Animals aggregate to reduce predation risk, facilitate mating, and access resources with patchy distributions. During a long-term study of turtle populations in the Santa Fe River (SFR) ecosystem in northern Florida, we observed a large aggregation of turtles at Gilchrist Blue Springs Park (GBSP) in August–October 2013 and again in March–May 2014. On 8 September 2013, we hand-captured 496 turtles of 5 species in GBSP. The Suwannee cooter (Pseudemys concinna suwanniensis) was the most abundant species in the sample, with 477 individuals representing all demographic groups. Density of this species was 530 turtles/ha and biomass was 2242 kg/ha. We hypothesize that hydrological changes in the SFR basin contributed to the temporary turtle aggregations at GBSP. The 113-km SFR originates as a tannin-stained blackwater stream, but receives input of clear water from ≥ 45 artesian springs in its lower 37 km. Heavy rainfall in the upper SFR basin from Tropical Storm Debby in June 2012 resulted in a large influx of tannic water that overwhelmed the capacity of the springs to dilute the river water. This storm in combination with additional episodes of heavy rainfall and declining spring flows led to an unusually long (34-mo) tannic period in the typically clear lower 37 km of the SFR. The resulting loss of most submerged aquatic macrophytes in the river due to insufficient sunlight may have been the stimulus that led the herbivorous P. c. suwanniensis to seek food in one of the few locations that had abundant submerged aquatic vegetation in 2013 and 2014. Turtles previously marked upriver (to 16 km) and downriver (to 4.6 km) from GBSP were in the aggregation, suggesting the individuals gathered at GBSP represented a large portion of the SFR P. c. suwanniensis population.