With the rapid development of aquaculture, disease-induced mortality has emerged as a significant hurdle, posing unpredictable risks. To mitigate this issue, aquaculturists often resort to antibiotics for disease management (Khan et al., 2020a; Sarkar et al., 2023). Common types include tetracyclines (TCs), β-lactams (BLCs), fluoroquinolones (FQs), phenicols, macrolides antibiotics, sulfonamides (SAs) and aminoglycosides (AGSs) antibiotics (Sun et al., 2021). TCs like oxytetracycline (OTC), tetracycline (TC), and gentamicin (GM) are commonly employed to address conditions such as white mouth, white skin, and tail rot in fish populations (Xu et al., 2021). BLCs antibiotics, encompassing penicillin and cephalosporin, play a role in combating bacterial gill rot and white beak disease (Chen et al., 2018). FQs, including enrofloxacin (ENR), ciprofloxacin (CIP) (Yousefi et al., 2024), and norfloxacin (NOR), are favored for their efficacy and broad spectrum of activity against pathogens like Aeromonas and Vibrio (Andrieu et al., 2015). Additionally, SAs and AGSs such as sulfadiazine (SD), sulfamethazine (SMZ), and sulfadimethoxine (SDM), along with aminoglycosides like neomycin (NEO), kanamycin (KAN), and tobramycin (TOB), remain integral to disease prevention and treatment strategies in aquaculture (Santos and Ramos, 2016). Nevertheless, the toxicity of antibiotics can exert a substantial influence on the ecosystem, as evidenced by the contamination of water and soil (Alsubih et al., 2022). Moreover, in the event of misuse, the presence of antibiotic residues and antimicrobial resistance can be instigated, thereby posing a considerable public health risk (Khan et al., 2021; Liu et al., 2020a; Lv et al., 2018; Sarkar et al., 2023). Therefore, efficient and accurate detection of antibiotic residues in aquatic products is crucial for ensuring food safety. Traditional methods for detecting antibiotic residues in aquatic foods include liquid chromatography (HPLC) (Zhang et al., 2019a), liquid or gas chromatography coupled with mass spectroscopy for antibiotic detection (LC/GC–MS) (Aldeek et al., 2015; Piatkowska et al., 2016; Ramadan et al., 2024), Enzyme-linked immunosorbent assay (ELISA) (Fernández et al., 2014; Panda et al., 2010), and other techniques (Bhavadharini et al., 2022; Jiang et al., 2021; Jiang et al., 2018). However, these methods are often limited by expensive equipment, complex pretreatment procedures, insufficient sensitivity, delicate instrumentation, and impracticality for multiple sample analyses (Wu et al., 2023). Thus, there is an urgent need for more effective strategies to analyze antibiotic residues.

Biosensors, featuring intelligent recognition elements, such as enzymes, antibodies, microorganisms, cells, and aptamers offer rapid and accurate monitoring based on colorimetric, electrochemical, and other transducer technologies, marking excellent frontiers in food safety (Khan, 2022; Lan et al., 2017; Sarkar et al., 2023). These biosensors leverage the high sensitivity and specificity of biological interactions to perform complex bio-analytical monitoring, such as microbiological detection and antibiotic residue analysis, addressing the food safety concerns (Lu et al., 2023a; Ravindran et al., 2023). Notably, the advent of biosensor platforms has made the analysis of diverse and trace amounts of antibiotics in aquatic foods more convenient and efficient (Chen et al., 2024; Khan et al., 2020b). Although traditional methods are highly accurate and reproducible, biosensors are constantly innovating towards this. In addition, biosensors have more advantages, such as fast analysis response, which is suitable for rapid screening in the field; high sensitivity, which can satisfy the demand for trace residue detection; and others, such as good portability, low cost, and high-throughput analysis. Thus, biosensors can achieve rapid, highly sensitive, and multiple antibiotic screening without the need for large-scale instruments and professional operators (Cháfer-Pericás et al., 2010; Quesada-González and Merkoçi, 2018; Youn et al., 2019).

Among the biosensors, enzyme biosensors demonstrate superior sensitivity and specificity to a wide range of targets. They have emerged as the increasingly prevalent choice for detecting antibiotic residues in foods. The concept of enzyme biosensors was pioneered by Clark et al., initially focusing on glucose sensing (Zhang et al., 2019b). The traditional enzyme biosensors leverage the natural enzymes' high selectivity and catalytic efficiency to monitor analytes through biochemical reactions (Alvarado-Ramirez et al., 2021). However, the limited availability of natural enzymes with specificity for particular substances has significantly restricted the range of analytes detectable by enzyme biosensors. Consequently, developing smart enzyme biosensor platforms that mimic enzyme-like properties, by integrating the biology recognition elements with catalytic transducers offers an excellent solution for antibiotic detection and food safety assurance (Gupta et al., 2022; Zhang et al., 2019a). Thus, in recent years, mimetic enzyme systems, designed as smart biosensors, boast enzyme-like activity, specific recognition ability, and efficient signal transduction capability, emerging as promising technologies due to their multifaceted advantages (Alvarado-Ramirez et al., 2021).

In this review, we highlight the innovations and advancements of various smart enzyme biosensors designed for detecting antibiotic residues in aquatic foods and related samples like fish pond water from 2014 to 2024. Our analysis specifically addresses the progression of enzyme biosensor technology, providing a systematic summary of their underlying working principles, specific recognition elements and techniques, detection performance, and their applications (Fig. 1). Moreover, the future trends in smart enzyme biosensor development were emphasized, featuring more robust and intelligent systems to enhance detection capabilities of the enzyme biosensors. It is noteworthy that this review provides a comprehensive understanding of the significant potential of enzyme biosensors in detecting antibiotics in aquaculture, providing insights for further development of novel enzyme biosensors for more targets in real samples.