Food and feed are frequently contaminated by numerous regulated and emerging mycotoxins. Humans and animals are thus exposed daily to mycotoxins through the oral route making of the gut the first and the more exposed tissue. Although many studies have evaluated and demonstrated the impact of mycotoxins on the intestinal epithelial cells (IECs) and on the brain cells, surprisingly only few studies have investigated their impact on cells of the enteric nerve system (ENS). In the present work, we measured the impact of major regulated and emerging mycotoxins (17 mycotoxins in total) on the proliferation and viability of ENS cells in vitro. On the 17 mycotoxins tested, 9 were found active with anti-proliferative and cytotoxic effects observed at doses ranging from 0.19 to 16.4 µM and 0.4 to 38.4 µM, respectively. Mechanistic approaches revealed that toxicity on ENS cells was related to i) alteration of the membrane permeability, ii) ROS production and/or iii) induction of apoptosis/necrosis. Importantly, toxic doses found on ENS cells were compared to toxic doses found on IECs in order to determine if toxicity toward ENS was selective or not. Finally, toxic doses on ENS cells were compared to PMTDI and range of exposure in humans and animals in order to evaluate if ENS’s cells are a realistic target of food-associated mycotoxins and if alterations of ENS participate in the global impact of these toxins on the gut and the full organism.