The deteriorating environmental quality requires a rapid in situ real-time monitoring of toxic compounds in environment including water and wastewater. One of the most toxic nitrogen-containing ions is nitrite ion, therefore, it is particularly important to ensure that nitrite ions are completely absent in surface and ground waters as well as in wastewater or, at least, their concentration does not exceed permissible levels. However, no selective ion electrode, which would enable continuous measurement of nitrite ion concentration in wastewater by bioelectrochemical sensor, is available. Microbial fuel cell (MFC)-based biosensor offers a sustainable low-cost alternative to the monitoring by periodic sampling for laboratory testing. It has been determined, that at low (0.01−0.1 mg·L−1) and moderate (1.0−10 mg·L−1) concentration of nitrite ions in anolyte-model wastewater, the voltage drop in MFC linearly depends on the logarithm of nitrite ion concentration of proving the potential of the application of MFC-based biosensor for the quantitative monitoring of nitrite ion concentration in wastewater and other surface water. Higher concentrations (100−1000 mg·L−1) of nitrite ions in anolyte-model wastewater could not be accurately quantified due to a significant drop in MFC voltage. In this case MFC can potentially serve as a bioelectrochemical early warning device for extremely high nitrite pollution.