Clostridium perfringens type A is the most common cause of poultry necrotic enteritis (NE). Of the four “major” toxins, type A strains produce only alpha toxin (CPA), which has long been considered a major factor in pathogenesis of NE. We investigated the virulence for poultry of type A strains from a variety of enteric sources. Newly-hatched Cornish×Rock chicks were fed a low protein diet for one week, a high protein diet for a second week, and then challenged with log-phase cultures of C. perfringens, mixed 3:4 (v/v) with high protein feed. Strain JGS4143 [genotype A, beta2 positive (cpb2 pos ), from a field case of NE] produced gross lesions compatible with NE in >85% of challenged birds. However, strains JGS1714 (enterotoxigenic genotype A, cpb2 pos , human food poisoning), JGS1936 (genotype A, cpb2 neg , bovine neonatal enteritis), JGS4142 (genotype A, cpb2 pos , bovine jejunal hemorrhage syndrome), JGS1473 (genotype A, cpb2 pos , chicken normal flora), JGS1070 (genotype C, cpb2 pos , porcine hemorrhagic enteritis), JGS1882 (genotype A, cpb2 pos , porcine neonatal enteritis), JGS1120 (ATCC 13124, genotype A, cpb2 neg , gas gangrene), JGS4151 (strain 13, genotype A, cpb2 pos , canine), and JGS4303 (SM101, enterotoxigenic genotype A, cpb2 neg , human food poisoning) failed to produce disease. In vivo passage failed to increase virulence of the non-NE strains. NE strains must have specific poultry-associated virulence attributes, such as the recently identified NetB and other factors, which allow for the development of disease.