Microbial acidification by N, S, Fe and Mn oxidation as a key mechanism for deterioration of subsea tunnel sprayed concrete.

The deterioration of fibre-reinforced sprayed concrete was studied in the Oslofjord subsea tunnel (Norway). At sites with intrusion of saline groundwater resulting in biofilm growth, the concrete exhibited significant concrete deterioration and steel fibre corrosion. Using amplicon sequencing and shotgun metagenomics, the microbial taxa and surveyed potential microbial mechanisms of concrete degradation at two sites over five years were identified. The concrete beneath the biofilm was investigated with polarised light microscopy, scanning electron microscopy and X-ray diffraction. The oxic environment in the tunnel favoured aerobic oxidation processes in nitrogen, sulfur and metal biogeochemical cycling as evidenced by large abundances of metagenome-assembled genomes (MAGs) with potential for oxidation of nitrogen, sulfur, manganese and iron, observed mild acidification of the concrete, and the presence of manganese- and iron oxides. These results suggest that autotrophic microbial populations involved in the cycling of several elements contributed to the corrosion of steel fibres and acidification causing concrete deterioration.

Low electric current in a bioelectrochemical system facilitates ethanol production from CO using CO-enriched mixed culture.

Fossil resources must be replaced by renewable resources in production systems to mitigate green-house gas emissions and combat climate change. Electro-fermentation utilizes a bioelectrochemical system (BES) to valorize industrial and municipal waste. Current electro-fermentation research is mainly focused on microbial electrosynthesis using CO2 for producing commodity chemicals and replacing petroleum-based infrastructures. However, slow production rates and low titers of metabolites during CO2-based microbial electrosynthesis impede its implementation to the real application in the near future. On the other hand, CO is a highly reactive gas and an abundant feedstock discharged from fossil fuel-based industry. Here, we investigated CO and CO2 electro-fermentation, using a CO-enriched culture. Fresh cow fecal waste was enriched under an atmosphere of 50% CO and 20% CO2 in N2 using serial cultivation. The CO-enriched culture was dominated by Clostridium autoethanogenum (≥89%) and showed electro-activity in a BES reactor with CO2 sparging. When 50% CO was included in the 20% CO2 gas with 10 mA applied current, acetate and ethanol were produced up to 12.9 ± 2.7 mM and 2.7 ± 1.1 mM, respectively. The coulombic efficiency was estimated to 148% ± 8% without an electron mediator. At 25 mA, the culture showed faster initial growth and acetate production but no ethanol production, and only at 86% ± 4% coulombic efficiency. The maximum optical density (OD) of 10 mA and 25 mA reactors were 0.29 ± 0.07 and 0.41 ± 0.03, respectively, whereas it was 0.77 ± 0.19 without electric current. These results show that CO electro-fermentation at low current can be an alternative way of valorizing industrial waste gas using a bioelectrochemical system.

Microbiome structure and function in parallel full-scale aerobic granular sludge and activated sludge processes.

Aerobic granular sludge (AGS) and conventional activated sludge (CAS) are two different biological wastewater treatment processes. AGS consists of self-immobilised microorganisms that are transformed into spherical biofilms, whereas CAS has floccular sludge of lower density. In this study, we investigated the treatment performance and microbiome dynamics of two full-scale AGS reactors and a parallel CAS system at a municipal WWTP in Sweden. Both systems produced low effluent concentrations, with some fluctuations in phosphate and nitrate mainly due to variations in organic substrate availability. The microbial diversity was slightly higher in the AGS, with different dynamics in the microbiome over time. Seasonal periodicity was observed in both sludge types, with a larger shift in the CAS microbiome compared to the AGS. Groups important for reactor function, such as ammonia-oxidising bacteria (AOB), nitrite-oxidising bacteria (NOB), polyphosphate-accumulating organisms (PAOs) and glycogen-accumulating organisms (GAOs), followed similar trends in both systems, with higher relative abundances of PAOs and GAOs in the AGS. However, microbial composition and dynamics differed between the two systems at the genus level. For instance, among PAOs, Tetrasphaera was more prevalent in the AGS, while Dechloromonas was more common in the CAS. Among NOB, Ca. Nitrotoga had a higher relative abundance in the AGS, while Nitrospira was the main nitrifier in the CAS. Furthermore, network analysis revealed the clustering of the various genera within the guilds to modules with different temporal patterns, suggesting functional redundancy in both AGS and CAS. KEY POINTS: • Microbial community succession in parallel full-scale aerobic granular sludge (AGS) and conventional activated sludge (CAS) processes. • Higher periodicity in microbial community structure in CAS compared to in AGS. • Similar functional groups between AGS and CAS but different composition and dynamics at genus level.

Biofilm colonization and succession in a full-scale partial nitritation-anammox moving bed biofilm reactor.

BACKGROUND: Partial nitritation-anammox (PNA) is a biological nitrogen removal process commonly used in wastewater treatment plants for the treatment of warm and nitrogen-rich sludge liquor from anaerobic digestion, often referred to as sidestream wastewater. In these systems, biofilms are frequently used to retain biomass with aerobic ammonia-oxidizing bacteria (AOB) and anammox bacteria, which together convert ammonium to nitrogen gas. Little is known about how these biofilm communities develop, and whether knowledge about the assembly of biofilms in natural communities can be applied to PNA biofilms. RESULTS: We followed the start-up of a full-scale PNA moving bed biofilm reactor for 175 days using shotgun metagenomics. Environmental filtering likely restricted initial biofilm colonization, resulting in low phylogenetic diversity, with the initial microbial community comprised mainly of Proteobacteria. Facilitative priority effects allowed further biofilm colonization, with the growth of initial aerobic colonizers promoting the arrival and growth of anaerobic taxa like methanogens and anammox bacteria. Among the early colonizers were known 'oligotrophic' ammonia oxidizers including comammox Nitrospira and Nitrosomonas cluster 6a AOB. Increasing the nitrogen load in the bioreactor allowed colonization by 'copiotrophic' Nitrosomonas cluster 7 AOB and resulted in the exclusion of the initial ammonia- and nitrite oxidizers. CONCLUSIONS: We show that complex dynamic processes occur in PNA microbial communities before a stable bioreactor process is achieved. The results of this study not only contribute to our knowledge about biofilm assembly and PNA bioreactor start-up but could also help guide strategies for the successful implementation of PNA bioreactors. Video Abstract.

Removal and release of microplastics and other environmental pollutants during the start-up of bioretention filters treating stormwater.

Untreated stormwater is a major source of microplastics, organic pollutants, metals, and nutrients in urban water courses. The aim of this study was to improve the knowledge about the start-up periods of bioretention filters. A rain garden pilot facility with 13 bioretention filters was constructed and stormwater from a highway and adjacent impervious surfaces was used for irrigation for ∼12 weeks. Selected plants (Armeria maritima, Hippophae rhamnoides, Juncus effusus, and Festuca rubra) was planted in ten filters. Stormwater percolated through the filters containing waste-to-energy bottom ash, biochar, or Sphagnum peat, mixed with sandy loam. Influent and effluent samples were taken to evaluate removal of the above-mentioned pollutants. All filters efficiently removed microplastics >10 µm, organic pollutants, and most metals. Copper leached from all filters initially but was significantly reduced in the biochar filters at the end of the period, while the other filters showed a declining trend. All filters leached nutrients initially, but concentrations decreased over time, and the biochar filters had efficiently reduced nitrogen after a few weeks. To conclude, all the filters effectively removed pollutants during the start-up period. Before being recommended for full-scale applications, the functionality of the filters after a longer period of operation should be evaluated.

The effects of electrochemical pretreatment and curing environment on strength and leaching of stabilized/solidified contaminated sediment.

Stabilization and solidification (S/S) is known to improve the structural properties of sediment and reduce contaminant mobility, enabling the utilization of dredged contaminated sediment. Further reduction of contaminants (e.g., tributyltin (TBT) and metals) can be done using electrochemical treatment prior to S/S and could potentially minimize contaminant leaching. This is the first study on how electrochemical pretreatment affects the strength and leaching properties of stabilized sediments. It also investigates how salinity and organic carbon in the curing liquid affect the stabilized sediment.The results showed that the electrolysis reduced the content of TBT by 22% and zinc by 44% in the sediment. The electrolyzed stabilized samples met the requirements for compression strength and had a reduced surface leaching of zinc. Curing in saline water was beneficial for strength development and reduced the leaching of TBT compared to curing in fresh water. The results indicate that pretreatment prior to stabilization could be beneficial in reducing contaminant leaching and recovering metals from the sediment. The conclusion is that a better understanding of the changes in the sediment caused by electrochemical treatment and how these changes interact with stabilization reactions is needed. In addition, it is recommended to investigate the strength and leaching behavior in environments similar to the intended in situ conditions.

Effect of anode material and dispersal limitation on the performance and biofilm community in microbial electrolysis cells.

In a microbial electrolysis cell (MEC), the oxidization of organic compounds is facilitated by an electrogenic biofilm on the anode surface. The biofilm community composition determines the function of the system. Both deterministic and stochastic factors affect the community, but the relative importance of different factors is poorly understood. Anode material is a deterministic factor as materials with different properties may select for different microorganisms. Ecological drift is a stochastic factor, which is amplified by dispersal limitation between communities. Here, we compared the effects of three anode materials (graphene, carbon cloth, and nickel) with the effect of dispersal limitation on the function and biofilm community assembly. Twelve MECs were operated for 56 days in four hydraulically connected loops and shotgun metagenomic sequencing was used to analyse the microbial community composition on the anode surfaces at the end of the experiment. The anode material was the most important factor affecting the performance of the MECs, explaining 54-80 % of the variance observed in peak current density, total electric charge generation, and start-up lag time, while dispersal limitation explained 10-16 % of the variance. Carbon cloth anodes had the highest current generation and shortest lag time. However, dispersal limitation was the most important factor affecting microbial community structure, explaining 61-98 % of the variance in community diversity, evenness, and the relative abundance of the most abundant taxa, while anode material explained 0-20 % of the variance. The biofilms contained nine Desulfobacterota metagenome-assembled genomes (MAGs), which made up 64-89 % of the communities and were likely responsible for electricity generation in the MECs. Different MAGs dominated in different MECs. Particularly two different genotypes related to Geobacter benzoatilyticus competed for dominance on the anodes and reached relative abundances up to 83 %. The winning genotype was the same in all MECs that were hydraulically connected irrespective of anode material used.

Resistance of aerobic granular sludge microbiomes to periodic loss of biomass.

Granular sludge is a biofilm process used for wastewater treatment which is currently being implemented worldwide. It is important to understand how disturbances affect the microbial community and performance of reactors. Here, two acetate-fed replicate reactors were inoculated with acclimatized sludge and the reactor performance, and the granular sludge microbial community succession were studied for 149 days. During this time, the microbial community was challenged by periodically removing half of the reactor biomass, subsequently increasing the food-to-microorganism (F/M) ratio. Diversity analysis together with null models show that overall, the microbial communities were resistant to the disturbances, observing some minor effects on polyphosphate-accumulating and denitrifying microbial communities and their associated reactor functions. Community turnover was driven by drift and random granule loss, and stochasticity was the governing ecological process for community assembly. These results evidence the aerobic granular sludge process as a robust system for wastewater treatment.

Weighted score ratios (WRS) give transparent weighting in multicriteria sustainability assessments - A case study on removal of pharmaceutical residues from wastewater.

Sustainability assessment using multicriteria analysis (MCA) is a structured way of including criteria from the three sustainability dimensions (environmental, economic, and social) when comparing different alternatives. A problem with the conventional MCA methods is that the consequences of the weights given to different criteria are not transparent. Here, we amend the simple additive weighting MCA method with weighted score ratios (WSRs), which are used during the sustainability assessment to show how the weights affect the valuation of the criteria (e.g., cost per kg CO2e). This enables comparisons to other sustainability assessments and reference values from society, which increases the transparency and can make weighting more objective. We applied the method to a comparison of technologies for removal of pharmaceutical residues from wastewater. Due to growing concern about the effects that pharmaceutical residues can have on our environment, implementations of advanced technologies are increasing. However, they entail high requirements of energy and resources. Therefore, many aspects must be considered to make a sustainable choice of technology. In this study, a sustainability assessment was performed of ozonation, powdered activated carbon and granular activated carbon for removal of pharmaceutical residues at a large wastewater treatment plant (WWTP) in Sweden. The outcome showed that powdered activated carbon is the least sustainable choice for the studied WWTP. Whether ozonation or granular activated carbon is most sustainable depends on how climate impact and energy use are valued. The total sustainability of ozonation is affected by how the electricity is assumed to be produced, whereas for granular activated carbon it depends on whether the carbon source is of renewable or fossil origin. Using WSRs allowed the participants in the assessment to make conscious choices on how they weighted different criteria in relation to how these criteria are valued in society at large.

Chemical Imaging of Pharmaceuticals in Biofilms for Wastewater Treatment Using Secondary Ion Mass Spectrometry.

The occurrence of pharmaceuticals in the aquatic environment is a global water quality challenge for several reasons, such as deleterious effects on ecological and human health, antibiotic resistance development, and endocrine-disrupting effects on aquatic organisms. To optimize their removal from the water cycle, understanding the processes during biological wastewater treatment is crucial. Time-of-flight secondary ion mass spectrometry imaging was successfully applied to investigate and analyze the distribution of pharmaceuticals as well as endogenous molecules in the complex biological matrix of biofilms for wastewater treatment. Several compounds and their localization were identified in the biofilm section, including citalopram, ketoconazole, ketoconazole transformation products, and sertraline. The images revealed the pharmaceuticals gathered in distinct sites of the biofilm matrix. While citalopram penetrated the biofilm deeply, sertraline remained confined in its outer layer. Both pharmaceuticals seemed to mainly colocalize with phosphocholine lipids. Ketoconazole concentrated in small areas with high signal intensity. The approach outlined here presents a powerful strategy for visualizing the chemical composition of biofilms for wastewater treatment and demonstrates its promising utility for elucidating the mechanisms behind pharmaceutical and antimicrobial removal in biological wastewater treatment.