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Biological invasions represent a significant threat to the ecological integrity and economic functionality of the Panama Canal watershed. This study reports, for the first time, the occurrence and distribution of four invasive mollusk species, the bivalves Corbicula fluminea and Potamocorbula amurensis, and the gastropods Melanoides tuberculata and Pomacea canaliculata—within Lake Gatun and its surrounding tributaries (La Arenosa and Playa Leona). Monthly sampling conducted throughout 2025 at depths of up to 3 m yielded 9,800 specimens. Taxonomic identification confirmed the presence of M. tuberculata (n = 3,000), P. canaliculata (n = 3,100), C. fluminea (n = 2,500), and P. amurensis (n = 1,200). These species exhibit high reproductive plasticity and rapid colonization rates, potentially altering benthic nutrient cycling and posing a significant biofouling risk to the Panama Canal’s hydraulic infrastructure. Our findings emphasize the urgent need for integrated management strategies and enhanced ballast water regulations to mitigate further ecological displacement in this globally significant transit corridor.