Choloepus didactylus is found in tropical moist lowland and montane forest. In Suriname and French Guiana it does not occur in mangroves. Two-toed sloths have nocturnal and solitary habits. Gestation length seems to be approximately 10 months (Eisenberg and Maliniak 1985) but estimates are quite variable.

There is little information on the diet of wild individuals (Chiarello 2008). A rather unusual observation in the Amazon of north-eastern Peru recorded an adult individual descending to the forest floor to feed on a human latrine (Heymann et al. 2011).

Choloepus didactylus ranges through Venezuela (the delta and south of the Río Orinoco) and the Guianas (French Guiana, Guyana, and Suriname) south into Brazil (Maranhão state west along the Rio Amazonas/Solimões) and west into the upper Amazon Basin of Ecuador, Peru, and Bolivia. Additional visual records from southeastern Pará State, Brazil, need to be confirmed. Its southern limit in the western Amazon of Brazil is unclear. It occurs in the southern departments of Colombia, with its northern limit being the departments of Meta and Guainía. It ranges from sea level up to 2,400 m asl (Britton 1941).

Choloepus didactylus is present in many protected areas.