In Venezuela, Antillean manatees (Trichechus manatus manatus) have declined mainly due to habitat loss and degradation, but other threats to the species persist. The objective of this work was to investigate the manatee distribution and conservation issues in Maracaibo Lake, Venezuela. Aiming to collect evidence on manatee presence, we conducted 404.14 hours of boat-based surveys and observations from the shore during 2003, 2004, 2009, and 2010. Environmental parameters were collected in areas where manatees were observed including pH, temperature, dissolved oxygen, salinity, transparency, and depth. Also, we recorded plants or algae that could serve as potential food for manatees. We developed 96 informal and opportunistic talks with local fishermen, to gather information about manatee occurrence, boat traffic, and people’s perception of the species. Finally, we reviewed published and unpublished documents containing information on manatees in Maracaibo Lake. The resulting database contains information on 39 sightings and 13 reports of dead manatees. The known causes of death were poaching, vandalism, boat collision, and entanglement in fishing nets. The Maracaibo Lake offers many suitable habitats for manatees, but also faces a wide array of conservation problems that represent critical threats to this endangered subspecies. Long-term monitoring of this population and potential sources of mortality is highly needed.