Parachlorometaxylenol stress caused multidrug-type antibiotic resistance genes proliferation via simultaneously reshaping microbial community and interfering metabolic traits during wastewater treatment process.

Despite biological wastewater treatment processes (e.g., sequencing batch reactors (SBR)) being able to reduce the dissemination of antibiotic resistance genes (ARGs), the variation of ARGs under exogenous pollutant stress is an open question. This work investigated the impacts of para-chloro-meta-xylenol (PCMX, typical antibacterial contaminants) on ARGs spread in long-term SBR operation. Although the SBR process inherently decreased ARGs abundance, the presence of PCMX substantially amplified both the prevalence (mainly multidrug) and abundance of total ARGs (1.17-fold of the control). Further analysis demonstrated that PCMX disintegrated sludge structures as well as increased membrane permeability, facilitating the release of mobile genetic elements and subsequent horizontal transfer of ARGs. In addition, PCMX selectively enriched potential ARG hosts, notably Nitrospira and Candidatus Accumulibacter, which predominantly served as multidrug ARG hosts. Concurrently, the self-adaptive functions of ARGs hosts in the PCMX-exposed SBR system were activated via quorum sensing, two-component regulatory system, ATP-binding cassette transporters, and bacterial secretion system. The upregulation of these metabolic pathways also contributed to the dissemination of ARGs.

Disinfectant polyhexamethylene guanidine triggered simultaneous efflux pump antibiotic- and metal-resistance genes propagation during sludge anaerobic digestion.

The environmental transmission of antibiotic resistance genes (ARGs) and metal resistance genes (MRGs) exerted devastating threats to global public health, and their interactions with other emerging contaminants (ECs) have raised increasing concern. This study investigated that the abundances of ARGs and MRGs with the predominant type of efflux pump were simultaneously increased (8.4-59.1%) by disinfectant polyhexamethylene guanidine (PHMG) during waste activated sludge (WAS) anaerobic digestion. The aggregation of the same microorganisms (i.e., Hymenobacter and Comamonas) and different host bacteria (i.e., Azoarcus and Thauera) were occurred upon exposure to PHMG, thereby increasing the co-selection and propagation of MRGs and ARGs by vertical gene transfer. Moreover, PHMG enhanced the process of horizontal gene transfer (HGT), facilitating their co-transmission by the same mobile genetic elements (20.2-223.7%). Additionally, PHMG up-regulated the expression of critical genes (i.e., glnB, trpG and gspM) associated with the HGT of ARGs and MRGs (i.e., two-component regulatory system and quorum sensing) and exocytosis system (i.e., bacterial secretion system). Structural equation model analysis further verified that the key driver for the simultaneous enrichment of ARGs and MRGs under PHMG stress was microbial community structure. The study gives new insights into the aggravated environmental risks and mechanisms of ECs in sludge digestion system, providing guidance for subsequent regulation and control of ECs.

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.

Plasticizers inhibit food waste anaerobic digestion performance by affecting microbial succession and metabolism.

The widely existed plastic additives plasticizers in organic wastes possibly pose negative influences on anaerobic digestion (AD) performance, the direct evidence about the effects of plasticizers on AD performance is still lacking. This study evaluated the influencing mechanism of two typical plasticizers bisphenol A (BPA) and dioctyl phthalate on the whole AD process. Results indicated that plasticizers addition inhibited methane production, and the inhibiting effects were reinforced with the increase of concentration. By contrast, 50 mg/L BPA exhibited the strongest inhibition on methane production. Physicochemical analysis showed plasticizers inhibited the metabolism efficiency of soluble polysaccharide and volatile fatty acids. Microbial communities analyses suggested that plasticizers inhibited the direct interspecies electron transfer participators of methanogenic archaea (especially Methanosarcina) and syntrophic bacteria. Furthermore, plasticizers inhibited the methane metabolisms, key coenzymes (CoB, CoM, CoF420 and methanofuran) biosynthesis and the metabolisms of major organic matters. This study shed light on the effects of plasticizers on AD performance and provided new insights for assessing the influences of plasticizers or plastic additives on the disposal of organic wastes.

Mitigated dissemination of antibiotic resistance genes by nanoscale zero-valent iron and iron oxides during anaerobic digestion: Roles of microbial succession and regulation.

Nanoscale zero-valent iron (ZVI) and the oxides have been documented as an effective approach for mitigating the dissemination of antibiotic resistance genes (ARGs) during anaerobic digestion (AD). However, the mechanism of ARGs dissemination mitigated by nanoscale ZVI and iron oxides remain unclear. Here, we investigated the influencing mechanisms of nanoscale ZVI and iron oxides on ARGs dissemination during AD. qPCR results indicated that nanoscale ZVI and iron oxides significantly declined the total ARGs abundances, and the strongest inhibiting effect was observed by 10 g/L nanoscale ZVI. Mantel test showed ARGs distribution was positively correlated with physiochemical properties, integrons and microbial community, among which microbial community primarily contributed to ARGs dissemination (39.74%). Furthermore, redundancy and null model analyses suggested the dominant and potential ARGs host was Fastidiosipila, and homogeneous selection in the determinism factors was the largest factor for driving Fastidiosipila variation, confirming the inhibition of Fastidiosipila was primary reason for mitigating ARGs dissemination by nanoscale ZVI and iron oxides. These results were related to the inhibition of ARGs transfer related functions. This work provides novel evidence for mitigating ARGs dissemination through regulating microbial succession and regulation induced by ZVI and iron oxides.

Revealing mechanisms of NH3 and N2O emissions reduction in the rapid bio-drying of food waste: Insights from organic nitrogen composition and microbial activity.

Inevitably, aerobic biological treatment processes generate emissions of ammonia (NH3) and greenhouse gas (GHGs) emissions, especially nitrous oxide (N2O). The rapid bio-drying process (RBD) for food waste (FW) alleviates issues arising from its substantial growth. However, its emissions of NH3 and N2O remain unknown, and the correlation with nitrogen components in the substrate remains unclear, significantly impeding its widespread adoption. Here, the nitrogen loss and its mechanisms in RBD were investigated, and the results are as follows: The total emission of NH3 and N2O were1.42 and 1.16 mg/kg FW (fresh weight), respectively, achieving a 98 % reduction compared to prior studies. Structural equation modeling demonstrates that acid ammonium nitrogen (AN) decomposition chiefly generates NH3 in compost (p < 0.001). Strong correlation (p < 0.001) exists between amino acid nitrogen (AAN) and AN. In-depth analysis of microbial succession during the process reveals that the enrichment of Brevibacterium, Corynebacterium, Dietzia, Fastidiosipila, Lactobacillus, Mycobacterium, Peptoniphilus, and Truepera, are conducive to reducing the accumulation of AN and AAN in the substrate, minimizing NH3 emissions (p < 0.05). While Pseudomonas, Denitrobacterium, Nitrospira, and Bacillus are identified as key species contributing to N2O emissions during the process. Correlation analysis between physicochemical conditions and microbial succession in the system indicates that the moisture content and NO3- levels during the composting process provide suitable conditions for the growth of bacteria that contribute to NH3 and N2O emissions reduction, these enrichment in RBD process minimizing NH3 and N2O emissions. This study can offer crucial theoretical and data support for the resource utilization process of perishable organic solid waste, mitigating NH3 and GHGs emissions.

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.

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.

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.

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.

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.

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.

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.

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.