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Resistance, tolerance, and persistence to antibiotics have mainly been studied at the level of a single microbial isolate. However, in recent years it has become evident that microbial interactions play a role in determining the success of antibiotic treatments, in particular by influencing the occurrence of persistence and tolerance within a population. Additionally, the challenge of resuscitation (the capability of a population to revive after antibiotic exposure) and pathogen clearance are strongly linked to the small size of the surviving population and to the presence of fluctuations in cell counts. Indeed, while large population dynamics can be considered deterministic, small populations are influenced by stochastic processes, making their behaviour less predictable. Our study argues that microbe-microbe interactions within a community affect the mode, tempo, and success of persister resuscitation and that these are further influenced by noise. To this aim, we developed a theoretical model of a three-member microbial community and analysed the role of cell-to-cell interactions on pathogen clearance, using both deterministic and stochastic simulations. Our findings highlight the importance of ecological interactions and population size fluctuations (and hence the underlying cellular mechanisms) in determining the resilience of microbial populations following antibiotic treatment., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Marco Fondi reports financial support was provided by Italian Ministry of University and Research (Grant code: 2022Z88RK4). If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Author(s).)

Cattle manure or its digestate, which often contains antibiotic residues, can be used as an organic fertilizer and copper (Cu) as a fungicide in agriculture. Consequently, both antibiotics and Cu are considered soil contaminants. In this work, microcosms were performed with soil amended with either manure or digestate with Cu and an antibiotic (sulfamethoxazole, SMX) co-presence and the planting of Lactuca sativa. After the addition of the organic amendments, a prompt increase in the microbial activity and at the same time of the sul1 and intI1 genes was observed, although ARGs generally decreased over time. In the amended and spiked microcosms, the microbial community was able to remove more than 99% of SMX in 36 days and the antibiotic did not bioaccumulate in the lettuce. Interestingly, where Cu and SMX were co-present, ARGs (particularly sul2) increased, showing how copper had a strong effect on resistance persistence in the soil. Copper also had a detrimental effect on the plant-microbiome system, affecting plant biomass and microbial activity in all conditions except in a digestate presence. When adding digestate microbial activity, biodiversity and lettuce biomass increased, with or without copper present. Not only did the microbial community favour plant growth, but lettuce also positively influenced its composition by increasing bacterial diversity and classes (e.g., Alphaproteobacteria) and genera (e.g., Bacillus), thus indicating a good-quality soil. KEY POINTS: • Cattle digestate promoted the highest microbial activity, diversity, and plant growth • Cattle digestate counteracted detrimental contaminant effects • Cu presence promoted antibiotic cross-resistance in soil., Competing Interests: Declarations The authors declare no competing interests. The authors declare that they agree with the content and that all gave explicit consent to submit and that they obtained consent from the responsible authorities at the institute/organization where the work has been carried out before the work is submitted and the research study was performed without involving human participants and/or animals., (© 2024. The Author(s).)

Microbial Fuel Cells (MFCs) transform organic matter into electricity through microbial electrochemical reactions catalysed on anodic and cathodic half-cells. Terrestrial MFCs (TMFCs) are a bioelectrochemical system for bioelectricity production as well as soil remediation. In TMFCs, the soil is the ion-exchange electrolyte, whereas a biofilm on the anode oxidises organic matter through electroactive bacteria. Little is known of the overall microbial community composition in a TMFC, which impedes complete exploitation of the potential to generate energy in different soil types. In this context, an experiment was performed to reveal the prokaryotic community structure in single chamber TMFCs with soil in the presence and absence of a municipal waste compost (3% w/v). The microbial community was assessed on the anode and cathode and in bulk soil at the end of the experiment (54 days). Moreover, TMFC electrical performance (voltage and power) was also evaluated over the experimental period, varying the external resistance to improve performance. Compost stimulated soil microbial activity, in line with a general increase in voltage and power. Significant differences were observed in the microbial communities between initial soil conditions and TMFCs, and between the anode, cathode and bulk soil in the presence of the compost. Several electroactive genera (Bacillus, Fulvivirga, Burkholdeira and Geobacter) were found at the anode in the presence of compost. Overall, the use of municipal waste compost significantly increased the performance of the MFCs in terms of electrical power and voltage generated, not least thanks to the selective pressure towards electroactive bacteria on the anode., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Electroactive bacteria (EAB) are natural microorganisms (mainly Bacteria and Archaea ) living in various habitats (e.g., water, soil, sediment), including extreme ones, which can interact electrically each other and/or with their extracellular environments. There has been an increased interest in recent years in EAB because they can generate an electrical current in microbial fuel cells (MFCs). MFCs rely on microorganisms able to oxidize organic matter and transfer electrons to an anode. The latter electrons flow, through an external circuit, to a cathode where they react with protons and oxygen. Any source of biodegradable organic matter can be used by EAB for power generation. The plasticity of electroactive bacteria in exploiting different carbon sources makes MFCs a green technology for renewable bioelectricity generation from wastewater rich in organic carbon. This paper reports the most recent applications of this promising technology for water, wastewater, soil, and sediment recovery. The performance of MFCs in terms of electrical measurements (e.g., electric power), the extracellular electron transfer mechanisms by EAB, and MFC studies aimed at heavy metal and organic contaminant bioremediationF are all described and discussed.

The degradation and bioaccumulation of selected antibiotics such as the sulfonamide sulfamethoxazole (SMX) and the fluoroquinolones enrofloxacin (ENR) and ciprofloxacin (CIP) were investigated in soil microcosm experiments where Lactuca sativa was grown with manure or digestate (1%) and spiked with a mixture of the three antibiotics (7.5 mg/kg each). The soil, rhizosphere and leaf phyllosphere were sampled (at 0 and 46 days) from each microcosm to analyze the antibiotic concentrations, main resistance genes (sul1, sul2, qnrS, aac-(6')-Ib-crand qepA), the intI1and tnpA mobile genetic elements and the microbial community structure.Overall results showed that SMX and CIP decreased (70-85% and 55-79%, respectively), and ENR was quite persistent during the 46-day experiment. In plant presence, CIP and ENR were partially up-taken from soil to plant. In fact the bioaccumulation factors were > 1, with higher values in manure than digestate amended soils. The most abundant gene in soil was sul2 in digestate- and aac-(6')-Ib-cr in the manure-amended microcosms. In soil, neither sulfamethoxazole-resistance (sul1 and sul2), nor fluoroquinolone-resistance (aac-(6')-Ib-cr, qepA and qnrS) gene abundances were correlated with any antibiotic concentration. On the contrary, in lettuce leaves, the aac-(6')-Ib-cr gene was the most abundant, in accordance with the fluoroquinolone bioaccumulation. Finally, digestate stimulated a higher soil microbial biodiversity, introducing and promoting more bacterial genera associated with antibiotic degradation and involved in soil fertility and decreased fluoroquinolone bioaccumulation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Some livestock farms rely on anaerobic digestion (AD) technology for manure disposal, thus obtaining energy (biogas) and fertilizer (digestate). Mixtures of antibiotics used for animal health often occur in organic waste and their possible synergistic/antagonistic effects on microorganisms involved in AD are still poorly studied. This work focuses on the effects of adding ciprofloxacin, alone (5 mg L -1 ) and in combination with sulfamethoxazole (2.5-5-10 mg L -1 ), on AD efficiency and microbial community structure. The experiment consisted of 90-day cattle manure batch tests and antibiotic removal percentages were assessed. Adding antibiotics always promoted CH 4 and H 2 production compared to untreated controls; however, CH 4 production was lowered with the highest ciprofloxacin (CIP) concentrations. The overall results show antibiotic degradation caused by acidogenic Bacteria , and CH 4 was mainly produced through the hydrogenotrophic-pathway by methanogenic Archaea . Shifts in microbial community abundance (DAPI counts) and composition (Illumina-MiSeq and FISH analyses) were observed.

The anionic surfactant sodium lauryl ether sulfate (SLES) is the main component of most commercial foaming agents (FAs) used in the excavation of highway and railway tunnels with Earth pressure balance-tunnel boring machines (EPB-TBMs). Several hundreds of millions of tons of spoil material, consisting of soil mixed with FAs, are produced worldwide, raising the issue of their handling and safe disposal. Reducing waste production and reusing by-products are the primary objectives of the "circular economy," and in this context, the biodegradation of SLES becomes a key question in reclaiming excavated soils, especially at construction sites where SLES degradation on the spot is not possible because of lack of space for temporary spoil material storage. The aim of the present work was to apply a bacterial consortium (BC) of SLES degraders to spoil material excavated with an EPB-TBM and coming from a real construction site. For this purpose, the BC capability to accelerate SLES degradation was tested. Preliminary BC growth, degradation tests, and ecotoxicological evaluations were performed on a selected FA. Subsequently, a bioaugmentation experiment was conducted; and the microbial abundance, viability, and SLES concentrations in spoil material were evaluated over the experimental time (0.5, 3, 6, 24, 48, and 144 h). Moreover, the corresponding aqueous elutriates were extracted from all the soil samples and analyzed for SLES concentration and ecotoxicological evaluations with the bacterium Aliivibrio fischeri . The preliminary experiments showed the BC capability to grow under 14 different concentrations of the FA. The maximum BC growth rates and degradation efficiency (100%) were achieved with initial SLES concentrations of 125, 250, and 500 mg/L. The subsequent bioaugmentation of the spoil material with BC significantly (sixfold) improved the degradation time of SLES (DT 50 1 day) compared with natural attenuation (DT 50 6 days). In line with this result, neither SLES residues nor toxicity was recorded in the soil extracts showing the spoil material as a by-product promptly usable. The bioaugmentation with BC can be a very useful for cleaning spoil material produced in underground construction where its temporary storage (for SLES natural biodegradation) is not possible., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Rolando, Barra Caracciolo, Grenni, Mariani, Rauseo, Spataro, Garbini, Visca and Patrolecco.)

Anaerobic digestion (AD) treatment of cattle manure and slurry makes it possible to produce biogas, a renewable and storable biofuel, as well as digestate, a residual organic matter that can be used to replace chemical fertilizers. On the other hand, the intense use of antibiotics (e.g., sulfamethoxazole) in animal husbandry practices is showing increasing negative impacts resulting from the release of still metabolically active molecules into agroecosystems. In the present study, cattle manure collected from an AD plant-feeding tank was used as feedstock for AD experiments in which some batches were spiked with 5 mg L -1 of sulfamethoxazole (SMX). Adding the antibiotic affected the microbial community dynamic; in particular, the efficiency of the acidogenic and acetogenic phases of the process corresponded to higher CH 4 and H 2 production than in the control. SMX was also degraded, and at the end of the experiment (69 days), just 20% of its initial concentration was found. The relative abundance (ARG/16S) of resistance genes sul1 , sul2 , and the proxy intI1 initially found in the ingestate decreased during the AD in both the spiked and control batches, suggesting that this process lowers the likelihood of antibiotic resistance genes spreading., (Copyright © 2020 Mazzurco Miritana, Massini, Visca, Grenni, Patrolecco, Spataro, Rauseo, Garbini, Signorini, Rosa and Barra Caracciolo.)

Due to their widespread use in industrial applications in recent decades, Polychlorobiphenyls (PCBs) and heavy metals (HMs) are the most common soil contaminants worldwide, posing a risk for both ecosystems and human health. In this study, a poplar-assisted bioremediation strategy has been applied for more than 4 years to a historically contaminated area (PCBs and HMs) in Southern Italy using the Monviso poplar clone. This clone was effective in promoting a decrease in all contaminants and an increase in soil quality in terms of organic carbon and microbial abundance. Moreover, a significant shift in the structure and predicted function of the belowground microbial community was also observed when analyzing both DNA and cDNA sequencing data. In fact, an increase in bacterial genera belonging to Proteobacteria able to degrade PCBs and resist HMs was observed. Moreover, the functional profiling of the microbial community predicted by PICRUSt2 made it possible to identify several genes associated with PCB transformation (e.g., bphAa , bphAb , bphB , bphC ), response to HM oxidative stress (e.g., catalase, superoxide reductase, peroxidase) and HM uptake and expulsion (e.g., ABC transporters). This work demonstrated the effectiveness of the poplar clone Monviso in stimulating the natural belowground microbial community to remove contaminants and improve the overall soil quality. It is a practical example of a nature based solution involving synergic interactions between plants and the belowground microbial community., (Copyright © 2020 Barra Caracciolo, Grenni, Garbini, Rolando, Campanale, Aimola, Fernandez-Lopez, Fernandez-Gonzalez, Villadas and Ancona.)

The anionic surfactant Sodium Lauryl Ether Sulfate (SLES) is the principal component of several commercial foaming products for soil conditioning in the tunneling industry. Huge amounts of spoil material are produced during the excavation process and the presence of SLES can affect its re-use as a by-product. Anionic surfactants can be a risk for ecosystems if occurring in the environment at toxic concentrations. SLES biodegradability is a key issue if the excavated soil is to be reused. The aim of this study was to identify bacteria able to degrade SLES, so that it could potentially be used in bioaugmentation techniques. Enrichment cultures were performed using bacterial populations from spoil material collected in a tunnel construction site as the inoculum. A bacterial consortium able to grow in a few hours with SLES concentrations from 125 mg/L to 2 g/L was selected and then identified by Next Generation Sequencing analysis. Most of bacteria identified belonged to Gamma- Proteobacteria (99%) and Pseudomona s (ca 90%) was the predominant genus. The bacterial consortium was able to degrade 94% of an initial SLES concentration of 250 mg/L in 9 h. A predictive functional analysis using the PICRUSt2 software showed the presence of esterase enzymes, responsible for SLES degradation. The bacterial consortium selected could be useful for its possible seeding (bioaugmentation) on spoil material from tunneling excavation., (Copyright © 2020 Rolando, Grenni, Rauseo, Pescatore, Patrolecco, Garbini, Visca and Barra Caracciolo.)