Food-borne Salmonella infections can produce symptoms from mild gastroenteritis to severe systemic disease and death, representing an important public health issue in U.S. livestock and livestock products, which have been implicated as frequent sources of Salmonella contamination. Concerns have been raised about the spread of antibiotic resistance in Salmonella strains, particularly those that originate from food animal sources, as a result of prophylactic and therapeutic antimicrobial use in these species. Longitudinal comparisons of Salmonella serovars isolated from porcine tissues submitted to the Iowa State University Veterinary Diagnostic Laboratory in 2003 and 2008 were conducted to evaluate changes in serovar dynamics and antimicrobial resistance. Incidence of recovered group C Salmonella enterica serovar Choleraesuis var. Kunzendorf decreased between 2003 and 2008, while recovery of group B strains Salmonella Typhimurium var. 5-(formerly, Copenhagen), Salmonella Agona, Salmonella Derby, Salmonella Heidelberg, and Salmonella Typhimurium increased. Significant changes in resistance interpretation were seen in Salmonella Derby with regard to spectinomycin and sulfadimethoxine; in Salmonella Heidelberg with regard to florfenicol, spectinomycin, and sulfadimethoxine; and in Salmonella Choleraesuis var. Kunzendorf, Salmonella Typhimurium, Salmonella Typhimurium var. 5-, and Salmonella Agona with regard to spectinomycin. Only 2 of 293 isolates in 2003 and 5 of 395 isolates in 2008 were resistant to enrofloxacin. Utilizing antibiotics approved for use in food animals to evaluate antimicrobial resistance provides more specific information on the selection pressure exerted on Salmonella populations through the use of these drugs.