Crop contamination by soil-borne pathogenic microorganisms often leads to serious infection outbreaks. Plant protection requires disinfection of agricultural lands. The chemical and the physical disinfection procedures have several disadvantages, including an irreversible change in the soil ecosystem. Plasma, the “fourth state of matter” is defined as an ionized gas containing an equal number of negatively and positively charged particles. Cold-plasma technology with air or oxygen as the working gas generates reactive oxygen species, which are found to efficiently eradicate bacteria. In this study, we examined the effect of atmospheric plasma corona discharges on soil bacteria viability. Soil that was exposed to plasma for 60 s resulted in bacterial reduction by two orders of magnitude, from 1.1 × 105 to 2.3 × 103 cells g−1 soil. Exposure for a longer period of 5 min did not lead to further significant reduction in bacterial concentration (a final reduction of only 2.5 orders of magnitude). The bacterial viability was evaluated using a colorimetric assay based on the bacterial hydrogenases immediately after exposure and at selected times during 24 h. The result showed no recovery in the bacterial viability. Plasma discharged directly on bacteria that were isolated from the soil resulted in a reduction by four orders of magnitude in the bacterial concentration compared to untreated isolated bacteria: 2.6 × 10−3 and 1.7 × 10−7, respectively. The plasma-resistant bacteria were found to be related to the taxonomic phylum Firmicutes (98.5%) and comprised the taxonomic orders Bacillales (95%) and Clostridiales (2%). To our knowledge, this is the first study of soil bacteria eradication using plasma corona discharges.

Summary This study focuses on a semi‐single‐chamber microbial electrolysis cell (MEC) based on an anode made of carbon cloth (CCp) combined with stainless‐steel (SS) and encapsulated in a dialysis bag with different molecular weight cut‐offs of 2, 14, and 50 kDa (D2‐, D14‐, and D50‐COMBp, respectively). The encapsulated anode was inoculated with Geobacter sulfurreducens in a small volume directly into the dialysis bag. While the MEC with the non‐encapsulated anode was inoculated into the main chamber volume. The hydrogen evolution rates obtained in the MECs based on D2‐COMBp, D14‐COMBp, and D50‐COMBp anodes were 0.134 ± 0.005, 0.144 ± 0.014, and 0.160 ± 0.009 m3m−2d−1, respectively; while the COMBp led to only 0.122 ± 0.004 m3m−2d−1. When the MEC utilizing D50‐COMBp was fed with wastewater, the current density was only 12.4 ± 0.21 A m−2, compared to feeding with acetate (15.7 ± 0.63 A m−2). The encapsulated anode in the presence of wastewater included 70% of G. sulfurreducens, while the non‐encapsulated included only 58%.

One of the promising technologies that can inactivate microorganisms without heat is pulsed electric field (PEF) treatment. The aim of this study was to examine the influence of PEF treatment (2.9 kV cm−1, 100 Hz, 5000 pulses in trains mode of 500 pulses with a pulse duration of 10 µs) on Saccharomyces cerevisiae eradication and resealing in different conditions, such as current density (which is influenced by the medium conductivity), the sort of medium (phosphate buffered saline (PBS) vs. yeast malt broth (YMB) and a combined treatment of PEF with the addition of preservatives. When the S. cerevisiae were suspended in PBS, increasing the current density from 0.02 to 3.3 A cm−2 (corresponding to a total specific energy of 22.04 to 614.59 kJ kg−1) led to an increase of S. cerevisiae eradication. At 3.3 A cm−2, a total S. cerevisiae eradication was observed. However, when the S. cerevisiae in PBS was treated with the highest current density of 3.3 A cm−2, followed by dilution in a rich YMB medium, a phenomenon of cell membrane resealing was observed by flow cytometry (FCM) and CFU analysis. The viability of S. cerevisiae was also examined when the culture was exposed to repeating PEF treatments (up to four cycles) with and without the addition of preservatives. This experiment was performed when the S. cerevisiae were suspended in YMB containing tartaric acid (pH 3.4) and ethanol to a final concentration of 10% (v/v), which mimics wine. It was shown that one PEF treatment cycle led to a reduction of 1.35 log10, compared to 2.24 log10 when four cycles were applied. However, no synergic effect was observed when the preservatives, free SO2, and sorbic acid were added. This study shows the important and necessary knowledge about yeast eradication and membrane recovery processes after PEF treatment, in particular for application in the liquid food industry.

Pulsed electric fields (PEFs) technology was reported to be useful as a disinfection method in the liquid food industry. This technology may lead to membrane permeabilization and bacterial death. However, resuscitation of viable but non-culturable cells and sublethally injured microorganisms in food was reported to be associated with foodborne outbreaks. The main aim of this study was to investigate the possible recovery of injured PEF-treated bacteria. The PEF treatment of Staphylococcus aureus and Pseudomonas putida led to a reduction of 3.2 log10 and 4.8 log10, respectively. After 5 h, no colony forming units (CFUs) were observed when the bacteria were suspended in phosphate buffer saline (PBS); and for 24 h, no recovery was observed. The PEF-treated S. aureus in brain-heart infusion (BHI) medium were maintained at 1.84 × 104 CFU mL−1 for about 1.5 h. While P. putida decreased to zero CFU mL−1 by the 4th hour. However, after that, both bacteria recovered and began to multiply. Flow cytometry analysis showed that PEF treatment led to significant membrane permeabilization. Mass spectrometry analysis of PEF-treated P. putida which were suspended in BHI revealed over-expression of 22 proteins, where 55% were related to stress conditions. Understanding the recovery conditions of PEF-treated bacteria is particularly important in food industry pasteurization. To our knowledge, this is the first comprehensive study describing the recovery of injured PEF-treated S. aureus and P. putida bacteria.

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.