ABSTRACT. Twelve species from six fungal genera were found to be associated with corn (Zea mays L.) grain samples collected from three main regions of Saudi Arabia. The average frequencies of the most common genera were Aspergillus (11.4%), Fusarium (9.5%), Penicillium (5.1%), and Alternaria (5.8%). Fifteen isolates of Aspergillus flavus were screened by HPLC for their ability to produce aflatoxins (AF). The percentage of aflatoxigenic A. flavus isolates was 53%. Eight isolates produced AF, at concentrations ranging 0.7-2.9 ppb. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) molecular markers were used to genetically characterize isolates of A. flavus and to discriminate between the aflatoxigenic and non-aflatoxigenic isolates. RAPD and ISSR analysis revealed a high level of genetic diversity in the A. flavus population, which was useful for genetic characterization. The clustering in the RAPD and ISSR dendrograms obtained was unrelated to geographic origin. The RAPD and ISSR markers could not discriminate between aflatoxigenic and non-aflatoxigenic isolates, but the ISSR primers were somewhat better.

Aspergillus spp. associated with cashew from the regions of Riyadh, Dammam, and Abha were isolated and three different culture media were used to qualitatively measure aflatoxin production by Aspergillus via UV light (365 nm), which was expressed as positive or negative. The obtained data showed that six isolates of A. flavus and four isolates of A. parasiticus were positive for aflatoxin production, while all isolates of A. niger were negative. Five commercially essential oils (thyme, garlic, cinnamon, mint, and rosemary) were tested to determine their influence on growth and aflatoxin production in A. flavus and A. parasiticus by performing high-performance liquid chromatography (HPLC). The results showed that the tested essential oils caused highly significant inhibition of fungal growth and aflatoxin production in A. flavus and A. parasiticus. The extent of the inhibition of fungal growth and aflatoxin production was dependent on the type and concentration of essential oils applied. The results indicate that cinnamon and thyme oils show strong antimicrobial potential. PCR was used with four sets of primer pairs for nor-1, omt-1, ver-1, and aflR genes, enclosed in the aflatoxin biosynthetic pathway. The interpretation of the results revealed that PCR is a rapid and sensitive method.

ABSTRACT. Twelve species belonging to six fungal genera were found to be associated with wheat (Triticum aestivum L.) grain samples collected from three main regions in Saudi Arabia. The most common genera (average frequency) were Aspergillus (14.3%), Fusarium (29.1%), Penicillium (9.3%), and Alternaria (8.2%). Nineteen isolates of Aspergillus flavus were screened for their ability to produce aflatoxins using HPLC. Thirteen isolates produced aflatoxins ranging from 0.5 to 2.6 µg/kg. Inter-simple sequence repeats (ISSR), and random amplified polymorphic DNA (RAPD) molecular markers were used, with the aim of genetically characterizing strains of A. flavus to discriminate between aflatoxigenic and non-aflatoxigenic isolates. RAPD and ISSR analysis revealed a high level of genetic diversity in the A. flavus population, useful for genetic characterization. Clustering based on RAPD and ISSR dendograms was unrelated to geographic origin. RAPD and ISSR markers were not suitable to discriminate aflatoxigenic and nonaflatoxigenic isolates, but ISSR primers were better compared to RAPD.

The use of potent fungal mixed cultures is a promising technique for the biodegradation of crude oil. Four isolates of fungi, namely, Alternaria alternata (AA-1), Aspergillus flavus (AF-3), Aspergillus terreus (AT-7), and Trichoderma harzianum (TH-5), were isolated from date palm soil in Saudi Arabia. The mixed fungal of the four isolates have a powerful tool for biodegradation up to 73.6% of crude oil (1%, w/v) in 14 days. The fungal consortium no. 15 containing the four isolates (1:1:1:1) performed significantly better as a biodegradation agent than other consortium in a variety of environmental factors containing crude oil concentration, incubation temperature, initial pH, biodegradation time and the salinity of the medium. The fungal consortium showed better performance in the biodegradation of normal alkanes (n-alkanes) than that of the polycyclic aromatic hydrocarbons (PAHs); the biodegradation efficiency of normal alkanes of the fungal consortium (67.1%) was clearly high than that of the PAHs (56.8%).

Maize is a significant staple crop and utilized in Saudi Arabia as food and feed, but maize is often infected with Aspergillus flavus in tropical and subtropical climates, especially during storage. This study intended at a polyphasic approach, consisting of microscopic morphological, biochemical, and molecular characterizations that were applied to 29 of A. flavus isolates of stored maize, with the goal of characterization and identification of aflatoxigenic and non-aflatoxigenic A. flavus isolates. The technique of real-time PCR (RTi-PCR) was used to detection of A. flavus in stored maize samples, the findings have been very accurate. Centered on macroscopic morphological (primarily colony color and morphology of conidia) and microscopic (morphology of conidia and size) characteristics. Results have shown 23 A. flavus isolates (80%) were categorized as the dark green of colonies also all isolates were rough conidia. The isolates have been two different groups, 16 isolates (62%) had sclerotium-forming and the remaining 13 isolates (38%) had no sclerotium-forming isolates. To the identification of aflatoxigenic isolates of A. flavus in stored maize, we utilized the qualitative methods (easy and inexpensive) like UV test, yellow pigmentation, and ammonia vapor and quantitative method as HPLC (accurate and expensive). the accuracy methods to the identification aflatoxigenicity isolates, vary, and classified in the following descending order: HPLC (100%) > UV method (81%) > yellow pigmentation (YP) and ammonia vapor (AV) (63%). The profile of Aflatoxigenicity of A. flavus isolates by HPLC has been involved in two types first of 11 isolates (38%) have been aflatoxigenic isolates while 18 isolates (62%) were non-aflatoxigenic isolates. The expression of six aflatoxins (AFs) genes ( aflD , aflM , aflO , aflP , aflR , and aflQ ) was estimated using PCR and RT-PCR. PCR of all genes did not correspond to the aflatoxigenic isolates. The transcriptional analysis of aflO and aflQ was a beneficial marker for discriminating aflatoxigenic from non-aflatoxigenic A. flavus isolates. Also, qRT-PCR indicated that non-aflatoxigenic isolates had a high incidence of defect or downregulation in late AF-genes contrast with early AF-genes. therefore, these non-aflatoxigenic isolates could be critical factors for an efficient and competent strategy for the control of aflatoxin contamination pre-harvest can be considered.

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The efficacy of four concentrations of aqueous extracts of 11 local plants in the management of Aspergillus flavus and aflatoxin contamination was investigated by measuring the dry weight of A. flavus. The extracts of Allium sativum gave the best results, decreasing the dry weight of the fungus, followed by Aloe vera, whereas 20% Coriandrum sativum extract had no significant effect on the fungal dry weight. Aqueous 20% extracts of the herb Thymus vulgaris and the rhizome of Zingiber officinalis most strongly inhibited aflatoxin production for B1 (79.1%), followed by the leaf extracts of Olea europaea and Eucalyptus globulus (75.0%), although the effect on A. flavus growth was moderate. The herb T. vulgaris and extract of Ocimum basilicum leaf showed the strongest inhibition of B2 (76.2%). Conversely, the leaf extracts of Zizyphus spina and Cassia italica produced only marginal effects on the percentage of inhibition of aflatoxins B1 and B2. No positive correlation was observed between mycelial growth and aflatoxin production in A. flavus.

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