Antibiotics are causing widespread concern as one of the emerging contaminants. There is the abuse of antibiotics in high-density open aquaculture, and the tailwater is often discharged into surrounding rivers. At the same time, the water replenishment of open aquaculture ponds from nearby rivers containing antibiotic contamination from different sources may result in cross-contamination. However, it is still unclear which pollution intensity is greater in rivers or in open aquaculture. So in this paper, the levels of 20 antibiotics (i.e., Fluoroquinolones (FQs), Sulfonamides (SAs), Tetracyclines (TCs), Macrolides (MLs) and Lincosamides (LCs)) in rivers and high-density open aquaculture ponds were investigated in the Baini River basin in the suburbs of Guangzhou, China. The results showed that norfloxacin (NFX) was the predominant antibiotic in river and aquaculture water, with concentrations ranging from 6.12 to 156.04 ng/L and from 7.47 to 82.62 ng/L in both aquatic systems, respectively. As for the pollution intensity of antibiotics, the annual pollution contribution (28.64 kg/a) of the river water supply to open aquaculture is higher than that (10.81 kg/a) of open aquaculture to the river, which means river pollution has a greater impact on aquaculture ponds. The risk quotient (RQ) showed that the ecological risk of lincomycin (LIN), erythromycin (ERY), sulfamethoxazole (SMX), norfloxacin (NFX), ciprofloxacin (CFX) and chlortetracycline (CTC) in rivers and aquaculture environments had high ecological risks from 1.21 to 1.81. Water interactions with contaminated rivers will result in a corresponding increase in the ecological risk of antibiotics in the aquaculture environment. Overall, according to the results, the risk of polluted rivers to open aquaculture cannot be ignored, and it is recommended that open aquaculture should use these water sources with caution, and that the water quality evaluation of aquaculture water should be increased with monitoring indicators for emerging contaminants such as antibiotics.