Roads disrupt wildlife movement by impacting species diversity, population ranges, and mortality rates. To improve wildlife connectivity, the California Department of Fish and Wildlife (CDFW) and Department of Transportation (Caltrans) have each identified “wildlife barriers,” regions where roads are thought to impede movement. These are candidate sites for many millions of dollars worth of mitigation measures, such as wildlife crossing structures. To evaluate these barriers, we compared terrestrial wildlife observations within barrier areas to adjacent control regions. We calculated counts of live observations and roadkills for each barrier-control pair and computed the difference (barrier minus control). Differences were assigned a direction (+, –, 0) and tested for significance (p<0.05) with pairwise Wilcoxon tests. We found that a majority of both Caltrans- and CDFW-identified barriers had significantly higher mammal observations compared to control regions (p<0.05), but no difference in the number of reptile or amphibian observations. Among CDFW barriers, 68% had a significantly higher number of mammal and reptile species than controls. Among Caltrans barriers, 64% had significantly higher reptile roadkills than controls. These results suggest that certain barriers may be worthy of mitigation action, whereas others may need re-evaluation before significant investments of limited state or other funds.