This work aims to contribute to the definition of a protocol for the safe release of Lontra longicaudis in the wild. It is used information from the database of the Animal Refuge Conservationist Breeding Center of the Instituto Ekko Brasil/Projeto Lontra. A proposed enclosure is presented, containing a minimum area of 60 m2, for an individual, with a tank occupying at least 40% of the enclosure, with a depth of 1,5 m, a feeding area of 2 m2, in addition to research actions, social mobilization, environmental education, and public policies. The research seeks to create a protocol adapted to the conditions of the Lontra longicaudis species, developed for the southern region of Brazil. It can serve as a subsidy for the definition of a protocol for this purpose for the neotropical otter. Such a protocol, associated with the eight otter birth successes at the Conservationist Animal Refuge, all reaching adulthood, can serve as an important support for the need to reintroduce otters into places where the species has gone extinct.
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