A previous investigation of the safety of Brucella abortus strain RB51 (sRB51) in various nontarget species suggested that Richardson's ground squirrels (Spermophilus richardsonii) may develop persistent infections when orally inoculated with the vaccine. In the present study, sRB51, B. abortus strain 19 (s19), and virulent B. abortus strain 9941 (s9941) were administered orally to Richardson's ground squirrels to further characterize B. abortus infection in this species. Six groups of nongravid ground squirrels were orally inoculated with 6×10⁸ colony forming units (cfu) sRB51 (n=10), 2.5×10⁴ cfu s19 (n=10), 2.5×10⁷ cfu s19 (n=6), 1.3×10⁶ cfu s9941 (n=5), 2.1×10⁸ cfu s9941 (n=5), or vaccine diluent (control; n=4). One of five animals in the lower-dose s19 group and two of three animals in the higher-dose s19 group showed persistence of bacteria in various tissues at 14 wk postinoculation (PI). At 18 wk PI, one of five animals in the sRB51 group and one of five animals in the high-dose s9941 group were culture positive. Although we did detect some persistence of B. abortus strains at 18 wk, we found no evidence of pathology caused by B. abortus strains in nonpregnant Richardson's ground squirrels based on clinical signs, gross lesions, and microscopic lesions.