Seed of 41 accessions of Crotalaria representing 35 species were analyzed for the concentration of total pyrrolizidine alkaloids (PAs) and for oral toxicity to 1-week-old chicks. All accessions were assayed for the presence of monocrotaline and spectabiline by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). High concentrations of PA were found in the seed of Crotalaria spectabilis Roth # CVTSP (3.85%), C. retusa L. (2.69%), C. alata Leveille (1.60%), C. quinquefolia L. (1.19%), and C. argyrolobioides Bak. (1.01%). The seed of Crotalaria argyrolobioides produced toxic signs but no deaths when fed at 10 mg/g of body weight, whereas the seed of the other four species were 100% lethal when fed to chicks in one dose at 2 to 6 mg/g of body weight. Seed of other species contained less than 0.6% PA and were nontoxic to chicks fed one dose at 10 mg/g of body weight. Monocrotaline was identified in 17 accessions and spectabiline in 7. No species contained both monocrotaline and spectabiline.