The presence of contaminants in water generates a great concern worldwide. As contaminants, we can refer different classes of chemicals, such as pharmaceuticals, personal care products, heavy metals, and also microorganisms, such as waterborne pathogens. Some of the chemical compounds have the potential to bioaccumulate in the aquatic biota. Hence, the development of simple and portable methods for the detection of contaminants in the aquatic environment can improve their monitoring and, consequently, the study of their environmental impact. In this context, the development of paper-based analytical tools and also of biosensor devices has been exploited for quantitative and semiquantitative analysis of several contaminants in different water matrices. The association of these two analytical strategies can provide the implementation of low-cost, portable, and easily handled methods for detecting chemical and biological contaminations in water. In this chapter, we provide a review of the developed paper-based analytical biosensors, highlighting the features of the paper-based (paper substrate and fabrication procedures) and biosensor devices (transducers and biorecognition elements). Moreover, the application of the referred paper-based biosensors for the detection of different water contaminants (pathogens, pharmaceuticals, and heavy metals) in environmental and wastewater samples is discussed.