Aflatoxins are potent carcinogens and produced by almost all Aspergillus parasiticus isolates and about 35% of Aspergillus flavus isolates. Chemical methods are used for detection of aflatoxins in food and feed. These methods cannot detect aflatoxinogenic fungi in samples, which contain undetectable amounts of aflatoxins. The objective of this research work was to ascertain the importance of molecular and microbiological methods in detection of aflatoxinogenic fungus A. parasiticus in food and feed samples in Jordan. Specific media for the detection of aflatoxins showed the prevalence of A. parasiticus (6–22%) in contaminated food and feed samples. HPLC method confirmed the presence of aflatoxins B1, B2, G1, and G2 in food sample contaminated with A. parasiticus. Primer set OmtBII-F and OmtBII-R amplified DNA fragment of 611 base pairs from genomic DNA of aflatoxinogenic A. parasiticus isolated from food and feed samples but could not amplify DNA fragment of nonaflatoxinogenic A. flavus. The results of this study showed the prevalence of aflatoxinogenic A. parasiticus in food and feed samples in Jordan and give further evidence of suitability of microbiological and molecular methods in detection of aflatoxins, which are reliable low-cost approach to determine food and feed biosafety.
The Cauliflower Mosaic Virus 35S promoter sequence, CaMV P-35S, is one of several commonly used genetic targets to detect genetically modified maize and is found in most GMOs. In this research we report the finding of an alternative P-35S sequence and its incidence in GM maize marketed in Jordan. The primer pair normally used to amplify a 123 bp DNA fragment of the CaMV P-35S promoter in GMOs also amplified a previously undetected alternative sequence of CaMV P-35S in GM maize samples which we term V3. The amplified V3 sequence comprises 386 base pairs and was not found in the standard wild-type maize, MON810 and MON 863 GM maize. The identified GM maize samples carrying the V3 sequence were found free of CaMV when compared with CaMV infected brown mustard sample. The data of sequence alignment analysis of the V3 genetic element showed 90% similarity with the matching P-35S sequence of the cauliflower mosaic virus isolate CabbB-JI and 99% similarity with matching P-35S sequences found in several binary plant vectors, of which the binary vector locus JQ693018 is one example. The current study showed an increase of 44% in the incidence of the identified 386 bp sequence in GM maize sold in Jordan’s markets during the period 2009 and 2012.
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