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721500.9815High-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
705510.9804Characterization of antibiotic resistance genes and bacterial community in selected municipal and industrial sewage treatment plants beside Poyang Lake. Sewage treatment plants (STPs) are significant reservoirs of antibiotic resistance genes (ARGs) and antibiotic-resistant bacteria (ARB). Municipal STPs (MSTPs) and industrial STPs (ISTPs) are the two most important STP types in cities. In this study, the ARGs, mobile genetic elements (MGEs), and bacterial communities of selected STPs, including two MSTPs and one ISTP, in the vicinity of Poyang Lake were comprehensively investigated through high-throughput qPCR and high-throughput Illumina sequencing. The results showed that the profiles of ARGs, MGEs and bacteria differed between the ISTP and the two MSTPs, most likely due to differences in influent water quality, such as the Pb that characterized in the ISTP's influent. The longer hydraulic retention times (HRTs) of the two MSTPs than of the ISTP may also have accounted for the different profiles. Thus, a prolonged HRT in the CASS process seems to allow a more extensive removal of ARGs and bacteria in ISTPs with similar treatment process. By providing comprehensive insights into the characteristics of ARGs, MGEs and the bacterial communities of the selected MSTPs and ISTP, our study provides a scientific basis for controlling the propagation and diffusion of ARGs and ARB in different types of STPs.202032092547
727620.9798Antibiotic 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
800130.9796Exploring resistomes and microbiomes in pilot-scale microalgae-bacteria wastewater treatment systems for use in low-resource settings. Antibiotic resistance genes (ARGs) released into the environment are an emerging human and environmental health concern, including ARGs spread in wastewater treatment effluents. In low-to-middle income countries (LMICs), an alternate wastewater treatment option instead of conventional systems are low-energy, high-rate algal ponds (HRAP) that use microalgae-bacteria aggregates (MABA) for waste degradation. Here we studied the robustness of ARG removal in MABA-based pilot-scale outdoor systems for 140 days of continuous operation. The HRAP system successfully removed 73 to 88 % chemical oxygen demand and up to 97.4 % ammonia, with aggregate size increasing over operating time. Fourteen ARG classes were identified in the HRAP influent, MABA, and effluent using metagenomics, with the HRAP process reducing total ARG abundances by up to 5-fold from influent to effluent. Parallel qPCR analyses showed the HRAP system significantly reduced exemplar ARGs (p < 0.05), with 1.2 to 4.9, 2.7 to 6.3, 0 to 1.5, and 1.2 to 4.8 log-removals for sul1, tetQ, bla(KPC), and intl1 genes, respectively. Sequencing of influent, effluent and MABAs samples showed associated microbial communities differed significantly, with influent communities by Enterobacteriales (clinically relevant ARGs carrying bacteria), which were less evident in MABA and effluent. In this sense, such bacteria might be excluded from MABA due to their good settling properties and the presence of antimicrobial peptides. Microalgae-bacteria treatment systems steadily reduced ARGs from wastewater during operation time, using sunlight as the energetic driver, making them ideal for use in LMIC wastewater treatment applications.202337080313
762140.9796Pre-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
713350.9796Prevalence of antibiotic resistance genes in bacterial communities associated with Cladophora glomerata mats along the nearshore of Lake Ontario. The alga Cladophora glomerata can erupt in nuisance blooms throughout the lower Great Lakes. Since bacterial abundance increases with the emergence and decay of Cladophora, we investigated the prevalence of antibiotic resistance (ABR) in Cladophora-associated bacterial communities up-gradient and down-gradient from a large sewage treatment plant (STP) on Lake Ontario. Although STPs are well-known sources of ABR, we also expected detectable ABR from up-gradient wetland communities, since they receive surface run-off from urban and agricultural sources. Statistically significant differences in aquatic bacterial abundance and ABR were found between down-gradient beach samples and up-gradient coastal wetland samples (ANOVA, Holm-Sidak test, p < 0.05). Decaying and free-floating Cladophora sampled near the STP had the highest bacterial densities overall, including on ampicillin- and vancomycin-treated plates. However, quantitative polymerase chain reaction analysis of the ABR genes ampC, tetA, tetB, and vanA from environmental communities showed a different pattern. Some of the highest ABR gene levels occurred at the 2 coastal wetland sites (vanA). Overall, bacterial ABR profiles from environmental samples were distinguishable between living and decaying Cladophora, inferring that Cladophora may control bacterial ABR depending on its life-cycle stage. Our results also show how spatially and temporally dynamic ABR is in nearshore aquatic bacteria, which warrants further research.201728192677
679060.9795Overlooked dissemination risks of antimicrobial resistance through green tide proliferation. Green tides, particularly those induced by Enteromorpha, pose significant environmental challenges, exacerbated by climate change, coastal eutrophication, and other anthropogenic impacts. More concerningly, these blooms may influence the spread of antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) within ecosystems. However, the manner in which Enteromorpha blooms affect the distribution and spread of antimicrobial resistance (AMR) remains uncertain. This study investigated ARG profiles, dynamic composition, and associated health risks within the Enteromorpha phycosphere and surrounding seawater in typical bays (Jiaozhou, Aoshan, and Lingshan) in the South Yellow Sea. The Enteromorpha phycosphere exhibited significantly higher ARG abundance (p < 0.05) but lower diversity compared to the surrounding seawater. Source-tracking and metagenomic analyses revealed that the phycosphere was the main contributor to the resistome of surrounding seawater. Moreover, resistant pathogens, especially ESKAPE pathogens, with horizontal gene transfer (HGT) potential, were more abundant in the phycosphere than in the surrounding seawater. The phycosphere released high-risk ARGs to the surrounding seawater during Enteromorpha blooms, posing serious health and ecological AMR risks in marine environments. This study highlights the significant role of Enteromorpha blooms in ARG spread and associated risks, urging a reassessment of AMR burden from a public health perspective.202539488061
811170.9795Effect of alkaline-thermal pretreatment on biodegradable plastics degradation and dissemination of antibiotic resistance genes in co-compost system. Biodegradable plastics (BDPs) are an eco-friendly alternative to traditional plastics in organic waste, but their microbial degradation and impact on antibiotic resistance genes (ARGs) transmission during co-composting remain poorly understood. This study examines how alkaline-thermal pretreatment enhances BDPs degradation and influences the fate of ARGs and mobile genetic elements (MGEs) in co-composting. Pretreatment with 0.1 mol/L NaOH at 100℃ for 40 minutes increased the surface roughness and hydrophilicity of BDPs while reducing their molecular weight and thermal stability. Incorporating pretreated BDPs film (8 g/kg-TS) into the compost reduced the molecular weight of the BDPs by 59.70 % during the maturation stage, facilitating compost heating and prolonging the thermophilic stage. However, incomplete degradation of BDPs releases numerous smaller-sized microplastics, which can act as carriers for microorganisms, facilitating the dissemination of ARGs across environments and posing significant ecological and public health risks. Metagenomic analysis revealed that pretreatment enriched plastic-degrading bacteria, such as Thermobifida fusca, on BDPs surfaces and accelerated microbial plastic degradation during the thermophilic stage, but also increased ARGs abundance. Although pretreatment significantly reduced MGEs abundance (tnpA, IS19), the risk of ARGs dissemination remained. Three plastic-degrading bacteria (Pigmentiphaga sp002188465, Bacillus clausii, and Bacillus altitudinis) were identified as ARGs hosts, underscoring the need to address the risk of horizontal gene transfer of ARGs associated with pretreatment in organic waste management.202539970645
721880.9794Mass-immigration shapes the antibiotic resistome of wastewater treatment plants. Wastewater treatment plants (WWTPs) are the hotspots for the spread of antibiotic resistance genes (ARGs) into the environment. Nevertheless, a comprehensive assessment of the city-level and short-term daily (inter-day) variations of ARG profiles in the whole process (influent-INF, activated sludge-AS and effluent-EF) of WWTPs is still lacking. Here, 285 ARGs and ten mobile gene elements were monitored in seven WWTPs in Xiamen for seven days via high-throughput qPCR. The average daily load of ARGs to WWTPs was about 1.32 × 10(20) copies/d, and a total of 1.56 × 10(18) copies/d was discharged to the environment across the entire city. Stochastic processes were the main force determining the assembly of ARG communities during sampling campaign, with their relative importance ranked in the order of INF > EFF > AS. There're little daily variations in ARG richness, abundance, β-diversity composition as well as assembly mechanisms. The results of SourceTracker, variation partitioning analysis, and hierarchical partitioning analysis indicated that bacteria and ARGs from upstream treatment processes played an increasingly dominant role in shaping ARG communities in AS and EFF, respectively, suggesting the importance of mass-immigration of bacteria and ARGs from the source on ARG transport in wastewater treatment processes. This emphasizes the need to revise the way we mitigate ARG contamination but focus on the source of ARGs in urban wastewater.202437914134
727190.9794Identification of critical control points for antibiotic resistance discharge in sewers. Disrupting the spread of clinically relevant antibiotic resistance genes (ARGs) is one of the key components for the success of the One Health strategy. While waste water treatment plants (WWTPs) represent a final control point for daily discharges of antibiotic resistance genes (ARGs) to the aquatic environment, a decentralized upstream monitoring of wastewater feeds of selected urban drainage areas for bla(CTX-M32), bla(CTX-M15), bla(OXA48), bla(CMY-2), mecA, bla(NDM-1), bla(KPC3), vanA, and mcr-1 representing clinically relevant ARGs has been performed. Besides hospitals, also retirement homes were found to be responsible for high levels of ARG discharges compared to housing area sewer systems. The monitoring combines qPCR-based quantifications, flow volume-based analyses, and multiple antibiotic resistance analyses of isolates. As result of the study, local actions at identified critical control points could help to prevent contaminations of larger volumes of wastewaters. This strategy will support a more cost-effective treatment compared to central actions at WWTPs, only. A polluter-pays principle should be applied by this monitoring strategy.202235051461
6395100.9794Risk control of antibiotics, antibiotic resistance genes (ARGs) and antibiotic resistant bacteria (ARB) during sewage sludge treatment and disposal: A review. Sewage sludge is an important reservoir of antibiotics, antibiotic resistance genes (ARGs), and antibiotic resistant bacteria (ARB) in wastewater treatment plants (WWTPs), and the reclamation of sewage sludge potentially threats human health and environmental safety. Sludge treatment and disposal are expected to control these risks, and this review summarizes the fate and controlling efficiency of antibiotics, ARGs, and ARB in sludge involved in different processes, i.e., disintegration, anaerobic digestion, aerobic composting, drying, pyrolysis, constructed wetland, and land application. Additionally, the analysis and characterization methods of antibiotics, ARGs, and ARB in complicate sludge are reviewed, and the quantitative risk assessment approaches involved in land application are comprehensively discussed. This review benefits process optimization of sludge treatment and disposal, with regard to environmental risks control of antibiotics, ARGs, and ARB in sludge. Furthermore, current research limitations and gaps, e.g., the antibiotic resistance risk assessment in sludge-amended soil, are proposed to advance the future studies.202336933744
7328110.9794Metagenomic analysis of bacterial community composition and antibiotic resistance genes in a wastewater treatment plant and its receiving surface water. The presence of pathogenic bacteria and the dissemination of antibiotic resistance genes (ARGs) may pose big risks to the rivers that receive the effluent from municipal wastewater treatment plants (WWTPs). In this study, we investigated the changes of bacterial community and ARGs along treatment processes of one WWTP, and examined the effects of the effluent discharge on the bacterial community and ARGs in the receiving river. Pyrosequencing was applied to reveal bacterial community composition including potential bacterial pathogen, and Illumina high-throughput sequencing was used for profiling ARGs. The results showed that the WWTP had good removal efficiency on potential pathogenic bacteria (especially Arcobacter butzleri) and ARGs. Moreover, the bacterial communities of downstream and upstream of the river showed no significant difference. However, the increase in the abundance of potential pathogens and ARGs at effluent outfall was observed, indicating that WWTP effluent might contribute to the dissemination of potential pathogenic bacteria and ARGs in the receiving river.201627340885
7327120.9794Exploring the microbiome, antibiotic resistance genes, mobile genetic element, and potential resistant pathogens in municipal wastewater treatment plants in Brazil. Wastewater treatment plants (WWTPs) have been widely investigated in Europe, Asia and North America regarding the occurrence and fate of antibiotic resistance (AR) elements, such as antibiotic resistance genes (ARGs), mobile genetic elements (MGEs) and antibiotic resistant bacteria and pathogens. However, monitoring data about AR elements in municipal WWTPs in Brazil are scarce. This study investigated the abundance of intI1, five ARGs (sul1, tetA, blaTEM, ermB and qnrB) and 16S rRNA in raw and treated wastewater of three WWTPs, using different sewage treatments named CAS (Conventional activated sludge), UASB/BTF (UASB followed by biological trickling filter) and MAS/UV (modified activated sludge with UV disinfection stage). Bacterial diversity and the presence of potentially pathogenic groups were also evaluated, and associations between genetic markers and the bacterial populations were presented. All WWTPs decreased the loads of genetic markers finally discharged to receiving water bodies and showed no evidence of being hotspots for antimicrobial resistance amplification in wastewater, since the abundances of intI1 and ARGs within the bacterial population were not increased in the treated effluents. UASB/BTF showed a similar performance to that of the CAS and MAS/UV, reinforcing the sanitary and environmental advantages of this biological treatment, widely applied for wastewater treatment in warm climate regions. Bacterial diversity and richness increased after treatments, and bacterial communities in wastewater samples differed due to catchment areas and treatment typologies. Potential pathogenic population underwent considerable decrease after the treatments; however, strong significant correlations with intI1 and ARGs revealed potential multidrug-resistant pathogenic bacteria (Aeromonas, Arcobacter, Enterobacter, Escherichia-Shigella, Stenotrophomonas and Streptococcus) in the treated effluents, although in reduced relative abundances. These are contributive results for understanding the fate of ARGs, MGEs and potential pathogenic bacteria after wastewater treatments, which might support actions to mitigate their release into Brazilian aquatic environments in the near future.202235724791
7014130.9793Correlations among Antibiotic Resistance Genes, Mobile Genetic Elements and Microbial Communities in Municipal Sewage Treatment Plants Revealed by High-Throughput Sequencing. Municipal sewage treatment plants (MSTPs) are environmental pools for antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs), which is cause for growing environmental-health concerns. In this study, the effects of different wastewater treatment processes on microbial antibiotic resistance in four MSTPs were investigated. PCR, q-PCR, and molecular cloning integrally indicated that the tetracycline resistance (tet) genes significantly reduced after activated-sludge treatment. Illumina high-throughput sequencing revealed that the broad-spectrum profile of ARGs and mobile element genes (MGEs) were also greatly decreased by one order of magnitude via activated sludge treatment and were closely associated with each other. Correlations between ARGs and bacterial communities showed that potential ARB, such as Acinetobacter, Bacteroides, and Cloaibacterium, were removed by the activated-sludge process. Sedimentation processes cannot significantly affect the bacterial structure, resulting in the relative abundance of ARGs, MGEs, and ARB in second-clarifier effluent water being similar to activated sludge. A comprehensive study of ARGs associated with MGEs and bacterial structure might be technologically guided for activated sludge design and operation in the MSTPs, to purposefully control ARGs carried by pathogenic hosts and mobility.202336834289
7233140.9793Distribution, sources, and potential risks of antibiotic resistance genes in wastewater treatment plant: A review. Irrational use of antibiotics produces a large number of antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs). Wastewater treatment plants (WWTPs) act as important sources and sinks of ARGs, and play an important role in their generation, treatment, and dissemination. This study summarizes the types, concentrations, and factors of ARGs in WWTPs, investigates the sources of ARGs in wastewater, compares the removal efficiencies of different treatment processes on ARGs, and analyzes the potential risks of ARGs accumulation in effluent, sludge and their emission into the air. The results show that the main ARGs detected in the influent of WWTPs are the genes resistant to macrolides (ermB, ermF), tetracyclines (tetW, tetA, tetC), sulfonamides (sul1, sul2), and β-lactams (bla(OXA), bla(TEM)). The concentrations of ARGs in the influent of the WWTPs are 2.23 × 10(2)-3.90 × 10(9) copies/mL. Wastewater quality and microbial community are the dominant factors that affect the distribution characteristics of ARGs. The accumulation of ARGs in effluent, sludge, and aerosols pose potential risks to the regional ecological environment and human health. Based on these results, research trends with respect to ARGs in WWTPs are also prospected.202235921944
7351150.9793Dynamics of integron structures across a wastewater network - Implications to resistance gene transfer. Class 1 and other integrons are common in wastewater networks, often being associated with antibiotic resistance genes (ARGs). However, the importance of different integron structures in ARG transfer within wastewater systems has only been implied, especially between community and hospital sources, among wastewater treatment plant compartments, and in receiving waters. This uncertainty is partly because current clinical class 1 integron qPCR assays (i.e., that target human-impacted structures, i.e., clintI1) poorly delineate clintI1 from non-impacted class 1 integron structures. They also say nothing about their ARG content. To fill these technical gaps, new real-time qPCR assays were developed for "impacted" class 1 structures (called aint1; i.e., anthropogenic class 1 integrons) and empty aint1 structures (i.e., carry no ARGs; called eaint1). The new assays and other integron assays then were used to examine integron dynamics across a wastewater network. 16S metagenomic sequencing also was performed to characterise associated microbiomes. aint1 abundances per bacterial cell were about 10 times greater in hospital wastewaters compared with other compartments, suggesting aint1 enrichment with ARGs in hospital sources. Conversely, the relative abundance of eaint1 structures were over double in recycled activated sludge compared with other compartments, except receiving waters (RAS; ∼30% of RAS class 1 structures did not carry ARGs). Microbiome analysis showed that human-associated bacterial taxa with mobile integrons also differed in RAS and river sediments. Further, class 1 integrons in RAS bacteria appear to have released ARGs, whereas hospital bacteria have accumulated ARGs. Results show that quantifying integron dynamics can help explain where ARG transfer occurs in wastewater networks, and should be considered in future studies on antibiotic resistance in the environment.202134673462
7326160.9793Antibiotic resistance genes associated with size-segregated bioaerosols from wastewater treatment plants: A review. The antibiotic-resistant pollution in size-segregated bioaerosols from wastewater treatment plants (WWTPs) is of increasing concern due to its public health risks, but an elaborate review is still lacking. This work overviewed the profile, mobility, pathogenic hosts, source, and risks of antibiotic resistance genes (ARGs) in size-segregated bioaerosols from WWTPs. The dominant ARG type in size-segregated bioaerosols from WWTPs was multidrug resistance genes. Treatment units that equipped with mechanical facilities and aeration devices, such as grilles, grit chambers, biochemical reaction tanks, and sludge treatment units, were the primary sources of bioaerosol antibiotic resistome in WWTPs. Higher enrichment of antibiotic resistome in particulate matter with an aerodynamic diameter of <2.5 μm, was found along the upwind-downwind-WWTPs gradient. Only a small portion of ARGs in inhalable bioaerosols from WWTPs were flanked by mobile genetic elements. The pathogens with multiple drug resistance had been found in size-segregated bioaerosols from WWTPs. Different ARGs or antibiotic resistant bacteria have different aerosolization potential associated with bioaerosols from various treatment processes. The validation of pathogenic antibiotic resistance bacteria, deeper investigation of ARG mobility, emission mechanism of antibiotic resistome, and development of treatment technologies, should be systematically considered in future.202438128715
7143170.9793Simulated discharge of treated landfill leachates reveals a fueled development of antibiotic resistance in receiving tidal river. Around 350 million tons of solid waste is disposed of in landfills every year globally, with millions of cubic meters of landfill leachates released into neighboring environment. However, to date, little is known about the variations of antimicrobial resistance (AMR) in on-site leachate treatment systems and its development in leachate-receiving water environment. Here, we quantified 7 subtypes of antibiotic resistance genes (ARGs), 3 types of culturable antibiotic resistant bacteria (ARB) and 6 subtypes of mobile genetic elements (MGEs) in the effluents from a combined leachate treatment process, including biological treatment (MBR), physical separation (UF), ultraviolet (UV) disinfection and advanced oxidation process (AOP). The contents of ARGs, ARB and MGEs were generally enriched by the MBR, but then decreased significantly along with the tertiary treatment process. However, in the effluent-receiving water samples, the abundance of dominant ARGs (i.e. ermB, sul1, bla(TEM)) increased by 1.5 orders of magnitude within 96 h, alongside a general increase of MGEs (~10.0 log(10)(copies/mL) and total ARB (~1100 CFU/mL). Structural correlation analyses reveal that target ARGs were closely associated with MGEs, particularly in effluent-receiving samples (Procrustes test; M(2) = 0.49, R = 0.71, P = 0.001); and occurrences of ARB were majorly affected by ARG's distribution and environmental conditions (e.g. nitrogen speciation) in effluent and recipient groups, respectively. This study indicates that current treatment technologies and operation protocols are not feasible in countering the development of AMR in effluent-receiving water environment, particularly in tidal rivers that are capable of retaining contaminants for a long residence time.201829501852
7134180.9792Elevated levels of antibiotic resistance in groundwater during treated wastewater irrigation associated with infiltration and accumulation of antibiotic residues. Treated wastewater irrigation (TWW) releases antibiotics and antibiotic resistance genes (ARGs) into the environment and might thus promote the dissemination of antibiotic resistance in groundwater (GW). We hypothesized that TWW irrigation increases ARG abundance in GW through two potential mechanisms: the contamination of GW with resistant bacteria and the accumulation of antibiotics in GW. To test this, the GW below a real-scale TWW-irrigated field was sampled for six months. Sampling took place before, during and after high-intensity TWW irrigation. Samples were analysed with 16S rRNA amplicon sequencing, qPCR of six ARGs and the class 1 integron-integrase gene intI1, while liquid chromatography tandem mass spectrometry was performed to detect antibiotic and pharmaceutical residues. Absolute abundance of 16S rRNA in GW decreased rather than increased during long-term irrigation. Also, the relative abundance of TWW-related bacteria did not increase in GW during long-term irrigation. In contrast, long-term TWW irrigation increased the relative abundance of sul1 and intI1 in the GW microbiome. Furthermore, GW contained elevated concentrations of sulfonamide antibiotics, especially sulfamethoxazole, to which sul1 confers resistance. Total sulfonamide concentrations in GW correlated with sul1 relative abundance. Consequently, TWW irrigation promoted sul1 and intI1 dissemination in the GW microbiome, most likely due to the accumulation of drug residues.202234555761
7234190.9792Urban and agriculturally influenced water contribute differently to the spread of antibiotic resistance genes in a mega-city river network. The widespread of water borne antibiotic resistance genes (ARGs) represents a growing threat to the health of millions of people. Our study detected the relative abundances of 10 ARG subtypes in the Shanghai river network, where the major ARG components were strB, sul1, and ermB. These ARGs were significantly enriched by the combined sewage, tail water from urban wastewater treatment plant and runoff from agricultural areas, which reached the Suzhou (SZ), Dianpu (DP), and Huangpu (HP) River, respectively (one-way ANOVA, P < 0.01). The target ARGs were distributed in varying patterns across different rivers. bla(CTX-M) and bla(TEM) contributed to the increase of total ARGs in the rivers influenced by urban sources, particularly in the SZ River, whose distribution of ARGs was significantly related to that of the confluence of the whole river network (Mantel test, P < 0.01). The bacterial community was closely structured with ARGs and potential pathogenic bacteria's association with target ARGs became significant in downstream samples (Procrustes test, P = 0.03). Water near urban wastewater fallouts was observed to have the highest content of intl1 in the DP River, whose downstream samples' intl -ARG relationship fitted the same regression model as that of the network confluence (R = 0.84, P < 0.001). The amelioration of river water quality does not reduce ARGs, but may affect their distributional patterns in the river network in Shanghai.201931009830