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811300.8736Fate 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
714910.8729Municipal Solid Waste Treatment System Increases Ambient Airborne Bacteria and Antibiotic Resistance Genes. Landfill and incineration are the primary disposal practices for municipal solid waste (MSW) and have been considered as the critical reservoir of antibiotic resistance genes (ARGs). However, the possible transmission of ARGs from the municipal solid waste treatment system (MSWT system) to ambient air is still unclear. In this study, we collected inside and ambient air samples (PM(10) and PM(2.5)) and potential source samples (leachate and solid waste) in the MSWT system. The results showed that the MSWT system contributed to the increased ambient airborne bacteria and associated ARGs. Forty-one antibiotic-resistant bacteria (ARB) harboring bla(TEM-1) were isolated, and the full-length nucleotide sequences of the bla(TEM-1) gene (harbored by identical bacillus) from air (downwind samples) were 100% identical with those in the leachate and solid waste, indicating that the MSWT system was the important source of disperse bacteria and associated ARGs in the ambient air. The daily intake (DI) burden level of ARGs via PM inhalation was comparable with that via ingestion of drinking water but lower than the DI level via ingestion of raw vegetables. The antibiotic-resistant opportunistic pathogen Bacillus cereus was isolated from air, with a relatively high DI level of Bacillus via inhalation (10(4)-10(6) copies/day) in the MSWT system. This study highlights the key pathway of airborne ARGs to human exposure.202032208626
811020.8727Removal 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
798930.8725Feasibility of sulfate-calcined eggshells for removing pathogenic bacteria and antibiotic resistance genes from landfill leachates. High abundance of human pathogen and antibiotic resistance genes (ARGs) in landfill leachate has become an emerging threat against human health. Therefore, sulfate- and calcination-modified eggshells as green agricultural bioresource were applied to test the feasibility of removing pathogenic bacteria and ARGs from leachate. The highest removal of Escherichia coli (E. coil) and gentamycin resistant gene (gmrA) from artificial contaminated landfill leachate was achieved by the application of eggshell with combined treatment of sulfate and calcination. The 16S and gmrA gene copies of E. coil declined significantly from 1.78E8±8.7E6 and 4.12E8±5.9E6 copies mL(-1) to 1.32E7±2.6E6 and 2.69E7±7.2E6 copies mL(-1), respectively, within 24h dynamic adsorption equilibrium process (p<0.05). Moreover, according to the Langmuir kinetic model, the greatest adsorption amount (1.56×10(9) CFU E. coil per gram of modified eggshells) could be obtained at neutral pH of 7.5. The optimal adsorption eggshells were then screened to the further application in three typical landfill leachates in Nanjing, eastern China. Significant decrease in species and abundance of pathogenic bacteria and ARGs (tet, sul, erm, qnr, and ampC) indicated its great efficiency to purify landfill leachates. This study demonstrated that sulfate-calcined eggshells can be an environmentally-friendly and highly efficient bioadsorbent to the management of reducing dissemination risk of pathogen and ARGs in landfill leachate.201728343745
812940.8724Pesticide contamination and antimicrobial resistance: Two threats to the Neotropical Otter (Lontra longicaudis) in the Peñas Blancas River Basin, Costa Rica. The effects of synthetic pesticides on antibiotic-resistance genes (ARGs) in bacterial communities from contaminated waters are unclear. Otters in the Peñas Blancas basin encounter various anthropogenic residues, including pesticides. In 2022, we analyzed the presence of pesticides in six water samples and ARGs in eight otter fecal samples. Thirteen pesticides (herbicides, insecticides, fungicides, and multi-target) and seven ARGs (qnrS, tetA, tetB, tetQ, tetW, sulI, sulII) were detected. Regulated pesticides such as chlorpyrifos and ethoprophos, along with diazinon, diuron, imidacloprid, and terbutryn were found. These pesticides have been implicated in promoting antimicrobial resistance (AMR) in bacteria, particularly when combined with sub-lethal doses of antibiotics. Elevated levels of ethoprophos (0.67 ng/L) and a fecal sample containing four ARGs (tetA, tetB, sulI, and sulII) came from the upper basin. Our findings reveal pesticide application practices in the region, and highlight the potential risk of pesticide exposure to wildlife, including development of AMR.202540473152
810950.8716The 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
782360.8711Gamma-ray irradiation as an effective method for mitigating antibiotic resistant bacteria and antibiotic resistance genes in aquatic environments. The prevalence of antibiotic resistance genes (ARGs) in municipal wastewater treatment plants (MWTPs) has emerged as a significant environmental concern. Despite advanced treatment processes, high levels of ARGs persist in the secondary effluent from MWTPs, posing ongoing environmental risks. This study explores the potential of gamma-ray irradiation as a novel approach for sterilizing antibiotic-resistant bacteria (ARB) and reducing ARGs in MWTP secondary effluent. Our findings reveal that gamma-ray irradiation at an absorbed dose of 1.6 kGy effectively deactivates all culturable bacteria, with no subsequent revival observed after exposure to 6.4 kGy and a 96-h incubation in darkness at room temperature. The removal efficiencies for a range of ARGs, including tetO, tetA, bla(TEM-1), sulI, sulII, and tetW, were up to 90.5% with a 25.6 kGy absorbed dose. No resurgence of ARGs was detected after irradiation. Additionally, this study demonstrates a considerable reduction in the abundances of extracellular ARGs, with the transformation efficiencies of extracellular tetracycline and sulfadiazine resistance genes decreasing by 56.3-81.8% after 25.6 kGy irradiation. These results highlight the effectiveness of gamma-ray irradiation as an advanced and promising method for ARB sterilization and ARG reduction in the secondary effluent of MWTPs, offering a potential pathway to mitigate environmental risks associated with antibiotic resistance.202438367438
783670.8706Efficient Degradation of Intracellular Antibiotic Resistance Genes by Photosensitized Erythrosine-Produced (1)O(2). Intracellular antibiotic resistance genes (iARGs) constitute the important part of wastewater ARGs and need to be efficiently removed. However, due to the dual protection of intracellular DNA by bacterial membranes and the cytoplasm, present disinfection technologies are largely inefficient in iARG degradation. Herein, we for the first time found that erythrosine (ERY, an edible dye) could efficiently degrade iARGs by producing abundant (1)O(2) under visible light. Seven log antibiotic-resistant bacteria were inactivated within only 1.5 min, and 6 log iARGs were completely degraded within 40 min by photosensitized ERY (5.0 mg/L). A linear relationship was established between ARG degradation rate constants and (1)O(2) concentrations in the ERY photosensitizing system. Surprisingly, a 3.2-fold faster degradation of iARGs than extracellular ARGs was observed, which was attributed to the unique indirect oxidation of iARGs induced by (1)O(2). Furthermore, ERY photosensitizing was effective for iARG degradation in real wastewater and other photosensitizers (including Rose Bengal and Phloxine B) of high (1)O(2) yields could also achieve efficient iARG degradation. The findings increase our knowledge of the iARG degradation preference by (1)O(2) and provide a new strategy of developing technologies with high (1)O(2) yield, like ERY photosensitizing, for efficient iARG removal.202337531556
811280.8704Fate 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
801990.8700In-feed antibiotic use changed the behaviors of oxytetracycline, sulfamerazine, and ciprofloxacin and related antibiotic resistance genes during swine manure composting. The dynamics of oxytetracycline (OTC), sulfamerazine (SM1), ciprofloxacin (CIP) and related antibiotic resistance genes (ARGs) during swine manure composting were compared between manure collected from swine fed a diet containing these three antibiotics (T(D)) and manure directly spiked with these drugs (T(S)). The composting removal efficiency of OTC (94.9 %) and CIP (87.8 %) in the T(D) treatment was significantly higher than that of OTC (83.8 %, P <  0.01) and CIP (83.9 %, P <  0.05) in the T(S) treatment, while SM1 exhibited no significant difference (P >  0.05) between the two treatments. Composting effectively reduced the majority of ARGs in both T(D) and T(S) types of manure, especially tetracycline resistance genes (TRGs). Compared with the T(S) treatment, the abundance of some ARGs, such as tetG, qepA, sul1 and sul2, increased dramatically up to 309-fold in the T(D) treatment. The microbial composition of the composting system changed significantly during composting due to antibiotic feeding. Redundancy analysis suggested that the abundance of ARGs had a considerable impact on alterations in the physicochemical parameters (C/N, pH and temperature) and bacterial communities (Actinobacteria, Proteobacteria and Firmicutes) during the composting of swine manure.202133254754
7876100.8699Sulfamethoxazole 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
7874110.8699Phenacetin promoted the rapid start-up and stable maintenance of partial nitrification: Responses of nitrifiers and antibiotic resistance genes. Phenacetin (PNCT) belongs to one of the earliest synthetic antipyretics. However, impact of PNCT on nitrifying microorganisms in wastewater treatment plants and its potential microbial mechanism was still unclear. In this study, PN could be initiated within six days by PNCT anaerobic soaking treatment (8 mg/L). In order to improve the stable performance of PN, 21 times of PNCT aerobic soaking treatment every three days was conducted and PN was stabilized for 191 days. After PN was damaged, ten times of PNCT aerobic soaking treatment every three days was conducted and PN was recovered after once soaking, maintained over 88 days. Ammonia oxidizing bacteria might change the dominant oligotype to gradually adjust to PNCT, and the increase of abundance and activity of Nitrosomonas promoted the initiation of PN. For nitrite-oxidizing bacteria (NOB), the increase of Candidatus Nitrotoga and Nitrospira destroyed PN, but PN could be recovered after once aerobic soaking illustrating NOB was not resistant to PNCT. KEGG and COG analysis suggested PNCT might disrupt rTCA cycle of Nitrospira, resulting in the decrease of relative abundance of Nitrospira. Moreover, PNCT did not lead to the sharp increase of absolute abundances of antibiotic resistance genes (ARGs), and the risk of ARGs transmission was negligible.202438744392
7922120.8696Simultaneous removal and high tolerance of norfloxacin with electricity generation in microbial fuel cell and its antibiotic resistance genes quantification. Norfloxacin (NFLX) is a synthetic antibiotic widely used in the treatment of infectious diseases. In this work, the performance of microbial fuel cells (MFCs) toward NFLX degradation, electricity production and the antibiotics resistances genes (ARGs) generation was investigated. NFLX degradation efficiency and COD removal reached 65.5% and 94.5% respectively. The increase in NFLX concentration (128 mg/L) had no significant influence on NFLX degradation efficiency, COD removal and MFCs voltage output while the electricity was successfully generated. The quantitative polymerase chain reaction (qPCR) indicated low absolute abundances of ARGs (mdtk, mdtm, and pmra) compared with the traditional wastewater treatment plants (WWTPs). Anodic bacteria can survive in the presence of high NFLX concentration and sustain its degradation and electricity production. In terms of NFLX degradation, COD removal, diminished ARGs generation and simultaneous energy production, MFC seems to be a promising technology for antibiotics wastewater treatment with a potential to overcome the ARGs challenge.202032097778
3483130.8696Abundance and diversity of antibiotic resistance genes and bacterial communities in the western Pacific and Southern Oceans. This study investigated the abundance and diversity of antibiotic resistance genes (ARGs) and the composition of bacterial communities along a transect covering the western Pacific Ocean (36°N) to the Southern Ocean (74°S) using the Korean icebreaker R/V Araon (total cruise distance: 14,942 km). The relative abundances of ARGs and bacteria were assessed with quantitative PCR and next generation sequencing, respectively. The absolute abundance of ARGs was 3.0 × 10(6) ± 1.6 × 10(6) copies/mL in the western Pacific Ocean, with the highest value (7.8 × 10(6) copies/mL) recorded at a station in the Tasman Sea (37°S). The absolute abundance of ARGs in the Southern Ocean was 1.8-fold lower than that in the western Pacific Ocean, and slightly increased (0.7-fold) toward Terra Nova Bay in Antarctica, possibly resulting from natural terrestrial sources or human activity. β-Lactam and tetracycline resistance genes were dominant in all samples (88-99%), indicating that they are likely the key ARGs in the ocean. Correlation and network analysis showed that Bdellovibrionota, Bacteroidetes, Cyanobacteria, Margulisbacteria, and Proteobacteria were positively correlated with ARGs, suggesting that these bacteria are the most likely ARG carriers. This study highlights the latitudinal profile of ARG distribution in the open ocean system and provides insights that will help in monitoring emerging pollutants on a global scale.202235085628
7054140.8696Effective removal of antibiotic resistance genes and potential links with archaeal communities during vacuum-type composting and positive-pressure composting. As a major reservoir of antibiotics, animal manure contributes a lot to the augmented environmental pressure of antibiotic resistance genes (ARGs). This might be the first study to explore the effects of different ventilation types on the control of ARGs and to identify the relationships between archaeal communities and ARGs during the composting of dairy manure. Several ARGs were quantified via Real-time qPCR and microbial communities including bacteria and archaea were analyzed by High-throughput sequencing during vacuum-type composting (VTC) and positive-pressure composting (PPC). The total detected ARGs and class I integrase gene (intI1) under VTC were significantly lower than that under PPC during each stage of the composting (p<0.001). The relative abundance of potential human pathogenic bacteria (HPB) which were identified based on sequencing information and correlation analysis decreased by 74.6% and 91.4% at the end of PPC and VTC, respectively. The composition of archaeal communities indicated that methane-producing archaea including Methanobrevibacter, Methanocorpusculum and Methanosphaera were dominant throughout the composting. Redundancy analysis suggested that Methanobrevibacter and Methanocorpusculum were positively correlated with all of the detected ARGs. Network analysis determined that the possible hosts of ARGs were different under VTC and PPC, and provided new sights about potential links between archaea and ARGs. Our results showed better performance of VTC in reducing ARGs and potential HPB and demonstrated that some archaea could also be influential hosts of ARGs, and caution the risks of archaea carrying ARGs.202031892399
8105150.8695Refluxing 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
7897160.8695Enhanced 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
7160170.8694High-throughput profiling of antibiotic resistance genes in the Yellow River of Henan Province, China. Profiling antibiotic resistance genes (ARGs) in the Yellow River of China's Henan Province is essential for understanding the health risks of antibiotic resistance. The profiling of ARGs was investigated using high-throughput qPCR from water samples in seven representative regions of the Yellow River. The absolute and relative abundances of ARGs and moble genetic elements (MGEs) were higher in summer than in winter (ANOVA, p < 0.001). The diversity and abundance of ARGs were higher in the Yellow River samples from PY and KF than the other sites. Temperature (r = 0.470 ~ 0.805, p < 0.05) and precipitation (r = 0.492 ~ 0.815, p < 0.05) positively influenced the ARGs, while pH had a negative effect (r = - 0.462 ~ - 0.849, p < 0.05). Network analysis indicated that the pathogenic bacteria Rahnella, Bacillus, and Shewanella were the possible hub hosts of ARGs, and tnpA1 was the potential MGE hub. These findings provide insights into the factors influencing ARG dynamics and the complex interaction among the MGEs, pathogenic bacteria and environmental parameters in enriching ARGs in the Yellow River of Henan Province.202439080455
7904180.8694Effect of the coexposure of sulfadiazine, ciprofloxacin and zinc on the fate of antibiotic resistance genes, bacterial communities and functions in three-dimensional biofilm-electrode reactors. Three-dimensional biofilm electrode reactors (3D-BERs) with high treatment efficiency were constructed to treat wastewater containing sulfadiazine (SDZ) and ciprofloxacin (CIP) coexposure with Zinc (Zn). The results showed that coexposure to target antibiotics and Zn increased the absolute and relative abundances of target antibiotic resistance genes (ARGs). Additionally, the target ARG abundances were higher on cathode of 3D-BER compared with ordinary anaerobic reactor while the abundances of total ARGs were decreased in the effluent. Meanwhile, redundancy analysis results revealed that the composition of bacteria carrying ARGs was greatly influenced in the cathode by the accumulation of Zn and antibiotic, which dominated the changes of ARG abundances. Additionally, ARGs with their host bacteria revealed by network analysis were partially deposited on electrode substrates when being removed from wastewater. Thus, 3D-BER exhibits capability of simultaneously eliminating antibiotic and Zn, and greatly reduces the risks of ARGs spread.202031677404
7988190.8694Electrokinetic treatment at the thermophilic stage achieves more effective control of heavy metal resistance in swine manure composting. Excessive heavy metals (HMs) and metal resistance genes (MRGs) in manure pose significant environmental and human health risks. Our previous work proved enhanced control of antibiotic resistance and quality of swine manure composting with electrokinetic technology (EK). As a continuous study, EK treatments were further employed at typical stages of composting. The humification level increased significantly in EK treatments applied at the thermophilic stage (EK1) and throughout the whole composting period (EK2). The immobilization efficiency of heavy metals increased by 3.02 %-20.90 % for EK1, and 3.86 %-20.56 % for EK2, compared with the EK treatment applied at maturity stage (EK3). EK1 showed the highest ability to remove MRGs (29.38 %-87.13 %), while the abundance of potential host bacteria increased in EK2, raising potential transmission risk of MRGs. Furthermore, there was an elevated presence of bacteria associated with membrane transport as a response mechanism to HMs stress in EK1. Considering economic factors and environmental effects, EK treatment during the thermophilic stage was more effective in compost maturation, HMs passivation, as well as control of HMs resistance. This study provides an effective method to address HMs-related contamination with highly efficient maturation in swine manure composting.202540543370