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Open Access Article
1 - Molecular identification of Aspergillus parasiticus and flavus in forage samples by Duplex PCR method and comparison with culture and direct examination
ناهید پدرام، منصور بیات، محمدحسن شاهحسینی، سعید بکائی، محمد قهری .Aspergillosis is one of infectious disease that it causes difficulties for human and poultry. The main aim of this study is the diagnosis of Aspergillus parasiticus and flavus in forage samples by Duplex PCR method and comparison with culture and direct examination. Aft MoreAspergillosis is one of infectious disease that it causes difficulties for human and poultry. The main aim of this study is the diagnosis of Aspergillus parasiticus and flavus in forage samples by Duplex PCR method and comparison with culture and direct examination. After extracting DNAs of standard strains of Aspergillus parasitcus and flavus along with primers of each fungus and PCR testing, genes of each fungus were amplified and PCR product was cloned using TA cloning by pTZ57R plasmid. After optimizing Duplex PCR (D-PCR) monoplex PCR tests, 50 forage samples by Duplex PCR method, culture and direct were tested.In the optimized PCR test, 343bp and 413bp products of Aspergillus parasiticus and flavus were respectively amplified. Fifty samples of forage by Duplex PCR method, culture and direct were tested. 13 samples were positive only by Duplex -PCR and 15 samples by both tests direct and culturing were positive and 26 samples were negative with three methods. Results of McNemar's test is equal about the three methods tests for diagnosing Aspergillus parasiticus and flavus (p=0.824). Molecular methods such as D-PCR are faster techniques than direct testing and culturing for diagnosis of Aspergillus parasiticus and flavus but there aren't difference between three methods for diagnosing Aspergillus parasiticus and flavus but with D-PCR we could diagnosis Aspergillus parasiticus. Manuscript profile -
Open Access Article
2 - جداسازی و شناسایی مولکولی گونههای آسپرژیلوس خوراک دام
سمیرا رنجبر راظیه نظری میترا نوری -
Open Access Article
3 - Isolation & Identification of toxigenic Aspergillus flavus, isolated from Rafsanjans, Pistachio by molecular study of nor-1 gene expression effective in the Aflatoxin biosynthetic pathway
مژگان سقازاهAflatoxins are highly toxic and carcinogenic secondary fungal metabolites and have been detected in various food commodities including pistachio nuts. Since Iran is one of the largest producers of pistachios in the world, the aim of this study is investigation of the ex MoreAflatoxins are highly toxic and carcinogenic secondary fungal metabolites and have been detected in various food commodities including pistachio nuts. Since Iran is one of the largest producers of pistachios in the world, the aim of this study is investigation of the expression of nor-1 gene, one of the most essential genes for producing of Aflatoxin, by molecular method in Aspergillus flavus isolated from Rafsanjan’s Pistachio. Randomly, 1000 Pistachios, were collected from 10 dried fruits wholesales in Rafsanjan city. The species of A. flavus were identified morphologically (Macroscopy & Microscopy). In the next step RNA of the A. flavus was extracted. RT-PCR technique was used for amplifying of nor-1 gene. Consequently, 41 Aspergillus flavus species were isolated from pistachioes. Overall, 9 samples (21.4%) were positive for nor-1 gene expression. Aflatoxin contamination in different kinds of foods have been reported in different part of the world. It is a threat for public health, so using surveillance methods, with molecular techniques is necessary to reduction of food contamination specially nuts with toxigenic fungi. Manuscript profile -
Open Access Article
4 - Molecular study of aflR gene in the Aspergillus flavus, isolated from Pistachios of Rafsanjan by RT-PCR
مژگان سقازاه بیتا نجمیAflatoxins are carcinogenic secondary metabolites produced primarily by the fungal species Aspergillus flavus and Aspergillus parasiticus. Foods and feeds especially in warm climates, are susceptible to invasion by aflatoxigenic Aspergillus species and the subsequent pr MoreAflatoxins are carcinogenic secondary metabolites produced primarily by the fungal species Aspergillus flavus and Aspergillus parasiticus. Foods and feeds especially in warm climates, are susceptible to invasion by aflatoxigenic Aspergillus species and the subsequent production of aflatoxins during preharvesting, processing, transportation or storage. Since Iran is one of the largest producers of pistachios in the world, the aim of this study is investigation of the expression of aflR gene, one of the most essential genes for producing of Aflatoxin, by molecular method in toxigenic Aspergillus flavus isolated from Rafsanjan’s Pistachio, First time in Iran. Randomly, 1000 Pistachios, were collected from 10 dried fruits wholesales in Rafsanjan city. The species of A. flavus were identified morphologically. In the next step RNA of the A. flavus was extracted. RT-PCR technique was used for amplifying of aflR gene. Consequently, 41 Aspergillus flavus species were isolated from pistachioes. Overall, 9 samples (21.4%) were positive for aflR expression gene. Toxigenic fungus like Aspergillus flavus can grow on nuts which appropriately stored, such as pistachio, corn, peanuts, nuts, wheat and etc. So a molecular method is necessary for exact and rapid detection of these toxigenic fungi. The genes which participate in toxin maturity pathway, like aflR, can be used for valid probing and also were used for this study. Manuscript profile -
Open Access Article
5 - Effects of Calcium Oxide and Radiation on Aspergillus flavus Population and Aflatoxins Concentrations in Corn Grains
E. Baigane ابوالفضل Fadavi H. koohsari< p >Aspergillus flavus is one of the hazardous fungi which appear in poor storage conditions. This mold produces dangerous toxins of aflatoxins in corn grains. Hence the investigation of reduction possibility of its growth and toxins is important. In this researc More< p >Aspergillus flavus is one of the hazardous fungi which appear in poor storage conditions. This mold produces dangerous toxins of aflatoxins in corn grains. Hence the investigation of reduction possibility of its growth and toxins is important. In this research the effects of calcium oxide (0, %0.5 and %1) and gamma ray (0, 5, 10, 15 and 20 KGy) on growth of Aspergillus flavus and levels of aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1 and aflatoxin G2 (AFG2) were investigated. Analysis of variance showed significant effects of irradiation (p < 0.0001), calcium oxide (p < 0.001) and their interactions (p < 0.0001). Aspergillus flavus and AFB1 and AFB2 toxins decreased with increasing irradiation. Further reduction of AFB1 and AFB2 toxins was observed during accompaniment of 0.5% calcium oxide with irradiation. However, simultaneous application of 1% calcium oxide with radiation prevented and reduced more Aspergillus flavus. AFG1 and AFG2 toxins were not detected in any of the samples. Consequently, with considering 10 KGy standard food irradiation limits, it is recommended that 0.5% calcium oxide concentration before storage and irradiation intensity of 10 KGy after storage were applied for corn grains to control mold growth and production of aflatoxin toxins. Manuscript profile -
Open Access Article
6 - Effect of ethanolic leaf extract of Pinus eldarica on inhibiting the growth of Aspergillus flavus and Aspergillus niger in corn biomass under in vitro conditions
بهروز Valipour Barenji رامین Salamatdoust NobarThis experiment was conducted as completely randomized design (CRD) with factorial arrangement (4×3) including three levels of extract and four incubation times (10, 20, 30 and 40 days) to evaluate the effect of alcoholic extract of pine tree leaves under in vitro MoreThis experiment was conducted as completely randomized design (CRD) with factorial arrangement (4×3) including three levels of extract and four incubation times (10, 20, 30 and 40 days) to evaluate the effect of alcoholic extract of pine tree leaves under in vitro conditions to inhibit the growth of Aspergillus flavus and Aspergillus Niger on the biomass of wet corn. Pine tree leaves were collected in autumn and their extract prepared using ethanol and used at concentrations of 0, 1 and 2 percent. Mass commercial corn was purchased from the factory. Ten grams of corn were mixed with 90 ml of saline solution and stirred several times, then serially diluted and the volume of 0.1 ml was placed on the culture medium. The samples were incubated at 25oC for one week and the growth of fungi were checked every 24 hours from the second day of incubation. The fungus colonies were counted visually. The results indicated that 1 and 2% of pine leaf extract effectively reduced the number of fungi colonies at days 10 and 20 of incubation and the growth of fungi were completely inhibited on days 30 and 40 of incubation. The results were similar for both species of fungi. According to the results, pine leaf extract at the level of 1% is recommended for inhibition of fungi growth. Manuscript profile -
Open Access Article
7 - بررسی اثر پلاسمای تخلیه سد دی الکتریک تک قطبی در میکروب زدائی و کیفیت پودر سیر
اشرف السادات حسینی سهیلا عبدی مریم مصلحی شاد -
Open Access Article
8 - Isolation Acinetobacteria spp. from frozen food and study them effects on some pathogenic fungi
fatemeh shahdadi sedigheh kariminejadIn recent years, producing new antifungal drugs has been considered due to the resistance of fungal toxins and their pathogenicity. In this research, the effect of inhibitory growth of Acinetobacter isolated from frozen foods against pathogenic fungi of Aspergillus nige MoreIn recent years, producing new antifungal drugs has been considered due to the resistance of fungal toxins and their pathogenicity. In this research, the effect of inhibitory growth of Acinetobacter isolated from frozen foods against pathogenic fungi of Aspergillus niger (A.niger) and A. flavus has been investigated. A total of 15 frozen food samples were collected from Jiroft city stores and tested for isolation and identification of potential Acinetobacter. A total of 28 bacterial isolates were obtained from frozen food that, from these, two isolates were identified Acinetobacter by biochemical methods. These bacteria were isolated from frozen hamburger and vegetables. The pathogenic fungi of Aspergillus niger and A. flavus were prepared from the center of the Iranian fungus and bacterial collection center. Then, the antagonistic effects of two isolates of Acinetobacter on these fungi were investigated. The results showed no inhibition of A. niger growth by two isolates of Acinetobacter isolated from frozen hamburger and vegetables, but isolates of frozen vegetables showed an inhibitory effect on A. flavus, and its inhibition zone was 11 mm. Finally, the inhibitory bacteria were selected for molecular identification. Based on the results of sequencing blast and phylogenetic tree drawing, the isolate studied was Acinetobacter baumannii. Manuscript profile -
Open Access Article
9 - Effect of Echinacea pupurea essential oil on growth and Aspergillus flavous mycotoxin production
Arezoo Malakouti Anousheh Sharifan Alireza BasiriFoodborne diseases contaminated with mycotoxins are a concern for human health. The use of bioactive compounds to improve food safety and preservation has been considered to reduce the economic losses and risks of these factors. The aim of this study was to use purple c MoreFoodborne diseases contaminated with mycotoxins are a concern for human health. The use of bioactive compounds to improve food safety and preservation has been considered to reduce the economic losses and risks of these factors. The aim of this study was to use purple coneflower (Echinacea purpurea) essential oil as a bio preservative to control the growth of Aspergillus flavus and to prevent the production of aflatoxin B1. For this purpose, minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values were calculated using a disk diffusion method and then the effect of different concentrations of the essential oil on aflatoxin B1 production was evaluated. The amount of aflatoxin B1 production in the YES (yeast extract sucrose) culture media was measured by HPLC. The results of this study showed that MIC and MFC were 10000 ppm and 15000 ppm, respectively. The results of the growth inhibition zone showed that increasing concentration of the essential oil significantly (p<0.05) increased the diameter of the inhibition zone. It was also founded that aflatoxin B1 production was significantly (p<0.05) dependent on purple coneflower concentration so that by increasing of the essential oil concentration the aflatoxin production significantly (p<0.05) decreased. Accordingly, the aflatoxin B1 production by A. flavus was prevented using concentration of 15000 ppm of purple coneflower essential oil. Overall, it can be concluded that purple coneflower essential oil can control the production of aflatoxin by A. flavus to an acceptable level Manuscript profile -
Open Access Article
10 - Comparison of the effect of watery and alcoholic Celery ( Apium graveolens) extraction on the growth of Aspergillus flavus,Trichophyton rubrum and Candida albicans : in vitro
robab ebrahimibarough seyed jamal hashemi roshanak daei Sadegh Khodavisi peghah ardi shima parsayNowadays, a large number of antifungal drugs are produced in the world, but the main issue of drug resistance to these treatments is BASHDV, therefore, new findings or materials that have antifungal effect have always been considered for researchers in different fields. MoreNowadays, a large number of antifungal drugs are produced in the world, but the main issue of drug resistance to these treatments is BASHDV, therefore, new findings or materials that have antifungal effect have always been considered for researchers in different fields.Therefore, this study the antifungal effects of celery extract on the yeast fungi of Candida albicans, Dermatophyte (Trichophyton rubrum) and sproblons (Aspergillus flavus) have been investigated in laboratory conditions. In this experimental study, watey and alcoholic extracts of celery were prepared by maceration method.Antifungal activity of alcoholic and watery extract of the celery plant was evaluated using application diffusion in disk and well diffusion for 3 types of Aspergillus flavus, Trichophyton, and Candida albicans with three replications. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MFC) by Microdilishin method were determined. The results were analyzed using one-way anova. The results showed that the alcoholic extract of the celery plant in comparison with the watery extract has a more inhibitory effect on the laboratory conditions. As a result, the alcoholic extract of the celery plant has a greater antifungal effect. Manuscript profile -
Open Access Article
11 - Assessment of antifungal activity of Rosemary oil extract and its effect on AFL1 gene expression in Aspergillus flavus by Real-Time PCR
Mojtaba Mohammadi Seyyed Jamal Hashemi Sasan Rezaei Mansour BayatBackground & Objectives: Rosemary is a very important medicinal herb which its effect on toxin-causing and pathogenic fungi is not studied very well. This study was aimed to investigate the antifungal effect of the extract of Rosemary extract on various fungal group MoreBackground & Objectives: Rosemary is a very important medicinal herb which its effect on toxin-causing and pathogenic fungi is not studied very well. This study was aimed to investigate the antifungal effect of the extract of Rosemary extract on various fungal groups including Aspergillus flavus, Candida albicans, Epidermophyton floccussom, Trichophyton verrucosum and its effect on AFL1 gene expression in A. flavus using real-time PCR method. Achieving an effective herbal medicine can be significant due to the limited amount of antifungal drugs and the prevalence of antifungal drug resistance. Materials & Methods: First of all A. flavus and C. albicans were cultured on sabouraud dextrose agar (SDA) media and T. verrucosum and E. floccosum were cultured on Mycocell agar media with 0.5 McFarland turbidity standard. Antifungal property of the rosemary extract was investigated using disk diffusion test. Then, the effective Rosemary extract concentration was evaluated using 10 standard tubes and sabouraud dextrose broth. Finally, the effect of Rosemary extract on AFL1 gene expression was examined. Results: Our results indicated that Rosemary extract has an inhibitory effect on various types of fungi so that the mean diameter of the inhibition zone was measured as about 16 to 18 mm. The effective MCI for C. albicans was observed as approximately 4 to 6 mg / L, for A. flavus as 3 to 5 mg /L and for E. floccosum and T. verrucosum as 4 to 6 mg /L. RT-PCR analysis confirmed the inhibitory effect of Rosemary extracts on aflatoxin- producing AFL1 gene expression at the molecular level, very well. Conclusion: The extract of Rosemary can have a considerable inhibitory effect on fungal growth, AFL1 gene expression and aflatoxin production in A. flavus. Manuscript profile -
Open Access Article
12 - Relationship between aflO and aflQ gene expression with aflatoxin B1 by Aspergillus species isolated from animal feed
Nooshin Sohrabi Morteza TaghizadehBackground & Objectives: Aflatoxins are produced as toxic secondary metabolites by some Aspergillus species. Since these fungal species can contaminate food and animal feed, microbial contamination monitoring and control is important. In the current study, the MoreBackground & Objectives: Aflatoxins are produced as toxic secondary metabolites by some Aspergillus species. Since these fungal species can contaminate food and animal feed, microbial contamination monitoring and control is important. In the current study, the correlation between expression of two aflatoxin biosynthetic pathway genes aflO and aflQ and was investigated. Materials & Methods: Sixty-seven different isolates of aflatoxin-producing Aspergillus species selected and the presence. expression of aflO and aflQ gene sequences were assayed by PCR and quantitative Real-Time RT-PCR methods, respectively. In addition, the amount of aflatoxin B1 by high-performance reverse chromatography (RP-HPLC) using a fluorescent detector. Results: Forthy one, out of 67 isolated species had both aflO and aflQ genes. Seventeen samples out of 26 (65%) had aflatoxin B1 levels higher than 5 ng/g and 6 samples (23%) produced high levels of this toxin (more than 100 ng / g) as well. Conclusion: Considering the relative correlation between the expression of selected aflatoxin genes and AFB1 produced in feedstuffs, it seems that the investigation of other aflatoxin-producing genes can lead to a better correlation and complete the results of this study. Manuscript profile -
Open Access Article
13 - Comparison the level of Aflatoxin in different varieties of internal and imported rice in different collection seasons and effect of cooking methods on the level of toxins
Mahmood NajafianBackground and Objectives: Nowadays, rice is the foodstuff for half the populations, worldwide. Rice is exposed to fungal and aflatoxin contaminations like other cereal. This study was aimed to investigate and to compare the amount of aflatoxin in rice samples in Rasht MoreBackground and Objectives: Nowadays, rice is the foodstuff for half the populations, worldwide. Rice is exposed to fungal and aflatoxin contaminations like other cereal. This study was aimed to investigate and to compare the amount of aflatoxin in rice samples in Rasht city and effects of cooking on the level of toxin. Material and Methods: This cross-sectional study was carried out on 72 samples of consumed rice from domestic and imported productions bought from six stores in Rasht city in both summer and winter. At first aflatoxin was extracted using 80% methanol in three different types of samples including raw, boiled and water cooked. Then aflatoxin content was determined in each sample using the ELISA technique. Results: Domestic samples were less contaminated than imported ones. Samples collected in the summer were less contaminated than winter. Also, in all cases, the cooked rice was less contaminated than seen in raw rice. This reduction rate of contamination was more effective in cooked water rice than in boiled. Conclusion: The results showed that all rice was contaminated in different levels. Therefore the needs for constant control and supervision over the contamination of rice must be considered. Since the contamination rate of imported rice was more than domestic one, for the purpose of reduction imported rice, the basic steps in order to increase domestic production of rice should be supported. Manuscript profile -
Open Access Article
14 - The inhibitory effect of mistletoe extract on the growth and sporulation of Aspergillus flavus in laboratory conditions
Siamak SalahiNearly 20 to 45 percent of the world's grains were exposed to mycotoxins produced by the fungus. Aspergillus flavus is one of the fungi that cause widespread contamination of crops and warehouse products. In this research, the antifungal activity of different extracts o MoreNearly 20 to 45 percent of the world's grains were exposed to mycotoxins produced by the fungus. Aspergillus flavus is one of the fungi that cause widespread contamination of crops and warehouse products. In this research, the antifungal activity of different extracts of the semi-parasitic mistletoe (Viscum album) plant on the growth and sporulation of Aspergillus flavus in laboratory conditions based on completely random design with three replications was evaluated. Various solvents including acetone, ethanol, methanol and water were used to extract mistletoe leaves and buds. Antifungal activity of the extracts was performed by diffusion method on the surface of agar. Each of the extracts including 0 (control), 2, 20, 200 and 2000 microliters per 20 milliliters were added to medium and colony growth and sporulation of fungi were evaluated. All experimental treatments showed a significant effect on mycelium growth and sporulation of Aspergillus flavus compared to control at 5% level. Methanol extract in 2000 microliters had the most effect on fungi growth and sporulation, and mistletoe water extract showed the least effect. Moreover, acetone extract in 2000 microliters, inhibited sporulation by 99 %. It can be concluded that the methanol extract of mistletoe can be a more effective antifungal agent to inhibit the growth and sporulation of Aspergillus flavus than other solvents Manuscript profile
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