Giant otters live in social groups, consisting of a mating pair and one or two litters. Groups are territorial and mark their territories often with scent-marks. Our objectives were to evaluate the frequencies of marking and over-marking according to the social status of the individuals and to define the different postures used during the marking. We observed four groups, totaling 25 individuals (five alpha males, four alpha females, seven adult females, one adult male and eight juveniles) with group size ranging between four and 13 individuals. The study was conducted between July 2006 and July 2007 in the Vermelho River and in a stretch of the Miranda River, in the Southern Pantanal. We observed the groups for a total of 2006 min and recorded 95 events of marking totaling 84.9 min. Time spent marking varied between groups and ranged from 4.3 to 44.7 min. The alpha males marked more frequently (62% of marking events, 55 min) than the alpha females (17% of marking events, 13.6 min). Of the 59 events of scent-marking by the alpha males, 32 over-marked the marks of other individuals from the group. Of the 16 events of scent-marking of the alpha females, five over-marked that of other females from the same group. When scent-marking, alpha males used the ‘stepping’ posture most frequently (63%), then ‘fore-paw rubbing’ (24%), ‘latrine use’ (7%), and ‘body rubbing’ (6%). Alpha females used the ‘stepping’ posture most frequently (65%), then ‘latrine use’ (19%) and ‘fore-paw rubbing’ (12%), with only one event of ‘body rubbing’ observed during marking. Subordinate females used the ‘stepping’ posture (76%) and ‘latrine use’ (24%) during marking. Scent-marking can play many roles in mammals and for giant otters, and the main roles appear to be communication of social and sexual status and territorial defense.