Entry pathways determined the effects of MPs on sludge anaerobic digestion system: The views of methane production and antibiotic resistance genes fates.

Sludge is one of the primary reservoirs of microplastics (MPs), and the effects of MPs on subsequent sludge treatment raised attention. Given the entry pathways, MPs would exhibit different properties, but the entry pathway-dependent effect of MPs on sludge treatment performance and the fates of antibiotic resistance genes (ARGs), another high-risk emerging contaminant, were seldom documented. Herein, MPs with two predominant entry pathways, including wastewater-derived (WW-derived) and anaerobic digestion-introduced (AD-introduced), were used to investigate the effects on AD performance and ARGs abundances. The results indicated that WW-derived MPs, namely the MPs accumulated in sludge during the wastewater treatment process, exhibited significant inhibition on methane production by 22.8%-71.6%, while the AD-introduced MPs, being introduced in the sludge AD process, slightly increased the methane yield by 4.7%-17.1%. Meanwhile, MPs were responsible for promoting transmission of target ARGs, and polyethylene terephthalate MPs (PET-MPs) showed a greater promotion effect (0.0154-0.0936) than polyamide MPs (PA-MPs) (0.0013-0.0724). Compared to size, entry pathways and types played more vital roles on MPs influences. Investigation on mechanisms based on microbial community structure revealed characteristics (aging degree and types) of MPs determined the differences of AD performance and ARGs fates. WW-derived MPs with longer aging period and higher aging degree would release toxics and decrease the activities of microorganisms, resulting in the negative impact on AD performance. However, AD-introduced MPs with short aging period exhibited marginal impacts on AD performance. Furthermore, the co-occurrent network analysis suggested that the variations of potential host bacteria induced by MPs with different types and aging degree attributed to the dissemination of ARGs. Distinctively from most previous studies, the MPs with different sizes did not show remarkable effects on AD performance and ARGs fates. Our findings benefited the understanding of realistic environmental behavior and effect of MPs with different sources.

Advancing chronic toxicity risk assessment in freshwater ecology by molecular characterization-based machine learning.

The continuously increased production of various chemicals and their release into environments have raised potential negative effects on ecological health. However, traditional labor-intensive assessment methods cannot effectively and rapidly evaluate these hazards, especially for chronic risk. In this study, machine learning (ML) was employed to construct quantitative structure-activity relationship (QSAR) models, enabling the prediction of chronic toxicity to aquatic organisms by leveraging the molecular characteristics of pollutants, namely, the molecular descriptors, fingerprints, and graphs. The limited dataset size hindered the notable advantages of the graph attention network (GAT) model for the molecular graphs. Considering computational efficiency and performance (R2 = 0.78; RMSE = 0.77), XGBoost (XGB) was used for reliable QSAR-ML models predicting chronic toxicity using small- or medium-sized tabular data and the molecular descriptors. Further kernel density estimation analysis confirmed the high accuracy of the model for pollutant concentrations ranging from 10-3 to 102 mg/L, effectively aligning with most environmental scenarios. Model interpretation showed SlogP and exposure duration as the primary influential factors. SlogP, representing the distribution coefficient of a molecule between lipophilic and hydrophilic environments, had a negative effect on the toxicity outcomes. Additionally, the exposure duration played a crucial role in determining the chronic toxicity. Finally, the chronic toxicity data of bisphenol A validated the robustness and reliability of the model established in this research. Our study provided a robust and feasible methodology for chronic ecological risk evaluation of various types of pollutants and could facilitate and increase the use of ML applications in environmental fields.

Pathogenic Bacteria Are the Primary Determinants Shaping PM2.5-Borne Resistomes in the Municipal Food Waste Treatment System.

Bioaerosol pollution poses a substantial threat to human health during municipal food waste (FW) recycling. However, bioaerosol-borne antibiotic-resistant genes (ARGs) have received little attention. Herein, 48 metagenomic data were applied to study the prevalence of PM2.5-borne ARGs in and around full-scale food waste treatment plants (FWTPs). Overall, FWTP PM2.5 (2.82 ± 1.47 copies/16S rRNA gene) harbored comparable total abundance of ARGs to that of municipal wastewater treatment plant PM2.5 (WWTP), but was significantly enriched with the multidrug type (e.g., AdeC/I/J; p < 0.05), especially the abundant multidrug ARGs could serve as effective indicators to define resistome profiles of FWTPs (Random Forest accuracy >92%). FWTP PM2.5 exhibited a decreasing enrichment of total ARGs along the FWTP-downwind-boundary gradient, eventually reaching levels comparable to urban PM2.5 (1.46 ± 0.21 copies/16S rRNA gene, N = 12). The combined analysis of source-tracking, metagenome-assembled genomes (MAGs), and culture-based testing provides strong evidence that Acinetobacter johnsonii-dominated pathogens contributed significantly to shaping and disseminating multidrug ARGs, while abiotic factors (i.e., SO42-) indirectly participated in these processes, which deserves more attention in developing strategies to mitigate airborne ARGs. In addition, the exposure level of FWTP PM2.5-borne resistant pathogens was about 5-11 times higher than those in urban PM2.5, and could be more severe than hospital PM2.5 in certain scenarios (<41.53%). This work highlights the importance of FWTP in disseminating airborne multidrug ARGs and the need for re-evaluating the air pollution induced by municipal FWTP in public health terms.

From air to airway: Dynamics and risk of inhalable bacteria in municipal solid waste treatment systems.

Municipal solid waste treatment (MSWT) system emits a cocktail of microorganisms that jeopardize environmental and public health. However, the dynamics and risks of airborne microbiota associated with MSWT are poorly understood. Here, we analyzed the bacterial community of inhalable air particulates (PM10, n = 71) and the potentially exposed on-site workers' throat swabs (n = 30) along with waste treatment chain in Shanghai, the largest city of China. Overall, the airborne bacteria varied largely in composition and abundance during the treatment (P < 0.05), especially in winter. Compared to the air conditions, MSWT-sources that contributed to 15 ∼ 70% of airborne bacteria more heavily influenced the PM10-laden bacterial communities (PLS-SEM, ß = 0.40, P < 0.05). Moreover, our year-span analysis found PM10 as an important media spreading pathogens (104 ∼ 108 copies/day) into on-site workers. The machine-learning identified Lactobacillus and Streptococcus as pharynx-niched featured biomarker in summer and Rhodococcus and Capnocytophaga in winter (RandomForest, ntree = 500, mtry = 10, cross = 10, OOB = 0%), which closely related to their airborne counterparts (Procrustes test, P < 0.05), suggesting that MSWT a dynamic hotspot of airborne bacteria with the pronounced inhalable risks to the neighboring communities.

The crystalline structure transition and hydrogen bonds shift determining enhanced enzymatic digestibility of cellulose treated by ultrasonication.

Global energy issue raised the necessity to develop second-generation biofuels, and biorefinery of cellulosic biomass becomes a promising solution. Various pretreatments were used to overcome the cellulose nature of recalcitrance and improve the enzymatic digestibility, but the lack of mechanism understanding hindered the development of efficient and cost-effective technologies of cellulose utilization. Using structure-based analysis, we demonstrate that the improved hydrolysis efficiency caused by ultrasonication was ascribed to the changed cellulose properties rather than the increased dissolubility. Further, isothermal titration calorimetry (ITC) analysis suggested that enzymatic digestion of cellulose is an entropically favored reaction driven by hydrophobic forces other than an enthalpically favored reaction. The changes in cellulose properties and thermodynamic paramenters due to ultrasonication accounted for the improved accessibility. Ultrasonication-treated cellulose showed porous, rough and disordered morphology, accompanying with the loss of crystalline structure. Despite the unaffected unit cell structure, ultrasonication expanded the crystalline lattice by increasing grain sizes and average cross-sectional area, resulting in the transformation from cellulose I to cellulose II, with the decreased crystallinity, better hydrophilicity and increased enzymatic bioaccessibility. Furthermore, FTIR combined with two-dimensional correlation spectroscopy (2D-CoS) verified that the sequential shift of hydroxyl group and intramolecular/intermolecular hydrogen bonds, the functional groups governing cellulose crystal structure and stability, accounted for the ultrasonication-induced transition of cellulose crystalline structure. This study provides a comprehensive picture of cellulose structure and property response caused by mechanistic treatments and will open up avenues to develop novel pretreatments for efficient utilization.

Non-negligible health risks caused by inhalation exposure to aldehydes and ketones during food waste treatments in megacity Shanghai.

Aldehydes and ketones in urban air continue to receive regulatory and scientific attention for their environmental prevalence and potential health hazard. However, current knowledge of the health risks and losses caused by these pollutants in food waste (FW) treatment processes is still limited, especially under long-term exposure. Here, we presented the first comprehensive assessment of chronic exposure to 21 aldehydes and ketones in urban FW-air environments (e.g., storage site, mechanical dewatering, and composting) by coupling substantial measured data (383 samples) with Monte Carlo-based probabilistic health risk and impact assessment models. The results showed that acetaldehyde, acetone, 2-butanone and cyclohexanone were consistently the predominant pollutants, although the significant differences in pollution profiles across treatment sites and seasons (Adonis test, P < 0.001). According to the risk assessment results, the estimated cancer risk (CR; mean range: 1.6 × 10-5-1.12 × 10-4) and non-cancer risk (NCR; mean range: 2.98-22.7) triggered by aldehydes and ketones were both unacceptable in most cases (CR: 37.8%-99.3%; NCR: 54.2%-99.8%), and even reached the limit of concern to CR (1 × 10-4) in some exposure scenarios (6.18%-16.9%). Application of DALYs (disability adjusted life years) as a metric for predicting the damage suggested that exposure of workers to aldehydes and ketones over 20 years of working in FW-air environments could result in 0.02-0.14 DALYs per person. Acetaldehyde was the most harmful constituent of all targeted pollutants, which contributed to the vast majority of health risks (>88%) and losses (>90%). This study highlights aldehydes and ketones in FW treatments may be the critical pollutants to pose inhalation risks.

Genome-centric metagenomics revealed functional traits in high-solids anaerobic co-digestion of restaurant food waste, household food waste and rice straw.

High-solids anaerobic co-digestion (HS-AcoD) of food waste (FW) and other organic wastes is an effective option to improve the biogas production and system stability compared to mono-digestion. However, the clean and sustainable HS-AcoD strategy for FW and associated microbial functional traits have not been well explored. Here, HS-AcoD of restaurant food waste (RFW), household food waste (HFW) and rice straw (RS) were performed. Results showed that the maximum synergy index (SI) of 1.28 were achieved when the volatile solids ratio of RFW, HFW and RS was 0.45:0.45:0.1. HS-AcoD alleviated the acidification process by regulating metabolism associated with hydrolysis and volatile fatty acids formation. The synergistic relationship between syntrophic bacteria and Methanothrix sp., and the enhanced metabolic capacity associated with the acetotrophic and hydrogenotrophic pathways dominated by Methanothrix sp., provided a further explanation of the synergistic mechanism. These findings advance the knowledge about microbial mechanisms underlying the synergistic effect of HS-AcoD.

Surfactant aggravated the antibiotic's stress on antibiotic resistance genes proliferation by altering antibiotic solubilization and microbial traits in sludge anaerobic fermentation.

The excessive application of antibiotics and surfactants resulted in their massive accumulation in waste activated sludge (WAS), but the co-occurrent impacts of antibiotics and surfactants on the antibiotic resistant genes (ARGs) fates have seldom reported. This work mainly revealed the roles and critical mechanisms of sodium dodecyl benzene sulfonate (SDBS) on the sulfadiazine (SDZ) stressing for ARGs distribution during WAS anaerobic fermentation. High-throughput qPCR and metagenomic analysis revealed that SDBS aggravated the SDZ selective pressure, and accelerated the proliferation of ARGs. The total abundance of ARGs was increased from 8.81 × 1010 in SDZ to 1.17 × 1011 copies/g TSS in the SDBS/SDZ co-occurrence system. Specifically, the absolute abundances of ermF (MLSB), mefA (MLSB), tetM-01 (tetracycline), tetX (tetracycline), sul2 (sulfonamide) and strB (aminoglycoside) were risen from 4.60 × 108-7.44 × 109 copies/g TSS in the SDZ reactor to 1.02 × 109-4.63 × 1010 copies/g TSS in SDBS/SDZ reactor. SDBS was contributed to the SDZ solubilization and simultaneously effective in disintegrating extracellular polymeric substances and improving cell membrane permeability, which would facilitate the SDZ transport and its interactions with ARGs hosts. Consequently, the microbial community structure was evidently altered, and the typical ARGs hosts (i.e., Saccharimonadales and Ahniella) were greatly enriched. Also, the cell signal transduction systems (i.e., glnL, glrK and pilG), oxidative stress response (i.e., frmA and recA) and bacterial secretion systems (i.e., VirB4), which were related with ARGs propagation, were all provoked in the co-occurred SDBS/SDZ reactor compared with that of sole SDZ. PLS-PM analysis suggested that the bacterial community was the predominant factor that determined the ARGs fates, followed by mobile genetic elements and metabolic pathway. This work demonstrated the interactive effects of surfactants and antibiotics on the ARGs fates in WAS fermentation systems and gave insightful implications on the ecological risks of different exogenous pollutants.

Enhanced anaerobic digestion performance of food waste by zero-valent iron and iron oxides nanoparticles: Comparative analyses of microbial community and metabolism.

The effects of zero-valent iron (ZVI) and iron oxides nanoparticles on anaerobic digestion (AD) performance of food waste (FW) were comparably clarified in this study. Results indicated that the nanoparticles supplement effectively enhanced the methane yields. As observed, these nanoparticles accelerated organics transformation and alleviated acidification process. Also, the enriched total methanogens and functional bacteria (e.g., Proteiniphilum) were consistent with the promotion of oxidative phosphorylation, citrate cycle, coenzymes biosynthesis and the metabolisms of amino acid, carbohydrate, methane. Additionally, these nanoparticles stimulated electron transfer potential via enriching syntrophic genera (e.g., Geobacter, Syntrophomonas), primary acetate-dependent methanogens (Methanosaeta, Methanosarcina) and related functions (pilus assembly protein, ferredoxins). By comparison, ZVI nanoparticle presented the excellent performance on methanogenesis. This study provides comprehensive understanding of the methanogenesis facilitated by ZVI and iron oxides nanoparticles through the enhancement of key microbes and microbial metabolisms, while ZVI is an excellent option for promoting the methane production.

Sulfite altered permanganate effects on acetate-enriched short-chain fatty acids production during sludge anaerobic fermentation.

Anaerobic fermentation is a promising method for waste activated sludge (WAS) treatment, but ineffective solubilization and hydrolysis limit its application. The current study examined the function of sodium sulfite (SDS) in potassium permanganate (PP)-conditioned WAS fermentation for short-chain fatty acids (SCFAs) biosynthesis. The presence of SDS in the PP system (PP/SDS) reduced the positive effects of PP on total SCFAs yield (2755 versus 3471 mg COD/L), while effectively increasing the proportion of acetate (from 41 to 81 %). Not only did SDS decrease the promoting effects of PP on WAS solubilization and hydrolysis efficiency by 5-42 %, it also shifted microbial metabolic pathways to favor acetate production. In addition, the amino acid metabolism with acetate as end product was enhanced. Moreover, PP/SDS inhibited methanogenesis, resulting in an accumulation of acetate in high quantities. Thus, the current study a provided insight and direction for effective WAS treatment with acetate-enriched SCFAs production.

The role of pretreatments in handling antibiotic resistance genes in anaerobic sludge digestion - A review.

Sludge is among the most important reservoirs of antibiotic resistance genes (ARGs), which would cause potential environmental risks with the sludge utilization. Currently, anaerobic digestion (AD) is effective to simultaneously realize the resource recovery and pollutants removal, including antibiotic resistance genes (ARGs), and various pretreatments are used to enhance the performance. Recently, plentiful publications have focused on the effects of pretreatment on ARGs removal, but the contradictory results are often obtained, and a comprehensive understanding of the research progress and mechanisms is essential. This study summarizes various pretreatment techniques for improving AD efficiency and ARGs reduction, investigates promising performance in ARGs removal when pretreatments combined with AD, and analyzes the potential mechanisms accounting for ARGs fates. The results showed that although thermal hydrolysis pretreatment showed the best performance in ARGs reduction during the pretreatment process, the significant rebound of ARGs would occur in the subsequent AD process. Conversely, ozone pretreatment and alkali pretreatment had no significant effect on ARGs abundance in the pretreatment stage, but could enhance ARGs removal by 15.6-24.3 % in the subsequent AD. Considering the efficiency and economic effectiveness, free nitrous acid pretreatment would be a promising and feasible option, which could enhance methane yield and ARGs removal by up to 27 % and 74.5 %, respectively. Currently, the factors determining ARGs fates during pretreatment and AD processes included the shift of microbial community, mobile genetic elements (MGEs), and environmental factors. A comprehensive understanding of the relationship between the fate of ARGs and pretreatment technologies could be helpful for systematically evaluating various pretreatments and facilitating the development of emerging and effective pretreatment techniques. Moreover, given the effectiveness, economic efficiency and environmental safety, we called for the applications of modern analysis approaches such as metagenomic and machine learning on the optimization of pretreatment conditions and revealing underlying mechanisms.

The co-occurrent microplastics and nano-CuO showed antagonistic inhibitory effects on bacterial denitrification: Interaction of pollutants and regulations on functional genes.

The wide occurrence of microplastics (MPs) and nanoparticles resulted in their inevitable coexistence in environment. However, the joint effects of these two types of particulate emerging contaminants on denitrification have seldomly been investigated. Herein, non-biodegradable polyvinyl chloride, polypropylene, polyethylene and biodegradable polyhydroxyalkanoate (PHA) MPs were chosen to perform the co-occurrent effects with nano copper oxide (nano-CuO). Both the nano-CuO and MPs inhibited the denitrification process, and biodegradable PHA-MPs showed severer inhibition than non-biodegradable MPs. However, the presence of MPs significantly alleviated the inhibition of nano-CuO, suggesting an antagonistic effect. Other than MPs decreasing copper ion release from nano-CuO, MPs and nano-CuO formed agglomerations and induced lower levels of oxidative stress compared to individual exposure. Transcriptome analysis indicated that the co-occurrent MPs and nano-CuO induced different regulation on denitrifying genes (e. g. nar and nor) compared to individual ones. Also, the expressions of genes involved in denitrification-associated metabolic pathways, including glycolysis and NADH electron transfer, were down-regulated by nano-CuO or MPs, but exhibiting recovery under the co-occurrent conditions. This study firstly discloses the antagonistic effect of nano-CuO and MPs on environmental process, and these findings will benefit the systematic evaluation of MPs environmental behavior and co-occurrent risk with other pollutants.

Explaining the resistomes in a megacity's water supply catchment: Roles of microbial assembly-dominant taxa, niched environments and pathogenic bacteria.

Antibiotic resistance genes (ARGs) in drinking water sources suggest the possible presence of resistant microorganisms that jeopardize human health. However, explanations for the presence of specific ARGs in situ are largely unknown, especially how their prevalence is affected by local microbial ecology, taxa assembly and community-wide gene transfer. Here, we characterized resistomes and bacterial communities in the Taipu River catchment, which feeds a key drinking water reservoir to a global megacity, Shanghai. Overall, ARG abundances decreased significantly as the river flowed downstream towards the reservoir (P < 0.01), whereas the waterborne bacteria assembled deterministically (|ßNRI| > 2.0) as a function of temperature and dissolved oxygen conditions with the assembly-dominant taxa (e.g. Ilumatobacteraceae and Cyanobiaceae) defining local resistomes (P < 0.01, Cohen's D = 4.22). Bacterial hosts of intragenomic ARGs stayed at the same level across the catchment (60 ∼ 70 genome copies per million reads). Among them, the putative resistant pathogens (e.g. Burkholderiaceae) carried mixtures of ARGs that exhibited high transmission probability (transfer counts = 126, P < 0.001), especially with the microbial assembly-dominant taxa. These putative resistant pathogens had densities ranging form 3.0 to 4.0 × 106 cell/L, which was more pronouncedly affected by resistome and microbial assembly structures than environmental factors (SEM, std-coeff ß = 0.62 vs. 0.12). This work shows that microbial assembly and resistant pathogens play predominant roles in prevelance and dissemination of resistomes in receiving water, which deserves greater attention in devisng control strategies for reducing in-situ ARGs and resistant strains in a catchment.

Distinct effects of typical sludge pretreatment approaches on the antibiotic resistance genes variations, associated bacterial community dynamics and metabolic activities during anaerobic fermentation process.

Anaerobic fermentation is effective for waste activated sludge (WAS) disposal to realize resource generation and pollutants reduction, and various pretreatments were commonly applied to improve the performance. This work mainly investigated the effects of typical WAS pretreatment approaches on the antibiotic resistance genes (ARGs, as emerging contaminants) removal during anaerobic fermentation processes and unveiled the underlying mechanisms. The results indicated that all the pretreatment strategies exhibited evident effects on the overall ARGs removal with the order of Fe2+ activated persulfate (PS/Fe2+) > pH 10 > Ultrasonication > Heat, and showed selective removal tendency for the specific ARGs (namely easily removed (aadA1 and sul1) and persistent ARGs). Mechanistic analysis demonstrated that the pretreatments disrupted the extracellular polymeric substances (EPS) and rose the cell membrane permeability (particularly for PS/Fe2+ and Heat). Then the increased ARGs release benefitted the subsequent reduction of mobile genetic elements (MGEs) and extracellular ARGs (especially for PS/Fe2+ and pH10), resulting the ARGs attenuation. Pretreatments significantly shifted the microbial community structure and the abundances of potential ARGs hosts (i.e., Sulfuritalea, and Denitratisoma). Also, the different pretreatments exhibited distinct effects on the microbial metabolic traits related with ARGs proliferation (i.e., ABC transporters, two-component system and bacterial secretion systems), which also contributed to the ARGs attenuations during WAS fermentation. The partial least-squares path modeling (PLS-PM) analysis indicated that the bacterial community (total effects = 0.968) was key factor determining ARGs fates.

Reductive Stress Boosts the Horizontal Transfer of Plasmid-Borne Antibiotic Resistance Genes: The Neglected Side of the Intracellular Redox Spectrum.

The dissemination of plasmid-borne antibiotic resistance genes (ARGs) among bacteria is becoming a global challenge to the "One Health" concept. During conjugation, the donor/recipient usually encounter diverse stresses induced by the surrounding environment. Previous studies mainly focused on the effects of oxidative stress on plasmid conjugation, but ignored the potential contribution of reductive stress (RS), the other side of the intracellular redox spectrum. Herein, we demonstrated for the first time that RS induced by dithiothreitol could significantly boost the horizontal transfer of plasmid RP4 from Escherichia coli K12 to different recipients (E. coli HB101, Salmonella Typhimurium, and Pseudomonas putida KT2440). Phenotypic and genotypic tests confirmed that RS upregulated genes encoding the transfer apparatus of plasmid RP4, which was attributed to the promoted consumption of intracellular glutamine in the donor rather than the widely reported SOS response. Moreover, RS was verified to benefit ATP supply by activating glycolysis (e.g., GAPDH) and the respiratory chain (e.g., appBC), triggering the deficiency of intracellular free Mg2+ by promoting its binding, and reducing membrane permeability by stimulating cardiolipin biosynthesis, all of which were beneficial to the functioning of transfer apparatus. Overall, our findings uncovered the neglected risks of RS in ARG spreading and updated the regulatory mechanism of plasmid conjugation.

Size-dependent effects of microplastics on antibiotic resistance genes fate in wastewater treatment systems: The role of changed surface property and microbial assemblages in a continuous exposure mode.

Microplastics (MPs) were continuously transported to wastewater treatment systems and accumulated in sludge constantly, potentially affecting systems function and co-occurrent contaminants fate. However, previous studies were based on acute exposure of MPs, which could not reflect the dynamics of MPs accumulation. Herein, this study firstly raised a more realistic method to evaluate the practical impacts of MPs on systems purification efficiency and antibiotic resistance genes (ARGs) fate. Continuous exposure of MPs did not pose negative effects on nutrients removal, but significantly changed the occurrence patterns of ARGs. ARGs abundances increased by 42.8 % and 54.3 % when exposed to millimeter-size MPs (mm-MPs) polyamide and polyethylene terephthalate, but increased by 31.3 % and 39.4 % to micron-size MPs (µm-MPs), respectively. Thus, mm-MPs posed severer effects on ARGs than µm-MPs. Further, mm-MPs surface properties were obviously altered after long-term exposure (higher specific surface area and O-containing species), which benefited microbes attachment. More importantly, more taxa linkages and changed topological properties (higher average degree and average weight) of co-occurrent network were observed in sludge with mm-MPs than with µm-MPs, as well as totally different potential host bacteria of ARGs. Rough surface of MPs and closer relations between ARGs and bacteria taxa contributed to the propagation of ARGs, which accounted for the observed higher ARGs abundances of mm-MPs. This study demonstrated that long-term accumulation of MPs in wastewater treatment systems affected ARGs fate, and mm-MPs caused severer risk due to their enrichment of ARGs. The results would promote the understanding of MPs real environmental behavior and influences.

Para-chloro-meta-xylenol reshaped the fates of antibiotic resistance genes during sludge fermentation: Insights of cell membrane permeability, bacterial structure and biological pathways.

The occurrence of para-chloro-meta-xylenol (PCMX, as largely consumed antimicrobial chemicals) in waste activated sludge (WAS) would pose environmental risks for WAS utilization. This study revealed that PCMX principally prompted the abundances and diversity of antibiotic resistance genes (ARGs), particularly for the multidrug- genes (i.e., acrB and mexW), and reshaped the resistance mechanism categories during WAS fermentation process. The genotype and phenotype results indicated that PCMX upregulated abundances of transposase and increased cell permeability via disrupting WAS structure, which further facilitated the horizontal transfer of ARGs. The network and correlation analysis among ARGs, mobile genetic elements (MGEs) and genera (i.e., Sphingopyxis and Pseudoxanthomonas) verified that PCMX enriched the potential ARGs hosts associated with multidrug resistance mechanism. Also, PCMX upregulated the genes involved in ARGs-associated metabolic pathways, such as two-component (i.e., phoP and vcaM) and quorum sensing systems (i.e., lasR and cciR), which determined the ARGs proliferation via multidrug efflux pump and outer membrane proteins, and facilitated the recognition between ARGs hosts. Variance partitioning analysis (VPA) implied that the shift of microbial community contributed predominantly to the dissemination of ARGs. These findings unveiled the environmental behaviors and risks of exogenous pollutants in WAS with insightful understanding, which could guide the WAS utilization for resource recovery.

Size-dependent effects of polystyrene microplastics on anaerobic digestion performance of food waste: Focusing on oxidative stress, microbial community, key metabolic functions.

Polystyrene (PS) microplastics (MPs) are widely existed in food waste (FW) due to the usage of plastic food-packaging. However, the effects and mechanisms of PS MPs with different sizes on anaerobic digestion (AD) performance of FW have not been comprehensively studied yet. Herein, the impacts of different PS MPs sizes (1 mm, 100 µm and 1 µm) with 20, 200 particles/g-TS were investigated. Results showed that 20 particles/g-TS PS MPs decreased cumulative methane production by 1.46-18.11 %, while the higher levels (200 particles/g-TS) significantly inhibited by 9.14-33.08 % (p < 0.05) compared with control group. The inhibiting effects were enhanced as particle size smaller. Physicochemical analysis indicated that MPs prolonged organic matter hydrolysis, weakened the volatile fatty acids metabolism and inhibited methanogenesis-related microorganisms (Synergistetes, Proteiniphilum and Methanosarcina). Small-sized MPs could induce more reactive oxygen species causing cell toxicity and suppressed key enzymes (α-glucoside, protease, acetate kinases and F420) activities, thereby restraining methane production. The analyses of acetyl-CoA synthase and methyl-coenzyme M reductase functional genes illustrated that small-sized MPs negatively affected acetoclastic methanogenesis pathways. Overall, these results provide new insights into the size-dependent effects on AD performance induced by PS MPs.

Unveiling the risks and critical mechanisms of polyhexamethylene guanidine on the antibiotic resistance genes propagation during sludge fermentation process.

This study mainly investigated the environmental risks of polyhexamethylene guanidine (PHMG) occurred in waste activated sludge (WAS) on the antibiotic resistance genes (ARGs) spread during anaerobic fermentation, and disclosed the critical mechanisms. The total ARGs abundance was increased by 32.2-46.4% at different stressing levels of PHMG. The main resistance mechanism categories of ARGs shifted to the target alternation and efflux pump. PHMG disintegrated WAS structure and increased the cell permeability, which benefitted the mobile genetic elements (MGEs) release and horizontal transfer of ARGs. Besides, PHMG induced the enrichment of potential ARGs hosts (i.e., Burkholderia, Bradyrhizobium and Aeromonas). Moreover, PHMG upregulated the metabolic pathways (i.e., two-component system, quorum sensing, and ATP-binding cassette transporters) and critical genes expression (i.e., metN, metQ, rpfF, rstA and rstB) related with ARGs generation and dissemination. Structural equation model analysis revealed that microbial community structure was the predominant contributor to the ARGs propagation.

Potential of biogas residue biochar modified by ferric chloride for the enhancement of anaerobic digestion of food waste.

Biogas residue biochar (BRB) and BRB modified by ferric chloride (BRB-FeCl3) were applied to promote anaerobic digestion (AD) of food waste (FW), related mechanisms were also proposed in this study. Results indicated BRB-FeCl3 showed higher specific surface area, more abundant functional groups and impregnate iron than BRB, and they respectively increased 22.50% and 12.79% cumulative methane yields compared with control group because of accelerated volatile fatty acids (VFAs) transformation, which were confirmed by enhanced metabolism of glycolysis, fatty acid degradation and pyruvate. BRB, especially BRB-FeCl3 facilitated the growth of Syntrophomonas, Methanofollis, Methanoculleus and Methanosarcina, which further promoted the methanogenesis by enhancing the metabolic activities of methanol, dimethylamine and methylamine pathways, thereby causing more metabolically diverse methanogenic pathways. Metagenomics analysis revealed BRB, especially BRB-FeCl3 promoted the relative abundances of functional genes involved in direct interspecies electron transfer (DIET). Present study explored the enhancement mechanisms and feasibility of BRB-FeCl3 for AD process.