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Environmental pollutants vigilance is one of the main problems that the aquaculture industry has to face with the objective to ensure the quality of their products and prevent entrance in the food chain that finally may arrive to the consumer. Contaminants such as hormones, antibiotics or biocides are especially relevant due to their toxicity, pharmacological effect or hormonal activity that can be considered harmful for the final consumer. The contaminants can be detected in the environment where the food is growing, and their concentration can be found (i.e., seawater) in the range of µg·L−1, ng·L−1 or even in lower concentrations. Thus, sensitive and selective methods for their monitoring are required to avoid their arrival in the food chain. Here, the development of a multiplexed amperometric biosensor is described, based on the use of specific antibodies to reach the necessary detectability to measure the targeted contaminants directly in seawater. The multiplexed immunosensor allows the detection of four relevant pollutants, such as el Irgarol 1051, sulfapyridine, chloramphenicol and estradiol, reaching an IC50 of 5.04 ± 0.29, 3.45 ± 0.29, 4.17 ± 0.44 and 5.94 ± 0.28 µg·L−1, directly measured in seawater. [ABSTRACT FROM AUTHOR]