Landmark-guided navigation is a common behavioral strategy for way-finding, yet prior studies have not examined how animals collect sensory information to discriminate landmark features. We investigated this question in animals that rely on active sensing to guide navigation. Four echolocating bats (Eptesicus fuscus) were trained to use an acoustic landmark to find and navigate through a net opening for a food reward. In experimental trials, an object serving as a landmark was placed adjacent to a net opening and a distractor object next to a barrier (covered opening). The location of the opening, barrier, and objects were moved between trials, but the spatial relationships between the landmark and opening, and between the distractor and barrier were maintained. In probe trials, the landmark was placed next to a barrier, while the distractor was placed next to the opening, to test whether the bats relied on the landmark to guide navigation. Vocal and flight behaviors were recorded with an array of ultrasound microphones and high-speed infrared motion-capture cameras. All bats successfully learned to use the landmark to guide navigation through the net opening. Probe trials yielded an increase in both the time to complete the task and number of net crashes, confirming that the bats relied largely on the landmark to find the net opening. Further, landmark acoustic distinctiveness influenced performance in probe trials and sonar inspection behaviors. Analyses of the animals’ vocal behaviors also revealed differences between call features of bats inspecting landmarks compared with distractors, suggesting increased sonar attention to objects used to guide navigation.

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