Molecular and HPLC-based approaches for detection of aflatoxin B1 and ochratoxin A released from toxigenic Aspergillus species in processed meat

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Abstract Background Meat-products are considered an enriched media for mycotoxins. This study aimed to investigate the prevalence of toxigenicAspergillusspecies in processed meat samples, HPLC-quantitative measurement of aflatoxin B1and ochratoxin A residues, and molecular sequencing ofaflR1andpksgenes. One hundred and twenty processed beef meat specimens (basterma, sausage, and minced meat;n = 40 for each) were collected from Ismailia Province, Egypt. Samples were prepared for total mold count, isolation, and identification ofAspergillusspecies. All samples were analyzed for the production of both Aflatoxin B1and Ochratoxin A mycotoxins by HPLC. Molecular identification ofAspergillus flavusandAspergillus ochraceuswas performed using PCR amplification of the internal transcribed spacer (ITS) region; furthermore, theaflR1andpksgenes were sequenced. Results The total mold count obtained from sausage samples was the highest one, followed by minced meat samples. The prevalence ofA. flavuswas (15%), (7.5%), and (10%), while the prevalence ofA. ochraceuswas (2.5%), (10%), and (0%) in the examined basterma, sausage, and minced meat samples, respectively. Using PCR, the ITS region was successfully amplified in all the testedA. flavusandA. ochraceusstrains. Aflatoxin B1was detected in six basterma samples (15%). Moreover, the ochratoxin A was detected only in four sausage samples (10%). TheaflR1andpksgenes were amplified and sequenced successfully and deposited in the GenBank with accession numbers MF694264 and MF694264, respectively. Conclusions To the best of our knowledge, this is the first report concerning the HPLC-Molecular-based approaches for the detection of aflatoxin B1and ochratoxin A in processed beef meat in Egypt. The production of aflatoxin B1and ochratoxin A in processed meat constitutes a public health threat. Aflatoxin B1is commonly associated with basterma samples. Moreover, ochratoxin A was detected frequently in sausage samples. The routine inspection of mycotoxins in processed meat products is essential to protect human consumers.

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