In Colombia the use of glyphosate commercial formulations (Roundup™) for spraying have left deleterious effects on animals and humans. Much of this spraying takes place at the Orinoco basin, habitat of one of the most exported ornamental fish in Colombia, Cardinal neon. To evaluate the effect of Roundup Activo™ four experimental treatments were carried out with 0 mg/L (T1), 0.1 mg/L (T2), 1 mg/L (T3) and 5 mg/L (T4) during 30 days of exposure. The fishes were processed for high-resolution optical microscopy. The main finding of Roundup Activo™ exposure was an increase in mast cells number in brain blood vessels and some neuronal nuclei of the preoptic and posterior diencephalic areas, including hypothalamus. A correlation between concentrations and mast cells number was observed, with the largest mast cells number in T4 treatment. Mast cells presence is a stress benchmark, suggesting the beginning of allergic, inflammatory and apoptotic events. Presence of mast cells in these brain areas may lead to alterations on reproduction, visual and olfactory information integration among other processes. These alterations may result in diminished survival, affecting the conservation of this species in its natural habitat.