Over the past 15 yr, Salmonella enterica ssp. arizonae (IIIa) 56:z4,z23:- has repeatedly been isolated from individual Crotalus willardi rattlesnakes with progressively debilitating osteomyelitis at the Knoxville Zoological Gardens. In April 2004, the serotype was linked with a fatal case of septicemia in another Crotalus species in this collection. Although the association of IIIa 56:z4,z23:- with disease in this colony of C. willardi is well established, prior disease or isolation of this serotype outside of the C. willardi colony had not been documented previously, and the serotype's distribution throughout the remainder of the Crotalus collection had yet to be determined. Forty-one fecal samples were obtained from each individual (n = 36) or exhibit group (n = 5) of crotalid snakes, representing nine species, housed at the zoo. Salmonella spp. were isolated from every sample, with 21 different serotypes. The 21 serotypes were distributed among S. enterica ssp. I (24%), IIIa (9%), and IIIb (67%). Although not recovered in the primary study, S. arizonae 56:z4,z23:- was recovered from additional samples taken from two C. willardi willardi. Although the overall recovery rate of this serotype from feces has been low, it seems that its distribution among the Crotalus collection at Knoxville Zoological Gardens remains largely restricted to the C. willardi species.