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811300.9732Fate of antibiotic resistance genes in mesophilic and thermophilic anaerobic digestion of chemically enhanced primary treatment (CEPT) sludge. Anaerobic digestion (AD) of chemically enhanced primary treatment (CEPT) sludge and non-CEPT (conventional sedimentation) sludge were comparatively operated under mesophilic and thermophilic conditions. The highest methane yield (692.46±0.46mL CH(4)/g VS(removed) in CEPT sludge) was observed in mesophilic AD of CEPT sludge. Meanwhile, thermophilic conditions were more favorable for the removal of total antibiotic resistance genes (ARGs). In this study, no measurable difference in the fates and removal of ARGs and class 1 integrin-integrase gene (intI1) was observed between treated non-CEPT and CEPT sludge. However, redundancy analysis indicated that shifts in bacterial community were primarily accountable for the variations in ARGs and intI1. Network analysis further revealed potential host bacteria for ARGs and intI1.201728797965
811210.9717Fate of antibiotic resistance bacteria and genes during enhanced anaerobic digestion of sewage sludge by microwave pretreatment. The fate of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) were investigated during the sludge anaerobic digestion (AD) with microwave-acid (MW-H), microwave (MW) and microwave-H2O2-alkaline (MW-H2O2) pretreatments. Results showed that combined MW pretreatment especially for the MW-H pretreatment could efficiently reduce the ARB concentration, and most ARG concentrations tended to attenuate during the pretreatment. The subsequent AD showed evident removal of the ARB, but most ARGs were enriched after AD. Only the concentration of tetX kept continuous declination during the whole sludge treatment. The total ARGs concentration showed significant correlation with 16S rRNA during the pretreatment and AD. Compared with unpretreated sludge, the AD of MW and MW-H2O2 pretreated sludge presented slightly better ARB and ARGs reduction efficiency.201626970692
810520.9704Refluxing mature compost to replace bulking agents: A low-cost solution for suppressing antibiotic resistance genes rebound in sewage sludge composting. Antibiotic resistance genes (ARGs) rebounding during composting cooling phase is a critical bottleneck in composting technology that increased ARGs dissemination and application risk of compost products. In this study, mature compost (MR) was used as a substitute for rice husk (RH) to mitigate the rebound of ARGs and mobile genetic elements (MGEs) during the cooling phase of sewage sludge composting, and the relationship among ARGs, MGEs, bacterial community and environmental factors was investigated to explore the key factor influencing ARGs rebound. The results showed that aadD, blaCTX-M02, ermF, ermB, tetX and vanHB significantly increased 4.76-32.41 times, and the MGEs rebounded by 38.60% in the cooling phase of RH composting. Conversely, MR reduced aadD, tetM, ermF and ermB concentrations by 59.49-98.58%, and reduced the total abundance of ARGs in the compost product by 49.32% compared to RH, which significantly restrained ARGs rebound. MR promoted secondary high temperature inactivation of potential host bacteria, including Ornithinibacter, Rhizobiales and Caldicoprobacter, which could harbor aadE, blaCTX-M02, and blaVEB. It also reduced the abundance of lignocellulose degrading bacteria of Firmicutes, which were potential hosts of aadD, tetX, ermF and vanHB. Moreover, MR reduced moisture and increased oxidation reduction potential (ORP) that promoted aadE, tetQ, tetW abatement. Furthermore, MR reduced 97.36% of total MGEs including Tn916/1545, IS613, Tp614 and intI3, which alleviated ARGs horizontal transfer. Overall finding proposed mature compost reflux as bulking agent was a simple method to suppress ARGs rebound and horizontal transfer, improve ARGs removal and reduce composting plant cost.202539798649
810930.9697The fate of antibiotic resistance genes and their influential factors in swine manure composting with sepiolite as additive. Manures are storages for antibiotic resistance genes (ARGs) entering the environment. This study investigated the effects of adding sepiolite at 0%, 2.5%, 5%, and 7.5% (CK, T1, T2, and T3, respectively) on the fates of ARGs during composting. The relative abundances (RAs) of the total ARGs in CK and T3 decreased by 0.23 and 0.46 logs, respectively, after composting. The RAs of 10/11 ARGs decreased in CK, whereas they all decreased in T3. The reduction in the RA of the total mobile genetic elements (MGEs) was 1.26 times higher in T3 compared with CK after composting. The bacterial community accounted for 47.93% of the variation in the abundances of ARGs. Network analysis indicated that ARGs and MGEs shared potential host bacteria (PHB), and T3 controlled the transmission of ARGs by reducing the abundances of PHB. Composting with 7.5% sepiolite is an effective strategy for reducing the risk of ARGs proliferating.202235063626
805440.9695Effects of nanoscale zero-valent iron on the performance and the fate of antibiotic resistance genes during thermophilic and mesophilic anaerobic digestion of food waste. The effects of nanoscale zero-valent iron (nZVI) on the performance of food waste anaerobic digestion and the fate of antibiotic resistance genes (ARGs) were investigated in thermophilic (TR) and mesophilic (MR) reactors. Results showed that nZVI enhanced biogas production and facilitated ARGs reduction. The maximum CH(4) production was 212.00 ± 4.77 ml/gVS with 5 g/L of nZVI in MR. The highest ARGs removal ratio was 86.64 ± 0.72% obtained in TR at nZVI of 2 g/L. nZVI corrosion products and their contribution on AD performance were analyzed. The abundance of tetracycline genes reduced significantly in nZVI amended digesters. Firmicutes, Chloroflexi, Proteobacteria and Spirochaetes showed significant positive correlations with various ARGs (p < 0.05) in MR and TR. Redundancy analysis indicated that microbial community was the main factor that influenced the fate of ARGs. nZVI changed microbial communities, with decreasing the abundance bacteria belonging to Firmicutes and resulting in the reduction of ARGs.201931505392
805850.9691Effects of biochars on the fate of antibiotics and their resistance genes during vermicomposting of dewatered sludge. It is currently still difficult to decrease the high contents of antibiotics and their corresponding antibiotic resistance genes (ARGs) in sludge vermicompost. To decrease the environmental risk of vermicompost as a bio-fertilizer, this study investigated the feasibility of biochar addition to decrease the levels of antibiotics and ARGs during vermicomposting of dewatered sludge. To achieve this, 1.25% and 5% of corncob and rice husk biochars, respectively, were added to sludge, which was then vermicomposted by Eisenia fetida for 60 days. The sludge blended with corncob biochar showed increased decomposition and humification of organic matter. Higher biochar concentration promoted both the number and diversity of bacteria, and differed dominant genera. The level of antibiotics significantly decreased as a result of biochar addition (P < 0.05), and tetracycline was completely removed. Relative to the control without addition of biochars, ermF and tetX genes significantly decreased with corncob biochar treatment (P < 0.05). Rice husk biochar (5%) could effectively decrease sul-1 and sul-2 genes in vermicompost (P < 0.05). However, the abundance of the intI-1 gene increased with biochar concentration. This study suggests that biochar addition can lessen the antibiotic and ARG pollution in sludge vermicompost, depending on the type and concentration of biochars.202032388093
794060.9691Microplastics affect the ammonia oxidation performance of aerobic granular sludge and enrich the intracellular and extracellular antibiotic resistance genes. Microplastics (MPs) and antibiotic resistance genes (ARGs), as emerging pollutants, are frequently detected in wastewater treatment plants, and their threats to the environment have received extensive attentions. However, the effects of MPs on the nitrification of aerobic granular sludge (AGS) and the spread patterns of intracellular and extracellular ARGs (iARGs and eARGs) in AGS were still unknown. In this study, the responses of AGS to the exposure of 1, 10 and 100 mg/L of typical MPs (polyvinyl chloride (PVC), polyamide (PA), polystyrene (PS) and polyethylene (PE)) and tetracycline were focused on in 3 L nitrifying sequencing batch reactors. 10 mg/L MPs decreased the nitrification function, but nitrification could recover. Furthermore, MPs inhibited ammonia-oxidizing bacteria and enriched nitrite-oxidizing bacteria, leading partial nitrification to losing stability. PVC, PA and PS stimulated the secretion of extracellular polymeric substances and reactive oxygen species. PE had less negative effect on AGS than PVC, PA and PS. The abundances of iARGs and eARGs (tetW, tetE and intI1) increased significantly and the intracellular and extracellular microbial communities obviously shifted in AGS system under MPs stress. Potential pathogenic bacteria might be the common hosts of iARGs and eARGs in AGS system and were enriched in AGS and MPs biofilms.202133387747
789870.9689Effects of graphite and Mn ore media on electro-active bacteria enrichment and fate of antibiotic and corresponding resistance gene in up flow microbial fuel cell constructed wetland. This study assessed the influence of substrate type on pollutants removal, antibiotic resistance gene (ARG) fate and bacterial community evolution in up-flow microbial fuel cell constructed wetlands (UCW-MFC) with graphite and Mn ore electrode substrates. Better COD removal and higher bacterial community diversity and electricity generation performance were achieved in Mn ore constructed UCW-MFC (Mn). However, the lower concentration of sulfadiazine (SDZ) and the total abundances of ARGs were obtained in the effluent in the graphite constructed UCW-MFC (s), which may be related to higher graphite adsorption and filter capacity. Notably, both reactors can remove more than 97.8% of ciprofloxacin. In addition, significant negative correlations were observed between SDZ, COD concentration, ARG abundances and bacterial a-diversity indices. The LEfse analysis revealed significantly different bacterial communities due to the substrate differences in the two reactors, and Geobacter, a typical model electro-active bacteria (EAB), was greatly enriched on the anode of UCW-MFC (Mn). In contrast, the relative abundance of methanogens (Methanosaeta) was inhibited. PICRUSt analysis results further demonstrated that the abundance of extracellular electron transfer related functional genes was increased, but the methanogen function genes and multiple antibiotic resistance genes in UCW-MFC (Mn) anode were reduced. Redundancy analyses indicated that substrate type, antibiotic accumulation and bacterial community were the main factors affecting ARGs. Moreover, the potential ARG hosts and the co-occurrence of ARGs and intI1 were revealed by network analysis.201931442759
811780.9686Composting of oxytetracycline fermentation residue in combination with hydrothermal pretreatment for reducing antibiotic resistance genes enrichment. Hydrothermal pretreatment can efficiently remove the residual antibiotics in oxytetracycline fermentation residue (OFR), but its effect on antibiotic resistance genes (ARGs) during composting remains unclear. This study compared the shifts in bacterial community and evolutions in ARGs and integrons during different composting processes of OFRs with and without hydrothermal pretreatment. The results demonstrated that hydrothermal pretreatment increased the bacterial alpha diversity at the initial phase, and increased the relative abundances of Proteobacteria and Actinobacteria but decreased that of Bacteroidetes at the final phase by inactivating mycelia and removing residual oxytetracycline. Composting process inevitably elevated the abundance and relative abundance of ARGs. However, the increase in ARGs was significantly reduced by hydrothermal pretreatment, because the removal of oxytetracycline decreased their potential host bacteria and inhibited their horizontal gene transfer. The results demonstrated that hydrothermal pretreatment is an efficient strategy to reduce the enrichment of ARGs during the OFR composting.202033099099
787690.9685Sulfamethoxazole impact on pollutant removal and microbial community of aerobic granular sludge with filamentous bacteria. In this study, sulfamethoxazole (SMX) was employed to investigate its impact on the process of aerobic granule sludge with filamentous bacteria (FAGS). FAGS has shown great tolerance ability. FAGS in a continuous flow reactor (CFR) could keep stable with 2 μg/L of SMX addition during long-term operation. The NH(4)(+), chemical oxygen demand (COD), and SMX removal efficiencies kept higher than 80%, 85%, and 80%, respectively. Both adsorption and biodegradation play important roles in SMX removal for FAGS. The extracellular polymeric substances (EPS) might play important role in SMX removal and FAGS tolerance to SMX. The EPS content increased from 157.84 mg/g VSS to 328.22 mg/g VSS with SMX addition. SMX has slightly affected on microorganism community. A high abundance of Rhodobacter, Gemmobacter, and Sphaerotilus of FAGS may positively correlate to SMX. The SMX addition has led to the increase in the abundance of the four sulfonamide resistance genes in FAGS.202336871701
8110100.9683Removal of chlortetracycline and antibiotic resistance genes in soil by earthworms (epigeic Eisenia fetida and endogeic Metaphire guillelmi). The impacts of two ecological earthworms on the removal of chlortetracycline (CTC, 0.5 and 15 mg kg(-1)) and antibiotic resistance genes (ARGs) in soil were explored through the soil column experiments. The findings showed that earthworm could significantly accelerate the degradation of CTC and its metabolites (ECTC) in soil (P < 0.05), with epigeic Eisenia fetida promoting degradation rapidly and endogeic Metaphire guillelmi exhibiting a slightly better elimination effect. Earthworms alleviated the abundances of tetR, tetD, tetPB, tetG, tetA, sul1, TnpA, ttgB and intI1 in soil, with the total relative abundances of ARGs decreasing by 35.0-44.2% in earthworm treatments at the 28th day of cultivation. High throughput sequencing results displayed that the structure of soil bacteria community was modified apparently with earthworm added, and some possible CTC degraders, Aeromonas, Flavobacterium and Luteolibacter, were promoted by two kinds of earthworms. Redundancy analysis demonstrated that the reduction of CTC residues, Actinobacteria, Acidobacteria and Gemmatimonadetes owing to earthworm stimulation was responsible for the removal of ARGs and intI1 in soil. Additionally, intI1 declined obviously in earthworm treatments, which could weaken the risk of horizontal transmission of ARGs. Therefore, earthworm could restore the CTC-contaminated soil via enhancing the removal of CTC, its metabolites and ARGs.202133798888
7877110.9679External circuit loading mode regulates anode biofilm electrochemistry and pollutants removal in microbial fuel cells. This study investigated the effects of different external circuit loading mode on pollutants removal and power generation in microbial fuel cells (MFC). The results indicated that MFC exhibited distinct characteristics of higher maximum power density (P(max)) (named MFC-HP) and lower P(max) (named MFC-LP). And the capacitive properties of bioanodes may affect anodic electrochemistry. Reducing external load to align with the internal resistance increased P(max) of MFC-LP by 54.47 %, without no obvious effect on MFC-HP. However, intermittent external resistance loading (IER) mitigated the biotoxic effects of sulfamethoxazole (SMX) (a persistent organic pollutant) on chemical oxygen demand (COD) and NH(4)(+)-N removal and maintained high P(max) (424.33 mW/m(2)) in MFC-HP. Meanwhile, IER mode enriched electrochemically active bacteria (EAB) and environmental adaptive bacteria Advenella, which may reduce antibiotic resistance genes (ARGs) accumulation. This study suggested that the external circuit control can be effective means to regulate electrochemical characteristics and pollutants removal performance of MFC.202439153696
8056120.9678Antibiotic resistance gene profiles and evolutions in composting regulated by reactive oxygen species generated via nano ZVI loaded on biochar. In this study, nano zero-valent iron loaded on biochar (BC-nZVI) was analyzed for its effects on antibiotic resistance genes (ARGs) in composting. The results showed that BC-nZVI increased reactive oxygen species (ROS) production, and the peak values of H(2)O(2) and OH were 22.95 % and 55.30 % higher than those of the control group, respectively. After 65 days, the relative abundances of representative ARGs decreased by 56.12 % in the nZVI group (with BC-nZVI added). An analysis of bacterial communities and networks revealed that Actinobacteria, Proteobacteria, and Firmicutes were the main hosts for ARGs, and BC-nZVI weakened the link between ARGs and host bacteria. Distance-based redundancy analysis showed that BC-nZVI altered the microbial community structure through environmental factors and that most ARGs were negatively correlated with ROS, suggesting that ROS significantly affected the relative abundance of ARGs. According to these results, BC-nZVI showed potential for decreasing the relative abundance of ARGs in composting.202337611721
7942130.9678Insight into effects of polyethylene microplastics in anaerobic digestion systems of waste activated sludge: Interactions of digestion performance, microbial communities and antibiotic resistance genes. 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.202235944782
8055140.9673Effects of nano-zerovalent iron on antibiotic resistance genes during the anaerobic digestion of cattle manure. This study investigated the effects of adding nano-zerovalent iron (nZVI) at three concentrations (0, 80, and 160 mg/L) on the methane yield and the fate of antibiotic resistance genes (ARGs) during the anaerobic digestion (AD) of cattle manure. The addition of nZVI effectively enhanced the methane yield, where it significantly increased by 6.56% with 80 mg/L nZVI and by 6.43% with 160 mg/L nZVI. The reductions in the abundances of ARGs and Tn916/1545 were accelerated by adding 160 mg/L nZVI after AD. Microbial community analysis showed that nZVI mainly increased the abundances of bacteria with roles in hydrolysis and acidogenesis, whereas it reduced the abundance of Acinetobacter. Redundancy analysis indicated that the changes in mobile genetic elements made the greatest contribution to the fate of ARGs. The results suggest that 160 mg/L nZVI is a suitable additive for reducing the risks due to ARGs in AD.201931247529
8017150.9672Dose-Dependent Effect of Tilmicosin Residues on ermA Rebound Mediated by IntI1 in Pig Manure Compost. The impact of varying antibiotic residue levels on antibiotic resistance gene (ARG) removal during composting is still unclear. This study investigated the impact of different residue levels of tilmicosin (TIM), a common veterinary macrolide antibiotic, on ARG removal during pig manure composting. Three groups were used: the CK group (no TIM), the L group (246.49 ± 22.83 mg/kg TIM), and the H group (529.99 ± 16.15 mg/kg TIM). Composting removed most targeted macrolide resistance genes (MRGs) like ereA, ermC, and ermF (>90% removal), and reduced ermB, ermX, ermQ, acrA, acrB, and mefA (30-70% removal). However, ermA increased in abundance. TIM altered compost community structure, driving succession through a deterministic process. At low doses, TIM reduced MRG-bacteria co-occurrence, with horizontal gene transfer via intI1 being the main cause of ermA rebound. In conclusion, composting reduces many MRG levels in pig manure, but the persistence and rebound of genes like ermA reveal the complex interactions between composting conditions and microbial gene transfer.202541011454
7897160.9672Enhanced removal of antibiotic and antibiotic resistance genes by coupling biofilm electrode reactor and manganese ore substrate up-flow microbial fuel cell constructed wetland system. Manganese ore substrate up-flow microbial fuel cell constructed wetland (UCW-MFC(Mn)) as an innovative wastewater treatment technology for purifying antibiotics and electricity generation with few antibiotic resistance genes (ARGs) generation has attracted attention. However, antibiotic purifying effects should be further enhanced. In this study, a biofilm electrode reactor (BER) that needs direct current driving was powered by a Mn ore anode (UCW-MFC(Mn)) to form a coupled system without requiring direct-current source. Removal efficiencies of sulfadiazine (SDZ), ciprofloxacin (CIP) and the corresponding ARGs in the coupled system were compared with composite (BER was powered by direct-current source) and anaerobic systems (both of BER and UCW-MFC were in open circuit mode). The result showed that higher antibiotic removal efficiency (94% for SDZ and 99.1% for CIP) in the coupled system was achieved than the anaerobic system (88.5% for SDZ and 98.2% for CIP). Moreover, electrical stimulation reduced antibiotic selective pressure and horizontal gene transfer potential in BER, and UCW-MFC further reduced ARG abundances by strengthening the electro-adsorption of ARG hosts determined by Network analysis. Bacterial community diversity continuously decreased in BER while it increased in UCW-MFC, indicating that BER mitigated the toxicity of antibiotic. Degree of modularity, some functional bacteria (antibiotic degrading bacteria, fermentative bacteria and EAB), and P450 enzyme related to antibiotic and xenobiotics biodegradation genes were enriched in electric field existing UCW-MFC, accounting for the higher degradation efficiency. In conclusion, this study provided an effective strategy for removing antibiotics and ARGs in wastewater by operating a BER-UCW-MFC coupled system.202337437616
8053170.9669Increasing the removal efficiency of antibiotic resistance through anaerobic digestion with free nitrous acid pretreatment. Swine manure is a significant reservoir for antibiotic resistance. Anaerobic digestion (AD) is a common biological process used to treat swine manure but still faces low efficiencies in biogas production and antibiotic resistance removal. It is here shown that AD with free nitrous acid pretreatment (FNA) was effective in reducing antibiotic resistance genes (ARGs) in swine manure. FNA pretreatment (nitrite =250 mg N/L, pH=5.0, temperature=20 ± 1 °C) simultaneously reduced antibiotics (Tetracyclines, Quinones and Sulfonamides), inactivated antibiotics resistance bacteria (ARB) by 0.5-3 logs, and decreased ARGs tet, sul and qnr by 1-2, 1-3 and 0.5 logs, respectively. In the following AD step, the total residual ARGs was reduced to ~3.49 × 10(7) gene copies/g dry total solids (TS), ~1 log lower than that in the AD without pretreatment (3.55 ×10(8) gene copies/g dry TS). Microbial community and network analyses revealed that the ARG removal was mainly driven by the direct FNA effect on reducing ARGs and antibiotics, not related to ARB. Besides, the FNA pretreatment doubled biochemical methane production potential from swine manure. Together these results demonstrate that AD with FNA pretreatment is a useful process greatly facilitating swine manure management.202235816802
8059180.9668Particle size of zero-valent iron affects the risks from antibiotic resistance genes in waste activated sludge during anaerobic digestion. Zero-valent iron (ZVI) is the promising enhancer for sludge anaerobic digestion (AD) performance and for mitigating the proliferation of antibiotic resistance genes (ARGs). However, concerns about its size effects in shifting the behavior and risk of ARGs in sludge, during the AD process. Here, the metagenomics-based profile of ARGs, along with their potential (pathogenic) hosts in sludge were investigated, during mesophilic AD enhanced by ZVI with three different sizes. Results showed that the size of ZVI affected the profiles of ARGs, with nano-ZVI (nZVI, 50 nm) demonstrating the most significant reduction in abundance (by 45.0 %) and diversity (by 8.6 %) of total ARGs, followed by micron-ZVI (150 μm) and iron scrap (1 mm). Similar trends were also observed for high-risk ARGs, pathogens, and potential pathogenic hosts for ARGs. Notably, nZVI achieved the greatest reductions in the abundance of risk ARGs and potential pathogenic hosts (superbugs) by 58.8 % and 53.9 %, respectively. Correlation and redundancy analyses revealed that, the size of ZVI induced concentration differences in ammonium nitrogen, pH, carbonaceous matters, iron, and potential microbial hosts were the main reasons for the variation in the risk of ARGs. Moreover, the down-regulation of genes involved in oxidative stress contributed to the lower risk of ARGs in the three ZVI groups, especially in nZVI. This study provides insights into AD processes of solid wastes using ZVI enhancers.202540043404
7941190.9668Microplastics accelerate nitrification, shape the microbial community, and alter antibiotic resistance during the nitrifying process. Microplastics (MPs) and antibiotic resistance genes (ARGs) are both emerging pollutants that are frequently detected in wastewater treatment plants. In this study, the effects of various MPs, including polyethylene (PE), polyvinyl chloride (PVC), and biodegradable polylactic acid (PLA), on nitrification performance, dominant microbial communities, and antibiotic resistance during nitrification were investigated. The results revealed that the addition of MPs increased the specific ammonia oxidation rate and specific nitrate production rate by 15.2 % - 15.5 % and 8.0 % - 11.6 %, respectively, via enrichment of nitrifying microorganisms, Nitrospira and Nitrosomonas. Moreover, ARGs were selectively enriched in nitrifying sludge and microplastic biofilms under stress from different MPs. Compared with PE-MPs (23.9 %) and PVC-MPs (21.4 %), exposure to PLA-MPs significantly increased intI1 abundance by 51.6 %. The results of the variance decomposition analysis implied that MPs and the microbial community play important roles in the behavior of ARGs. Network analysis indicated that Nitrosomonas and potentially pathogenic bacteria emerged as possible hosts, harboring ARGs and intI1 genes in the nitrifying sludge and microplastic biofilms. Critically, PLA-MPs were found to enrich both ARGs and potential pathogenic bacteria during nitrification, which should be considered in their promotion of application processes due to their biodegradability.202539740624