Although never reported in Alabama, 50% of horses had low detectable synergistic hemolysis inhibition (SHI) titers (1:8 to 1:64) and 2.6% of horses had titers ≥1:128. Increasing SHI titers in horses was associated with contact with ruminants. Further evaluation of cross-reaction with phospholipase D exotoxin from C. p. biovar ovis is warranted. Additional research regarding the serological diagnosis of pigeon fever is needed.