associated with a high attributable mortality rate. Because Candida species to attach to vascular catheter material, early detection of organisms in pus or aspiration of the thrombus in catheter, remove of catheter and treatment with antibiotics (anti-bacterial and anti-fungal) will be an important role in disease control and prevention of septicemia. Morphological and molecular epidemiologic studies of isolates of Candida species recovered from a given patient reveal strong genetic relatedness between isolates from the skin and those in the bloodstream.

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A case of fungus ball type pansinusitis caused by Schizophillum commune

Mehdi Nazeri, Mojtaba Mohammadi Ardehali, Maryam Moazeni, Seyed Jamal Hashemi, Ali Akbar Fallahi, Hassan Ehteram, Sassan Rezaie

Department of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Schizophillum commune has been increasingly reported from allergic bronchopulmonary mycosis (ABPM) as well as fungus ball, brain abscess and several cases of maxillary or allergic fungal sinusitis. In the present study, we reported a case of fungus ball type pansinusitis from a 32-year-old woman In Iran. According to computed tomography (CT) scan, fungus ball type pan-sinusitis was likely to be the first diagnosis. Mycological examination revealed hyaline hyphae with small projection and also clamp connection structures on PDA medium. To identify the obtained isolate properly, molecular analysis of the internal transcribed spacer region was performed and indicated that the causing agent of the infection is Schizophillum commune. The patient was completely recovered after surgical endoscopic operation and consequent post- operation MRI revealed clearance of sinuses. Keywords: Schizophillum commune, DNA sequencing, Fungus ball, Pansinusitis

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Dermatophytosis due to Microsporum fulvum, a new case from Iran

Sadegh Nouripour-Sisakht, Ali Rezaei-Matehkolaei, Hossein Mirhendi, Mohammad Javad Najafzadeh, Kazuo Satoh, Bahram Ahmadi

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Microsporum fulvum is a ubiquitous geophilic dermatophyte that has rarely been considered as the agent of human dermatophytosis. Until now, there was no report regarding the human infection due to M. fulvum in Iran and here we present the first proven report of dermatophytosis by this fungus throughout the country, identified by sequence based methods. Direct microscopy of skin scrapings showed a tinea corporis infection and morphological study of the recovered colony from culture resulted in the identification of M. gypseum. To determine the main agent of the infection the isolate was subjected to molecular analysis. Restriction digestion of the internal transcribed spacers (ITS) and sequencing was indicative of M. fulvum as the causative agent. To verify the phenotypically identified isolates of M. gypseum and moreover, to highlight the role of M. fulvum in human dermatophytosis the sequence based analysis is recommended. Keywords: Microsporum fulvum, Dermatophytosis, Tinea corporis, ITS

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First case of disseminated phaeohyphomycosis in an immunocompetent individual due to Alternaria malorum

Mohsen Geramishoar, Hossein Mirhendi, Mohammad Javad Fatemi, Hamid Bateni, Mahboubeh Hajabdolbaghi, Bahram Ahmadi, Hamid Badali

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

A 27-year-old Iranian, previously health male presented with sub-cutaneous necrotic lesions with a localized dermatosis affecting the anterior chest, neck and face. These lesions consisted of singular, well-defined verrucous plaques which gradually developed and disseminated over time. The dermatosis was followed by the development of necrotic swollen lesions localized on the hard palate. The patient did not recall any history of trauma or puncture at any of the sites of infection. While histopathological examination of periodic acid-Schiff (PAS) stained material revealed irregular, unbranched, septate hyphae, direct examination (KOH 10%) of lesion samples demonstrated the presence of septate indistinct brownish hyphae. Alternaria malorum was isolated (CBS 126589) and its identity was confirmed by sequencing of the internal transcribed spacer (ITS rDNA). Since the palate lesion reoccurred after 10 years and the patient ’ s condition did not improve with amphotericin B combination therapy, the lesion was surgical excised and he underwent antifungal therapy with amphotericin B and itraconazole. There was no dehiscence or fistula formation or any evidence of relapse of fungal infection during a one year follow-up and the patient was successfully cured. In vitro antifungal susceptibility tests revealed that the MIC values for those antifungals employed in this case were amphotericin B (0. 125μg/ml), fluconazole (32μg/ml), itraconazole (0.125μg/ml), voriconazole (1μ g/ml), and posaconazole (0. 063μg/ml). The MECs for caspofungin and anidulafungin were 0.25 μ g/ml and 0.016μg/ml, respectively. However, treatment of A. malorum infections with the latter agents remains to be evaluated. Keywords: Alternaria malorum, Pleosporales, ITS rDNA, In vitro susceptibility

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Study of the role of Candida species in urinary tract infections with special attention to the predisposing conditions

Elaheh Mahmmodi, Farzad Katiraee

Department of Pathobiology, Faulty of Veterinary Medicine, University of Tabriz, Tabriz, Iran Email: [email protected]

Introduction and Objectives: The most common human fungal pathogens are Candida species. Studies show that the incidence of urinary tract infections caused by Candida pathogens has increased in recent decades. The purpose of this research was to study the prevalence of Candida urinary tract infections among other causes of urinary tract infections as well as the prevalence percentage of different Candida species in causing this infection. Materials and Methods: In the current study, 500 specimens from patients with positive urine culture test were evaluated. Of these, 479 cases of bacterial infection, 19 cases of candidiasis and 2 cases of mixed infections caused by bacteria and fungi were diagnosed. Candida species were diagnosed based on germ tube test, the colony color on CHROMagar Candida medium, intracellular beta-glycosidase enzyme activity and carbohydrate assimilation pattern. Results: According to the results obtained, 33.3%, 28.6%, 14.3%, 9.2% and 9.5% of the isolates were C. albicans, C. glabrata, C. tropicalis, C. kruzei and C. kefir, respectively. Also, an isolate with a 4.7% incidence of C. dubliniensis was identified. The patients were also examined regarding to predisposing factors in this experiment. Interestingly, the presence of one or more predisposing factors was proved in all the patients of which diabetes was the most prevalent factor by 38% frequency. Concerning blood culture and HIV, all patients were negative. Fourteen patients had proteinuria and 13 patients had glycosuria. Conclusion: Based on obtained results, Candida albicans was the most prevalent species, and in general, urinary tract fungal infections were less prevalent than bacterial urinary infections. Keywords: Urinary tract infections, Candida species, Predisposing factors

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Distribution of Malassezia species in patients with pityriasis versicolor compared to healthy individuals in Yazd, Iran

Abbas Ali Jafari, Hossein Zarrinfar, Farzanenh Mirzaei, Farzad Katiraee

Department of Pathobiology, Faulty of Veterinary Medicine, University of Tabriz, Tabriz, Iran Email: [email protected]

Introduction and Objectives: Malassezia species as part of skin microflora of humans and other warm-blooded vertebrates are lipophilic yeasts that associate with various human diseases, especially pityriasis versicolor (PV). The objective of this study was to identify Malassezia species on the scraped skin of patients with PV in Yazd, Iran, using morphological, biochemical and physiological methods. We also compared the results of PV patients with normal healthy volunteers. Materials and Methods: A total of 200 subjects, including 100 patients (with skin lesion) and 100 healthy volunteers as controls referred to Yazd Central Laboratory, were evaluated for Malassezia species by morphological and biochemical methods. Results: In PV lesions, the most commonly isolated species was M. globosa (38. 3%), followed in frequency by M. furfur (29. 4%), M. sympodialis (14. 9%), M. pachydermatis (9. 6%) and M. slooffiae (5. 3%). Also, in healthy skin the most commonly isolated species was M. furfur (37. 2%), followed in frequency by M. globosa (25. 6%), M. sympodialis (16. 3%), M. pachydermatis (13. 9%) and M. slooffiae (4. 6%). Conclusion: Highest prevalence of PV in our study was observed in 20–39 year-old group, suggesting that the peak of the infection is coincided with ages and increasing sebum production in the highest level. M. globosa was the most commonly isolated Malassezia species on the patients group and M. furfur in normal individuals. We couldn’t find any significant differences of Malassezia species distribution between groups. The rate of isolation of Malassezia species on patients was higher than normal individuals. Keywords: Malassezia species, Pityriasis versicolor, Yazd

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Survey the bacterial, fungal contamination in three dental impression materials

Abbas Ali Jafari, Mohammad Hossein Lotfikamran, Abbas Falah Tafti, Esmaeil Khair Khah

Department of Medical Parasitology and Mycology, Medical School, Shahid Sadoughi University of Yazd Medical Sciences, Yazd, Iran Email: [email protected]

Introduction and Objectives: Infection control for preventing cross infection in dentistry and dental laboratories is critical. Since the dental impression materials have close contact with oral mucosa, saliva and blood, these materials can be dangerous for immunosupressed patients, which are susceptible to low virulent microorganisms. The aim of present study is determination of bacterial and fungal contamination in three dental impression materials including Iranian and foreign alginates and speedex. Materials and Methods: Five specimens from 3 dental impression materials and 2 accumulation silicones (Speedex) were randomly collected from new un-used packs. All samples were homogenized in (1 ml) Tween 80 and cultured onto specific bacteriologic and mycological media. All bacterial and fungal colonies were counted and diagnosed based on bacterial and fungal diagnostic tests. Results: Totally, 75% of all samples were shown bacterial and 55% fungal contamination. Iranian alginate and speedex (putty) were the most contaminated samples (p= 0.01). Species of bacteria such as Micrococcus, Staphylococcus, Bacillus, Corynebacteria, Citrobacter, Actinomycetes and Neisseria and few fungi like Aspergillus, Penicillium and Candida were isolated from samples. Conclusion: The dental impression materials act as a source of microbial contamination in dentistry, which can distribute directly and indirectly in impression process as well as by aerosols existed in the environment of dental laboratory and dentistry. Therefore informing the dentistry society and also production factories of these diseases and pollutant resources is very important. Keywords: Bacterial, Fungal, Impression

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Electron microscope studies of actinomycete isolates Kn10 and 115 with antifungal effect against Microsporum gypseum

Naser Keikha, Gholamhosein Shahidi Bonjar, Seyyed Amin Ayatollahi Mousavi

Department of Medical Mycology and Parasitology, Faculty of Medicine, Kerman Medical University, Kerman, Iran Email: [email protected]

Introduction and Objectives: Human cutaneous infections are caused by a homogeneous group of creatine-consuming fungi called dermatophytes. These ubiquitous fungi are the most common infectious agents to humans of most populations globally. Microsporum gypseum is a cause of baldness in recent years, particularly in Iran. Cases occur sporadically due to M. gypseum in puppies and soil then transmitted to humans. As these pathogenic dermatophytes are eukaryotes, chemical treatment with antifungal drugs may also affect host tissue cells. In this study, we tried to find new antifungal agents against the pathogen and investigate their morphological criteria by transmission and scanning electron microscopes. Materials and Methods: To find new antifungal agents against that pathogen, approximately 100 Actinomycetes isolated from soil different area of the Kerman province. The antifungal effects of Actinomycetes were studied by culturing them on specific media such as, SCC and PDA. Electron microscopic studies (TEM and SEM) performed according to the relevant protocols. Results: The Actinomycete isolates with antifungal activity against M. gypseum, the isolates Kn10 and 115 which had the most antagonistic in vitro against fungi were used in this study. Images produced by Electron Microscope revealed specific forms of spores and mycelia together with branch of Actinomycetes filamentous. Conclusion: The findings of this research show the morphological criteria of streptomycete antagonists with antifungal effects against M. gypseum. This is the early step in search for new antifungals against the pathogen. Development of such principles highlights a long way to reality of production of antifungal antibiotics for human use. Electromicrographs of spore chains and mycelia and rDNA characterizations are in progress to reveal the taxonomical position of each active antagonist. Keywords: DermatophyteS, Actinomycetes, Streptomyces, Antifungal, Electron Microscopy, Microsporum gypseum

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A review on the role of fungi in atopic dermatitis

Mehdi Taheri Sarvtin, Zohreh Hajheydari, Mohammad Taghi Hedayati

Department of Medical Mycology and Parasitology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran Email: [email protected]

Introduction and Objectives: Atopic dermatitis (AD) is a chronic or relapsing inflammatory skin disease which affects 10–20% of children and 1-3% of adults. Attention has recently been focused on microbial agents, particularly fungi as aggravating factors of AD. Fungi can be found throughout the world. Some of fungal genera are as the normal flora, and some of them are abundantly found in the environment. The purpose of this study was to review the recent knowledge concerning the pathogenesis of AD in relation to the role of fungi. Materials and Methods: A comprehensive literature search of published studies from 1966 until 2011 in PubMed/MEDLINE and Iranian databases regarding role of fungi in AD was performed. Results: Malassezia, Candida, Alternaria and Cladosporium were the most common fungi which the most studies show their role in atopic dermatitis. Frequency of Candida and Malassezia-colonization were higher in AD patients than in control group; moreover the presence of type I hypersensitivity reaction to Malassezia, Candida, Alternaria and Cladosporium were seen in a significant percentage of patients with AD. Conclusion: Fungi are an important source of allergens which can trigger cutaneous inflammation in patients with AD. Therefore, we suggest that clinicians pay attention to screen and treatment of fungal hyper colonization and type I hypersensitivity reaction to fungi in patients with AD especially in patients who do not respond to traditional treatment. Keywords: Atopic dermatitis, Alternaria alternata, Candida, Cladosporium herbarum, Malassezia

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A Survey on oral Candida flora in patients with atopic dermatitis and compared with healthy subjects

Mehdi Taheri Sarvtin, Mohammad Taghi Hedayati, Tahereh Shokohi

Department of Medical Mycology and Parasitology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran Email: mehditaheri. [email protected]

Introduction and Objectives: Atopic dermatitis (AD) is a chronic, itching, inflammatory skin disease which is associated with asthma, IgE-mediated food allergy and hay fever. The increasing prevalence and severity of atopic diseases including AD over the last three decades has been attributed to decreased exposure to microorganisms during early life. Candida species are common commensals of the oral cavity, intestinal tract and vagina. The purpose of this study was to examine the presence of Candida albicans and other species of Candida in the oral cavity of patients with atopic dermatitis. Materials and Methods: All samples were taken by swab and were cultured on Sabouraud dextrose agar contain chloramphenicole medium. The Candida species was identified by genotyping using ITS–PCR and sequence analysis of the D1/D2 domain of the 26S rRNA gene. The date was analyzed using Independent t- test and Chi-Square Tests. Results: In this study 100 (44 male and 22 female) patients with atopic dermatitis and 50 (22 male and 28 female) healthy people as control group were participated. 31% of patients and 24% of healthy people were colonized by Candida species (P > 0.05). 23% of patients and 6% of healthy people were colonized by Candida albicans (P < 0.05). Conclusion: The colonization by Candida albicans in patients was significantly higher than control subjects. Therefore, it is necessary to examine the type and amount of Candida colonization in patients. Antifungal treatment is recommended in cases of increased Candida colonization. Keywords: Atopic dermatitis, Candida flora, Candida albicans

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Gastrointestinal basidiobolomycosis: first culture-proven case in Iran

Ali Naseri, Nona Zabolinejad, Mohammad Hassan Aelami, Marjan Joudi, Seyed Ali Alamdaran

Department of Medical Mycology and Parasitology, Imam Reza Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Email: [email protected]

A 3-year-old boy with one-month history of abdominal pain was admitted to the Doctor Sheikh Hospital, Mashhad University of Medical Sciences, Mashhad, Iran. After physical examination ultrasound and computerized tomography was done and the patient was subjected to laparotomy. Following, the resected material was prepared for histological and mycological investigations. Direct microscopic examination was carried out using 15% KOH preparations. Clinical materials were culture on Sabouraud dextrose agar with chloramphenicol. Cultures were incubated in 27 °C aerobically. The cultures were identified on the basis of colony morphology and microscopic characteristics. On physical examination, the patient’s abdomen was soft and lax, but he had a tender palpable mass. Hyaline, broad, sparsely septate hyphal elements were observed in direct examination of KOH preparations. Cultures inoculated with portions of clinical material yielded Basidiobolus after 7 days. The colonies were flat, thin, waxy and gray to yellowish in color initially and later became cerebriform in center and radial fissures in the outer zone with a pale reveres. Microscopic characteristics of fungus was the formation of broad hyphae and intercalary in hyphae, thick-walled with protuberances of gametangial remains (beaks) zygospores. In tissue sections, broad, thin-walled and septate hyphae which were surrounded by strongly eosinophilic materials were observed. This isolate was identified as B. ranarum. The first culture- proven case of GIB in Iran is reported. This case emphasizes the need to consider GI basidiobolomycosis in children presenting with abdominal mass and eosinophilia. Keywords: Basidiobolus ranarum, Gastrointestinal basidiobolmycosis, Mashhad

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The prevalence of candidiasis in pregnant patients with urinary tract infection referred to Mehrin clinical laboratory

Zahra Bolandghamat Pour, Nahid Shoaie, Golriz Zaker, Marzieh Saghafi

Mehrin Clinical Laboratory, Tehran, Iran Email: [email protected]

Introduction and Objectives: Urinary tract infection is a common disease in pregnancy period and can harm both the fetus and the mother. This study has been done to determine UTI and candiduria prevalence in pregnant women. Materials and Methods: during six month of study, 968 midstream clean catch urine were obtained from women aged 17-42. From 968 urine specimen taken, 326 were pregnant. Urine analysis and culture were done for all of samples. Urine samples were cultured on EMB, SDA and Blood a gar medium. Results: In this research, abundant of species is Candida albicans 4.76%, Candida sp 23.8%, GBS 23.8%, Entrobacteriaceae 19.06% and Entrococci 14.28%. In addition, the prevalence of Staphylococci coagulase negative and coagulase positive was 14.28% and 7.14%, respectively. The frequency of Candida sp was significantly higher than C. albicans. Conclusion: It can be concluded that Candida species are predominant microorganisms which were isolated of pregnant women with UTI. To treat patients with these kinds of infections, at the first step, it is necessary to identify causes of infection. For this reason, isolation and characterization of microorganisms in these women was carried out. Keywords: UTI, Pregnancy, Candida species

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Genetic diversity among Iranian Candida albicans isolated from nails and skin

Farzad Katiraee, Sahar Shadi, Yashar Farshi, Omid Fakhri, Payman Zare

Department of Pathobiology, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran Email: [email protected]

Introduction and Objectives: Candida albicans is the most prevalent yeast species that causes fungal infection. Also, C. albicans is one of the most frequent etiologic agents of dermatomycosis and onychomycosis. In spite of the new antifungals and health care, the incidence of mycoses has increased markedly, especially in immunocompromised individuals. The aim of this study is assessing the genotypic diversity and genetic relationship of C. albicans isolated from nail and skin. Materials and Methods: The genetic diversity of 50 clinical and reference strains of C. albicans from nails and skin clinical disorders were studied through randomly amplified polymorphic DNA (RAPD) techniques. C. albicans isolates were diagnosed by conventional and PCR- RFLP method. RAPD-PCR was optimized using three primers of arbitrary sequences. Cluster analysis of PCR product on agarose gel was performed using UPGMA method. Results: C. albicans isolates generated up to15 bands and were grouped in 27 genotypes depending on the primers used. RAPD profiles identified 7-9 clusters of genotype, based on primer used, among the 50 isolates. Conclusion: Our results revealed moderate genetic diversity among nail and skin isolates, and no association with their origin and the clinical characteristics. RAPD was suitable for fingerprinting analysis and was found to generate the reproducible fingerprints, yielding well- resolved banding patterns. RAPD fingerprinting can be used to genotype C. albicans. For obtaining reliable results by RAPD method, using more than one primer and comparative analysis of these primers are appropriate. Keywords: Candida albicans, RAPD, Genotyping

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Identification of the significant medical dermatophytes from the clinical samples of the skin clinics of the Tonekabon through the Real Time ITS – PCR sequencing method

Ayatolah Nasrollahi Omran, Seyedeh Sahar Yaghoubian, Ali Nazemy, Sajad Amani

Department of Microbiology, Islamic Azad University, Tonekabon Branch, Mazandaran, Iran Email: [email protected]

Introduction and Objectives: Gaining access to a rapid and accurate laboratory technique to identify the dermatophytes in the level of genus and species is very important. Direct microscopic identification and culture of the components of the fungus from the clinical samples is a rapid identification method but lacks the specificity and sensitivity. In this research, we study the direct and rapid diagnosis of these fungi using the seminested Real-Time PCR Method in skin sample of the patient. Materials and Methods: In this study, 30 samples of crust from patients were collected and for identifications of Dermatophytes, seminested PCR was done. DNA extraction by phenol- chloroform method was done and for first step PCR, ITS 1-4 universal primer was used. First step PCR products for Realtime-PCR by ITS 86-4 universal primer were used. PCR products for identification of dermatophytes were sequenced. Results: The results for the 30 samples PCR-Sequensing fungus, Trichophyton rubrum and 8 as the dermatophyte and non-dermatophyte molds were identified as one Candida bombi. The six colonies of dermatophytes: T. rubrum, T. mentagrophytes and T. verrucosum was isolated by culture. Conclusion: This method is useful in the quick identification of the pathogenic samples and this case is for the purpose of the fast treatment of the patients with the proper drug and the accurate dose of drug and the appropriate period of the drug consumption. Keywords: Dermatophytes, Clinical sample, Seminested Ral-Time PCR, Tonekabon

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Candida vaginitis due to consumption of azithromycin

Zahra Nasrollahi, Maryam Roudbari, Shahla Roudnbar Mohammadi, Yadegari Mohammad Hossein, Fatemeh Nikoomanesh

Department of Mycology, Faculty of Medicine, Tarbiat Modares University, Tehran, Iran Email: [email protected]

Introduction and Objectives: Genitourinary infections are the most usual disorders of the patients who visit a gynecologist. The three infections most commonly associated with vaginal discharge in adult women are bacterial vaginosis, vulvovaginal candidiasis and trichomoniasis. Microscopic examination findings, potassium hydroxide (KOH) preparations, wet mount and additional cultures and testing can be provided para-clinical examinations. In some cases gynecologist routinely prescribe according to his experience seeing some predominant symptoms and follows treatment of bacteria vaginosis with an ignorance of yeast infections and also the careless medical method of using antibiotics can cause Candida overgrowth. This study was performed to evaluate the effect of consumption of azithromycin in risk of Candida vaginitis in 23 women with vaginitis infections. Materials and Methods: This descriptive-analytic research was performed on 23 women with vaginitis infections women taking various antibiotics who were referred to a clinic in Tehran- Iran 2012. One group received antibacterial drugs including clindomicin, doxicilin, spiromycin and azithromycin and the other group received only azithromycin. Patients present with vaginal discharge, itching and pruritus. In the present study, we investigated the effect of azithromycin on vaginitis caused by Candida albicans. The sample of secretions was taken from both endocervix and exocervix entering into sterilized physiological serum. A microscopic examination serving KOH showed numerous budding yeasts at samples and chrome agar culture revealed C. albicans by growth as light to medium green colonies and most apparent after 24 h of incubation at 30°C. Results: Based on clinical results, 19 persons (82%) were received various antibacterial drugs including azithromycin and 4 persons (18%) received only azithromycin overally had C. albicans infection. Based on clinical results sensitivity for detecting vaginal candidiasis has been weak and para-clinical examination has not been done. It is claimed that emerging candidiasis has been due to consumption of azithromycin as its side effect of the first group (82%) comparing with the other one (18%) as a control. Conclusion: The results of this study show that para-clinical examinations especially a simple microscopic examination should be considered along with clinical survey in order to diagnose of the cause of vaginal infections accurately. Para-clinical experiment helps to prevent and reduce the side effects of medications. There is also a doubt which claims that the patients in this study have had candidiasis primarily. To eliminate this doubt, using Para-clinical experiment would be strongly recommended. However, using some antibiotics such as azithromycin cause vulvo- vaginal candidiasis secondary to cure bacterial vaginosis. Regarding that, azithromycin should be prescribed with caution following checking the patients for Candida vaginitis during the period of the treatment frequently. Keywords: Candida albicans, Vaginitis, Azithromycin

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Actinomycosis in Iran

Roshanak Daie-Ghazvini, Seyed Jamal Hashemi, Ensieh Zibafar, Mohsen Geramishoar, Sadegh Khodaveisi

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and objectives: Actinomycosis is an indolent, slowly progressive infection caused by anaerobic or microaerophilic bacteria, primarily of a genus Actinomyces that colonize the mouth, colon and vagina. Mucosal disruption may lead to infection at virtually any site in the body. The aim of this study was to underline different features of actinomycosis and to represent total data about etiologic agents, clinical, diagnostic and therapeutic approach these infections. Materials and Methods: From 1972 to 2012, the data obtained from all of case reports studies related to actinomycosis cases in Iran were analyzed. Diagnosis was performed based on microscopic examinations and cultures of specimens on specific media obtained from the patients. Results: Ninety three cases with actinomycosis (57 males, 36 females) were identified during this study. Findings represented oral-servicofacial (21 cases); thoracic (7 cases); abdominal (17 cases); disseminated forms (21 cases) and others (27 cases). Most patients were successfully treated with penicillin although some cases needed surgery along with antibiotic therapy. Conclusion: Since some clinical features of actinomycosis are mimic lesions with malignant appearance, the differential diagnosis of invasive forms must be included. Therefore, this report emphasizes the importance of differential diagnosis of actinomycosis from similar diseases by clinicians. Keywords: Actinomycosis, Oral-servicofacial, Thoracic, Abdominal, Disseminated

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Prevalence of candidiasis in Vali-Asr hospital of Birjand during 1998-2012

Toktam Bakhshi, Zohreh Bakhshi , Hossein Mashmool

Birjand University of Medical Sciences, Vali Asr Hospital, Microbiology Lab, Birjand, Iran Email: [email protected]

Introduction and Objectives: Candidiasis is one of the most common opportunistic infection in humans that is caused by Candida. The aim of this study was to investigate the prevalence of Candidiasis in Vali Asr hospital of Birjand during 1378-1390 and their association with age and sex. Materials and Methods: In this retrospective study, using the information recorded in microbiology lab during years 1378-1390 in Birjand hospital, the incidence of candidiasis was studied according to age and sex. Results: Total number of cases was 13217 and Candida was isolated from 215 samples (1.62%). Maximum numbers of samples were urine (84.65%) and the rest of the samples, respectively contained: feces (6.04%), sputum (4.65%), throat (2.32%), tracheal tube (1.39%), eye discharge (0.46%), wound (0.46%), and vaginal sample (0.46%). Conclusion: The most of the samples were related to children ward. The mean age of patients was 25 years old; but most of the sample was related to the age group 0-4. Chi –square analysis indicated that there are no differences between men and women at risk of Candidiasis. (P value>0.05) and statistical test showed that there was a significant interaction between age and type of sample (p value <0.05). Keywords: Prevalence, Candidiasis, Opportunistic infection

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Molecular strain-typing of Microsporum canis prevalent in Japan and Iran by randomly amplified polymorphic DNA.

Azar Sabokbar, Shahnaz Shahnavaz, Amir Bakhtiari

Department of Microbiology, Karaj Branch, Islamic Azad University, Karaj, Iran Email: [email protected]

Introduction and Objectives: The aim of the current study was to evaluate the genetic polymorphisms and presence of various molecular profiles in Microsporum canis isolates using RAPD (random amplified polymorphic DNA) technique. M. canis is a zoophile dermatophyte fungus which is widely distributed throughout the world and receives major consideration as a common cause of scalp in dogs and cats as well as in human. Materials and Methods: Molecular techniques are simple, rapid, and reproducible that has utilities for identification of dermatophyte fungi to the species level. A total of 32 strains of M. canis, screened for genetic variation by randomly amplified polymorphic DNA (RAPD) analysis, with four completely random nucleotide sequences (A, B, C, D) as a primer (14-12). Results: The results showed primer A, D yielded identical polymorphism profiles as compared with primer B, C. Interestingly, statistical analysis of the data, obtained from RAPD marker system, resulted in no correlation between clinical isolates and their host as well as their geographic regions. Therefore, it was clarified that type of host and territorial circumstances cannot determine molecular variation within the species. Among 32 specimens only 12 species didn't appropriately recognize with 4 distinct primers and were excluded. Conclusion: Epidemiological studies of human and animal infections contagious strains of M. canis have remained unresolved due to a lack of polymorphic molecular markers. So molecular typing could be particularly beneficial in solving epidemiological question revealing infective routes and reservoirs of contaminated species and might help considerably in the treatment and prophylaxis dermatophytes which lead to saving financial resources. Keywords: Dermatophytosis, PCR-RAPD, ITS region, Microsporum canis

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A case of sinusitis caused by Nattrassia mangiferae in Iran

Mandana Bakhshizadeh, Hamid Reza Hashemian, Hossein Zarrinfar, Mohammad Javad Najafzadeh

Department of Medical Parasitology and Mycology, Ghaem Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Email: [email protected]

Nattrassia mangiferae is a plant pathogen that is also known as cause of superficial, subcutaneous and disseminated infections in humans. Reports of human infections are extremely rare in Iran. They are characterized by development of dark colored filamentous hyphae in the involved tissues and their colonies are narrow branching and wide subhyaline to dark brown hyphae which fragmented to one-septate or no-septate arthroconidia. To our knowledge, this study describe the first case of phaeohyphomycosis sinusitis caused by N. mangiferae in an immunocompetent 20-year-old man who presented with maxillary sinusitis, frequent headaches in the forehead, nasal congestion, post-nasal discharge (PND) and dyspnea. The diagnostic workup includes computed tomography scan followed by direct microscopic examination and culture of tissue biopsy and sinus discharge specimens. We observed bilateral obstruction of maxillary sinus and mucosal hypertrophy in CT scan; and presence of septate, branched, and subhyaline to dark colored hyphae in direct examination; and rapid growth of the olivaceous grey to black colonies with aerial strands of greyish black mycelia with one-septate to no-septate arthroconidia on mycological media. The patient was treated with Amphotericin B that less than two weeks improved gradually. Keywords: Nattrassia mangiferae, Phaeohyphomycosis, Sinusitis

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Molecular phylogeny of the NADPH oxidase pathway in the filamentous fungus Aspergillus fumigatus, a human pathogen

Azita Dilmaghani, Florence Chapeland-Leclerc, Jp Latgé, Philippe Silar, Gwenaël Ruprich- Robert

Biotechnology Medical, Institut de Génétique et Microbiologie, France Email: [email protected]

Reactive oxygen species (ROS) have for a long time been considered as deleterious by-products of aerobic metabolism. However, studies on NADPH oxidases (Nox), a large family of enzymes dedicated to ROS production, have recently shed light on the many important biological roles of ROS. Nox are plasma membrane enzymes that utilize NADPH to generate superoxide (O2-). They have been implicated in defense mechanisms against pathogens in animals and plants, in regulating symbiosis and pathogeny in fungi, but also in developmental and morphogenetic processes in animals, plants, slime molds and fungi. The presence of at least one or two Nox homologues in all available genome of filamentous fungi oxidase-generated ROS play important roles in fungal physiology and differentiation. Recent studies have shown that ROS are key players of signaling during all stages of fungal development. The exact function (s) of ROS and the role of Nox in fungi are still not completely understood. The role of the Nox enzymes is unknown in human pathogens such as Aspergillus fumigatus, an opportunistic fungus that contains a single Nox enzyme of the Nox1 family. Furthermore, NoxR, the homologue of the gene encoding the regulatory subunit p67phox that activates the Nox genes remains also unstudied. In order to characterize the Nox pathway, we have generated a deletion of the AfNoxA gene. Phenotypic characterization of the invalidated mutants, including asexual and sexual spore production will be presented. To complete this study, the deletion of the AfNoxR, encoding the regulatory protein will be also realized. Keywords: Aspergillus, NADPH, Human Pathogen, Phylogeny

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Fungal peritonitis: the disease should be considered in patient with cancer (a case report)

Seyed Reza Aghili, Ghasem Jan Babaei, Tahereh Shokohi, Samaneh Afshar, Bahar Salmanian

Department of Medical Mycology and Parasitology, Faculty of Medicine, Mazandaran University of Medical Sciences Sari, Iran Email: [email protected]

A 52-year-old woman with a history of cervical cancer, presented to Tehran Imam Khomeini Hospital. On presentation she complained of mid-epigastric abdominal pain, which had persisted for 2 weeks. The patient described she had anorexia over the previous 2 weeks, lost 4. 5 kg during this time and the pain radiated to both the upper right and upper left quadrants. The patient denied history of fever, chills, and change in bowel habits. The patient's medical history included treatment and management for cervical cancer. She had completed radiation treatment before his presentation at our clinic and his current status in the follow-up for cervical cancer was stable. Poorly controlled hypertension before the initiation of dialysis was also notable in the patient's medical history. Vital signs upon presentation included: temperature, 36.9°C (normal range 36.5-37.2°C); pulse, 106 bpm (normal range 60-100 bpm); blood pressure, 96/78 mmHg (normal range 100/60-130/80 mmHg); and respiratory rate, 15 breaths per minute (normal range 10-16 breaths per minute). Abdominal examination revealed decreased bowel sounds and epigastric tenderness with diffuse rebound. Laboratory tests revealed a normal white blood cell count, low hematocrit level, low blood urea nitrogen level, and a high level of creatinine consistent with his renal and cervical disease. Analysis of the peritoneal fluid collected showed a slightly hazy appearance. An initial Gram stain of the dialysate effluent revealed that there were yeast blastoconidia and no bacteria present; this finding was confirmed by subsequent culture of the peritoneal fluid. Based on morphological characteristics of colonies on SDA and CHROM agar, germ tube test and urease test, Candida albicans was distinguished. It seems that tumor-related local factors permit fungi to cross the gut wall and to enter the peritoneum, where the host immune status determines whether the infection spreads or not. Following the growth of fungal in gastrointestinal tract or peritoneal cavity, inflammation and weakening of the immune system occurs in peritonitis. Because of lack of specific clinical and difficulty of isolation of pathogenic organisms from clinical specimens, treatment is very difficult. Approach to this disease has changed in recent years. In 2005 the International Society of Peritoneal Dialysis guidelines advocated that peritoneal fluid in the peritoneal dialysis catheter and peritoneal fluid should be examined in terms of the identification of fungal elements. So in malignant patients with inflammation of peritoneum, examination of peritoneal fluid for fungal element, (direct microscopic exam and culture) is necessary. Keywords: Fungal peritonitis, Malignancy, Ascites, Candidiasis, cancer

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Candida species in vulvovaginal candidiasis patients from the town Taibad in Iran; identified by PCR-RFLP method

Habibeh Zabardast, Maryam Moazeni, Parivash Kordbacheh, Roshanak Daie-Ghazvini, Keramat Allah Noori Jalyani, Sassan Rezaie

Division of Molecular Biology, Department of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Vulvovaginal candidiasis (VVC) is caused by overgrowth of members of the genus Candida in the vagina and is described by curd-like vaginal discharge, itching, and erythema. It affects 75% of all women at least once in their lifetime, especially during childbearing years. Although several Candida species have been implicated in VVC and recurrent vulvovaginal candidiasis (RVVC), Candida albicans is still the predominant etiological agent, causing 85– 95% of these infections. However, in the present study, we aimed at an epidemiological survey on the distribution of Candida species among women from the border town, Taibad. Moreover any possible alternation in this distribution is mainly concerned. Materials and Methods: From September 2010 to March 2011 (about 6 months), 300 clinical cervicovaginal samples were collected from women referred to laboratories as well as medical centers all over Taibad. Fungal direct examination and culture were performed afterward. PCR- RFLP was performed on yeast colonies obtained from positive cultures with the use of ITS1/ITS4 primers which amplify the ITS1-5, 8S-ITS2 regions of fungal rRNA genes. Digestion of the PCR products with one restriction enzyme, MspI, allowed discrimination of medically important Candida species. Results: Among 300 suspicious patients, 150 were proven to suffer from vulvovaginal candidiasis by conventional as well as PCR-RFLP methods. In 0.7% of cases two different kind of Candida species were isolated as etiologic agents from one patient (C. albicans/C. glabrata). The obtained restriction pattern of each Candida species was completely discriminatory, except for 3.97% of cases which were identified by sequencing afterward. The results indicated that C. albicans (53.64%) was the most frequent species followed by C. glabrata (28.47%), C. Krusei (11.92%), C. Tropicalis (1.32%) and C. guilliermondii (0.66%). Conclusion: In conclusion, PCR-RFLP method was successfully applied for identification of clinical Candida species within the Taibad town. According to our obtained results, C. albicans is the predominant causative agent of vulvovaginal candidiasis. Keywords: Candida, Vulvovaginal candidiasis, PCR-RFLP, Taibad

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Detection of invasive candidiasis in patients with hematological malignancies and bone marrow transplant recipients

Mohsen Ashrafi, Mojtaba Nabili, Ghasem Jan Babaie, Mohamad Taghi Hedayati, Kamran Ali Moghaddam, Tahereh Shokohi

Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran- Social Security Organization, Golestan, Iran Email: [email protected]

Introduction and Objectives: Invasive Candidiasis (IC) is a significant cause of morbidity and mortality in patients with hematologic malignancies and bone marrow transplant recipients. Basically, quick and early diagnosis prevents IC progress. We established a real-time PCR assay for the detection of Invasive Candidiasis and identification of causing Candida species. Materials and Methods: From 2009 to 2011, 72 patients with hematologic malignancies and marrow transplant recipients were evaluated for IC in Sari and Tehran. Ten milliliter bloods for Real Time PCR and blood culture were collected twice from patient. DNA was extracted from blood using glass beads and QIAamp DNA Blood Mini Kit. The primers and hybridization probes were designed to potentiate the specific sequence of 18S rRNA genes using Light Cycler system and Fluorescence Resonance Energy Transfer (FERT). Melting temperature (Tm) analysis performed to differentiate the different between Candida species simultaneously. The patients were evaluated for IC based on European Organization for Research and Treatment of Cancer/ Mycoses Study Group (EORTC/MSG) Consensus value definitions. Results: The female -to- male ratio was 1: 2; the mean age was 32.1 years. The most common malignancies in these patients were AML; 20 (27.8%), ALL; 19 (26.4%), Hodgkin’s lymphoma (11.3%) and other patients had non-Hodgkin’s lymphoma, multiple myeloma, thalasemia and chronic lymphocytic leukemia that received chemotherapy. Out of 72 patients, eleven patients (15. 3%) had positive real time PCR /probe results. The etiologic agents were C. krusei (5 cases), C. tropicalis (3 cases), C. parapsilosis (2 cases), and C. albicans (1 case). According to the criteria defined by EORTC/MSG who met clinical signs and host factor criteria of IC, 1 (1. 38%) and 10 (13. 8%) patients were stratified into probable and possible groups, respectively. Conclusion: The established Real-time PCR/FRET probe assay is a useful method for the quick identification of Candida species for the management of patients suffering from hematologic malignancies and bone marrow transplant recipients, in the risk of Candidiasis. Keywords: Invasive candidiasis, Hematological malignancy, Real-time PCR

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Detection of Malassezia isolated from patients with pityriasis versicolor and seborrheic dermatitis in Ahvaz using Nested- PCR

Maryam Azish, Ali Zarei Mahmoudabadi, Majid Zarrin

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: The species of the genus Malassezia are lipophilic and dimorphism yeasts that are regarded as part of normal flora of the skin of the humans and warm blooded animals. All species are identified by morphologic and physiologic features, but these phenotypic methods are time-consuming and lacked sufficient distinction. Thus using molecular techniques, identification of the species is done faster and more correct than phenotypic methods. The purpose of this study was to determine the frequency of common Malassezia species in patients infected with pityriasis versicolor and seborrheic dermatitis using of the Nested - PCR in the city of Ahvaz. Materials and Methods: In total, 75 samples of Malassezia, 45 samples isolated from patients with pityriasis versicolor and 30 samples isolated from peoples who that affected with seborrheic dermatitis, were included in the study. Isolates identified by Nested-PCR method. In the first stage, the ITS region from the ribosomal DNA was reproduced by using primers ITS4-R and ITS1F-N. The product of the first step was used as DNA and using three special pair primers of Mf-F, 5.8SR and M. gl-F, 5. 8SR and M. rt-F and M. rt-R, detected the inner part of the first phase. Results: Of the 45 scotch tape sample that were microscopically positive, 37 (82.2%) isolates identified with three special pair primers and did not detect eight isolate. The most common isolates in frequency sequence were Malassezia furfur (51.3%) M. globosa (35.2%), and M. restricta (13.5%), respectively. From 30 patients that affected with seborrheic dermatitis, 15 cases (65.2%) M. restricta, in 6 cases (26.1%) M. globosa and in 2 cases (8.7%) M. furfur was detected and did not detect seven isolate. Wholly 15 (20%) were not detected in pityriasis versicolor and seborrheic dermatitis patients were other of Malassezia species not surveyed in our study. Conclusion: The most common species isolated from pityriasis versicolor lesions, was M. furfur and then M. globosa. The most common species isolated from seborrheic dermatitis lesions, was Malassezia restricta. The Nested-PCR, is rapid and repeatable method for identification of important Malassezia species and this method is recommended to be used on more patients. Keywords: ITS rDNA, Nested-PCR, Malassezia, Pityriasis versicolor, Seborrheic dermatitis

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Identification of Candida species isolated from oral colonization in Iranian HIV-positive patients by PCR-RFLP

Seyyed Amin Ayatollahi Mousavi, Samira Salari, Sasan Rezaie, Naser Shahabi Nejad, Sanaz Hadizadeh, Hossein Kamyabi, Hossein Aghasi

Department of Medical Mycology and Parasitology, School of Medicine, Medical University of Kerman, Kerman, Iran Email: [email protected]

Introduction and Objectives: The incidence of opportunistic infections due to Candida albicans and other Candida spp. has been increasing. Rapid identification of candidiasis is important for the clinical management of immunocompromised patients. Polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) is a rapid, sensitive and specific method for detection of clinically important fungi. The purpose of this study was to identify Candida spp. isolated from the oral cavities of HIV-infected patients in southeastern Iran (Kerman), by using PCR-based restriction enzyme digestion. Materials and Methods: We identified 96 Candida isolates obtained from 139 Iranian patients infected with the human immunodeficiency virus (HIV), between April 2009 and April 2010 by using PCR-RFLP assay. Universal primers for the internal transcribed spacer (ITS) region (ITS1–ITS4) of the fungal rRNA genes were used for this assay. Results: We successfully identified the different Candida spp. by using the restriction enzyme MspI. C. albicans was the most commonly identified species (82. 2%), followed by C. glabrata (7. 29%), C. parapsilosis and C. kefyr (both 4. 1%), and C. tropicalis (2%). Conclusion: PCR-RFLP is a highly sensitive, specific and direct method for fungal detection and can be used for fungal epidemiological studies in HIV-positive and other immunocompromised patients. Keywords: Candida spp, PCR-RFLP, Candida albicans

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Study of fungal colonization of burn wound in patients admitted to the burn center of Zare' hospital, Sari, 2011-2012

Nazanin Lotfi, Tahereh Shokohi, Seyed Zahra Nouranibaladezaei

Department of Medical Mycology and Parasitology, Invasive Fungi Research Center, Mazandaran University of Medical Sciences, Sari, Iran Email: [email protected]

Introduction and Objectives: Despite major advances in the care of burn patients, infection remains an important cause of morbidity and death. The incidence of fungal infection is reported to vary between 6 and 44% of all burns admissions. We performed this study to determine incidence of fungal colonization and to identify the fungal isolates in burn wound of the patients admitted to the burn center of Zare' Hospital, Sari, 2011-2012. Materials and Methods: In a cross-sectional study from June 2011 to March 2012, the 7 day post burn wound with sign of infection was included. Specimens for fungal culture were collected using swab from the subeschar infiltrations. In some burn patients, subschar tissue biopsies were taken for culture. The isolated fungi were identified as to the species level by conventional procedure and PCR-sequencing. Results: Of 136 patients who were admitted during the study period, 88 patients were clinically suspected to have fungal colonization in their burn wound. Two hundred thirty one sample were taken from these suspected burn wound. On investigation, 84 samples (36.4%) taken from 47 patients revealed fungal colonization. The most predominant isolated fungus was Candida spp., mostly non albicans species. Mixed infections due to Candida spp., Fusarium spp. and Zygomycete were observed on the wound of a burn patient. Conclusion: Fungal colonization is an important marker in management of fungal sepsis in burn patients. Non albicans Candida spp were the most predominant fungi colonized in burn wounds. Keywords: Burn, Candida, Fungal colonization

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Mycotic keratitis among patients referred to mycology laboratory, Emam Reza hospital of Mashhad, Iran

Fariba Berenji, Sara Fata, Hamid Khakshour, Akbar Derakhshan

Parasitology and Mycology lab, Emam Reza Educational, Research and Treatment Center, Mashhad University of Medical Sciences, Mashhad 91379, Iran Email: [email protected]

Introduction and Objectives: Bacteria, amoebae and fungi usually cause keratitis. Mycotic keratitis is the infection of cornea due to different types of fungi including molds and yeasts. Predisposing factors are trauma (vegetable matter), surgery and prolong use of steroids. In order to investigate the prevalence of mycotic keratitis among the patients suffering from infection of cornea/this study was undertaken over a period of 8 years. Materials and Methods: We retrospectively reviewed the medical records of patients who referred to Mycology lab of Emam Reza Hospital of Mashhad. After admission of patient to the mycology lab, a questionnaire including personal and clinical information was completed for him / her. Sampling by ophthalmologist and preparation of fresh smear and culture performed for each patient. Results: The results showed that %57 of the patients were male and the others were female. Aspergillus spp. and Fusarium solani were isolated from 55% and 7% of the cultures respectively. Trauma due to penetration and plant particles was the common predisposing factor. The disease was mostly seen in farmers. Conclusion: It is concluded that fungal keratitis should be considered in the differential diagnosis of keratitis especially if predisposing factor is trauma due to plant matter. Keywords: Mycotic Keratitis, Aspergillus, Fusarium

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Candidiasis prevalence and relationship with clinical and para clinical observations in women supported by selected health centers of Tabriz

Fahimeh Sehhatie, Asieh Namazi, Mohammad Adibpour, Jalil Babapour

Instructor of Nursing and Midwifery Faculty, Tabriz University of Medical Sciences Email: [email protected]

Introduction and Objectives: Vulvavaginitis of candidiasis with bacterial vaginosis are 90% of infections. Midwifes have first role in evaluation, control and prevention with patient education in vagina health. The aims of this study were to determine candidiasis prevalence and relationship with clinical and para-clinical observations in women referred to Tabriz health centers. Materials and Methods: This is a cross- sectional study, in which 1000 women who had the inclusion criteria’s were selected by random sampling from 12 selected health centers of Tabriz. A questionnaire was used to obtain their personal and reproductive information, check list for clinical observations and Wife test, direct observation, and Sabouraud agar culture methods were used to diagnose this infection. pH of their vaginal discharges was determined. Data were analyzed by using SPSS version 13.5, and frequency, mean and standard deviation, χ2 and fisher exact test. Results: The prevalence of candidiasis was 25.2 %. Findings showed that the majority of clinical observations include abnormal discharge volume, clear, appearance, consistency of discharge. pH of vagina were statistically related to candidiasis (p<0.01). With direct observation, rate of infection was 50.7%. Wife test was negative in most patients, culture method was positive in 34.1% of patients. pH of vagina was lower than 4.5 in 18.9%. Conclusion: According to high prevalence of candidiasis infection, it seems necessary to pay more attention to these infections, and more efforts should be done for prevention. Midwives and other health professionals have an important role in giving more information to women about infections and improving their quality of life. Keywords: Candidiasis, Clinical and Para clinical observations

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An evaluation of direct microscopy examination and culture sensitivity in diagnosis of dermatophytosis

Seyed Jamal Hashemi, Roshanak Daie-Ghazvini, Ensieh Zibafar, Mohsen Geramishoar, Zeinab Borjian Borujeni, Leila Hosseinpour, Mahdi Zareei, Hossein Mirhendi

Department of Medical Mycology and Parasitology, School of Public Health and Institute of Public Health Research, Tehran University of Medical Sciences, Tehran, Iran Emil: [email protected]

Introduction and Objectives: Diseases of skin and its appendixes (hair and nail) are caused by two groups of infectious and non-infectious agents. The former agents consist of fungi, bacteria, viruses and others (e. g insects). The most important fungi in these diseases are dermatophytes, Malassezia, Candida and non-dermatophytes. Each one of infectious agents have specific characteristics and particular treatment, also, in the fungal diseases each causative genus (even, sometimes each causative species) have different sensitivity to antifungal therapy; therefore, identifying causative agent is necessary. Thus, applying a sensitive and confidential laboratory method for rapid and non-expensive diagnosis can be useful and it is necessary to prefer and give priority to specific method based on laboratory documents. The common and available laboratory diagnosis methods are DME and culture, thus, the aim of this study is to determine the priority of these two methods to diagnosis (not to identify) dermatophytosis Materials and Methods: This study has been accomplished by checking the information of referred persons to the laboratory of department of medical mycology, school of public health and institute of public health research, Tehran University of Medical sciences in Iran, to diagnose of clinical suspected dermatophytosis. Results: From the beginning to the end of 2011, information survey of 1601 persons were revealed that 301 persons of them were infected by dermatophytes, the DME and culture results of these patients have been divided to several groups and stated in below. G1: DME positive and culture positive = 186 (61/8%) cases, G2: DME positive and culture negative = 110 (36/5%) cases, G3: DME negative and culture positive = 5 (1/7%) cases, G4: DME positive or culture positive (total of cases) = 301 (100%) cases. Conclusion: Although culture is necessary for identification of dermatophytes, but, the important matter is only diagnosis of disease, the DME- at least in our laboratory- has high sensitivity (36/5% more than culture). In conclusion, to achieve two aims (diagnosis and identifying), both methods (DME and culture) are necessary. Keywords: Direct, Microscopy, Culture, Diagnosis, Dermatophytosis

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The effects of vaginal candidiasis on newborn

Sima Rasti, Mohammad Ali Asadi, Mitra Behrashi, Afsaneh Taghriri, Gholamabbas Mosavie

Department of Parasitology, Kashan University of Medical Sciences Email: [email protected]

Introduction and Objectives: Vaginal candidiasis associated with an increased incidence of adverse outcomes of pregnancy. Considering various reports regarding Premature Rupture of Membrane (PROM) and Low Birth Weights (LBW) in candidiasis, this research was carried out on the pregnant women from Kashan. Materials and Methods: This follow –up study was carried out on pregnant women hospitalized for preterm or term delivery in Kashan obstetrics and gynecology. 150 pregnant women were studied for candidiasis. Clinical signs, PROM and LBW were recorded. Vaginal secretions were cultured in Sabouraud dextrose agar (SDA) and SDA with chloramphenicol and cycloheximide. The data were analyzed by X2 and Fischer exact tests. Results: Of 150 pregnant women, 32.7 % were positive for Candida albicans. 35% patients with preterm labor and 31.8% with term labor were positive for C. albicans. No significant relation was found between delivery and PROM and candidiasis (P=0.71 and P<0.1). 17.2% pregnant women that had Low birth weights newborns infected with C. albicans but the rate of infection in whom had appropriate gestational age newborns was 36.4% (P=0.08). Conclusion: In spite of high prevalence of candidiasis in pregnant women, no significant relation was found between PROM and LBW and candidiasis. Keywords: PROM, LBW, Candidiasis, Kashan

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Identification of dermatophytes by AP-PCR

Zahra Salehi, Majid Zarrin, Ali Zarei Mahmoudabadi

Department of Medical Mycology, School of Medicine, and Infectious Diseases and Tropical Research Centre, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Dermatophytes are closely related keratinophilic fungi that invade the keratinized tissues in humans causing dermatophytosis. Identification of dermatophytes with conventional methods is time consuming and sometimes problematic because of similarities and variability of species. Genetic amplification has made rapid and perfect identification of dermatophytes possible. The aim of this research is evaluation of AP- PCR method for identification of dermatophytes and to find a suitable approach for a rapid distinguish of dermatophytes. Materials and Methods: Fifty-two isolates from 10 species of dermatophytes include: T. mentagrophytes (10), T. verrucosum (9), T. rubrum (5), T. tonsurans (3), T, violaceum (2), T. schoenleinii (1), M. gypseum (8), M. canis (4), M. ferrugineum (2), E. floccosum (8) were collected and confirmed with microscopic, macroscopic and biochemical tests. Molecular identification was done using AP-PCR (arbitrarily primed PCR) with random primers OPAA11, OPU15, OPAA17 and OPD18. These primers amplified bands of different sizes in species of dermatophytes DNA. Results: All species of dermatophytes were recognized with a distinct DNA band patterns on gel agarose. The best random primer identified for each dermatophyte. The range of obtained bands was between 250 to 2000 bp for all dermatophyte. The best primers for recognizing species were OPAA11 and OPAA17. When OPD18 and OPU15 primers were used more bands observed than OPAA11 and OPPA17 primers. Conclusion: In laboratory, it is always difficult to distinguish T. mentagrophytes from T. rubrum but using this method, the distinction of the two species of fungus can easily done. AP-PCR of dermatophytes with random use of the following primers OPAA11, OPU15, OPAA17 and OPD18 is a useful and rapid method for distinguishing dermatophytes. Keywords: Dermatophyte, Random primer, AP-PCR

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Onychomycosis in Mashhad

Fariba Berenji, Abdolmajid Fata, Ali Naseri, Robabe Izadi Jahanparvar, Majid Ganjbakhsh

Parasitology and Mycology Lab, Emam Reza Educational, Research and Treatment Center, Mashhad University of Medical Sciences, Mashhad 91379, Iran Email: [email protected]

Introduction and Objectives: Onychomycosis is a chronic infection of the nails. Knowledge of the epidemiological and mycological characteristics is an important tool for controlling this infection. The aim of this study was to document the descriptive epidemiological features of onychomycosis in 404 patients referred to parasitology and mycology laboratory of Emam Reza educational, research and treatment center of Mashhad (2005-2010). Materials and Methods: During 5 years (Mar 2005-Mar 2010), 404 patients with nail lesions suspected of having onychomycosis, were examined in mycology laboratory of educational, research and cure center of Emam Reza of Mashhad. The specimens were obtained from clinically abnormal nails, by scrapping of the nail bed, the underside of the nail plate. Fresh smears with 10% KOH were prepared and examined directly under the microscope. For fungal cultures, all samples were inoculated on each of two isolation media (1) Sabouraud dextrose agar (SDA) (2) SDA with 5% chloramphenicol and cycloheximide. For positive sample of Candida CHROMagar was used. The culture tubes were incubated at 25°C and 37°C and examined daily for six weeks. Results: From 404 suspected cases of onychomycosis 107 were positive in direct smear and 90 cases in culture. (Direct smear man 34/6%, Woman 65/4%, culture man 33/4%, and woman 76/6%). Females were affected more frequently than males. Fingernails (61%) were affected more frequently than toenails (39%). Candida albicans (36 cases) was the prevalent yeast, C. glabrata (17 cases), C. parapsilosis (2 cases). Dermatophytes were isolated in 5 cases, yeasts in 2 cases. Moulds were mainly Aspergillus spp. (23 cases), Penicillium (5 cases), Acremonium (2 cases). Conclusion: This study demonstrated that C. albicans, Aspergillus spp. and C. glabrata dermatophytes and Penicillium are the most prevalent agent causing onychomycosis in our region. Female were more infected as they have more risk exposure like more contact to water and detergents and using nail polishes. 10% of patients are under 10 years old which can be reasoned by habit of sucking finger. Keywords: Onychomycosis, Fingernails, Candida albicans, Mashhad

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Study the relationship of Candida species with Lactobacillus species in patients with fungal vaginitis and healthy individuals.

Parvin Dehghan, Kamiar Zomorodian, Mahboobeh Kharazi, Maryam Yazdani, Mostafa Chadeganipour, Keyvan Pakshir, Mohammad Javad Rahimi, Mahboobeh Pishva

Department of Medical Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran Email: [email protected]

Introduction and Objectives: Vaginal candidiasis is the second cause of vaginal infections and almost 75% of healthy females infected by these yeasts at least once during their reproductive age. The present study aimed to determine the relationship between the presence of different species of Lactobacilli and overgrowth of Candida in patients with vaginal candidiasis and healthy individuals. Materials and Methods: The study conducted on 120 patients suspected with vaginal candidiasis and 21 healthy individuals. Following the clinical examination by a gynecologist, samples were taken from the vaginal discharges for culturing on SDA and MRS agar, gram staining, and determining the pH. The Candida species were differentiated by CHROMagar Candida and rapid-ID 32 tests. The Lactobacillus species were identified using the biochemical test (API-CH 50). Results: Of the examined patients, 42 women (35%) were identified as vaginal candidiasis based on clinical symptoms and positive-culture. The most frequently isolated species identified was C. albicans (57. 1%), followed by C. glabrata (11. 9%), C. krusei (9. 5%), C. kefyr (7. 1%), C. parapsilosis (2. 4%), C. (2. 4%) and C. rugosa (2. 4%). Of the 21 healthy controls, the culture of the vaginal swabs only yielded positive results in 5 cases (23. 8%) which were all identified as C. albicans. L. acidophilus and L. delbrueckii were the most frequent isolated species in both groups. No significant association was found between the number and species of lactobacillus and vaginal candidiasis. A significant relation was found between the pH of vaginal discharge and itching. Conclusion: To the best our knowledge, there is no previous report on distribution of both vaginal Lactobacillus and Candida species in healthy women and patient in Iran. As a significant association was found between the growth of Lactobacilli and pH as well as pH and Candida colonization, these two organisms might have a mutual interaction. Further studies are still needed to reveal such interaction. Keywords: Candida, Lactobacillus, Patients, Vaginitis, Relationship

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Identification of dermatophytes species by PCR-RFLP during 2011-2012 in Isfahan

Rasoul Mohammadi, Hossein Mirhendi, Mehdi Abastabar, Shahla Shadzi, Parinaz Pourfathi, Niloufar Jalalizand

Department of Medical Mycology, Tarbiat Modares University, Tehran, Iran Email: [email protected]

Introduction and Objectives: Dermatophytes are keratinophilic fungi capable of causing dermatophytosis. These organisms are among the most common fungal infections in humans. These species cause superficial infections and rarely progress to more invasive disease. Dermatophytes are classified in three anamorphic genera: Epidermophyton, Microsporum and Trichophyton. Identification of dermatophytes Include clinical features, culture characteristics, microscopic morphology and physiological test results. Molecular techniques are highly accurate and rapid but they are costly and sometimes difficult to implement. In this study, we applied PCR-RFLP method in order to develop a fast and reliable assay to identify dermatophytes on a large number of samples. Materials and Methods: One hundred and thirty dermatophytes species were isolated from patients in Esfahan during 2011-2012. All isolates were evaluated by direct microscopy and culture. DNA extraction of isolates was performed by Phenol- chloroform method. PCR amplification was applied in a final volume of 50µl and ITS1-5. 8S-ITS2 region was amplified using ITS1 and ITS4 primers. PCR products were digested by restriction enzyme MvaI and digested products were visualized in 2% agarose gel electrophoresis in TBE buffer and photographed by ultraviolet photography. Sequence analysis was done for suspicious isolates. Results: Isolates were obtained from sole 42 (32. 3%), nail 33 (25. 3%), groin 28 (21. 5%), skin 12 (9. 2%), hand 8 (6. 1%), hair 6 (4. 6%) and wound 1 (0. 8%). We identified 62 (47. 7%) T. interdigitale, 25 (19. 2%) E. floccosum, 22 (17%) T. rubrum, 11 (8. 5%) M. canis, 4 (3%) T. erinacei, 2 (1. 5%) T. violaceum, 1 (0. 7%) T. tonsurans, 1 (0. 7%) M. gypseum, 1 (0. 7%) T. schoenleinii, respectively and also we could identify 1 (0. 7%) Chrysosporium parvum. It was obtained from sole of a 33 years old man. Eighty two (63%) patients were male and 48 (36. 9%) were female. The most number of patients related to (21-30) age bracket and the least related to (0-10). Conclusion: PCR-RFLP identification of dermatophytes by MvaI is rapid and efficient and we can use it in clinical laboratories. Keywords: Identification, Dermatophytes, PCR-RFLP, Isfahan

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Molecular identification of Candida species isolated from onychomycosis in Iran

Rasoul Mohammadi, Hossein Mirhendi, Mohammad Ghahri, Mohammad Hossein Yadegari, Shahla Shadzi, Parinaz Pourfathi, Niloufar Jalalizand

Department of Medical Mycology, Tarbiat Modares University, Tehran, Iran Email: dr. [email protected]

Introduction and Objectives: Onychomycosis is a chronic fungal infection of nails caused by dermatophytes, yeasts and moulds. Nail infections caused by Candida species are normally associated with chronic paronychia or chronic mucocutaneous candidiasis. Onychomycosis due to Candida (candidal onychomycosis) is increasingly found in individuals having defective immunity consequential to aging, diabetes mellitus, vascular diseases, HIV infection and drug therapies, chronic exposure to moisture and chemicals including smoke, detergents, soap, etc. Diagnosis of onychomycosis is made by direct microscopic examination and fungal culture but in recent decades molecular techniques have been used to improve onychomycosis diagnosis. In this study we investigated the incidence and identification of Candida species that caused onychomycosis by PCR-RFLP method. Materials and Methods: We isolated three hundred and seventy two Candida species caused onychomycosis in patients through 2009-2011. Yeast genomic DNA was extracted using Whatman FTA-filters and ITS1-ITS2 region was amplified by PCR method. PCR products were digested by the restriction enzyme MspI. For secondary screening, in C. parapsilosis group, SADH gene was amplified and PCR products were digested by the restriction enzyme NlaIII to discriminate C. parapsilosis, C. orthopsilosis and C. metapsilosis. Results: We identified 154 (41.4%) C. albicans, 95 (25.5%) C. parapsilosis, 46 (12.3%) C. tropicalis, 37 (9.9%) C. kefyr, 25 (6.7%) C. krusei, 10 (2.7%) C. glabrata and 5 (1.3%) C. guilliermondii. There was no any C. orthopsilosis and C. metapsilosis among C. parapsilosis group. Totally 41.4% isolates were C. albicans and 58.6% isolates were non-albicans spp. Conclusion: Increasing of non-albicans species and resistance of them to antifungal drugs is an important topic of fungal infections that needed precise epidemiological surveys. Molecular methods like PCR-RFLP can be helpful to reach to this goal. Keywords: Onychomycosis, Candida

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Isolation of fungal agents from burn wounds and determining their susceptibility to antifungal drugs

Neda Kiasat, Ali Zarei Mahmoudabadi, Majid Zarrin, Mahsa Rajabi

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Burning wound provides a proper environment for growth of infectious agents. These infections have been known as the cause of more than 50% of deaths by burning. Saprophytic fungi are the most important causative agents. The aim of this study reviews the causes of fungal infections and determining their susceptibility to antifungal drugs in patients with burn injuries. Materials and Methods: In the present study 104 patients hospitalized due to burns in Taleghani hospital of Ahvaz were sampled. All samples were cultured on the Sabouraud dextrose agar containing chloramphenicole. Fungal agents were detected using morphology, microscopic appearance and additional tests for the detection of yeasts. Determination of drug sensitivity was done with disk diffusion method and sensitivity to common antifungal agents determined. Results: In our study fungal agents were detected in 15 cases (14. 42%). Fungi isolated from these patients, including Candida albicans 6 cases (40%), C. glabrata 2 cases (13. 33%), Aspergillus niger 2 cases (13. 33%), C. dubliniensis, A. fumigatus, A. flavus, Penicillium, and Mucor each one 1 case (6. 66%). Children with 13 cases (66. 66%) had the most cases of infection. Of fungal drug resistance, the isolated fungi were one hundred percent resistant to fluconazole. The greatest sensitivity respectively to clotrimazole (46. 6%), terbinafine (33. 33%), miconazole (26. 66%) and nystatin (20%) was observed. Conclusion: Recurring infections in each hospital to identify relevant studies of drug resistance and completely done until positive step in the direction taken to hospital infection control. Keywords: Burning, Fungal agents, Antifungal sensitivity

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Isolation of Malassezia species using modified CHROMagar and confirmation by PCR

Molood Orumishi, Ali Zarei Mahmoudabadi, Hatef Ajoudanifar

Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran, Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: The species of the genus Malassezia are lipophilic and dimorphism yeast that are regarded as part of normal flora of the skin of the humans and warm- blooded animals. These organisms are the causes of pityriasis versicolor and seborrheic dermatitis. All species are identified by morphologic and biochemical features. But these phenotypic methods are time consuming and lacked sufficient distinction. Thus, using molecular techniques, identification of the species is done faster and more correct than phenotypic methods. The purpose of this study was to separate and determine the frequency of Malassezia species using of the well-adjusted CHROMagar culture medium and using of the Nested-PCR in the city of Ahvaz. Materials and Methods: DNA extractions were done on the nine cultivated dimorphism yeast of Malassezia well-adjusted CHROMagar culture medium and then PCR by using primers18SF1 (forward) and 58SR (reverse) were done on the nine separated Malassezia DNA and (ITS1rDNA) was enhanced for all of the different parts of the isolated in the range between 280- 400 base-pairs. Results: Out of 30 cultivated-samples in the well-adjusted CHROMagar culture medium, 29 cases were positive cultured and 1 case was negative cultured. Out of 9 dandruff cases of pityriasis versicolor, 8 cases were bonded and 1 case was unbounded. From 9 recognized cases, 2 cases were Malassezia globosa and M. restricta. Conclusion: The most common species isolated from dandruff were M. globosa and M. restricta. Malassezia species had better and faster cultivation in the well adjusted CHROMagar culture medium in comparison with Dixon agar culture medium and Liming-Notman agar culture medium. It is recommended that the well adjusted CHROMagar culture-medium to be used for identification in most laboratories. Keywords: CHROMagar, PCR, ITS1rDNA, Malassezia, Pityriasis versicolor

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Superficial mycoses in patients who referred to clinic 2 Tabriz University of medical sciences in 2007-2011

Sanam Nami, Mohammad Adibpour, Maryam Hajizadeh

Department of Medical Parasitology and Mycology, School of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran Email: [email protected]

Introduction and Objectives: Superficial mycoses are defined as the development of fungal growth on epithelial tissues such as human hair, skin or nails, without noticeable invasion of living tissue and without apparently provoking an immune response by the host. The prevalence of superficial mycoses infections has risen to such a level that skin mycoses now affect more than 20-25% of the world's population, making them the most frequent form of infection. The aim of this study was to determine the prevalence of superficial fungal infections in patients who referred to clinic 2 Tabriz University of medical sciences in 2007-2011. Materials and Methods: A total of 663 skin and nails samples were positive for superficial fungal infections were collected from patients who referred to clinic 2 Tabriz University of medical sciences. Samples were examined directly with 10% KOH (potassium hydroxide) and cultures were done on Sabouraud dextrose agar (S). Results: Corynebacterium minutissimum remain the most commonly isolated organisms accounting for 306 cases (46. 15%) followed by Malassezia spp. 222 cases (33.48%), Candida 93 cases (14.03%), Malassezia ovalis 28 cases (4.22%) less frequent superficial fungal infections was saprophytes 14 cases (2.11%). Conclusion: Our study reveals the importance of C. minutissimum and the appreciable frequency of Malassezia spp. in patients who referred to clinic 2 Tabriz University of medical sciences Tabriz, Iran. Consideration of the current epidemiologic trends in the incidence of superficial fungal pathogens is of key importance to investigational effort, diagnosis and treatment. Keywords: Superficial mycoses, Skin, Tabriz

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β-tubulin restriction mapping for identification of medically relevant dermatophytes

Mahdi Abastabar, Hossein Mirhendi, Ali Rezaei-Matehkolaei, Koichi Makimura, Mohammad Reza Shidfar, Parivash Kordbacheh

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Email: [email protected]

Introduction and Objectives: Identification of dermatophytes to the species level relying on macro and microscopic properties of the colonies is time-consuming, questioned in many circumstances and requires considerable expertise. Recent advances in molecular biology introduced several rapid and accurate DNA based tools for identification of pathogenic fungi including dermatophytes and in this regard different genetic loci have been proposed as useful markers. In this study, we examined the potency of β-tubulin gene (BT2), a new genetic marker for differentiation of dermatophytes in an in silico and experimental restriction fragment length polymorphism (RFLP) assay. Materials and Methods: BT2 sequences of the clinically relevant dermatophyte species were retrieved from GenBank and subjected to virtual RFLP analysis. Consequently, the BT2 region of some clinical and standard strains was partially amplified using the primers T1 and Bt2b and undertaken to enzymatic digestion followed by agarose gel electrophoresis and final analysis. Results: In the first round of restriction digestion by the enzyme FatI, three species i. e. Microsporum gypseum, M. canis and T. verrucosum produced unique restriction maps while the remaining species including Trichophyton interdigitale, T. rubrum, T. tonsurans, T. schoenleinii, T. violaceum and Epidermophyton floccosum were identified with further restriction digestion by Alw21I, MwoI and HpyCH4V endonucleases. Conclusion: It was shown that a two-step BT2 restriction mapping is an accurate and fast tool for reliable differentiation of the clinically relevant species of pathogenic dermatophytes. Keywords: Dermatophytes, β-tubulin, identification, RFLP

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Use of rolling circle amplification (RCA) to rapidly identify species of Cladophialophora potentially causing human infection

Saham Ansari, Mitra Golehkheyli, Seyed Amir Yazdanparast, Hamid Badali

Department of Medical, Mycology and Parasitology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran Email: [email protected]

Introduction and Objectives: The genus Cladophialophora comprises etiologic agents of disease in immunocompetent patients, ranging from mild cutaneous colonization to cerebral encephalitis, in addition to saprobic species. Due to the high degree of phenotypic similarity between closely related species of the genus, identification problems are imminent. In the present study, we described rapid and sensitive rolling circle amplification (RCA) method based on species-specific padlock probes targeted for the internal transcribed spacer regions of rDNA. Materials and Methods: ITS regions of 12 Cladophialophora species were sequenced and subsequently 10 specific padlock probes were designed for the detection of single nucleotide polymorphisms. The majority of circularizable padlock probes were designed based on single nucleotide polymorphisms (SNPs), while for C. bantiana, C. immunda and C. devriesii were characterized by two or more nucleotides. Results: Individual species-specific probes correctly identified in all ten Cladophialophora species by visualization on 1.2% agarose gels used to verify specificity of probe-template binding; no cross reactivity was observed. Conclusion: Simplicity, sensitivity, robustness and low costs provide RCA a distinct place among isothermal techniques for DNA diagnostics. However restriction and specificity and sensitivity should be lowered and increased, respectively to be useful for a wide variety of clinical applications. Keywords: RCA, Cladophialophora species, Cerebral infection, Black yeasts

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Epidemiology of candidiasis in patients referred to the medical mycology laboratory, Afzalipoor faculty of medicine

Sanaz Hadizadeh, Seyyed Amin Ayatollahi Mousavi

Department of Medical Mycology and Parasitology, Afzalipoor Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran Email: [email protected]

Introduction and Objectives: Candidiasis is a mycosis caused by Candida species, which is of clinical importance due to the increase in resistance yeasts. Candida species are considered as the most important fungal human pathogens, causing a variety of clinical entities ranging from superficial, cutaneous to deep-seated, systemic infections. Generally, the normal adult human has a strong innate immunity to infection by Candida which lives as a commensally yeast on the body surface and mucous membranes. However, it is considered as opportunistic yeast, as well as an emerging pathogen especially in immunocompromized hosts. Materials and Methods: This retrospective study was conducted on all suspected patients in the medical mycology laboratory of Afzalipoor Kerman faculty of medicine for 4 years from March 2007 to March 2011. Direct smear (wet mount and KOH 10% - 20%), staining, culture and molecular procedures were performed to distinguish the species. Results: 511 (28.39%) patients out of 1800 cases were positive for yeast infections. The highest prevalence of infection belongs to the group of less than 25 years old (35.22%). Men and women were involved equally. The most common agents of the candidiasis were Candida albicans (18.4%), C. parapsilosis (13.11%) and C. tropicalis (9.6%), respectively. The most common site of infections was nails (19.17%) and skin (14.09%) and least one was faeces (0.19%). Conclusion: In the present study, the most affected group of age is 20-30 years old. Nails and skin are the most sites of infection. C. albicans is also the most common agent. Keywords: Epidemiology, Candidiasis, Candida albicans

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The survey of presence Candida Spp. individuals skin with clinical manifestation of Acne referred to dermatology clinic of Booalh Sina hospital in Sari

Ayat Nasrollahi Omran, Mohammad Taghi Hedayti, Taibeh Etezadi

Department of Medical Mycology, Faculty of Medical Sciences, Islamic Azad University of Tonekabon Branch, Tonekabon Iran, P. O. Box: Tonekabon 46815-559 Email: [email protected]

Introduction and Objectives: Candida is dimorphic yeast that some of their species are normal flora of human skin. Its pathogenesis factors include the ability to grow in different temperature and pH produce protease and phospholipase enzymes and switching to hyphae form and phenomenon of phenotype change. Candida is one of the etiologic agents of skin disease like cutaneous Candidiasis and its also opportunistic pathogen. The microbial colonization and inflammatory response due to extra cellular exudes and superficial antigens are one of the main etiologies of acnetic lesions. In the other hand it was proven that Candida is capable of producing skin disease and by considering that Mazandaran condition is suitable for skin lesions and Candida growth and also lack of any investigation in this field, this study was made to evaluate the presence of Candida in patients with acne and identify Candida species in inside and surface of acne. Materials and Methods: 125 patients (70 female and 55 male) enrolled in this study. The sample were collected from inside and surface of acne lesions and mounted by KOH to direct microscopic examination. Samples were also cultured on Sabouraud dextrose agar with chloramphenicole (SC) media. The isolated species were identified by morphologic and physiologic examination include: culturing and SCC media and CHROMagar Candida, germ tube test, chlamydospore forming on cornmeal agar, growth in 45οC and identification kit Hicandida. Results: Our finding showed that 57cases were positive for yeast cells in direct microscopic examination. In total, 45 samples were positive in cultures that 11cases isolated from inside acne, include: Candida parapsilosis, C. krusei, C. glabrata, C. dubliniensis, C. kefyr and 34 cases isolated from surface acne include: Candida krusei, C. parapsilosis, C. guilliermondii, C. dubliniensis, C. albicans, C. glabrata, C. kefyr, and C. zeylanoidea. In 5 cases Candida were detected in both inside and surface of acne lesions that in one case the species were identified. Conclusion: In this study a variety of Candida species were isolated from skin acnetic patients but by considering the limitation of the samples in this study that can influence our knowledge and there is no study in this field so it be suggested to design some more specific studies in different areas in the future. Keywords: Candida, Acne, Skin microflora

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A survey on fungal burden in the intensive care units environment in Ahvaz University hospitals, 2011

Nasrin Amirrajab, Ommolbanin Nikraftar, Forogh Ghalavandi, Hamide Bakhtiari Zadeh, Farzane Sharbati, Maryam Hosaini

Department of Laboratory Sciences, School of Paramedical Sciences, Tropical Medicine Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran and Department of Medical Mycology and Parasitology, Invasive Fungi Research Center, Mazandaran University of Medical Sciences, Sari, Iran Email: [email protected]

Introduction and Objectives: Nosocomial infections due to presence of opportunistic fungi are very important in the hospital setting, especially intensive care units (ICUs), because of the increasing numbers of immunosuppressed and severely ill patients. These groups are at the highest risk of suffering from potentially fatal invasive fungal diseases. The objective of this study was to determine the fungi in the air and medical equipment of 5 university hospitals ICUs in Ahvaz, 2011. Materials and Methods: 16 exposed petri dishes with Sabouraud’s dextrose agar plus chloramphenicole (SC) from the air and 132 wet swabs from environmental surface and touch surface of medical instruments were collected from 8 ICUs of 5 Ahvaz university hospitals. Swab samples were cultured on SC and all of the petri dishes were incubated for five to seven days at 27°C. The number of colonies was counted. The isolated fungal genera were identified by morphological features. Results: In the present study, the majority of the samples (63.5%) from the ICUs were positive and 450 colonies from 15 genera were isolated. The remaining samples (36.5%) were free of fungi. The most isolated fungal genera were Cladosporium spp (40%) from the air and the equipment Hospital І, with 119 isolated colonies (26.4%), was the most environmentally contaminated hospitals Conclusion: This study verifies the presence of fungal burden in the air and medical instruments of the ICUs. The fungal burden of indoor air and touch surfaces of medical equipments in ICUS can be the important source of fungal morbidity and mortality in the hospitals and should be considered to design control measures to improve the quality of indoor air of ICU as a potential source of nosocomial fungal infection. Keywords: Fungal burden, Intensive care unit (ICU), Nosocomial infections, Ahvaz

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Identification of fungal keratitis agents in the east of Mazandaran area by PCR and determination of their drug sensitivity

Shaghayegh Nikpour, Ayatollah Nasrollahi Omran, Saeid Mahdavi Omran, Azar Karmi

Department of Medical Mycology, Faculty of Medical Sciences, Islamic Azad University, Tonekabon Branch, Tonekabon, Iran Email: [email protected]

Introduction and Objectives: Fungal infection causes extensive infective ocular ulcer worldwide and can due to morbidity and blindness. Keratitis is prevalent in tropical and developing countries. For diagnosis and treatment fungal infection is one of the most challenging causes of corneal tumult and ulcer. The aim of this study is Identification of Fungal Keratitis Agents in the East area of Mazandaran by PCR and determination of their Drug sensitivity. Materials and Methods: Corneal scraping was collected from 38 patients suspense with keratitis from the east area of Mazandaran that 30 patient certainly recognized keratitis. Samples were used for two purposes. PCR assay was developed to amplify a portion of the fungal 18s ribosome gene. As well as by culture technique for diagnosis of keratitis agents, and the results were compared. To susceptibility of fungi isolated from keratitis to antifungal drugs amphotericin B, itraconazole, voriconazole and posaconazole based on the NCCLS method. Results: of 38 patients enrolled, 30 patients certainly recognized keratitis. 21 (70%) of 30 patients were men and 9 (30%) were women. They were 6 years old to 80 years old (average 10.4 ± 46.7 years old). Fungal culture were positive in 16 (53.3%) of 30 samples and they recognized by subculture 7 cases (43.75%) were Aspergillus niger and 9 cases (56.25%) were Fusarium sp. PCR assay were positive in 22 (73.3%) of 30 and 8 case were negative. And one (3%) of 30were culture positive and PCR were negative. In 7 (23%) specimens were PCR and culture negative,4 of these appear bacterial growth and 3 samples had shown no growth. Based on this study PCR and fungal culture results were matched in 16 (53%) cases. Posaconazole had the lowest MICs and amphotericin B had the highest MICs against A. niger. Itraconazole had no inhibited effect on Fusarium (MIC90 >8). Voriconazole and amphotericin B had the lowest MICs against Fusarium oxosporium. Conclusion: PCR is promising as a mean to diagnose fungal keratitis and offers some advantages over culture methods, including rapid analysis and the ability to analyze species from where they are collected. Keywords: Fungal Keratitis, PCR, MIC, Mazandaran

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Investigation, isolation and identification of cutaneous mycoses in laboratory animals

Seyed Nasser Shariat, Ali Pourahmadi

Razi Vaccine and Serum Research Institute, Karaj, Iran Email: [email protected]

Introduction and Objectives: Today, laboratory animals as important tool in biomedical researches have been used in scientific centers and universities. According to the Federation of European Laboratory Animal Science Association (FELASA) the issue of animal health is very important for certain groups of diseases and causative agents such as some fungi that they be monitored monthly based on once- three month. Materials and Methods: During two years, 20 laboratory animals (guinea pigs, rabbits, rats and mice) that had produced in the breeding section of Razi Institute were randomly selected. All of their suspicious body surface fungal lesions were reviewed, examined and cultured in direct method. Results: Out of 640 animals, 7 rabbits, 10 pigs, five rats and three rats with suspected fungal lesions were selected and the result of test indicated that no any fungal pathogens. Hair Samples from the healthy animals showed some fungal saprophytes including of Penicillium, Aspergillus niger, Rhizopus, yeast (spp), A. fumigatus, Alternaria, Cladosporium in the guinea pigs, rats, rabbits and rats. It should be mentioned that the temperature and humidity in the breeding room was recorded in accordance with the international standards. Conclusion: We suggest that the study to determine the animal body surface normal flora is necessary to be done. Keywords: Animal laboratory, Fungal disease, Cutaneous mycoses

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Frequency of vulvovaginal candidiasis in 19-65 years old women in Lar in 2011

Fatemeh Jourkesh, Zahra Salehi, Babak Moumeni

Larestan Faculty of Medical Sciences Hazrat Zeinab School of Nursing, Shiraz University of Medical Sciences Email: [email protected]

Introduction and Objectives: Vulvovaginal candidiasis is the second common fungal infection among women. Almost 75% of the women in their life have faced it. In the past three decades, the rate of attacking fungal infections has been increasing. The majority of cases of vulvovaginal candidiasis were diagnosed to be Candida albicans. The aim of this study was to investigate the prevalence of vulvovaginal candidiasis in pap smear samples non-pregnant women in 2011 year in Lar. Materials and Methods: In this descriptive study, 3897 Pap smear samples from nonpregnant women with symptoms of vulvovaginal candidiasis have been cleared by KOH 10%; presence of fungal elements was examined such as yeast and hyphae. The analysis of Candida species was done through germ tube and chlamydospore agar tests. Results: 3897 women ageing from 19-65 years were studied in the current study. In this study, among patients with symptoms of abnormal vaginal discharge, burning and itching of the 482 Candida species were separated from the culture medium. Between 69.3% (334) were Candida albicans and 30.7% (148) cases were non-albicans species, respectively. 6 cases of trichomoniasis were isolated from patients. Conclusion: the majority of cases isolated were fungi and therefore people should be strongly informed about fungal infection, symptoms and the precaution measured. Keywords: Vulvovaginal Candidiasis, Candida, Pap smear

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Assessment of oral candidal colonization in diabetic patients and aetiologic factors associated with this colonization.

Kamiar Zomorodian, Hooman Ebrahimi, Fatemeh Kavoosi, Gholamreza Pishdad, Keyvan Pakshir

Department of Medical Mycology and Parasitology, School of Medicine and Basic Sciences in Infectious Diseases Research Center, Shiraz University of Medical Sciences, Shiraz. Iran Email: [email protected]

Introduction and Objectives: It has been reported that poor glycaemic controls may predispose diabetic patients to oral candidiasis. In this study, oral candidal carriage state in diabetics and healthy controls, species diversity and antifungal susceptibility were determined. In addition, the influence of some local aetiologic and systemic factors such as age, smoking habits, wearing of denture, stomatitis symptoms, dry mouth, on candidal carriage rate were investigated. Methods and Materials: Study subjects included 24 patients with diabetes mellitus type I, 112 patients with diabetes mellitus type II and 104 age and sex matched non-diabetic controls. Total glycosylated haemoglobin (HbA1) and fasting blood sugar was measured to assess glycaemic control. After examination of the oral cavity by a dentist, specimens were collected by swabbing the oral mucosa or lesions (if present). Samples were inoculated onto CHROMagar-Candida plates. Morphologic analyses were used to identify potential yeast-positive cultures which were then further characterized using PCR-RFLP analysis. Antifungal susceptibility testing of the yeasts was done in accordance with CLSI document M44-A. Results: In both groups Candida albicans was the predominant species followed in frequency by C. dubliniensis and C. glabrata. We found a direct association between oral candidal colonization and diabetes. Moreover, a direct association was found between blood glucose level (HbA1C) and higher candidal colonization rate. However, no significant difference in the candidal carriage was found between diabetes type-I and type-II. Among the antifungals, the highest rates of resistance was found in itraconazole (6. 2%), followed by ketoconazole (4. 1%) and fluconazole (1. 4%) and nystatin (0. 7%). Discussion: We identified a high prevalence of C. dubliniensis in diabetic patients which might be misdiagnosed as C. albicans. Considering higher candidal colonization in diabetics than normal individuals, controlling the blood glucose and examination of the oral cavities in these patients at frequent intervals is highly recommended. Keywords: Diabetes, Candida, Antifungal

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Palate fistula due to sinonasal mucormycosis, A case report in diabetic patient

Batool Sadeghi-Nejad, Soheila Nickakhlagh

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Mucormycosis is an acute, fatal and opportunistic mycosis infection caused by organisms that belong to the order Mucorales (Rhizopus, Rhizomucor, Absidia, Mucor, Syncephalastrum) of class Zygomycetes. It can present in two clinical forms: localized or disseminated. The form of localized rhinocerebral mucormycosis is often lethal and is associated with ketoacidosis non- controlled diabetic and immunocompromised patients. This survey describes a clinical case of a 15 year-old, diabetic with hard palate fistula due to sinonasal mucormycosis. In CT scan paranasal sinusitis this case, no involvement of orbit or brain was observed. In the direct examination and culture of tissue specimens from sinonasal, the broad, branching, irregularly shaped with thickness wall, non-septate hyphae was observed and in the culture colonies rapidly growth white at first then becoming grayish that it is Rhizopus fungus. Patient was treated with a combination of surgical excision of involved tissues, the washing of sinuses by H2O2 and systemic amphotericin B. via intra venues for six weeks. Mucormycosis of the hard palate is an ominous sign. The authors point out the need to perform in a short time and adequate cure and surgical treatment of mucormycosis, because of the risk of intracranial extension and increasing of mortality. Keywords: Palate, Fistula, Sinonasal, Mucormycosis, Diabetic patient

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Molecular determination of dermatophyte species isolated from cutaneous infections

Arezoo Gheibi, Kambiz Diba, Khosro Hazrati, Davood Jabari

Department of Medical Mycology and Parasitology, School of Medicine, Urmia University of Medical Sciences Email: [email protected]

Introduction and Objectives: The most common cutaneous fungal infections are caused by dermatophyte fungi including Microsporum, Trichophyton and Epidermophyton species. This study identifies the epidemiologic trends and the predominant organisms causing dermatophytosis in Urmia, Iran. Materials and Methods: A total of 357 specimens were collected from clinically suspected dermatophytic lesions. All specimens were transported to the medical mycology lab. A morphological diagnosis was carried out including microscopic examination of KOH smears and use of Sabouraud agar medium with/ without antibiotics: chloramphenicol and cycloheximide. All dermatophyte isolates were identified in the level of species and also the most important species by the molecular method, PCR-RFLP. Results: From all tested 357 clinical specimens 30 dermatophytic infections were detected. The dermatophyte species: Trichophyton mentagrophytes (36%), Microsporum canis (32%), Microsporum gypseum (16%), T. rubrum (4%) and Epidermophyton floccosum (12%) were morphologically identified. Our results showed that some dermatophyte species including T. rubrum and T. mentagrophytes were mainly isolated from foot, where as E. floccosum was predominantly found in groin and hand scraping. Conclusion: In general, use of the conventional and molecular diagnostic methods enables us to identify the most common dermatophytic species as causal agents of various types of Tinea and underlies the epidemiological study of the frequent dermatophytic infections. Keywords: Dermatophytosis, Epidemiology, Molecular method

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Aspergillosis of the paranasal sinuses; report of 7 cases treated by endoscopic sinus surgery (ESS)

Batool Sadeghi-Nejad, Soheila Nikakhlagh

Department of Medical Mycology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Email: [email protected]

Introduction and Objectives: Nowadays, due to the increasing number of immunocompromised patients, the occurrence of mycosis infections have been increased and fungal sinusitis is usually observed in weakened immune system individuals but may appear in healthy people. Aspergillosis of the paranasal sinuses has classified in two original groups, non- invasive "aspergilloma" form and invasive form usually observed in the immunosuppressed patients. Materials and Methods: In this study 34 sinusitis patients attending department of ENT, Emam Khomeiny hospital, Ahvaz Jundishapur University of Medical sciences Ahvaz, Iran were evaluated. Fungal sinusitis was diagnosed based on clinical findings, functional endoscopic sinuses surgery (FESS) of nasal and paranasal sinuses, CTscan and MRI, mycological and histopathological examinations. Results: In histopathological and mycological examinations of tissue specimens endoscopic sinuses of nasal and paranasal, in 7 cases out of 34 cases was observed hyaline, branching septate hyphae with conidiophores and conidia in crusted fungus ball samples. In cultured, Aspergillus flavus and A. niger were isolated microscopically, stained smears from colonies with lacto phenol anillin blue showed conidiophores and the conidia of A. flavus and A. niger. In this study, fungal sinusitis was reported in individuals who were diabetic and immunosuppressed such as kidney transplantation, leukemia and record of nasal polyp surgery and allergic chronic sinusitis. All cases were successfully treated with surgical debridement involved tissue and systemic amphotericin B. Conclusion: We confirmed the importance of early clinical diagnosis and treatment with surgical debridement of infected tissue in nasal cavity and paranasal sinuses combined with amphotericin B in special patients such as diabetic individuals as soon as possible. Keywords: Fungal sinusitis, Aspergillosis, Paranasal sinus

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Epidemiological and microbiological study of fungal disease (inpatient and outpatient) in education hospitals, Kerman, 2012

Azadeh Karamii, Seyyed Amin Ayatollahi Mousavi, Mahboobeh Madani, Sanaz Hadizadeh

Department of Microbiology, Falavarjan Islamic Azad University, Falavarjan, Isfahan, Iran Email: [email protected]

Introduction and Objectives: Fungal disease can cause Acute and chronic lesions that depend on virulence and toxic products of the fungus, host defense and local authority are attacked. In addition, active infections are seen in the form of allergic disease. In this study, attention was paid to prevalence of fungal diseases in inpatient and outpatient in three hospitals in Kerman for six months (from September 2011 to March 2012). Materials and Methods: This descriptive study was performed on 175 patients with suspected fungal infections. Simultaneously direct smear and culture samples were taken for mycology studies and Candida species were identified by diagnostic methods, such as chromium agar and molecular methods Results: 127 cases (72/6%) of inpatient and outpatient hospital infected fungal disease. Fungal agents were isolated from patients, Candida, dermatophytes and saprophytes. Candida albicans that is most fungal specie was isolated of fungal infections. Conclusion: In general, human have a natural resistance to fungal infection and rarely is affected by these diseases. Patients with immunodeficiency diseases caused by deficient T cells that are occur opportunistic fungal infections frequently. Care and treatment as the necessary steps are important to prevent the risk of infection. Keywords: Fungal diseases, Fungal agents, Candida albicans, PCR-RFLP

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Evaluation of genetic diversity by use of DNA sequencing and PCR-AFLP with antifungal susceptibility profile for Hortaea werneckii

Massoud Zanjanchi Nikoo, Hamid Badali

Rasoul Akram Hospital, Social Security Organization, Rasht, Iran Email: [email protected]

Introduction and Objectives: Tinea nigra is a superficial mycosis caused by the melanized, yeast-like fungus Hortaea werneckii. The disorder has long-time been regarded as an infection and therefore H. werneckii was classified as an opportunistic organism. However, tinea nigra is sub clinical, only dead keratin cells on the skin being colonized. H. werneckii show differences in levels of disorder and resistance to antifungal agents. Therefore, it is important to be able to correctly identify the causative organism to the species level. AFLP analysis as an identification method for medically important H. werneckii was investigated. Antifungal therapy is based on the experience of isolated case reports which mostly involved amphotericin B (AmB), itraconazole and 5-flucytosine. Therefore, we have tested a total of 8 new and established antifungal drugs against 29 clinical isolates of this rare neurotropic fungus. The aim of this study is to determine the in vitro activity of eight existing and new antifungal drugs against H. werneckii. Materials and Methods: A collection of 29 isolates of H. werneckii were obtained from the CBS-KNAW Fungal Biodiversity Centre in Utrecht, The Netherlands. MICs were determined for AmB, FLU, ITC, VOC, POS, ISA, TER and MECs for CAS and ANID based on CLSI M38- A2 guidelines. Results: AFLP patterns exhibited very clear differences among H. werneckii with other related species. The resulting MIC90s for H. werneckii strains were as follows, in increasing order: POS (0.125 mg/L); ITC (0.25 mg/L); VOC (0.25 mg/L); ISA (0.25 mg/L); TER (0.5 mg/L); AmB (2 mg/L); ANID (4 mg/L); CAS (4 mg/L) and FLU (32 mg/L). The new agent is avuconazole, which is also available as an intravenous preparation, shows a good efficacy as the same as voriconazole. Conclusion: AFLP might prove to be a reliable method for the identification of H. werneckii against other related strains which is very essential in antifungal susceptibility methods. Posaconazole and itraconazole were the most active drugs with high in vitro activity against H. werneckii. From the echinocandins tested CAS and ANID demonstrated poor in vitro activity, those drugs might not be useful for treating a range of severe fungal infections, either alone or as part of a combination therapy regimen. However, their clinical effectiveness in the treatment of cerebral infection remains to be determined. Keywords: Hortaea werneckii, Antifungal, Genetic diversity

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Professional ethics in health system and primary health care

Abdolhassan Kazemi, Mostafa Kazemi, Najat Fayzollahi, Sadyar Atallo, Mohammad Masodi Far

Medical Ethics and History Research Center, Biotechnology research Center, Tabriz University of Medical Sciences, Tabriz, Iran Email: [email protected]

Traditionally, in professional ethics field, medical ethics has in the past well thought-out the rights of health care staffs and their framework. Previous medical ethics codes (1847 version) of the American Medical Association’s (AMA) Code integrated sections on the obligations of patients and of the public to the occupation. During professional experiences the WMA has assumed some policy agreements and declarations on the rights of physicians and health care system staff (and of course responsibility of them about patients), governments and other sections of societies for example NGOs, charities, etc. These agreements and declarations were included: • The 1984 Statement on Freedom to Attend Medical Meetings • The 1986 Declaration on Physician Independence and Professional Freedom • The 1995 Statement on Professional Responsibility for Standards of Medical Care • The 1997 Declaration Concerning Support for Medical Doctors Refusing to Participate in, or to Condone, the Use of Torture or Other Forms of Cruel, In human or Degrading Treatment • The 2003 Statement on Ethical Guidelines for the International Recruitment of Physicians Despite such support on behalf of physicians and other staffs of health care system is necessary, given the threats and confronts listed above, physicians sometimes need also to be reminded of the rights they enjoy. Public inspections in various societies and countries have constantly shown that health care system staffs are among the most highly considered and confidences professional groups. Usually physicians and other staffs of health care system receive higher than average salary (much higher in some societies). Physicians and other staffs of health system still have an enormous deal of clinical autonomy; many are financed in an exciting search for new information through contribution in workshops, training, research etc. Most significant, they provide services that are of great value to individual patients and to society in broad-spectrum. Few professions and employments have the capacity to be more satisfying than medicine and health care system occupations. Therefore, even in case of concerning complete medical ethical duties, responsibilities and elegances by physicians and other staffs of health care system "may be a small price to pay for all these privileges and satisfied occupation. Keywords: Physicians, Health system, Professional ethic, Primary health care

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