In the summer of 2023, ingestion of Astylus atromaculatus (pollen beetle) was linked to spontaneous fatal disease in grazing cattle and sheep in Argentina and Uruguay. While the disease was experimentally reproduced in sheep and guinea pigs in the 1970’s, no experimental reproductions have been attempted in cattle, and controversy exists as to whether this insect is indeed noxious to cattle and at which dose. Here, we demonstrate that A. atromaculatus causes acute fatal disease in Hereford calves at single oral dosages of 2.5, 4.5, 10.0, and 15.0 g of insect/kg body weight. Death or severe disease necessitating euthanasia occurred at 38 to 48 hours postinoculation regardless of the dose, suggesting that the single fatal dosage is likely <2.5 g/kg body weight (this dose representing approximately 850 mL of intact beetles in a 100 kg calf). Clinically, the disease was characterized by acute anorexia, prolonged recumbency, reluctance to move, listlessness/apathy, depression, ruminal hypomotility and tympany, hypothermia, bruxism with frothing at the mouth, and mucoid diarrhea progressing to death. Hematologic and biochemical alterations included hemoconcentration, stress/acute inflammatory leukogram, negative energy balance, and ketosis. The pathological hallmark of this experimental disease is acute necrotizing omaso-reticulo-rumenitis, fibrinohemorrhagic enteritis, and exfoliative colitis with intralesional chitinous insect fragments. While A. atromaculatus might contain a gastrointestinal toxin or pathogen, extensive toxicological testing failed to identify a causative toxin. Other pathomechanisms such as direct physical damage caused by insect fragments on the alimentary tract seem plausible, although further studies are needed to elucidate the pathogenesis of A. atromaculatus-associated disease.