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Neotropical otters Lontra annectens (Carnivora, Mustelidae) are widely distributed in Central and South America. Studies on the behavior of this species are rare, resulting in the fact that its ethology is one of the lesser known among the mustelids. The Neotropical otter is considered solitary and territorial but not aggressive, and it generally shows a shy and elusive behavior. Here we described the first two documented cases of attack by Neotropical otters on humans. The first occurred in Sacatepéquez, Guatemala, and the second, in La Guajira, Colombia. Respectively, adult female and male humans were injured with differences in the severity of the wounds, clinical course, and outcomes. We recommend further research on the agonistic behavior of Neotropical otters. A protocol for attention to injuries caused by Neotropical otters is suggested. Inglés