Simple SummaryIn recent years, the use of technologies to remotely monitor animal behaviour has increased. However, when they are used for the first time in a new species, validation is needed. Therefore, a tri-axial accelerometer was used to monitor lesser anteater (Tamandua tetradactyla) behaviour. First, the influence of using a vest on the animals’ behaviour was evaluated. Second, correlations between video recordings and accelerometer data were made, allowing the researchers to obtain summary measures for each behaviour and a threshold to discriminate activity/inactivity events. Sensitivity and precision variables revealed the robustness of accelerometer monitoring. Third, animals responded to a reduction in enclosure complexity by increasing their activity. The relevance of the accelerometer was determined comparing data (activity level and activity cycle) with video recordings. The results indicate that an accelerometer attached to a vest is a reliable technology for monitoring the activity pattern of T. tetradactyla under semi-controlled environmental conditions.Accelerometers are a technology that is increasingly used in the evaluation of animal behaviour. A tri-axial accelerometer attached to a vest was used on Tamandua tetradactyla individuals (n = 10) at Biodiversity Park. First, the influence of using a vest on the animals’ behaviour was evaluated (ABA-type: A1 and A2, without a vest; B, with a vest; each stage lasted 24 h), and no changes were detected. Second, their behaviour was monitored using videos and the accelerometer simultaneously (experimental room, 20 min). The observed behaviours were correlated with the accelerometer data, and summary measures (X, Y and Z axes) were obtained. Additionally, the overall dynamic body acceleration was calculated, determining a threshold to discriminate activity/inactivity events (variance = 0.0055). Then, based on a 24 h complementary test (video sampling every 5 min), the sensitivity (85.91%) and precision (100%) of the accelerometer were assessed. Animals were exposed to an ABA-type experimental design: A1 and A2: complex enclosure; B: decreased complexity (each stage lasted 24 h). An increase in total activity (%) was revealed using the accelerometer (26.15 ± 1.50, 29.29 ± 2.25, and 35.36 ± 3.15, respectively). Similar activity levels were detected using video analysis. The results demonstrate that the use of the accelerometer is reliable to determine the activity. Considering that the zoo-housed lesser anteaters exhibit a cathemeral activity pattern, this study contributes to easily monitoring their activities and responses to different management procedures supporting welfare programs, as well as ex situ conservation.