Bacteriologic cultures of 65 rectal mucosal samples and 335 fecal samples from 53 horses and 5 cattle shedding Salmonella were performed. Salmonella spp were isolated from 34 (52%) rectal mucosal samples, 21 (32%) concurrent fecal samples, and 150 (45%) total fecal samples. The use of rectal mucosal samples when compared with concurrently obtained fecal samples significantly (P less than 0.025) improved the ability to isolate Salmonella spp. Concurrent bacteriologic culture of rectal mucosal samples and fecal samples resulted in 39 (60%) isolations. Compared with a series of fecal samples, Salmonella was isolated significantly more often when rectal mucosa and feces were cultured concurrently. Salmonella was isolated from rectal mucosal samples when it was not isolated from feces.