Acinetobacter spp. increasingly have been wreaking havoc in hospitals and communities worldwide. Although much has been reported regarding Acinetobacter isolates responsible for nosocomial infections, little is known about these organisms in correctional facilities. In this study, we performed species identification, examined the antibiotic resistance profiles, and determined the mechanisms of resistance and clonal relationships of 123 Acinetobacter isolates obtained from inmates of 20 California correctional facilities (CCFs). We found that 57.7% of the isolates belong to A. baumannii, followed by isolates of Acinetobacter genomic species 3 (gen. sp. 3; 23.6%) and of Acinetobacter gen. sp. 13TU (10.6%). Multidrug-resistant (MDR) CCF isolates were found in only six CCFs. Additionally, DNA sequences of gyrA and parC genes were consistent with fluoroquinolone (FQ) susceptibility phenotypes. Furthermore, the presence of class 1 integrons was detected in 15 CCF isolates, all of which are MDR. Integron-associated gene cassettes encode several aminoglycoside modification enzymes, which correlate with most of the aminoglycoside-resistant phenotypes. Antimicrobial susceptibility testing in the presence of Phe-Arg-␤-naphthylamide dihydrochloride and 1-(1-naphthylmethyl)-piperazine indicated the involvement of efflux pumps in the FQ resistance of only a few CCF isolates. Finally, genetic profiling showed that there was no evidence of A. baumannii outbreaks in CCFs. Instead, our analyses revealed only limited clonal dissemination of mostly non-MDR A. baumannii strains in a few facilities. This study represents the first report to characterize phenotypic and molecular features of Acinetobacter isolates in correctional facilities, which provides a baseline for monitoring the antimicrobial resistance changes and dissemination patterns of these organisms in such specialized institutions.