Redox potential-induced regulation of extracellular polymeric substances in an electroactive mixed community biofilm.

Electroactive biofilms are promising in achieving efficient wastewater treatment and energy conversion in bioelectrochemical systems (BESs). Extracellular polymeric substances (EPS) are important for physical contact with electrode surface and extracellular electron transfer (EET) within biofilm. Redox potential is an important trigger for the regulation of EPS in microbial aggregates, but this yet is lacking for electroactive mixed community biofilms. This study first explored how redox potential affected EPS of electroactive mixed community biofilms, which were cultured in BES reactors with different anode potentials (-0.3 V, 0 V, +0.3 V, +0.6 V vs. SCE) using artificial brewery wastewater as substrate. The anode potential regulated biocurrent generation, overall EPS production, EPS composition and EPS redox properties. The biofilms poised at 0 V exhibited the highest current production (7.2 mA) and EPS redox capacity, while the +0.6 V biofilms had the lowest current production (1.2 mA) with lowest EPS redox capacity. The steady-state current exhibited a significant positive correlation with EPS redox capability, suggesting an important role of EPS in anode potential-dependent current production. Significant positive correlations between proteins or humic substances in EPS and EPS redox properties further verified that EPS redox feature raised from proteins and humic substances. This study provided a potential mechanism that anode potential determined the electroactivity of anode biofilm via regulating EPS composition and redox properties, and will facilitate the use of electroactive biofilms in bioelectrochemical applications.

Buffer capacity regulates the stratification of anode-respiring biofilm during brewery wastewater treatment.

Improving the buffer capacity of the electrolyte can enhance the anode performance in bioelectrochemical systems (BESs). To elucidate the mechanism underlying the facilitated BESs performance, this study used three different anode biofilms cultured with different concentrations of phosphate buffer (5, 50 and 100 mM) to investigate the biofilm response, in terms of the spatial structure of metabolic activity and microbial community, to different buffer capacities. Results showed that the electrochemical activities of the anode biofilms positively correlated with the buffer concentration. The spatial stratification of metabolic activity and microbial community of the anode biofilms were regulated by the buffer capacity, and the spatial microbial heterogeneity of the anode biofilm decreased as the buffer concentration increased. With increasing buffer capacity, Geobacter spp. were enriched in both the inner and outer layers of the biofilm, and the inhibition of methanogens growth improved the COD removal attributed to anode respiration. Additionally, the stimulation of EPS production in biofilms played a role in increasing the electrochemical performance of biofilms by buffer improvement. This study first revealed the regulation of buffer capacity on the stratification of anode biofilm during brewery wastewater treatment, which provided a deep insight into the relation of biofilm structure to its electrochemical properties.

Physiological potential of extracellular polysaccharide in promoting Geobacter biofilm formation and extracellular electron transfer.

Geobacter sulfurreducens biofilms have promising applications in renewable energy, pollutant bioremediation, and bioelectronic applications. Genetically manipulating G. sulfurreducens biofilms is an effective strategy to improve the capacity of extracellular electron transfer (EET). Extracellular polysaccharide, a sticky component surrounding microbes, plays an important role in EET. Herein, we constructed a mutant of G. sulfurreducens strain PCA overexpressing the gene GSU1501 (part of the ATP-dependent exporter of the polysaccharide biosynthesis gene operon), designated strain PCA-1501, to increase EET capacity. Experimental results showed that the overexpression of GSU1501 increased extracellular polysaccharide secretion by 25.5%, which promoted the formation of biofilm with higher thickness and viability, as well as the content of extracellular c-type cytochromes. Compared with the control strain, the mutant showed a higher capacity of Fe(III) oxide reduction and current generation (increased by 20.4% and 22.2%, respectively). Interestingly, the overexpression of GSU1501 hindered the pili formation by reducing the transcription level of pilA; a compensatory relationship between extracellular polysaccharide and pili in promoting biofilm formation deserves further investigation. This study provides a feasible method to promote the EET capacity of G. sulfurreducens biofilms, which benefit their bioelectrochemical applications.

Co-selection of antibiotic-resistant bacteria in a paddy soil exposed to As(III) contamination with an emphasis on potential pathogens.

The increased acquisition of antibiotic resistance by pathogens is a global health concern. The environmental selection of antibiotic resistance can be caused by either antibiotic residues or co-selecting agents such as toxic metal(loid)s. This study explored the potential role of As(III) as a co-selecting driver in the spread of antibiotic resistance in paddy soils. By applying high-throughput sequencing, we found that the diversity and composition of soil microbial communities was significantly altered by As(III) exposure, resulting in an increased proportion of potential pathogens (9.9%) compared to the control soil (0.1%). Meanwhile, a total of 46 As(III)-resistant isolates were obtained from As(III)-exposure soil, among which potential pathogens accounted for 54.3%. These As(III)-resistant bacteria showed a high incidence of resistance to sulfanilamide (100%) and streptomycin (88-93%). The association between antibiotic and As(III) resistances was further investigated in a potentially pathogenic isolate by whole-genome sequencing and a transcription assay. The results showed that As(III) and antibiotic resistance genes might co-occur in a mobile genomic island and be co-regulated by As(III), implying that antibiotic resistance could be co-selected by As(III) via co-resistance and co-regulation mechanisms. Overall, these results suggest that As(III) exposure provides a strong selective pressure for the expansion of soil bacterial resistome.

Stratified microbial structure and activity within anode biofilm during electrochemically assisted brewery wastewater treatment.

In a bioelectrochemical system (BES), microbial community of anode biofilm is crucial to BES performance. In this study, the stratified pattern of community structure and activity of an anode-respiring biofilm in a BES fueled with brewery wastewater was investigated over time. The anode biofilm exhibited a superior performance in the removal of ethanol to that of an open-circuit system. The electrical current density reached a high level of 0.55mA/cm2 with a Coulombic efficiency of 71.4%, but decreased to 0.18mA/cm2 in the late stage of operation. A mature biofilm developed a more active outer layer covering a less active inner core, although the activities of the outer and inner layers of biofilm were similar in the early stage. More Geobacter spp., typical exoelectrogens, were enriched in the outer layer than in the inner layer of biofilm in the early stage, while more Geobacter spp. were distributed in the inner layer than in the outer layer in the late stage. The inactive and Geobacter-occupied inner layer of biofilm might be responsible for the decreased electricity generation from wastewater in the late stage of operation. This study provides better understanding of the effect of anode biofilm structure on BES performance.