Pesticides, chemical substances extensively employed in agriculture to optimize crop yields, pose potential risks to human and environmental health. Consequently, regulatory frameworks are in place to restrict pesticide residue concentrations in water intended for human consumption. These regulations are implemented to safeguard consumer safety and mitigate any adverse effects on the environment and public health. Although gas chromatography- and liquid chromatography-mass spectrometry (GC–MS and LC–MS) are highly efficient techniques for pesticide quantification, their use is not suitable for real-time monitoring due to the need for sophisticated laboratory pretreatment of samples prior to analysis. Since they would enable analyte detection with selectivity and sensitivity without sample pretreatment, biosensors appear as a promising alternative. These consist of a bioreceptor allowing for specific recognition of the target and of a detection platform, which translates the biological interaction into a measurable signal. As early detection systems remain urgently needed to promptly alert and act in case of pollution, we review here the biosensors described in the literature for pesticide detection to advance their development for use in the field.