Wastewater treatment plants (WWTPs) are almost the only place where plastic fragments are artificially removed, resulting in mass accumulation of nanoplastics (NPs). In this research, four different concentrations (0 mg/L, 0.1 mg/L, 1 mg/L, 10 mg/L) of polystyrene nanoplastics (PS-NPs) were used to investigate the cell damage and nitrogen inhibition of activated sludge, exposed in a self-assembled SBR reactor for 30 days. Intracellular reactive oxides (ROS) and extracellular lactate dehydrogenase (LDH) increased with the rise of exposure concentration, and morphological analysis disclosed the creases, collapse, and even rupture of cell membranes. However, exposure damage (PS-NPs ≤ 1 mg/L) appeared to be reversible, attributed to that extracellular polymeric substances (EPS) secretion can thicken the three protective layers outside the membrane. PS-NPs did not disrupt the EPS chemical structure, but increased humic acid content. Prolonged exposure time (from 15 to 30 days) was directly related to the nitrogen inhibition. Due to the habitat changes under PS-NPs exposure, abundance and diversity of microorganisms in the original activated sludge decreased significantly, and the dominant phylum was occupied by Patescibacteria (PS-NPs = 10 mg/L). Changes in enzyme activities of AMO, NR, NIR, and NOR with exposure concentration may explain the conversion of nitrogen in SBR. This research broadens our horizons to understand the response mechanism of activated sludge bacteria to PS-NPs exposure individually and collectively.

Biochemical oxidation and reduction are key processes in treating biological wastewater and they require the presence of electron acceptors. The functional impact of electron acceptors on microbiomes provides strategies for improving the treatment efficiency. This research focused on two of the most important electron acceptors, nitrate and oxygen. Molecule ecological network, null model, and functional prediction based on high-throughput sequencing were used to analyze the microbiomes features and assembly mechanism. The results revealed nitrate via the homogeneous selection (74.0%) decreased species diversity, while oxygen via the homogeneous selection (51.1%) and dispersal limitation (29.6%) increased the complexity of community structure. Microbes that were more strongly homogeneously selected for assembly included polyphosphate accumulating organisms (PAOs), such as Pseudomonas and variovorax in the nitrate impacted community; Pseudomonas, Candidatus_Accumulibacter, Thermomonas and Dechloromonas, in the oxygen impacted community. Nitrate simplified species interaction and increased the abundance of functional genes involving in tricarboxylic acid cycle (TCA cycle), electron transfer, nitrogen metabolism, and membrane transport. These findings contribute to our knowledge of assembly process and interactions among microorganisms and lay a theoretical basis for future microbial regulation strategies in wastewater treatment.

The widespread presence and persistence of perfluorooctane sulfonate (PFOS) in wastewater treatment plants, as well as its toxicity and bioaccumulation potential, necessitates the investigation on their impact on bioreactor performance. A 48-day exposure test was adopted to study the effects of low (10 μg L-1) and high (1000 μg L-1) PFOS concentrations in a sequencing batch reactor on the performance, composition, and microbial community of activated sludge. The results suggested that adding PFOS at low and high concentrations lowered the removal efficiency of total nitrogen by 22.48% (p < 0.01) and 16.30% (p < 0.01) respectively, while enhanced that of total phosphorus by 1.87% (p > 0.05) and 7.07% (p < 0.05) respectively, compared with the control group. The addition of PFOS also led to the deterioration of activated sludge dewatering performance. Composition and spectroscopic measurements revealed that the PFOS dosage changed the composition of the activated sludge. Furthermore, the PFOS altered the structure and function of the activated sludge microbial community as well as key enzyme activities.

This study investigated the effects of adding biochar (BC) on the fate of ciprofloxacin (CIP) and its related antibiotic tolerance (AT) in activated sludge. Three activated sludge reactors were established with different types of BC, derived from apple, pear, and mulberry tree, respectively, and one reactor with no BC. All reactors were exposed to an environmentally relevant level of CIP that acted as a definitive selective pressure significantly promoting AT to four representative antibiotics (CIP, ampicillin, tetracycline, and polymyxin B) by up to two orders of magnitude. While CIP removal was negligible in the reactor without BC, the BC-dosed reactors effectively removed CIP (70–95% removals) through primarily adsorption by BC and biodegradation/biosorption by biomass. The AT in the BC-added reactors was suppressed by 10–99%, compared to that without BC. The BC addition played a key role in sequestering CIP, thereby decreasing the selective pressure that enabled the proactive prevention of AT increase. 16S rRNA gene sequencing analysis showed that the BC addition alleviated the CIP-mediated toxicity to community diversity and organisms related to phosphorous removal. Machine learning modeling with random forest and support vector models using AS microbiome data collectively pinpointed Achromobacter selected by CIP and strongly associated with the AT increase in activated sludge. The identification of Achromobacter as an important AT bacteria revealed by the machine learning modeling with multiple models was also validated with a linear Pearson's correlation analysis. Overall, our study highlighted Achromobacter as a potential useful sentinel for monitoring AT occurring in the environment and suggested BC as a promising additive in wastewater treatment to improve micropollutant removal, mitigate potential AT propagation, and maintain community diversity against toxic antibiotic loadings.

The extensive use of nano-TiO₂ has caused concerns regarding their potential environmental risks. However, the stress responses and self-recovery potential of nitrogen removal and greenhouse gas N₂O emissions after long-term nano-TiO₂ exposure have seldom been addressed yet. This study explored the long-term effects of nano-TiO₂ on biological nitrogen transformations in a sequencing batch reactor at four levels (1, 10, 25, and 50 mg/L), and the reactor's self-recovery potential was assessed. The results showed that nano-TiO₂ exhibited a dose-dependent inhibitory effect on the removal efficiencies of ammonia nitrogen and total nitrogen, whereas N₂O emissions unexpectedly increased. The promoted N₂O emissions were probably due to the inhibition of denitrification processes, including the reduction of the denitrifying-related N₂O reductase activity and the abundance of the denitrifying bacteria Flavobacterium. The inhibition of carbon source metabolism, the inefficient electron transfer efficiency, and the electronic competition between the denitrifying enzymes would be in charge of the deterioration of denitrification performance. After the withdrawal of nano-TiO₂ from the influent, the nitrogen transformation efficiencies and the N₂O emissions of activated sludge recovered entirely within 30 days, possibly attributed to the insensitive bacteria survival and the microbial community diversity. Overall, this study will promote the current understanding of the stress responses and the self-recovery potential of BNR systems to nanoparticle exposure.

In this study, the occurrence of pharmaceuticals, hormones and bacterial community structures was studied at a wastewater treatment plant in Finland having two different parallel treatment lines: conventional activated sludge (CAS) treatment with a sedimentation stage, and a membrane bioreactor (MBR). Influent and effluents were sampled seven times over a period of one year. The bacterial communities of the influent samples showed a high degree of similarity, except for the February sample which had substantially lower diversity. There was significant fluctuation in the species richness and diversity of the effluent samples, although both effluents showed a similar trend. A marked decrease in diversity was observed in effluents collected between August and November. The initiation of nitrogen removal as a result of an increase in temperature could explain the changes in microbial community structures. In overall terms, suspended solids, bacteria and total organic matter (COD and BOD) were removed to a greater extent using the MBR, while higher Tot-N, Tot-P and nitrate removal rates were achieved using the CAS treatment. Estrone (E1) concentrations were also consistently at a lower level in the MBR effluents (<0.1–0.68 ng/l) compared to the CAS effluents (1.1–12 ng/l). Due to the high variation in the concentrations of pharmaceuticals, no clear superiority of either process could be demonstrated with certainty. The study highlights the importance of long-term sampling campaigns to detect variations effectively.

The environmental risks of microplastics (MPs) have raised an increasing concern. However, the effects of MPs in anaerobic digestion (AD) systems of waste activated sludge (WAS), especially on the fate of antibiotic resistance genes (ARGs), have not been clearly understood. Herein, the variation and interaction of digestion performance, microbial communities and ARGs during AD process of WAS in the presence of polyethylene (PE) MPs with two sizes, PE MPs-180μm and PE MPs-1mm, were investigated. The results showed that the presence of PE MPs, especially PE MPs-1mm, led to the increased hydrolysis of soluble polysaccharides and proteins and the accumulation of volatile fatty acids. The methane production decreased by 6.1% and 13.8% in the presence of PE MPs-180μm and PE MPs-1mm, respectively. Together with this process, hydrolytic bacteria and acidogens were enriched, and methanogens participating in acetoclastic methanogenesis were reduced. Meanwhile, ARGs were enriched obviously by the presence of PE MPs, the abundances of which in PE MPs-180μm and PE MPs-1mm groups were 1.2–3.0 times and 1.5–4.0 times higher than that in the control by the end of AD. That was associated with different co-occurrence patterns between ARGs and bacterial taxa and the enrichment of ARG-hosting bacteria caused by the presence of PE MPs. Together these results suggested the adverse effects of PE MPs on performance and ARGs removal during AD process of WAS through inducing the changes of microbial populations.

Wastewater treatment plants have been identified as an important gathering spot for nanoplastics, possibly having unintended impacts on important biological nutrient removal processes. The underlying effects of long-term exposure of activated sludge to nanoplastics on nutrient removal and the mechanisms involved remain unclear. This study investigated the effect of polystyrene nanoplastics (Nano-PS) on the treatment performance and microbial community structure, and network in activated sludge. The results indicate that 1000 μg/L Nano-PS had chronic negative effects on the treatment performance in a continuous test over 140 days. Nano-PS had no significant impact in the earlier stages (0–50 days). However, as exposure time increased, the removal efficiencies of chemical oxygen demand, total phosphorous, and total nitrogen (TN) decreased by 2.7, 33.2, and 23.5%, respectively, in the later stages (87–132 days). These adverse impacts further manifested as a change in the topological characteristics, forming a smaller scale, lower complexity, and weaker transfer efficiency of the microbial network. Moreover, the scale and complexity of subnetwork-nitrogen removal bacteria and subnetwork-nitrifier were inhibited, leading to an increase in the effluent TN and NH₄⁺-N. The decreased modules and connectors (keystone taxa) likely caused the deterioration of treatment performance and functional diversity, which was consistent with the change in PICRUSt results. Less competition, denser nodes, and more complex module structures were induced as a strategy to mediate the long-term stress of nano-PS. To our knowledge, this is the first attempt to explore the long-term effects of nano-PS on the microbial interaction network of activated sludge, laying an experimental foundation for reducing the risks associated with nanoplastics.

Ecotoxicological assessment of landfill leachate has become a priority to determine its impacts on the ecosystem. Toxicity assays with microorganisms stand out due to their quick response, low cost and ease of testing. In this context, the present study evaluated the acute toxic effects of leachates from two landfills of different ages and modes of operation to bacterium Aliivibrio fischeri and activated sludge microorganisms and the ammonia nitrogen and humic substances (HS) sensitivity to these organisms. Reductions greater than 30% in leachate toxicity were observed after ammonia removal for A. fischeri and activated sludge microorganisms. After 97% removal of HS, the greater reductions in toxicity (44.28 to 79.82%) were verified for microbial species studied, indicating that the organic compounds (measured as chemical oxygen demand, total organic carbon and humic substances) were the primary pollutants responsible for the toxicity of the leachates. Concerning the organisms studied, A. fischeri showed greater sensitivity to the leachates’ pollutants compared to the activated sludge microorganisms. Nevertheless, a strong correlation was observed between A. fischeri and activated sludge microorganisms’ toxicity responses, suggesting that respirometry assay can be used to determine leachate toxicity.

Per- and polyfluoroalkyl substances (PFASs) have received increasing attention due to their widespread presence in diverse environments including wastewater treatment plants (WWTPs) and their potential adverse health effects. Perfluorooctanoic acid (PFOA) is one of the most detected forms of PFASs in WWTPs. However, there is still a paucity of knowledge about the effect of PFASs on microorganisms of the key component of WWTP, activated sludge. In this study, lab-scale microcosm experiments were established to evaluate the influences of PFOA on activated sludge microbes under aerobic and anaerobic conditions. The diversity, structure, and microbe-microbe interaction of microbial community were determined by 16S rRNA gene amplicon sequencing and co-occurrence network analysis. After 90 days of exposure to PFOA, activated sludge microbial richness decreased under both aerobic and anaerobic conditions. Specifically, under aerobic condition, Rhodopseudomonas (mean relative abundance 3.6%), Flavobacterium (2.4%), and Ignavibacterium (6.6%) were enriched in PFOA-spiked activated sludge compared with that in the unspiked sludge (2.6%, 0.1%, and 1.9%, respectively). By contrast, after 90 days of exposure to PFOA, Eubacterium (2.1%), Hyphomicrobium (1.8%), and Methyloversatilis (1.2%) were enriched under anaerobic condition, and more abundant than that in the control sludge (0.4%, 1.5%, and 0.6%, respectively). These genera were the potential PFOA-resistant members. In addition, Azospirillum and Sporomusa were the most connected taxa in PFOA-aerobic and PFOA-anaerobic networks, respectively. Prediction of the functional gene showed that PFOA inhibited some gene expression of sludge microbes, such as transcription, amino acid transport and metabolism, and energy production and conversion. In summary, continued exposure to PFOA induced substantial shifts of the sludge bacterial diversity and composition under both aerobic and anaerobic conditions.