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<p>Mycotoxin-contamination of food and feed is a distributed worldwide with serious health risks to animals and humans. Aflatoxins (AFs) and ochratoxin A (OTA) are among the most potent carcinogenic agents and several diseases of humans and animals. Therefore, the early detection of contamination of commodities by toxigenic molds should be useful. This study aimed to determine the role of polymerase chain reaction (PCR), high-performance liquid chromatography (HPLC) techniques, Ultraviolet detection (UV) and other rapid assays in distinguishing between toxigenic and non-toxigenic producer isolates. In this work, coconut medium (CAM) 20% was used for detection of toxigenic fungal strains by ultraviolet light (UV). Most isolates of <i>Aspergillus</i>, <i>Penicillium</i> and <i>Monascus</i> genera showed fluorescent rings under ultraviolet light. Furthermore, the detection of aflatoxigenic fungi was confirmed by ammonium hydroxide vapor test on yeast extract sucrose (YES) and Potato dextrose agar (PDA) agar media. <i>Aspergillus flavus</i> isolate 1 (AF1), <i>A. flavus</i> isolate 2 (AF2), <i>A. terrreus</i> (AT), <i>Eurotium chevalieri</i> (EC) and <i>A. ochraceus</i> (AO) appeared red color when their cultures exposed to ammonium hydroxide vapor. HPLC demonstrated the ability of <i>A. niger</i> (AN) and AO to produce OTA had been detected about 33.3 and 28.16 ng/ml respectively. AF1 produced AFB1 (3.17 ng/ml) and AFG2 (1.15 ng/ml). AF2 produced AFB1, AFG1, AFG2 (2.7, 36.0, 23 ng/ml) respectively. Five primer pairs of AFs and OTA were designed. Fungal DNA was extracted and submitted to PCR for amplifying aflatoxins and ochratoxin encoding genes. PCR products showed that AF1 that exhibited the presence of four genes of aflatoxins (AFF, Nor, Omt and AflR), AF2 (AFF and omt), <i>A. flavus</i> isolate 3 (AF3) (AFF and Nor), EC (AFF) and ochratoxin A gene of AO and AN (AoLC35). This work presented the various approaches and methods in the detection of mycotoxins. Hence, PCR protocol should be recommended as a routine technique to detect toxigenic molds food commodities.</p>

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Molecular detection and quantification of aflatoxins and ochratoxin A produced by toxigenic fungi in feedstuff

  • 期刊论文

  • 发表日期：

    2026年1月19日

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  • Mayada F. El-Fawal,
  • Amira A. El-Fallal,
  • Ahmed K. A. El-Sayed,
  • Hoda M. El-Gharabawy

摘要

Mycotoxin-contamination of food and feed is a distributed worldwide with serious health risks to animals and humans. Aflatoxins (AFs) and ochratoxin A (OTA) are among the most potent carcinogenic agents and several diseases of humans and animals. Therefore, the early detection of contamination of commodities by toxigenic molds should be useful. This study aimed to determine the role of polymerase chain reaction (PCR), high-performance liquid chromatography (HPLC) techniques, Ultraviolet detection (UV) and other rapid assays in distinguishing between toxigenic and non-toxigenic producer isolates. In this work, coconut medium (CAM) 20% was used for detection of toxigenic fungal strains by ultraviolet light (UV). Most isolates of Aspergillus, Penicillium and Monascus genera showed fluorescent rings under ultraviolet light. Furthermore, the detection of aflatoxigenic fungi was confirmed by ammonium hydroxide vapor test on yeast extract sucrose (YES) and Potato dextrose agar (PDA) agar media. Aspergillus flavus isolate 1 (AF1), A. flavus isolate 2 (AF2), A. terrreus (AT), Eurotium chevalieri (EC) and A. ochraceus (AO) appeared red color when their cultures exposed to ammonium hydroxide vapor. HPLC demonstrated the ability of A. niger (AN) and AO to produce OTA had been detected about 33.3 and 28.16 ng/ml respectively. AF1 produced AFB1 (3.17 ng/ml) and AFG2 (1.15 ng/ml). AF2 produced AFB1, AFG1, AFG2 (2.7, 36.0, 23 ng/ml) respectively. Five primer pairs of AFs and OTA were designed. Fungal DNA was extracted and submitted to PCR for amplifying aflatoxins and ochratoxin encoding genes. PCR products showed that AF1 that exhibited the presence of four genes of aflatoxins (AFF, Nor, Omt and AflR), AF2 (AFF and omt), A. flavus isolate 3 (AF3) (AFF and Nor), EC (AFF) and ochratoxin A gene of AO and AN (AoLC35). This work presented the various approaches and methods in the detection of mycotoxins. Hence, PCR protocol should be recommended as a routine technique to detect toxigenic molds food commodities.