Resumo. Abstract: Aim This study provides the first direct evidence about the location of a spawning site of Brycon orbignyanus (Valenciennes, 1850) in the Middle Uruguay River. Methods Sampling was carried out with a plankton net (500 µm) in November 2017 in the Uruguay River near the confluence with the Piratinim River, southern Brazil. Eggs were sorted in the field, and to confirm species identity we used the live ichthyoplankton technique. Results We captured 47 eggs of B. orbignyanus in the channel of the Uruguay River, characterized by fast flow, shallow depth (approximately 3 m) and rocky bottom, located a few kilometers upstream from its confluence with the Piratinim River. Eggs are spherical, well hydrated, semi-dense, transparent and non-adhesive, with a single membrane and large perivitelline space. Total diameter varied between 3,500 and 4,000 µm. Eggs were in early stages of embryonic development, and cells showed 8, 16 or 32 blastomeres. A total of 16 eggs hatched into larvae in the plastic bags. Larvae hatched approximately 18 hours after incubation, and took 14 days to become juvenile. Conclusion This seems to be the first record of a spawning site of B. orbignyanus in the La Plata River Basin, and points to the relevance of the Middle Uruguay River as a recruitment area for this endangered species.