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721500.9823High-throughput qPCR profiling of antimicrobial resistance genes and bacterial loads in wastewater and receiving environments. Wastewater treatment plants (WWTPs) are hot spots for the acquisition and spread of antimicrobial resistance (AMR). This regional-based study quantified antibiotic resistance genes (ARGs), mobile genetic elements (MGEs), and bacteria in hospital and community-derived wastewater and receiving environments, using high-throughput qPCR (HT-qPCR). This is the first study to apply Resistomap's Antibiotic Resistance Gene Index (ARGI) as a standardised metric to find the overall AMR level across different WWTPs. ARGI of WWTPs ranged from 2.0 to 2.3, indicating higher relative ARG levels than the mean European ARGI of 2.0, but lower than the global mean of 2.4. The highest diversity and abundance of ARGs were observed in untreated hospital and community wastewater. The reduction of total ARGs during wastewater treatment (0.2-2 logs) and bacteria (0.3-1.5 logs) varied spatio-temporally across the WWTPs. Despite a decrease in ARG and bacterial abundance in treated effluents, substantial loads were still released into receiving environments. Notably, ARG levels in coastal sediments were comparable to those in untreated wastewater, and most ARGs were shared between wastewater and receiving environments, highlighting the impact of wastewater discharge on these ecosystems. Sewage outfall exposure increased ARGs in shellfish, emphasising risks to shellfish hygiene. This study provides evidence to inform policymaking, emphasising advanced wastewater treatment methods and combined sewer overflow (CSO) management to mitigate ARG release, protecting water users and the food chain.202540127809
792410.9822Electro-peroxone pretreatment for enhanced simulated hospital wastewater treatment and antibiotic resistance genes reduction. Hospital wastewater is one of the possible sources responsible for antibiotic resistant bacteria spread into the environment. This study proposed a promising strategy, electro-peroxone (E-peroxone) pretreatment followed by a sequencing batch reactor (SBR) for simulated hospital wastewater treatment, aiming to enhance the wastewater treatment performance and to reduce antibiotic resistance genes production simultaneously. The highest chemical oxygen demand (COD) and total organic carbon (TOC) removal efficiency of 94.3% and 92.8% were obtained using the E-peroxone-SBR process. The microbial community analysis through high-throughput sequencing showed that E-peroxone pretreatment could guarantee microbial richness and diversity in SBR, as well as reduce the microbial inhibitions caused by antibiotic and raise the amount of nitrification and denitrification genera. Specially, quantitative real-time PCRs revealed that E-peroxone pretreatment could largely reduce the numbers and contents of antibiotic resistance genes (ARGs) production in the following biological treatment unit. It was indicated that E-peroxone-SBR process may provide an effective way for hospital wastewater treatment and possible ARGs reduction.201829550711
762120.9817Pre-chlorination in source water endows ARB with resistance to chlorine disinfection in drinking water treatment. Chlorine disinfection is widely used to ensure biosafety of drinking water. However, antibiotic resistance bacteria (ARB) and antibiotic resistance genes (ARGs) are often detected in treated drinking water. The impact of chlorine disinfection on the abundance of ARGs in drinking water is currently contradictory. Some studies suggested that chlorine disinfection could reduce the abundance of ARGs, while others had found that chlorine disinfection increased the abundance of ARGs. Pre-chlorination is widely applied in raw water to kill the algae cells in source water Pump Station. Different distances between the source water Pump Station and the drinking water treatment plants (DWTPs) resulted in different degrees of residual chlorine decay in the incoming raw water. This study found that the abundance of ARGs in drinking water would be increased during chlorine disinfection when the chlorine concentration in raw water was higher (> 0.2 mg/L). On the contrary, chlorine disinfection would decrease the abundance of ARGs in drinking water when the chlorine concentration in raw water was lower (< 0.09 mg/L). Pre-chlorination in source water with sub-lethal concentration could allow ARB to adapt to the chlorine environment in advance, endowing ARB with chlorine resistance, which resulted in ineffective removal of ARB and increased ARGs abundance during subsequent chlorine disinfection. High abundance of chlorine and antibiotics co-resistance bacteria in raw water was the main reason for the increase in ARGs abundance in chlorine treated drinking water. It should be noticed that, pre-chlorination treatment in source water would increase the difficulty of removing ARGs in subsequent chlorine disinfection process.202540398032
793330.9816Removal of antibiotic microbial resistance by micro- and ultrafiltration of secondary wastewater effluents at pilot scale. Low-pressure membrane filtration was investigated at pilot scale with regard to its removal of antimicrobial resistance genes (ARGs) in conventional secondary treated wastewater plant effluents. While operating microfiltration (MF) and ultrafiltration (UF) membranes, key operational parameters for antimicrobial resistance (AMR) studies and key factors influencing AMR removal efficiencies of low-pressure membrane filtration processes were examined. The main factor for AMR removal was the pore size of the membrane. The formation of the fouling layer on capillary membranes had only a small additive effect on intra- and extrachromosomal ARG removal and a significant additive effect on mobile ARG removal. Using feeds with different ARGs abundances revealed that higher ARG abundance in the feed resulted in higher ARG abundance in the filtrate. Live-Dead cell counting in UF filtrate showed intact bacteria breaking through the UF membrane. Strong correlations between 16S rRNA genes (as surrogate for bacteria quantification) and the sul1 gene in UF filtrate indicated ARBs likely breaking through UF membranes.202235598662
727840.9814Effects of snowmelt runoff on bacterial communities and antimicrobial resistance gene concentrations in an urban river in a cold climate region. Urban rivers are essential for human activities and ecosystems. Urban runoff is a major source of various pollutants in urban rivers. In this study, we investigated the effect of rainfall and snowmelt subsequently causing urban runoff in a cold climate region on bacterial community structures and antimicrobial resistance gene concentrations in an urban river in Sapporo city, Japan, which has an average snowfall of 4.8 m. Bacterial community structures of the river water were analyzed by next generation sequencing of bacterial 16S rRNA genes. The antimicrobial resistance genes, mphA and bla(IMP), were determined using quantitative polymerase chain reaction. Rainfall and snowmelt increased the effluent discharge rate of treated wastewater, and river water depth. Rainfall and snowmelt also increased Escherichia coli concentrations by 4-20 folds in the river, probably because of combined sewer overflows, urban runoff, or increased effluent discharge rate of treated wastewater to the river. Urban runoff and the subsequent discharge of treated wastewater decreased the bacterial alpha diversity and increased the species evenness of bacteria. Bacterial beta diversity analysis showed that the discharge of treated wastewater caused by rainfall and snowmelt changed the structure and diversity of the bacterial community in the river. The concentrations of the antimicrobial resistance gene mphA were related to the discharge of treated wastewater. In contrast, the antimicrobial resistance gene bla(IMP) appeared to be present in the upstream pristine environment. Results of this study should be informative for challenge to reduce the antimicrobial resistance bacteria due to combined sewer overflows by wastewater management authorities.202540042701
811250.9814Fate 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
730660.9814Impact of the particulate matter from wastewater discharge on the abundance of antibiotic resistance genes and facultative pathogenic bacteria in downstream river sediments. Wastewater treatment plants (WWTPs) are point sources for both, the release of antibiotic resistance genes (ARGs) and the discharge of antibiotics (ABs) into the environment. While it is well established that ARGs emission by WWTPs leads to an ARGs increase in receiving rivers, also the role of sub-inhibitory AB concentrations in this context is being discussed. However, the results obtained in this study suggest that, at environmental concentrations, ABs do not have an effect on resistance selection. Instead, we emphasize the significance of ARG transport and, in that respect, highlight the relevance of wastewater particles and associated microorganisms. We can show that ARGs (ermB, bla(TEM,)tetM, qnrS) as well as facultative pathogenic bacteria (FPB) (enterococci, Pseudomonas aeruginosa, Acinetobacter baumannii) inside the particulate fraction of WWTP effluent are very likely to remain in the riverbed of the receiving water due to sedimentation. Moreover, ARG and FPB abundances measured in the particulate fraction strongly correlated with the delta ARG and FPB abundances measured in the receiving river sediment (downstream compared to upstream) (R(2) = 0.93, p < 0.05). Apparently, the sheer amount of settleable ARGs and FPB from WWTP effluent is sufficient, to increase abundances in the receiving riverbed by 0.5 to 2 log units.201930308888
755370.9813Does the biological treatment or membrane separation reduce the antibiotic resistance genes from swine wastewater through a sequencing-batch membrane bioreactor treatment process. Swine wastes are the reservoir of antibiotic resistance genes (ARGs), which can potentially spread from swine farms to the environment. This study establishes a sequencing-batch membrane bioreactor (SMBR) for ARG removal from swine wastewater, and analyzes the effect of biological treatment and membrane separation on the ARG removal at different solid retention times (SRTs). The SMBR removed 2.91 logs (copy number) of ARGs at a short SRT (12 days). Raising the SRT reduced the removal rates of the detected genes by the biological treatment. Under the relative long SRT (30 days), ARGs and mobile genetic elements (MGEs) were maximized within the reactor and were well removed by membrane separation, with the average genes removal rate of 2.95 (copy number) and 1.18 logs (abundance). At the relatively low SRT, the biological treatment showed the dominant ARG removal effect, while the membrane separation took the advantages of ARG removal especially at the relatively long SRT. The ARG profile was related to the shift of the microbial community structure. The ARGs coexisted with the functional bacteria (ammonia oxidizing bacteria, nitrite oxidizing bacteria and denitrifiers), suggesting they are hosted by the functional bacteria.201829906721
785880.9812Photocatalytic Reactive Ultrafiltration Membrane for Removal of Antibiotic Resistant Bacteria and Antibiotic Resistance Genes from Wastewater Effluent. Biological wastewater treatment is not effective in removal of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs). In this study, we fabricated a photocatalytic reactive membrane by functionalizing polyvinylidene fluoride (PVDF) ultrafiltration (UF) membrane with titanium oxide (TiO(2)) nanoparticles for the removal of ARB and ARGs from a secondary wastewater effluent. The TiO(2)-modified PVDF membrane provided complete retention of ARB and effective photocatalytic degradation of ARGs and integrons. Specifically, the total removal efficiency of ARGs (i.e., plasmid-mediated floR, sul1, and sul2) with TiO(2)-modified PVDF membrane reached ∼98% after exposure to UV irradiation. Photocatalytic degradation of ARGs located in the genome was found to be more efficient than those located in plasmid. Excellent removal of integrons (i.e., intI1, intI2, and intI3) after UV treatment indicated that the horizontal transfer potential of ARGs was effectively controlled by the TiO(2) photocatalytic reaction. We also evaluated the antifouling properties of the TiO(2)-UF membrane to demonstrate its potential application in wastewater treatment.201829984583
721790.9812Emission and dispersal of antibiotic resistance genes through bioaerosols generated during the treatment of municipal sewage. Wastewater treatment plants act as socio-ecological couplers through the concentration, treatment, and subsequent environmental release of sewage collected from surrounding communities and are often considered hotspots for antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs). While studies have identified the release of ARB/ARGs in treated liquid sewage, little is known about potential dispersal through wastewater bioaerosol emissions. The aim of this study was to better define the contribution of WWTP bioaerosols to potential environmental distribution of ARB/ARGs. Bioaerosols were collected immediately upwind and downwind from the aeration tanks of a municipal wastewater treatment plant and liquid sludge samples were obtained from the aeration tanks. From the bioaerosol and liquid samples, qPCR assays identified 44 ARGs that confer resistance to a wide range of antibiotics. Comparison of the ARG profiles across samples showed that the downwind bioaerosol profile was 68% similar to the profile found in liquid sludge samples. Community 16S rRNA gene sequencing also showed that downwind bioaerosols had similar taxonomic profiles as those generated from liquid sludge while the upwind profiles showed a distinct difference. Preliminary ARG dispersion modeling estimated an ARG emission rate of ~10,620 genes per hour from the liquid sludge and indicated that the bioaerosols have the potential to be carried kilometers away from the WWTP source based on wind speed. The overall results from this study suggest that bioaerosols generated during WWTP processes can aid in the emission and dispersal of bacteria and ARGs, resulting in a possible route of human exposure and deposition into surrounding environments.201931181526
7932100.9811How multi-walled carbon nanotubes in wastewater influence the fate of coexisting antibiotic resistant genes in the subsequent disinfection process. Wastewater treatment plants (WWTPs) are important hubs for the spread of antibiotic resistance genes (ARGs). Engineered nanoparticles, which was inevitably released to WWTPs, could change environmentally sensitive of antibiotic resistant bacteria (ARB). This would influence the fate of ARGs in subsequent disinfection process and consequent health risk. In this study, the ARGs fate of the effluent in conventional sodium hypochlorite (NaClO) disinfection process was investigated as multi-walled carbon nanotubes (MWCNTs) existed in sequencing batch reactor (SBR). The results showed the existence of MWCNTs in SBR could enhance the removal efficiency of intracellular 16S rRNA gene and intI1, extracellular intI1, sul2 and tetX in the effluent by NaClO. This is mainly due to the variation of bacterial physiological status, bacterial population structure and the activation of NaClO under the role of MWCNTs. MWCNTs in SBR could increase in membrane permeability of bacterial cells, which would be conducive to the penetration of chlorination to cytoplasm. MWCNTs in SBR also could change the bacterial population structure and induce the chlorine-sensitive bacteria; thus the potential hosts of ARGs in the effluent would be more easily inactivated by NaClO. Moreover, the residual MWCNTs in the effluent could activate NaClO to generate various free radical, which would enhance the oxidizing capacity of chlorination.202235500623
7276110.9811Antibiotic resistance in urban and hospital wastewaters and their impact on a receiving freshwater ecosystem. The main objective of this study was to investigate the antibiotic resistance (AR) levels in wastewater (WW) and the impact on the receiving river. Samples were collected once per season over one year in the WW of a hospital, in the raw and treated WW of two wastewater treatment plants (WWTPs), as well as upstream and downstream from the release of WWTPs effluents into the Zenne River (Belgium). Culture-dependent methods were used to quantify Escherichia coli and heterotrophic bacteria resistant to amoxicillin, sulfamethoxazole, nalidixic acid and tetracycline. Six antibiotic resistance genes (ARGs) were quantified in both particle-attached (PAB) and free-living (FLB) bacteria. Our results showed that WWTPs efficiently removed antibiotic resistant bacteria (ARB) regardless of its AR profile. The ARGs levels were the highest in the hospital WW and were significantly reduced in both WWTPs. However, ARB and ARGs abundances significantly increased into the Zenne River downstream from the WWTPs outfalls. The variation in the relative abundance of ARGs through WW treatment differed depending on the WWTP, fraction, and gene considered. The sul1 and sul2 genes in PAB fraction showed significantly higher relative abundances in the effluent compared to the influent of both WWTPs. This study demonstrated that WWTPs could be hotspots for AR spread with significant impacts on receiving freshwater ecosystems. This was the first comprehensive study investigating at the same time antibiotics occurrence, fecal bacteria indicators, heterotrophic bacterial communities, and ARGs (distinguishing PAB and FLB) to assess AR levels in WW and impacts on the receiving river.201829730567
8545120.9810Role of anaerobic sludge digestion in handling antibiotic resistant bacteria and antibiotic resistance genes - A review. Currently, anaerobic sludge digestion (ASD) is considered not only for treating residual sewage sludge and energy recovery but also for the reduction of antibiotic resistance genes (ARGs). The current review highlights the reasons why antibiotic resistant bacteria (ARB) and ARGs exist in ASD and how ASD performs in the reduction of ARB and ARGs. ARGs and ARB have been detected in ASD with some reports indicating some of the ARGs can be completely removed during the ASD process, while other studies reported the enrichment of ARB and ARGs after ASD. This paper reviews the performance of ASD based on operational parameters as well as environmental chemistry. More studies are needed to improve the performance of ASD in reducing ARGs that are difficult to handle and also differentiate between extracellular (eARGs) and intracellular ARGs (iARGs) to achieve more accurate quantification of the ARGs.202133735726
7934130.9810Mitigated membrane fouling and enhanced removal of extracellular antibiotic resistance genes from wastewater effluent via an integrated pre-coagulation and microfiltration process. Antibiotic resistance genes (ARGs) have been regarded as an emerging pollutant in municipal wastewater treatment plant (WWTP) effluents due to their potential risk to human health and ecological safety when reused for landscape and irrigation. Conventional wastewater treatment processes generally fail to effectively reduce ARGs, especially extracellular ARGs (eARGs), which are persistent in the environment and play an important role in horizontal gene transfer via transformation. Herein, an integrated process of pre-coagulation and microfiltration was developed for removal of ARGs, especially eARGs, from wastewater effluent. Results show that the integrated process could effectively reduce the absolute abundances of total ARGs (tARGs) (>2.9 logs) and eARGs (>5.2 logs) from the effluent. The excellent performance could be mainly attributed to the capture of antibiotic resistant bacteria (ARB) and eARGs by pre-coagulation and co-rejection during subsequent microfiltration. Moreover, the integrated process exhibited a good performance on removing common pollutants (e.g., dissolved organic carbon and phosphate) from the effluent to improve water quality. Besides, the integrated process also greatly reduced membrane fouling compared with microfiltration. These findings suggest that the integrated process of pre-coagulation and microfiltration is a promising advanced wastewater treatment technology for ARGs (especially eARGs) removal from WWTP effluents to ensure water reuse security.201931085389
7922140.9810Simultaneous 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
7555150.9809Deciphering the factors influencing the discrepant fate of antibiotic resistance genes in sludge and water phases during municipal wastewater treatment. The discrepant fate of antibiotic resistance genes (ARGs) in sludge and water phases was investigated in a municipal wastewater treatment plant, and a lab-scale A(2)O-MBR was operated to provide background value of ARGs. The influencing factors of ARGs including microbial community, co-selection from heavy metals, biomass and horizontal gene transfer were concerned. Results showed that iA(2)O (inversed A(2)O) showed better ARGs reduction, and longer SRT (sludge retention time) increased ARGs relative abundance while reduced the gene copies of ARGs in the effluent, but significantly increased the ARGs in sludge phase. Compared to background value, the most enriched ARG was tetX in water phase, while it was intI1 in sludge phase. There existed higher abundance of multi-resistant bacteria in sludge phase, and microbial community determined the fate of ARGs in both water and sludge phase, while the direct effects from horizontal gene transfer should not be overlooked especially in water phase.201829909361
8007160.9809Distinguishing removal and regrowth potential of antibiotic resistance genes and antibiotic resistant bacteria on microplastics and in leachate after chlorination or Fenton oxidation. The prevalence of antibiotic resistance, as well as microplastics (MPs) as vectors for antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) has attracting growing attention. However, the fate of ARB/ARGs on MPs treated by chlorination and Fenton oxidation were poorly understood. Herein, the removal and regrowth of ARGs/ARB on MPs and in MPs-surrounding landfill leachate (an important reservoir of MPs and ARGs) after chlorination and Fenton oxidation were comparatively analyzed. Target ARGs on MPs were reduced obviously less than that in leachate, with the largest percentages reduction of 34.0-46.3% vs. 54.3-77.6% after chlorination and 92.1-97.3% vs. > 99.9% after Fenton oxidation, and similar removal patterns were observed for ARB. Moreover, a considerable regrowth of ARGs/ARB in leachate were found after 48 h of storage at the end of chlorination (5, 10, 20 and 50 mg/L), and a greater regrowth of ARGs and ARB occurred on MPs with up to 17 and 139 fold, respectively. In contrast, Fenton oxidation achieved a reduced regrowth of target ARGs/ARB. These findings indicated that the removal of ARGs/ARB on MPs were more difficult than that in leachate, and ARGs/ARB in leachate and especially on MPs exhibited a considerable potential for rapid regrowth after chlorination.202235158247
3542170.9809Fecal indicators, pathogens, antibiotic resistance genes, and ecotoxicity in Galveston Bay after Hurricane Harvey. Unprecedented rainfall after Hurricane Harvey caused a catastrophic flood in the southern coast of Texas, and flushed significant floodwater and sediments into Galveston Bay, the largest estuary along the Texas Gulf Coast. This study investigated the immediate and long-term (6 months post-Harvey) fecal indicators, pathogenic bacteria, antibiotic resistance genes (ARGs), and ecotoxicity in the Galveston Bay. Dramatic decrease of salinity profile to zero, increased levels of fecal indicator bacteria and pathogenic bacteria, and detection of various ARGs were observed in the water and sediment samples collected 2 weeks post-Harvey. High levels of Bla(TEM) and cytotoxicity measured by yeast bioluminescent assay (BLYR) were also observed especially near the river mouths. While Vibrio spp. was dominant in water, much higher abundance of fecal indicator bacteria and pathogen were detected in the sediments. A decreasing trend of Bla(TEM) and cytotoxicity was observed in March 2018 samples, suggesting the Bay has returned to its pre-hurricane conditions 6 months post-Harvey. Interestingly, the abundance of fecal indicator bacteria and pathogens were shifted dramatically according to high-streamflow and low-streamflow seasons in the Bay. The data are useful to construct the model of risk assessment in coastal estuaries system and predict the effects of extreme flooding events in the future.202133445049
7827180.9809Inactivation of antibiotic-resistant bacteria and antibiotic resistance genes by electrochemical oxidation/electro-Fenton process. Antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in the environment are of great concern due to their potential risk to human health. The effluents from wastewater treatment plants and livestock production are major sources of ARB and ARGs. Chlorination, UV irradiation, and ozone disinfection cannot remove ARGs completely. In this study, the potential of electrochemical oxidation and electro-Fenton processes as alternative treatment technologies for inactivation of ARB and ARGs in both intracellular and extracellular forms was evaluated. Results showed that the electrochemical oxidation process was effective for the inactivation of selected ARB but not for the removal of intracellular ARGs or extracellular ARGs. The electro-Fenton process was more effective for the removal of both intracellular and extracellular ARGs. The removal efficiency after 120 min of electro-Fenton treatment under 21.42 mA/cm(2) was 3.8 logs for intracellular tetA, 4.1 logs for intracellular ampC, 5.2 logs for extracellular tetA, and 4.8 logs for extracellular ampC, respectively in the presence of 1.0 mmol/L Fe(2+). It is suggested that electrochemical oxidation is an effective disinfection method for ARB and the electro-Fenton process is a promising technology for the removal of both intracellular and extracellular ARGs in wastewater.202032701499
7541190.9809The knock-on effects of different wastewater feeding modes: Change in microbial communities versus resistance genes in pilot-scale aerobic sludge granulation reactors. To explore the effects of wastewater feeding modes on the formation of aerobic granular sludge (AGS) and the complex relationships between resistance genes and bacteria, two pilot-scale sequencing batch reactors (SBRs) were established. The SBR with influent wastewater introduced uniformly through pipes at bottom was designated as BSBR, and the SBR with inlet wastewater flowing directly from top was TSBR. BSBR formed dense AGS due to uniform wastewater feeding at bottom, while TSBR failed to cultivate AGS. Metagenomic sequencing illustrated that rapid growth of AGS in BSBR was accompanied with increase of antibiotic resistance genes (ARGs) abundance, but ARGs diminished when the size of AGS was stable. The ARGs continued to elevate in TSBR, and abundance of metal resistance genes (MRGs) was always higher than that in BSBR. Two reactors had markedly different bacterial community, microbes in BSBR owned stronger activity, conferred greater potential to proliferate. AdeF in two systems had the most complex gene-bacteria relationships which would undergo HGT within bacterial genus. The different feeding modes of wastewater directly led to the changing size of sludge, which caused knock-on effects of variations in the abundance of microbial communities and resistance genes. This study provided promising suggestions for the rapid cultivation of AGS and control of resistance genes at pilot-scale.202337257591