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811300.9046Fate 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
811210.9038Fate 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
805420.9032Effects 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
810930.9032The 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
810540.9029Refluxing 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
774750.9022Hydrothermal pre-treatment followed by anaerobic digestion for the removal of tylosin and antibiotic resistance agents from poultry litter. Hydrothermal pretreatment (HPT) followed by anaerobic digestion (AD) is an alternative for harvesting energy and removing organic contaminants from sewage sludge and animal manure. This study investigated the use, in an energetically sustainable way, of HPT and AD, alone or combined, to produce methane and remove tylosin and antimicrobial resistance genes (ARG) from poultry litter (PL). The results showed that HPT at 80 °C (HPT80), followed by single-stage AD (AD-1S), led to the production of 517.9 ± 4.7 NL CH(4) kg VS(-1), resulting in 0.11 kWh kg PL(-1) of electrical energy and 0.75 MJ kg PL(-1) of thermal energy, thus supplying 33.6% of the energy spent on burning firewood at a typical farm. In this best-case scenario, the use of HPT alone reduced tylosin concentration from PL by 23.6%, while the process involving HPT followed by AD-1S led to the removal of 91.6% of such antibiotic. The combined process (HPT80 + AD-1S), in addition to contributing to reduce the absolute and relative abundances of ARG ermB (2.13 logs), intI1 (0.39 logs), sul1 (0.63 logs), and tetA (0.74 logs), led to a significant removal in the relative abundance of tylosin-resistant bacteria present in the poultry litter.202336648713
787660.9021Sulfamethoxazole 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
798870.9018Electrokinetic 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
798980.9016Feasibility 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
810890.9009Insights into the beneficial effects of woody peat for reducing abundances of antibiotic resistance genes during composting. Antibiotic resistance genes (ARGs) in manure endangered human health, while heavy metals in manure will pose selective pressure on ARGs. This study explored the effects on ARGs of adding woody peat during composting at different ratios (0 (CK), 5% (T1), and 15% (T2)). After composting, the relative abundances of 8/11 ARGs were 6.97-38.09% and 10.73-54.31% lower in T1 and T2, respectively, than CK. The bioavailable Cu content was 1.40% and 18.40% lower in T1 and T2, respectively, than CK. Network analysis showed that ARGs, mobile genetic elements (MGEs), and metal resistance genes possessed common potential host bacteria, such as Streptococcus, Dietzia, and Corynebacterium_1. Environmental factors, especially bioavailable Cu, and MGEs accounted for 80.75% of the changes in the abundances of ARGs. In conclusion, 15% Woody peat is beneficial to decrease the bioavailable Cu content and weaken horizontal gene transfer for controlling the spread of ARGs during composting.202134534940
8058100.9009Effects 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
7877110.9006External 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
7745120.9004Iron-modified biochar boosts anaerobic digestion of sulfamethoxazole pharmaceutical wastewater: Performance and microbial mechanism. The accumulation of volatile fatty acids (VFAs) caused by antibiotic inhibition significantly reduces the treatment efficiency of sulfamethoxazole (SMX) wastewater. Few studies have been conducted to study the VFAs gradient metabolism of extracellular respiratory bacteria (ERB) and hydrogenotrophic methanogen (HM) under high-concentration sulfonamide antibiotics (SAs). And the effects of iron-modified biochar on antibiotics are unknown. Here, the iron-modified biochar was added to an anaerobic baffled reactor (ABR) to intensify the anaerobic digestion of SMX pharmaceutical wastewater. The results demonstrated that ERB and HM were developed after adding iron-modified biochar, promoting the degradation of butyric, propionic and acetic acids. The content of VFAs reduced from 1166.0 mg L(-1) to 291.5 mg L(-1). Therefore, chemical oxygen demand (COD) and SMX removal efficiency were improved by 22.76% and 36.51%, and methane production was enhanced by 6.19 times. Furthermore, the antibiotic resistance genes (ARGs) such as sul1, sul2, intl1 in effluent were decreased by 39.31%, 43.33%, 44.11%. AUTHM297 (18.07%), Methanobacterium (16.05%), Geobacter (6.05%) were enriched after enhancement. The net energy after enhancement was 0.7122 kWh m(-3). These results confirmed that ERB and HM were enriched via iron-modified biochar to achieve high efficiency of SMX wastewater treatment.202337030222
8053130.9004Increasing 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
8055140.9002Effects 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
8110150.9002Removal 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
8049160.9000Microalgae simultaneously promote antibiotic removal and antibiotic resistance genes/bacteria attenuation in algal-bacterial granular sludge system. This study investigated the effects of microalgae growth on antibiotic removal and the attenuation of antibiotic resistance genes (ARGs)/ARGs host bacteria in algal-bacterial granular sludge (ABGS) system. In the presence of tetracycline (TC) and sulfadiazine (SDZ) mixture (2-4 mg/L), microalgae could grow on bacterial granular sludge (BGS) to form ABGS, with a chlorophyll-a content of 7.68-8.13 mg/g-VSS being achieved. The removal efficiencies of TC and SDZ by ABGS were as high as 79.0 % and 94.0 %, which were 4.3-5.0 % higher than those by BGS. Metagenomic analysis indicated that the relative abundances of TC/SDZ- related ARGs and mobile genetic elements (MGEs) in BGS were 56.1 % and 22.1 % higher than those in ABGS. A total of 26 ARGs were detected from the granules, and they were identified to associate with 46 host bacteria. 13 out of 26 ARGs and 13 out of 46 hosts were shared ARGs and hosts, respectively. The total relative abundance of host bacteria in BGS was 30.8 % higher than that in ABGS. Scenedesmus and Chlorella were the dominant microalgae that may reduce the diversity of ARGs hosts. Overall, ABGS is a promising biotechnology for antibiotic-containing wastewater treatment.202235777142
7861170.8999The removal of antibiotic resistant bacteria and genes and inhibition of the horizontal gene transfer by contrastive research on sulfidated nanoscale zerovalent iron activating peroxymonosulfate or peroxydisulfate. Antibiotic resistant bacteria (ARB) and the antibiotic resistance genes (ARGs) dissemination via plasmid-mediated conjugation have attracted considerable attentions. In this research, sulfidated nanoscale zerovalent iron (S-nZVI)/peroxymonosulfate (PMS) and S-nZVI/peroxydisulfate (PDS) process were investigated to inactivate ARB (Escherichia coli DH5α with RP4 plasmid, Pseudomonas. HLS-6 contains sul1 and intI1 on genome DNA sequence). S-nZVI/PMS system showed higher efficiency than S-nZVI/PDS on ARB inactivation. Thus, the optimal condition 28 mg/L S-nZVI coupled with 153.7 mg/L (0.5 mM) PMS was applied to remove both intracellular ARGs (iARGs) and ARB. The oxidative damage of ARB cell was systemically studied by cell viability, intracellular Mg(2+) levels, the changes of extracellular and internal structure, integrity of cell walls and membranes and enzymatic activities. S-nZVI/PMS effectively inactivated ARB (~7.32 log) within 15 min. These effects were greatly higher than those achieved individually. Moreover, removal efficiencies of iARGs sul1, intI1 and tetA were 1.52, 1.79 and 1.56 log, respectively. These results revealed that S-nZVI and PMS have a synergistic effect against ARB and iARGs. The regrowth assays illustrated that the ARB were effectively inactivated. By verifying the inhibitory impacts of S-nZVI/PMS treatment on conjugation transfer, this work highlights a promising alternative technique for inhibiting the horizontal gene transfer.202234482079
7831180.8998Integration of nanowire-confined electroporation of antibiotic-resistant bacteria and electroactivation of peracetic acid for eliminating intracellular resistance genes. Antimicrobial resistance is one of the most substantial challenges for global public health. To address the inefficient elimination of intracellular resistance genes (i-ARGs) in antibiotic-resistant bacteria (ARB) by peracetic acid (PAA) oxidation, we developed an integration strategy (NW-EP/EA) of nanowire-confined electroporation (NW-EP) of ARB cells and nanowire-confined electroactivation (NW-EA) of PAA with a sequential oxidation-reduction process. The locally enhanced electric field and electrocatalytic activity over NW tips prompted the formation of electroporation pores on ARB cells and the generation of reactive ⋅OH and RO⋅ radicals by PAA electroactivation. The NW-EP/EA with Pd-coated TiO(2)NW cathode with atomic H* evolution exhibited 0.6 -2.8-log higher i-ARG removal than the pristine TiO(2)NW cathode, especially achieving ∼5.0-log i-ARG removal (99.999 %) at 4.0 V and 2.0 mM PAA with ∼4.1-log synergistic effect and ∼10 times lower energy consumption as compared with the individual NW-EP (∼0.32-log and 52.1 %) and PAA (∼0.56-log and 74.4 %). For the sequential oxidation-reduction process, the electrooxidative activation of PAA on TiO(2)NW anode produced H(+) ions, ⋅OH and RO⋅ radicals for enlarging electroporation pores, and the generated H(+) ions promoted the evolution of atomic H* and electroreduction of PAA on subsequent Pd-TiO(2)NW cathode for further facilitating ARB cell damages, i-ARG leakage and degradation. The effective i-ARGs removal and HGT inhibition in tap water suggested the great application potentials of NW-EP/EA in the control of ARGs dissemination risks in drinking water.202540907311
7923190.8997Effect of ultrasonic and ozone pretreatment on the fate of enteric indicator bacteria and antibiotic resistance genes, and anaerobic digestion of dairy wastewater. In this study, the effect of ultrasound (US), ozone and US combined with ozone (US/ozone) pretreatments on the fate of enteric indicator bacteria and antibiotic resistance genes (ARGs), and anaerobic digestion (AD) of dairy wastewater was investigated. The pretreatment conditions included US power 200 W, ozone concentration 4.2 mg O(3)/L, and pretreatment time 0-30 min. The results showed that US/ozone pretreatment was effective in the inactivation of enteric indicator bacteria. Total coliforms and enterococci were reduced by 99% and 92% after 30 min US/ozone pretreatment. Pretreatments could not decrease ARGs in absolute concentration, but could decrease ARGs in relative abundance. In the subsequent AD process, methane production increased more than 10% with 20 min ozone or 20 min US/ozone pretreatments. Pretreatment-AD together obviously inhibited the enrichment of ARGs in relative abundance. This study provided a pretreatment way to enhance methane production and to prevent the enrichment of ARGs.202133186838