Vector control remains an important strategy in preventing rodent-borne diseases. Studies quantifying the impact of anticoagulant bait use on rodent populations are scarce in tropical settings. This study examined the impact of anticoagulant bait use on three measures of rodent activity in Singapore to inform rodent-borne disease control strategies. Using a controlled interrupted time-series analytical design with negative binomial and linear regression models, the average rodent activity levels were compared in the pre- and post-intervention periods. There was a 62.7% (Incidence Rate Ratio (IRR): 0.373, 95% CI: 0.224, 0.620) reduction in the number of rodents caught, a 25.8-unit (coefficient = -25.829, 95% CI: -29.855, -21.804) reduction in the number of 30 g/unit baits consumed and a 61.9% (IRR: 0.381, 95% CI: 0.218, 0.665) reduction in the number of marred bait stations relative to the pre-intervention period. There was a rise in all three outcome measures within four months after the post-intervention period. This study provided strong evidence that anticoagulant baits substantially reduces rodent activity. The population resurgence after the post-intervention period reinforces the importance of timing the resumption of control measures aimed at reducing rodent-borne disease transmission.