Highly sensitive standardized toxicity biosensors for rapid water quality warning.

Microbial electrochemical sensor (MES) using electroactive biofilm (EAB) as the sensing element represents a broad-spectrum technology for early warning of biotoxicity of water samples. However, its commercial application is impeded by limited sensitivity and repeatability. Here, we proposed a layered standardized EAB (SEAB) with enriched Geobacter anodireducens SD-1 in the inner layer and self-matched outer layer. The SEAB sensors showed a 2.3 times higher sensitivity than conventional EAB acclimated directly from wastewater (WEAB). A highly repeatable response sensitivity was concentrated at 0.011 ± 0.0006 A/m2/ppm in 4 replicated batches of SEAB sensors (R2 > 0.95), highlighting their potential for reliable toxicity monitoring in practical applications. In contrast, the sensing performance of all WEAB sensors was unpredictable. SEAB also exhibited a better tolerance towards low concentration of formaldehyde, with only a 4 % loss in viability. Our findings improved the sensitivity and reproducibility of standardized MES for toxicity early warning.

Understanding the Tail Current Behavior of Electroactive Biofilms Realizes the Rapid Measurement of Biochemical Oxygen Demand.

The use of microbial electrochemical sensors, with electroactive biofilms (EABs) as sensing elements, is a promising strategy to timely measure the biochemical oxygen demand (BOD) of wastewater. However, accumulation of Coulombic yield over a complete degradation cycle is time-consuming. Therefore, understanding the correlation between current output and EAB metabolism is urgently needed. Here, we recognized a tail stage (TS) on a current-time curve according to current increase rate─a period with the least electron harvesting efficiency. EAB adopted a series of metabolic compensation strategies, including slow metabolism of residual BOD, suspended growth, reduced cell activity, and consumption of carbon storage polymers, to cope with substrate deficiency in TS. The supplementary electrons provided by the decomposition of glycogen and fatty acid polymers increased the Coulombic efficiencies of TS to >100%. The tail current produced by spontaneous metabolic compensation showed a trend of convergent exponential decay, independent of BOD concentration. Therefore, we proposed the TS prediction model (TSPM) to predict Coulombic yield, which shortened BOD measurement time by 96% (to ∼0.5 h) with deviation <4 mg/L when using real domestic wastewater. Our findings on current output in TS give insights into bacterial substrate storage and consumption, as well as regulation in substrate-deficient environment, and provide a basis for developing BOD sensors.

Rapid Surface Modification of Stainless Steel 304L Electrodes for Microbial Electrochemical Sensor Application.

Electrogenic microorganisms serve as important biocatalysts for microbial electrochemical sensors (MESes). The electrical signal produced is based on the rate of electron transfer between the microbes and electrodes, which represents the biotoxicity of water. However, existing MESes require complex and sophisticated fabrication methods. Here, several low-cost and rapid surface modification strategies (carbon powder-coated, flame-oxidized, and acid-bleached) have been demonstrated and studied for biosensing purposes. Surface-modified MESe bioanodes were successfully applied to detect multiple model pollutants including sodium acetate, ethanol, thinner, and palm oil mill effluent under three different testing sequences, namely, pollutant incremental, pollutant dumping, and water dilution tests. The carbon powder-coated bioanode showed the most responsive signal profile for all the three tests, which is in line with the average roughness values (Ra) when tested with atomic force microscopy. The carbon powder-coated electrode possessed a Ra value of 0.844, while flame-oxidized, acid-bleached, and control samples recorded 0.323, 0.336, and 0.264, respectively. The higher roughness was caused by the carbon coating and provided adhesive sites for microbial attachment and growth. The accuracy of MESe was also verified by correlating with chemical oxygen demand (COD) results. Similar to the sensitivity test, the carbon powder-coated bioanode obtained the highest R2 value of 0.9754 when correlated with COD results, indicating a high potential of replacing conventional water quality analysis methods. The reported work is of great significance to showcase facile surface modification techniques for MESes, which are cost-effective and sustainable while retaining the biocompatibility toward the microbial community with carbon-based coatings.

Layered Design of a Highly Repeatable Electroactive Biofilm for a Standardized Biochemical Oxygen Demand Sensor.

Microbial electrochemical sensors are promising to monitor bioavailable organics in real environments, but their application is restricted by the unpredictable performance of the electroactive biofilm (EAB), which is randomly acclimated from environmental microflora. With a long-term stable EAB as a template, we successfully designed EAB (DEAB) by the sequential growth of Geobacter anodireducens and automatched microbes, achieving a reproducible high current than those naturally acclimated from wastewater (NEAB). Pre-inoculation of planktonic aerobes as oxygen bioscavengers was necessary to ensure the colonization of Geobacter in the inner layer, and the abundant Geobacter (50%) in DEAB guaranteed 4 times higher current density with a 15-fold smaller variation among 20 replicates than those of NEAB. The sensor constructed with DEAB exhibited a shorter measuring time and a precise biochemical oxygen demand (BOD) measurement with acetate, real domestic wastewater, and supernatant of anaerobic digestion. Here, we for the first time proposed an applicable strategy to standardize EABs for BOD sensors, which is also crucial to ensure a stable performance of all bioelectrochemical technologies.

[Construction and performance analysis of a microbial electrochemical sensor for monitoring heavy metals in water environment].

A microbial fuel cell (MFC)-based microbial electrochemical sensor was developed for real-time on-line monitoring of heavy metals in water environment. The microbial electrochemical sensor was constructed with staggered flow distribution method to optimize the parameters such as external resistance value and external circulation rate. The inhibition of concentration of simulated heavy metal wastewater on voltage under optimal parameters was analyzed. The results showed that the best performance of MFC electrochemical sensor was achieved when the external resistance value was 130 Ω and the external circulation rate was 1.0 mL/min. In this case, the microbial electrochemical sensors were responsive to 1-10 mg/L Cu2+, 0.25-1.25 mg/L Cd2+, 0.25-1.25 mg/L Cr6+ and 0.25-1.00 mg/L Hg2+ within 60 minutes. The maximum rejection rates of the output voltage were 92.95%, 73.11%, 82.76% and 75.80%, respectively, and the linear correlation coefficients were all greater than 0.95. In addition, the microbial electrochemical sensor showed a good biological reproducibility. The good performance for detecting heavy metals by the newly developed microbial electrochemical sensor may facilitate the real-time on-line monitoring of heavy metals in water environment.

Enhancing quorum sensing in biofilm anode to improve biosensing of naphthenic acids.

The feasibility of enhancing quorum sensing (QS) in anode biofilm to improve the quantifications of commercial naphthenic acid concentrations (9.4-94 mg/L) in a microbial electrochemical cell (MXC) based biosensor was demonstrated in this study. First, three calibration methods were systematically compared, and the charging-discharging operation was selected for further experiments due to its 71-227 folds higher electrical signal outputs than the continuous closed-circuit operation and cyclic voltammetry modes. Then, the addition of acylase (5 µg/L) as an exogenous QS autoinducer (acylase) was investigated, which further improved the biosensor's electrical signal output by ∼70%, as compared to the control (without acylase). The addition of acylase increased the relative expression of QS-associated genes (lasR, lasI, rhlR, rhlI, lasA, and luxR) by 7-100%, along with increased abundances of known electroactive bacterial genera, such as Geobacter (from 42% to 47%) and Desulfovibrio (from 6% to 11%). Furthermore, toxicities of different NAs concentrations measured with the Microtox bioassay test were correlated with corresponding electrical signals, indicating that MXC-biosensor can provide a dual platform for rapid assessment of both NA concentrations and NA-associated toxicity.

Enhancing the sensitivity of water toxicity detection based on suspended Shewanella oneidensis MR-1 by reversing extracellular electron transfer direction.

Water toxicity detection is of great significance to ensure the safety of water supply. With suspended electrochemically active bacteria (EAB) as the sensing element, a novel microbial electrochemical sensor (MES) has recently been reported for the real-time detection of water toxicity, but its practical applications need to further improve the sensitivity. Extracellular electron transfer (EET) is an important factor affecting MES performance. In the study, the EET of suspended EAB-based MES was optimized to further enhance the sensitivity. Firstly, by using a model EAB stain Shewanella oneidensis MR-1, it was revealed that the sensitivity was increased at most 2.7 times with inward EET (i.e., cathodic polarization). Then, a novel conjecture based on electron transfer and energy fluxes was proposed and testified to explain this phenomenon. Finally, three key operating parameters of inward EET were orthogonally optimized. The optimized parameters of inward EET included a potential of - 0.5 V, a cell density of 1.8 × 108 CFU/mL, and an electron acceptor concentration of 15 mM.

Construction and performance analysis of a microbial electrochemical sensor for monitoring heavy metals in water environment / 生物工程学报

A microbial fuel cell (MFC)-based microbial electrochemical sensor was developed for real-time on-line monitoring of heavy metals in water environment. The microbial electrochemical sensor was constructed with staggered flow distribution method to optimize the parameters such as external resistance value and external circulation rate. The inhibition of concentration of simulated heavy metal wastewater on voltage under optimal parameters was analyzed. The results showed that the best performance of MFC electrochemical sensor was achieved when the external resistance value was 130 Ω and the external circulation rate was 1.0 mL/min. In this case, the microbial electrochemical sensors were responsive to 1-10 mg/L Cu2+, 0.25-1.25 mg/L Cd2+, 0.25-1.25 mg/L Cr6+ and 0.25-1.00 mg/L Hg2+ within 60 minutes. The maximum rejection rates of the output voltage were 92.95%, 73.11%, 82.76% and 75.80%, respectively, and the linear correlation coefficients were all greater than 0.95. In addition, the microbial electrochemical sensor showed a good biological reproducibility. The good performance for detecting heavy metals by the newly developed microbial electrochemical sensor may facilitate the real-time on-line monitoring of heavy metals in water environment.