Neurotoxicity of mycotoxins produced in vitro by Penicillium roqueforti isolated from maize and grass silage.
Fungal growth in human foods and animal feeds causes profound damage indicating a general spoilage, nutritional losses, and the formation of mycotoxins. Thirty apparently contaminated maize and grass silage samples were analyzed for the presence of total fungi. Penicillium roqueforti were isolated from all (100%) moldy silage samples on general and selective culture media. Furthermore, P. roqueforti-positive samples culture media subjected to the toxin extraction and toxins of patulin, penicillic acid, mycophenolic acid, and roquefortin-C (ROQ-C) were identified by means of high-performance liquid chromatography method. Cytotoxicity of identified toxins was investigated on neuro-2a cells. Alamar blue reduction, neutral red uptake, and intracellular adenosine triphosphate (ATP) content assays indicated that patulin and ROQ-C exert the strongest and weakest toxicity, respectively. Reactive oxygen species (ROS) generation by the toxins-exposed cells was measured, and the results supported the mitochondrial and lysosomal dysfunction and ATP depletion in exposed cells. Our data suggest that P. roqueforti is the widely present mold in analyzed maize and grass silage samples, which is able to produce toxins that cause neurotoxicity. This finding may explain in part some neuronal disorders in animals, which are fed contaminated feedstuffs with mentioned fungus. Moreover, mitochondrial and lysosomal dysfunction, intracellular ATP depletion, and the excessive ROS generation were found as the mechanisms of cytotoxicity for P. roqueforti-produced toxins.
Department of Veterinary Pharmacology and Toxicology, Faculty of Veterinary Medicine, Urmia University, Urmia, Islamic Republic of Iran Department of Pharmacology and Toxicology, Faculty of Pharmacy, Urmia University of Medical Sciences, Urmia, Islamic Republic of Iran Department of Veterinary Pharmacology, Pharmacy and Toxicology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands hassanmalekinejad@yahoo.com.
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H
Malekinejad
Department of Neurosurgery, Urmia University of Medical Sciences, Urmia, Islamic Republic of Iran.
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J
Aghazadeh-Attari
Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Islamic Republic of Iran.
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A
Rezabakhsh
Department of Veterinary Pharmacology and Toxicology, Faculty of Veterinary Medicine, Urmia University, Urmia, Islamic Republic of Iran.
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M
Sattari
Department of Veterinary Pharmacology and Toxicology, Faculty of Veterinary Medicine, Urmia University, Urmia, Islamic Republic of Iran.
B
B
Ghasemsoltani-Momtaz
D000255:Adenosine Triphosphate / Q000378:metabolism; D000821:Animal Feed; D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D005506:Food Contamination; D051379:Mice; D009183:Mycotoxins / Q000633:toxicity*; D009498:Neurotoxins / Q000633:toxicity*; D010407:Penicillium; D006109:Poaceae; D017382:Reactive Oxygen Species / Q000378:metabolism; D003313:Zea mays
D009183:Mycotoxins; D009498:Neurotoxins; D017382:Reactive Oxygen Species; D000255:Adenosine Triphosphate
Cytotoxicity; intracellular ATP; mitochondria dysfunction; neuro-2a cells; silage