Microbial biosensors are bioanalytical devices that can measure the toxicity of pollutants or detect specific substances. This is the greatest advantage of microbial biosensors which use whole cells of microorganisms as powerful tools for measuring integral parameters of environmental pollution. This review explores the core principles of microbial biosensors including biofuel devices, emphasizing their capacity to evaluate biochemical oxygen demand (BOD), toxicity, heavy metals, surfactants, phenols, pesticides, inorganic pollutants, and microbiological contamination. However, practical challenges, such as sensitivity to environmental factors like pH, salinity, and the presence of competing substances, continue to hinder their broader application and long-term stability. The performance of these biosensors is closely tied to both technological advancement and the scientific understanding of biological systems, which influence data interpretation and device optimization. The review further examines cutting-edge developments, including the integration of electroactive biofilms with nanomaterials, molecular biology techniques, and artificial intelligence, all of which significantly enhance biosensor functionality and analytical accuracy. Commercial implementations and improvement strategies are also discussed, providing a comprehensive overview of the state-of-the-art in this field. Overall, this work consolidates recent progress and identifies both the potential and limitations of microbial biosensors, offering valuable insights into their future development for environmental monitoring.

Electropolymerized thionine was used as a redox-active polymer to create a two-mediated microbial biosensor for determining biochemical oxygen demand (BOD). The electrochemical characteristics of the conducting system were studied by cyclic voltammetry and electrochemical impedance spectroscopy. It has been shown that the most promising in terms of the rate of interaction with the yeast B. adeninivorans is the system based on poly(thionine), single-walled carbon nanotubes (SWCNT), and neutral red (k_int = 0.071 dm³/(g·s)). The biosensor based on this system is characterized by high sensitivity (the lower limit of determined BOD concentrations is 0.4 mgO₂/dm³). Sample analysis by means of the developed analytical system showed that the results of the standard dilution method and those using the biosensor differed insignificantly. Thus, for the first time, the fundamental possibility of effectively using nanocomposite materials based on SWCNT and the redox-active polymer poly(thionine) as one of the components of two-mediator systems for electron transfer from yeast microorganisms to the electrode has been shown. It opens up prospects for creating stable and highly sensitive electrochemical systems based on eukaryotes.

The growing threat of organic pollution to surface waters necessitates the development of rapid and scalable monitoring tools that transcend the limitations of the standard 5-day biochemical oxygen demand (BOD₅) test. This study presents a novel approach by developing a highly stable and rapid BOD biosensor based on the microorganism Paracoccus yeei, immobilized within a sol–gel-derived xerogel matrix synthesized on a polyhydroxybutyrate (PHB) substrate. The PHB-supported xerogel significantly enhanced microbial viability and sensor stability. This biosensor demonstrated a correlation (R² = 0.93) with the standard BOD₅ method across 53 diverse water samples from the Tula region, Russia, providing precise results in just 5 min. The second pillar of our methodology involved analyzing multi-year Landsat satellite imagery via the Global Surface Water Explorer to map hydrological changes and identify zones of potential anthropogenic impact. The synergy of rapid ground-truth biosensor measurements and remote sensing analysis enabled a comprehensive spatial assessment of water quality, successfully identifying and ranking pollution sources, with wastewater discharges and agro-industrial facilities constituting the most significant factors. This work underscores the high potential of PHB–xerogel composites as efficient immobilization matrices and establishes a powerful, scalable framework for regional environmental monitoring by integrating advanced biosensor technology with satellite observation.

Polyvinyl alcohol (PVA) hydrogels modified through radical polymerization under UV irradiation and Ce⁴⁺ ion treatment were investigated as a potential platform for developing highly sensitive biosensors for rapid biochemical oxygen demand analysis in water. These modifications enhance PVA physicochemical properties, including mechanical strength, stability, and biocompatibility, making it promising for immobilizing microorganisms in bioanalytical systems. A dual-mediator biosensor system using ferrocene (FC) and neutral red (NR) was developed with yeast Blastobotrys adeninivorans immobilized in modified PVA. The FC+NR–B. adeninivorans–PVA–Ce⁴⁺ system exhibited high sensitivity (linear range of 0.1–3.81 mgO₂/dm³), selectivity, and operational stability (up to 37 days service life), outperforming existing analogs. Testing with wastewater confirmed strong correlation with standard BOD₅, highlighting the potential for monitoring water quality. The described radical modification method is a simple and effective approach for creating sensitive and stable biosensors. It opens up new possibilities for environmental monitoring technology.