ABSTRACT Background and Objectives: One major source of antibiotic contamination in the sea is from aquaculture. We monitored the concentration of commonly used antibiotic classes and antibiotic resistance genes (tet(M), sul1, sul2 and sul3) in aquaculture farms in Peninsular Malaysia. Methods: Antibiotic residues and resistance genes were quantified using high-performance liquid chromatography and real-time PCR respectively. Risk quotients in European technical guidance document on risk assessment was used to assess the potential environmental risk. Results: We detected 23 antibiotics with tetracyclines, sulfonamides and quinolones were the most frequently detected classes, indicating a wide distribution of antibiotics in Malaysian aquaculture farms. The dendrogram and heatmap revealed three groups of antibiotic concentration patterns but with no differences in the types of antibiotics usage among aquaculture farms. The ARGs (10−3 copies/16S) were detected in >90% of the sites except for sul3. Ciprofloxacin, enrofloxacin, norfloxacin and lincomycin posed risks to cyanobacteria and algae in Kelantan, Perak and Pahang. Conclusion: Relative to Asian aquaculture farms, the residues detected here were at low or moderate levels except for quinolones. This study will be useful to develop effective management of aquaculture wastewater in order to mitigate antibiotic pollution and transmission of ARGs to humans through the food chain.