Traffic disturbances (i.e. pollution, light, noise, and vibrations) often extend into the area surrounding a road creating a 'road-effect zone'. Habitat within the road-effect zone is degraded or, in severe cases, completely unsuitable for wildlife, resulting in indirect habitat loss. This can have a disproportionate impact on wildlife in highly modified landscapes, where remaining habitat is scarce or occurs predominantly along roadside reserves. In this study, we investigated the road-effect zone for insectivorous bats in highly cleared agricultural landscapes by quantifying the change in call activity with proximity to three major freeways. The activity of seven out of 10 species of bat significantly decreased with proximity to the freeway. We defined the road-effect zone to be the proximity at which call activity declined by at least 20% relative to the maximum detected activity. The overall road-effect zone for bats in this region was 307 m, varying between 123 and 890 m for individual species. Given that this road-effect zone exceeds the typical width of the roadside verges (<50 m), it is possible that much of the vegetation adjacent to freeways in this and similar landscapes provides low-quality habitat for bats. Without accounting for the road-effect zone, the amount of habitat lost or degraded due to roads is underestimated, potentially resulting in the loss of wildlife, ecosystem services and key ecosystem processes (e.g. predator-prey or plant-pollinator interactions) from the landscape. We suggest all future environmental impact assessments include quantifying the road-effect zone for sensitive wildlife, in order to best plan and mitigate the impact of roads on the environment. Mitigating the effects of new and existing roads on wildlife is essential to ensure enough high-quality habitat persists to maintain wildlife populations.