Microbial electrolysis cells (MECs) are an emerging technology capable of harvesting part of the potential chemical energy in organic compounds while producing hydrogen. One of the main obstacles in MECs is the bacterial anode, which usually contains mixed cultures. Non-exoelectrogens can act as a physical barrier by settling on the anode surface and displacing the exoelectrogenic microorganisms. Those non-exoelectrogens can also compete with the exoelectrogenic microorganisms for nutrients and reduce hydrogen production. In addition, the bacterial anode needs to withstand the shear and friction forces existing in domestic wastewater plants. In this study, a bacterial anode was encapsulated by a microfiltration membrane. The novel encapsulation technology is based on a small bioreactor platform (SBP) recently developed for achieving successful bioaugmentation in wastewater treatment plants. The 3D capsule (2.5 cm in length, 0.8 cm in diameter) physically separates the exoelectrogenic biofilm on the carbon cloth anode material from the natural microorganisms in the wastewater, while enabling the diffusion of nutrients through the capsule membrane. MECs based on the SBP anode (MEC-SBPs) and the MECs based on a nonencapsulated anode (MEC control) were fed with Geobacter medium supplied with acetate for 32 days, and then with artificial wastewater for another 46 days. The electrochemical activity, chemical oxygen demand (COD), bacterial anode viability and relative distribution on the MEC-SBP anode were compared with the MEC control. When the MECs were fed with artificial wastewater, the MEC-SBP produced (at −0.6 V) 1.70 ± 0.22 A m⁻², twice that of the MEC control. The hydrogen evolution rates were 0.017 and 0.005 m³ m⁻³ day⁻¹, respectively. The COD consumption rate for both was about the same at 650 ± 70 mg L⁻¹. We assume that developing the encapsulated bacterial anode using the SBP technology will help overcome the problem of contamination by non-exoelectrogenic bacteria, as well as the shear and friction forces in wastewater plants.

The bacterial anode in microbial fuel cells was modified by increasing the biofilm’s adhesion to the anode material using kaolin and graphite nanoparticles. The MFCs were inoculated with G. sulfurreducens, kaolin (12.5 g·L⁻¹), and three different concentrations of graphite (0.25, 1.25, and 2.5 g·L⁻¹). The modified anode with the graphite nanoparticles (1.25 g·L⁻¹) showed the highest electroactivity and biofilm viability. A potential of 0.59, 0.45, and 0.23 V and a power density of 0.54 W·m⁻², 0.3 W·m⁻², and 0.2 W·m⁻² were obtained by the MFCs based on kaolin–graphite nanoparticles, kaolin, and bare anodes, respectively. The kaolin–graphite anode exhibited the highest Coulombic efficiency (21%) compared with the kaolin (17%) and the bare (14%) anodes. Scanning electron microscopy and confocal laser scanning microscopy revealed a large amount of biofilm on the kaolin–graphite anode. We assume that the graphite nanoparticles increased the charge transfer between the bacteria that are in the biofilm and are far from the anode material. The addition of kaolin and graphite nanoparticles increased the attachment of several bacteria. Thus, for MFCs that are fed with wastewater, the modified anode should be prepared with a pure culture of G. sulfurreducens before adding wastewater that includes non-exoelectrogenic bacteria.

The bacterial anode of microbial electrolysis cells (MECs) is the limiting factor in a high hydrogen evolution reaction (HER). This study focused on improving biofilm attachment to a carbon-cloth anode using an alginate hydrogel. In addition, the modified bioanode was encapsulated by a filter bag that served as a physical barrier, to overcome its low mechanical strength and alginate degradation by certain bacterial species in wastewater. The MEC based on an encapsulated alginate bioanode (alginate bioanode encapsulated by a filter bag) was compared with three controls: an MEC based on a bare bioanode (non-immobilized bioanode), an alginate bioanode, and an encapsulated bioanode (bioanode encapsulated by a filter bag). At the beginning of the operation, the Rct value for the encapsulated alginate bioanode was 240.2 Ω, which decreased over time and dropped to 9.8 Ω after three weeks of operation when the Geobacter medium was used as the carbon source. When the MECs were fed with wastewater, the encapsulated alginate bioanode led to the highest current density of 9.21 ± 0.16 A·m⁻² (at 0.4 V), which was 20%, 95%, and 180% higher, compared to the alginate bioanode, bare bioanode, and encapsulated bioanode, respectively. In addition, the encapsulated alginate bioanode led to the highest reduction currents of (4.14 A·m⁻²) and HER of 0.39 m³·m⁻³·d⁻¹. The relative bacterial distribution of Geobacter was 79%. The COD removal by all the bioanodes was between 62% and 88%. The findings of this study demonstrate that the MEC based on the encapsulated alginate bioanode exhibited notably higher bio-electroactivity compared to both bare, alginate bioanode, and an encapsulated bioanode. We hypothesize that this improvement in electron transfer rate is attributed to the preservation and the biofilm on the anode material using alginate hydrogel which was inserted into a filter bag.

Soil-borne pathogenic microorganisms are known to cause extensive crop losses. Agrobacterium tumefaciens, a member of the Proteobacteria, causes the neoplastic crown gall disease in plants. Plant protection is mainly based on toxic chemicals that are harmful to the environment. The use of cold atmospheric-pressure plasma is an attractive method for microbial eradication. Its antimicrobial mechanism includes the formation of large quantities of reactive oxygen species (ROS). The advantages of eradicating bacteria using cold plasma are not needed for chemicals, short treatment, and environmental temperatures. This study examined the impact of plasma corona discharge exposure on A. tumefaciens viability, membrane permeability, relative cell size, and ROS formation. The results showed that 90 s of plasma exposure led to a reduction by four orders of magnitude when the initial concentration was 1 × 10⁷ CFU/mL and in a dry environment. When the initial concentration was 1 × 10⁶ CFU/mL, 45 s of exposure resulted in total bacterial eradication. In a liquid environment, in an initial concentration of 2.02 × 10⁶ CFU/mL, there was no complete bacterial eradication even at the most prolonged examined exposure (90 s). The influence of plasma treatment on the membrane permeability of A. tumefaciens, and their possible recovery, were analyzed using flow cytometer analysis using propidium iodide (PI). When the plasma-treated bacteria were suspended in Luria–Bertani (LB) (rich medium), the PI-positive count of the plasma-treated bacteria after two hours was 12 ± 3.9%. At the 24th hour, this percentage was only 1.74 ± 0.6%, as the control (0.7 ± 0.1%). These results may indicate the repair of the plasma-treated bacteria that were suspended in LB. At the 24th hour, the relative cell size of the treated bacteria shifted to the right, to ~3 × 10⁴ forward side scatter (FSC), about 0.5-fold higher than the untreated cells. Measurement of the ROS showed that the intracellular fluorescence of the 90-s plasma-treated cells led to significant fluorescence formation of 32 relative fluorescence units (RFU)/cell (9 × 10⁴ fold, compared to the nontreated cells). This study showed that cold plasma is a useful method for A. tumefaciens eradication. The eradication mechanism involves ROS generation, membrane permeability, and changes in cell size.

Sorghum (Sorghum bicolor) has emerged as a promising lignocellulosic substrate for anaerobic digestion; however, its biogas production potential and energy recovery efficiency are strongly influenced by varietal characteristics and operating temperature. In this study, eleven sorghum varieties were evaluated for their biogas production potential, biodigestibility, and theoretical energy recovery efficiency under mesophilic (37 ℃) and thermophilic (55 ℃) conditions. The experiments were conducted in 250 mL batch-scale anaerobic digestors operated for 60 days. A clear difference in biogas yield was observed, with consistently higher yields in thermophilic reactors. Among the evaluated varieties, Combined Sorghum Variety-48 (CSV-48), Combined Sorghum Variety-49 Sweet sorghum (CSV-49SS), yellow sorghum, and white sorghum showed superior biogas productivity and higher total solids, volatile solids, and chemical oxygen demand removal efficiencies. Assuming 50% methane content, Bio-Compressed Natural Gas (Bio-CNG) potential ranged from 36.06 to 83.27 Nm³ t^− 1 FM under thermophilic conditions. Corresponding electricity and heat generation ranged from 146.3 to 337.9 kWh t^− 1 FM. Sorghum-derived Bio-CNG could replace approximately 33.9–78.3 L of diesel or 37.9–87.4 L of petrol per tonne of fresh biomass, reducing CO₂ emissions by 90.9–209.8 kg CO₂-eq. t^− 1 FM on diesel basis and 87.5–201.1 kg CO₂-eq. t^− 1 FM on a petrol basis. These findings highlight that identifying high-performing sorghum varieties with favourable lignocellulosic composition and a balanced C/N ratio can improve process stability, substrate biodegradability, and energy recovery efficiency. Using the identified high-yielding varieties for industrial purposes instead of random varieties can reduce operational variability in sorghum-based biogas systems.