Circadian use of time is an important, but often neglected, part of an animal's niche. We compared the activity patterns of the Neotropical otter Lontra longicaudis in two different areas in Brazil using camera traps placed at the entrance of holts. We obtained 58 independent photos in the Atlantic Forest (273 camera trap-days) and 46 photos in Pantanal (300 camera trap-days). We observed different kernel density probabilities on these two areas (45.6% and 14.1% overlap between the 95% and 50% density isopleths respectively). We observed the plasticity in Neotropical otter activity behaviour with different activity patterns in the two areas. In the Pantanal, the Neotropical otter selected daylight (Ivlev = 0.23) and avoided night (Ivlev = −0.44), while in the Atlantic Forest it selected dawn (Ivlev = 0.24) and night (Ivlev = 0.14), avoiding daylight (Ivlev = −0.33). We believe that this pattern can be due to human activity or shifts in prey activity.