Structural effects of sulfonamides on the proliferation dynamics of sulfonamide resistance genes in the sequencing batch reactors and the mechanism.

Antibiotic resistance genes (ARGs) can be easily promoted by antibiotics, however, the structural effects of antibiotics on the proliferation of ARGs dynamic and the associated mechanisms remain obscure in, especially, activated sludge sequencing batch reactors. In the present study, the effects of 9 sulfonamides (SAs) with different structures on the proliferation dynamic of sulfonamide resistance genes (Suls) in the activated sludge sequencing batch reactors and the corresponding mechanisms were determined (30 days), and the results showed that the largest proliferation value (∆AR) of Suls dynamic for SAs (sulfachloropyridazine) was approximately 2.9 times than that of the smallest one (sulfadiazine). The proliferation of Suls was significantly related to the structural features (minHBint6, SssNH, SHBd and SpMax2_Bhm) that represent the biological activity of SAs. To interpret the phenomenon, a mechanistic model was developed and the results indicated that the biodegradation of SAs (T1/2) rather than conjugative transfer frequency or mutation frequency tends to be the key process for affecting Suls proliferation. T1/2 was proved to be dependent on the interactions between SAs and receptors (Ebinding), the cleavage mode (bond dissociation energy), and the site of nucleophilic assault. Besides, the metagenomic analysis showed that SAs posed significant effect on antibiotic resistome and Tnp31 played a vital role in the proliferation of Suls. Overall, our findings provide important insight into a theoretical basis for understanding the structural effects of SAs on the proliferation of ARGs in SBR systems.

Insight into the evolution of microbial communities and resistance genes induced by sucralose in partial nitrification system with triclosan pre-exposure.

Sucralose (SUC), an artificial sweetener widely used in food, beverages and pharmaceuticals, is frequently detected in various environmental matrices. Triclosan (TCS) is commonly used as a disinfectant and often co-exists with SUC in sewage environments. This study investigated the effects of SUC (0.1-10 mg/L) on the transmission of intracellular and extracellular antibiotic resistance genes (ARGs) in the partial nitrification systems with and without TCS pre-exposure. The reactors operated for 150 days, and SUC did not affect ammonia oxidation performance, while TCS led to the maintenance of partial nitrification. The types and abundances of extracellular ARGs in sludge and free ARGs in water increased significantly after TCS pre-exposure when faced SUC stress, which might be caused by a decrease in α-Helix/(ß-Sheet + Random coil). SUC was more easily to enrich ARGs in partial nitrification systems with TCS pre-exposure, exacerbating the risk of ARGs transmission. The microbial community showed stronger relationships to cope with the direct stress of SUC, and the functional bacteria (Thauera and Nitrosomonas) in TCS pre-exposure system might be potential hosts of ARGs. This study might provide insights for better understanding the fates of SUC in partial nitrification systems and the ecological risks in wastewater containing TCS and SUC. ENVIRONMENTAL IMPLICATION: Sucralose (SUC) is often detected in the environment and considered as an emerging contaminant due to its soaring consumption and environmental persistence. Triclosan (TCS) is an antibacterial agent that often co-exists with SUC in personal care products and sewage environments. During 150 d, two partial nitrification reactors with and without TCS pre-exposure were established to study the effects of SUC on nitrification performance, antibiotic resistance genes (ARGs) and microbial communities. This study showed the refractory nature of SUC, and SUC led to the transmission of extracellular ARGs in partial nitrification system with TCS pre-exposure, exacerbating the risk of ARGs dissemination.

Fate and transport modelling for evaluating antibiotic resistance in aquatic environments: Current knowledge and research priorities.

Antibiotics have revolutionised medicine in the last century and enabled the prevention of bacterial infections that were previously deemed untreatable. However, in parallel, bacteria have increasingly developed resistance to antibiotics through various mechanisms. When resistant bacteria find their way into terrestrial and aquatic environments, animal and human exposures increase, e.g., via polluted soil, food, and water, and health risks multiply. Understanding the fate and transport of antibiotic resistant bacteria (ARB) and the transfer mechanisms of antibiotic resistance genes (ARGs) in aquatic environments is critical for evaluating and mitigating the risks of resistant-induced infections. The conceptual understanding of sources and pathways of antibiotics, ARB, and ARGs from society to the water environments is essential for setting the scene and developing an appropriate framework for modelling. Various factors and processes associated with hydrology, ecology, and climate change can significantly affect the fate and transport of ARB and ARGs in natural environments. This article reviews current knowledge, research gaps, and priorities for developing water quality models to assess the fate and transport of ARB and ARGs. The paper also provides inputs on future research needs, especially the need for new predictive models to guide risk assessment on AR transmission and spread in aquatic environments.

Different fates of extracellular and intracellular antibiotic resistance genes in flocs, granular and biofilm nitrification systems under the stress of acetaminophen.

The spread of antibiotic resistance genes (ARGs), including intracellular ARGs (i-ARGs) and extracellular ARGs (e-ARGs), has become a global problem that cannot be ignored. This study clarified the fates of e-ARGs and i-ARGs in floc sludge reactor (FS), granular sludge reactor (GS) and biofilm reactor (BF) under the stress of acetaminophen (APAP). The results showed that the risk of ARGs transmission, especially for e-ARGs, in FS and BF could increase with the increasing times of APAP treatment, except for that in GS. The fates of i-ARGs in three different systems were similar, which were mainly clustered as the efflux pumps mechanism. The secretion and disintegration of extracellular polymeric substances mainly affected the fates of e-ARGs. In the three systems, the complexity of network relationships between ARGs and microbial communities was FS, GS and BF. Partial least-squares path model analysis indicated that bacterial community directly contributed to the variations of e-ARGs and i-ARGs under APAP treatment in the three systems, playing a leading role. And i-ARGs and protein secondary structure showed direct effects on e-ARGs. This study indicated that e-ARGs in complex systems were more susceptible to be influenced, which should be paid more attention to prevent further propagation of ARGs.

Network complexity of bacterial community driving antibiotic resistome in the microbiome of earthworm guts under different land use patterns.

Recently, the study of antibiotic resistance in the soil animal microbiome has attracted extensive attention; however, the patterns of antibiotic resistance genes (ARGs) in soil and soil animals related to different land use types remain poorly studied. In the present study, soil and earthworms were collected from four different land-use types (farmland, hospital, park land, and mountain park), and 162 ARGs in the microbiomes of the soil and earthworms were quantified using high-throughput quantitative PCR. Our study showed that the abundance and number of ARGs were higher in soil samples than in earthworm guts, but earthworms as the living organisms created relatively isolated ambient surroundings, which allowed for a more heterogeneous ARGs profile. Meanwhile, land use significantly influenced the abundance, number and co-occurrence pattern of ARGs in the soil and earthworm samples. Furthermore, abiotic and biotic factors had significant effects on the ARGs profile, among which pH had a negative effect on the ARGs profiles of both soil and earthworm microbiomes, and bacterial network complexity had a positive effect on the earthworm ARGs profile. Our study provides new insights into the distribution and dispersal of ARGs in the soil animal gut microbiome under different land use patterns.

Abatement of antibiotics and resistance genes during catalytic ozonation enhanced sludge dewatering process: Synchronized in volume and hazardousness reduction.

Based on the efficiency of the catalytic ozonation techniques (HDWS+O3 and MnFe2O4 @SBC+O3) in enhancing the sludge dewaterability, the effectiveness in synchronized abatement antibiotics and antibiotic resistance genes (ARGs) was conducted to determine. The results revealed that catalytic ozonation conditioning altered the distribution of target antibiotics (tetracycline (TC), oxytetracycline (OTC), norfloxacin (NOR), ofloxacin (OFL)) in the dewatered filtrate, the dewatered sludge cake and the extra-microcolony/cellular polymers (EMPS/ECPS) layers, achieving the redistribution from solid-phase adsorption to liquid-phase dissolution. The total degradation rate was over 90% for TC and OTC, 72-78% for NOR and OFL; the abatement efficiency of eleven ARGs reached 1.47-3.01 log and 1.64-3.59 log, respectively, and more than four eARGs were eliminated. The effective abatement of the absolute abundance of Mobile genetic elements (MGEs) (0.91-1.89 log) demonstrated that catalytic ozonation conditioning could also significantly inhibit horizontal gene transfer (HGT). The abundance of resistant bacteria was greatly reduced and the signal transduction of the typical ARGs host bacteria was inhibited. The highly reactive oxidation species (ROS) generated were responsible for the abatement of antibiotics and ARGs. These findings provided new insights into the sludge conditioning for ideal and synchronized reduction in volume and hazardousness by catalytic ozonation processes in sludge treatment.

Co-exposure of microplastics and sulfamethoxazole propagated antibiotic resistance genes in sediments by regulating the microbial carbon metabolism.

The concerns on the carriers of microplastics (MPs) on co-existing pollutants in aquatic environments are sharply rising in recent years. However, little is known about their interactions on the colonization of microbiota, especially for the spread of pathogens and antibiotic resistance genes (ARGs). Therefore, this study aimed to investigate the influences on the propagation of ARGs in sediments by the co-exposure of different MPs and sulfamethoxazole (SMX). The results showed that the presence of MPs significantly enhanced the contents of total organic carbon, while having no effects on the removal of SMX in sediments. Exposure to SMX and MPs obviously activated the microbial carbon utilization capacities based on the BIOLOG method. The propagation of ARGs in sediments was activated by SMX, which was further promoted by the presence of polylactic acid (PLA) MPs, but significantly lowered by the co-exposed polyethylene (PE) MPs. This apparent difference may be attributed to the distinct influence on the antibiotic efflux pumps of two MPs. Moreover, the propagation of ARGs may be also dominated by microbial carbon metabolism in sediments, especially through regulating the carbon sources of carboxylic acids, carbohydrates, and amino acids. This study provides new insights into the carrier effects of MPs in sediments.

Determining the spatiotemporal variation, sources, and ecological processes of antibiotic resistance genes in a typical lake of the middle reaches of the Yangtze River.

Antibiotic resistance genes (ARGs) are emerging environmental pollutants, influenced by complex regulatory factors. River-lake systems act as natural reservoirs for ARGs and provide an ideal model for studying their regulatory mechanisms. This study employed high-throughput quantitative PCR, high-throughput sequencing, correlation analyses, and model predictions to investigate the dynamics of ARGs and their influencing factors in Liangzi Lake, located in the mid-reaches of the Yangtze River. The research specifically centered on three environmental components: lake water, sediment, and river water. Results indicated that the ARGs from eight major antibiotic classes, displaying distinct seasonal distribution patterns. In comparison to the sediment, the water phase demonstrated a higher diversity of ARGs, with the highest level of ARGs sharing observed between lake and river waters (approximately 83.7 %). Furthermore, seasonal variations significantly influenced the distributions of both ARGs and bacterial communities. The diversity of ARGs was highest during the summer and autumn, and specific bacterial species exhibited robust correlations with ARGs (including matA/mel, aac (6')-Ib-03, and blaROB). It is worth noting that environmental attributes and bacterial diversity had the most substantial impact on the dynamic changes in ARGs. Lastly, source tracking analysis pinpointed that sediment as the primary source of ARGs in lake water, constituting 45 % to 48 % of the total ARGs. Our study provides a comprehensive analysis of ARGs and their influencing factors in the river-lake system of the middle reaches of the Yangtze River, with Liangzi Lake as a representative case.

Metagenomics-assembled analysis revealed the characteristics of antibiotic resistome and community coalescence in the soils irrigated with different irrigation materials.

Antibiotic resistance has received widespread attention in recent years. Soil irrigation and fertilization are routine agricultural practices, but also lead to the spread of antibiotic resistance genes (ARGs) in soil-crop system such as via resistome coalescence. Despite community coalescence being ubiquitous and important in natural ecosystems, little research has been done to investigate resistome coalescence during soil irrigation activities. In this study, the characteristics of antibiotic resistome and community coalescence in the soils irrigated with different irrigation materials (wastewater, wastewater-river water, and wastewater-manure) have been revealed by utilizing microcosm experiments and high-throughput sequencing-based metagenomic assembly approaches. Results showed irrigation and coalescence changed soil quality and resistome. Totally, 789 unique ARGs were identified in the irrigation system, including some emerging ARGs. The abundance and diversity of ARGs increased in the coalesced soils, mainly due to the newly imported ARGs from irrigation materials. Relatively, the soils irrigated with wastewater and manure showed higher level of ARGs. Irrigation with the mixtures containing river water caused greater loss of indigenous taxa, while the community structure of mixing treatment with manure changed more dramatically. Interestingly, the succession of community in coalesced soils was influenced by transient competition for resources and ecological niche width, and the highest abundance and diversity of microorganisms and ARGs were found in the initial phase of coalescence, followed by a gradual succession towards the original community. With increasement of wastewater in the irrigation materials, the soil community showed a stepwise change rather than linear change. Notably, natural deposit of irrigation materials reduced their impacts on the ARGs in the coalesced soils. Findings provide new insights into the resistome coalescence during agricultural practices for reducing the spread risks of ARGs.

Performance of sewage sludge treatment for the removal of antibiotic resistance genes: Status and prospects.

Wastewater treatment plants (WWTPs) receive wastewater containing antibiotic resistant bacteria (ARB) and antibiotic resistant genes (ARGs), which are predominant contributors to environmental pollution in water and soil. Of these sources, sludge is a more significant contributor than effluent. Knowing how sludge treatment affects the fate of ARGs is vital for managing the risk of these genes in both human and natural environments. This review therefore discusses the sources and transmission of ARGs in the environment and highlights the risks of ARGs in sludge. The effects of co-existing constituents (heavy metals, microplastics, etc.) on sludge and ARGs during treatment are collated to highlight the difficulty of treating sludge with complex constituents in ARGs. The effects of various sludge treatment methods on the abundances of ARGs in sludge and in soil from land application of treated sludge are discussed, pointing out that the choice of sludge treatment method should take into account various potential factors, such as soil and soil biology in subsequent land application. This review offers significant insights and explores the abundances of ARGs throughout the process of sludge treatment and disposal. Unintentional addition of antibiotic residues, heavy metals, microplastics and organic matter in sludge could significantly increase the abundance and reduce the removal efficiency of ARGs during treatment, which undoubtedly adds a barrier to the removal of ARGs from sludge treatment. The complexity of the sludge composition and the diversities of ARGs have led to the fact that no effective sludge treatment method has so far been able to completely eliminate the ecological risk of ARGs. In order to reduce risks resulting by transmission of ARGs, technical and management measures need to be implemented.

Size matters: Powder biochar promotes the elimination of antibiotics resistance genes and potential hosts during swine manure composting.

Livestock manure faced acute environmental pollution and ecology risky caused by antibiotic resistance genes (ARGs). This study investigated the effects of biochar particle size including powder biochar (75 µm, PB), and granular biochar (2 mm, GB) on ARGs variation during the aerobic composting. The results showed that the total relative abundance (RA) of the ARGs decreased significantly in all the treatments after composting. While compared to the removal efficiency of total RA in the control (CK), PB decreased by 90.99 % and GB increased by 93.25 %, and both PB and GB removed MGEs completely. Sulfonamide antibiotic resistance genes were the main contributor of the ARGs rebounding. PB addition could hinder the rebounding of sulfonamide antibiotic resistance genes during the later stage of the composting. Co-occurrence network analysis showed that the addition of biochar (both types) increased the complexity of the microbial community the competition of inter-phylum, which was indicated by the higher number of edge and density and lower positive connection. The different ARGs removal efficiency in these two treatments might be that PB promoted the competition both inter-phylum and potential hosts-other microbes, resulted in fewer kinds and abundance of ARGs hosts, while GB increased the stability of ARGs hosts making it more resistant to environment changes. Totally, compared with the global adjustment strategy of microbial communities, more exclusive methods focusing on the controlling of ARGs hosts should be explored to decrease the ecological risk of composting products during composting process.

Deterioration of sludge characteristics and promotion of antibiotic resistance genes spread with the co-existing of polyvinylchloride microplastics and tetracycline in the sequencing batch reactor.

With the continuous increase in microplastics (MPs) and tetracycline (TC) entering wastewater treatment plants (WWTPs) along with sewage, the co-existence of MPs and TC in the biological treatment of wastewater has attracted extensive attention. This study investigated the effect of 1 mg/L polyvinyl chloride (PVC) MPs and 100 ng/L TC co-existing on sequencing batch reactors (SBRs) (S2) treating phenol wastewater in contrast to the control with TC alone (S1). The phenol removal efficiency was significantly inhibited by the co-existence of PVC MPs and TC. Sludge characteristics were also distinctively influenced. The decreased zone sludge velocity (ZSV) and increased sludge volume index (SVI) indicated that the combined effect of PVC MPs and TC deteriorated sludge settleability, which had positive and negative linear correlations with extracellular polymeric substances (EPS) content and the protein (PN)/polysaccharide (PS) ratio, respectively. Moreover, the decreased and increased relative abundances of potential phenol-degraders and antibiotic resistance gene (ARG) carriers may elucidate the inhibition of phenol removal and promotion of ARGs propagation with the co-occurrence of PVC MPs and TC. In addition, the enhanced potential ARGs hosts, loss of the EPS protective effect, and increased membrane permeability induced by reactive oxygen species (ROS) jointly promoted ARGs dissemination in the co-existence of PVC MPs and TC. Notably, the co-occurrence of ARGs and mobile genetic element (MGEs) indicated that the co-existence of PVC MPs and TC promoted the spread of some transposase-associated ARGs mediated by horizontal gene transfer (HGT).

The effects of didodecyl dimethyl ammonium chloride on microbial communities and resistance genes in floc, granular and biofilm denitrification sludge.

As a type of quaternary ammonium compounds, didodecyl dimethyl ammonium chloride (DADMAC C12) was frequently detected in wastewater treatment plants. Here, floc-based sequencing batch reactor (FSBR), granule-based SBR (GSBR) and biofilm SBR (BSBR) were fed with 0.5, 5 and 10 mg/L of DADMAC C12 for 120 d. Compared with floc sludge and granule sludge, biofilm had the strongest ability to resist the impact of DADMAC C12. Notably, in both FSBR and GSBR systems, 5 mg/L DADMAC C12 promoted denitrification sludge to become hydrophobic and compact due to an increase in α-Helix/(ß-Sheet+Random coil), consequently enhancing sludge granulation. Besides，high concentration of DADMAC C12 generally increased the abundances of MGEs in three denitrification systems, except extracellular MGEs in water. The variation of efflux pump ARGs was basically consistent with that of MGEs. The stimulation of DADMAC C12 also increased significantly the abundance of extracellular antibiotics deactivation ARGs in water in three denitrification systems. Besides, DADMAC C12 induced co-selection among various ARGs and promoted the proliferation and spread of sulfonamide ARGs in water.

Distribution and major driving elements of antibiotic resistance genes in the soil-vegetable system under microplastic stress.

Microplastics (MPs) and antibiotic resistance genes (ARGs) are both enriched in soil-vegetable systems as a consequence of the prolonged use of agricultural mulches. MPs can form unique bacterial communities and provide potential hosts for ARGs. Therefore, MPs stress may promote the spread of ARGs from soil to crops. Increasing ARGs pollution in soil-vegetable system. In our research, we investigated the distribution and major driving elements of antibiotic resistance genes in the soil-vegetable system under microplastic stress. The results showed that MPs treatment decreased the relative abundance of ARGs in non-rhizosphere soil. High concentrations of MPs promoted the enrichment of tetracycline antibiotic resistance genes in rhizosphere soil. MPs treatment promoted the enrichment of ARGs and mobile genetic elements (MGEs) in lettuce tissues, and the overall abundance of ARGs in root after 0.5 %, 1 %, and 2 % (w/w, dry weight) polyethylene (PE) administration was considerably higher compared to that in the untreated group (p < 0.05). At the same time, high PE concentrations promoted the spread of sulfa ARGs from root to leaf. MPs also impacted the bacterial communities in the soil-plant system, and the changes in ARGs as well as MGEs in each part of the soil-vegetable system were significantly correlated with the bacterial diversity index (p < 0.05). Correlation analysis and network analysis showed that bacterial communities and MGEs were the main drivers of ARGs variation in soil-lettuce systems.

A review of antibiotics in surface water and their removal by advanced electrocoagulation technologies.

The overuse and misuse of antibiotics have posed a serious threat to environment and human health, and even given rise to antibiotic resistance genes (ARGs). Antibiotics are ubiquitous in surface water worldwide with concentrations ranging from ng/L to µg/L level, being widely detected in rivers, lakes, seawater, and even drinking water. To address this thorny issue, numerous advanced technologies have been implemented to remove antibiotics. Advanced electrocoagulation (AEC) technologies, known as the combination of EC and other technologies capable of generating •OH in situ, have garnered considerable attention owing to their advances and high efficiency. This critical review investigated >120 relevant publications from the last few years (2017-2023) for the global distribution of commonly used antibiotics in surface water and their removal by various AEC technologies. Significant AEC technologies, such as combined electro-Fenton and EC (EF-EC) and combined electro-oxidation and EC (EO-EC), were reviewed. Their mechanism and characteristics were detailed. The major research results on removing antibiotics or the application potentials were elaborately described and discussed. Finally, the application trends of AEC technologies, as well as the challenges that may arise were prospected. The recommendations for controlling global antibiotic contamination in surface water were shared.

Two glyceraldehyde-3-phosphate dehydrogenases with distinctive roles in Pseudomonas syringae pv. tomato DC3000.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH or Gap) is a ubiquitously distributed enzyme that plays an essential role in the glycolytic and gluconeogenic pathways. However, additional roles have been described unrelated to its enzymatic function in diverse organisms, often linked to its presence in the cell surface or as a secreted protein. Despite being a paradigm among multifunctional/moonlighting proteins, little is known about its possible roles in phytopathogenic bacteria. In the present work we have studied three putative gap paralogous genes identified in the genome of Pseudomonas syringae pv. tomato (Pto) DC3000, an important model in molecular plant pathology, with the aim of determining their physiological and possible non-canonical roles in this bacterium and in the plant infection process. We have established that the Gap1 protein has a predominantly glycolytic activity, whereas the NADPH-dependent Gap2 main activity is gluconeogenic. The third paralogue lacks GAPDH activity in Pto but is indispensable for vitamin B6 metabolism and displays erythrose-4-phosphate dehydrogenase activity, thus referred as epd. Both Gap enzymes exhibit distinct functional characteristics depending on the bacterium physiological state, with Gap1 presenting a substantial role in motility, biosurfactant production and biofilm formation. On the other hand, solely Gap2 appears to be essential for growth on tomato plant. Furthermore, Gap1 and Gap2 present a distinctive transcriptional regulation and both have been identified exported outside the cells with different definite media compositions. This serves as compelling evidence of additional roles beyond their central metabolic functions.

Metagenomic analysis reveals the distribution, function, and bacterial hosts of degradation genes in activated sludge from industrial wastewater treatment plants.

For comprehensive insights into the bacterial community and its functions during industrial wastewater treatment, with a particular emphasis on its pivotal role in the bioremediation of organic pollutants, this study utilized municipal samples as a control group for metagenomic analysis. This approach allowed us to investigate the distribution, function, and bacterial hosts of biodegradation genes (BDGs) and organic degradation genes (ODGs), as well as the dynamics of bacterial communities during the industrial wastewater bioprocess. The results revealed that BDGs and ODGs associated with the degradation of benzoates, biphenyls, triazines, nitrotoluenes, and chlorinated aromatics were notably more abundant in the industrial samples. Specially, genes like clcD, linC, catE, pcaD, hbaB, hcrC, and badK, involved in the peripheral pathways for the catabolism of aromatic compounds, benzoate transport, and central aromatic intermediates, showed a significantly higher abundance of industrial activated sludge (AS) than municipal AS. Additionally, the BDG/ODG co-occurrence contigs in industrial samples exhibited a higher diversity in terms of degradation gene carrying capacity. Functional analysis of Clusters of Orthologous Groups (COGs) indicated that the primary function of bacterial communities in industrial AS was associated with the category of "metabolism". Furthermore, the presence of organic pollutants in industrial wastewater induced alterations in the bacterial community, particularly impacting the abundance of key hosts harboring BDGs and ODGs (e.g. Bradyrhizobium, Hydrogenophaga, and Mesorhizobium). The specific hosts of BDG/ODG could explain the distribution characteristics of degradation genes. For example, the prevalence of the Adh1 gene, primarily associated with Mesorhizobium, was notably more prevalent in the industrial AS. Overall, this study provides valuable insights into the development of more effective strategies for the industrial wastewater treatment and the mitigation of organic pollutant contamination.

Mass-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 × 1020 copies/d, and a total of 1.56 × 1018 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, ß-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.

Changes in community structure and microbiological risks in a small stream after receiving treated shale gas wastewater for two years.

Discharge of treated shale gas wastewater is becoming prevalent in the Sichuan Basin in China, and the resulting potential environmental impacts have raised concern. In this study, the responses of microbial community in the receiving water to discharge of treated shale gas wastewater were assessed during a two-year study period, covering two wet seasons and one dry season. The results showed that the discharge of treated shale gas wastewater had no significant effects on alpha diversity in the two wet seasons, but had significant effects in the dry season after 15 months of discharge. Obvious changes in microbial community structure were observed in all three seasons at the downstream sites near the wastewater outfall, as compared to the control site. Multimetric indices indicated that the impacts of wastewater discharge on microbial ecosystem occurred with the extension of the discharge period. Moreover, special attention was given to the microbiological risks associated with antibiotic resistance genes (ARGs), virulence factor genes (VFGs), and pathogenic antibiotic resistant bacteria (PARBs) in the dry season in sediments of the receiving water. At downstream sites near the outfall, five subtypes of ARGs and seven VFGs showed a significant increase in relative abundance. Forty-two PARBs carrying ARGs and VFGs were detected, and three PARBs (Pseudomonas aeruginosa, Pseudomonas stutzeri and Pseudomonas fluorescens) increased obviously in relative abundance at the downstream site near the outfall. In conclusion, long-term wastewater discharge had effects on the microbial community, and limited microbiological risks existed in the receiving waters.

Metagenomic analysis of antibiotic-resistance genes and viruses released from glaciers into downstream habitats.

Glaciers serve as effective reservoirs of antibiotic resistance genes (ARGs) and viruses for millions of years. Climate change and anthropogenic activity have accelerated the melting of glaciers, but the patterns of release of ARGs and viruses from melting glaciers into downstream habitats remain unknown. We analyzed 171 metagenomic samples from glaciers and their downstream habitats and found that the abundance and diversity of ARGs were higher in glaciers (polar and plateau glaciers) than downstream habitats (Arctic Ocean, Qinghai Lake, and Yangtze River Basin), with the diversity of viruses having the opposite pattern. Proteobacteria and Actinobacteria were the main potential hosts of ARGs and viruses, and the richness of ARGs carried by the hosts was positively correlated with viral abundance, suggesting that the transmission of viruses in the hosts could disseminate ARGs. Source tracking indicated that >18 % of the ARGs and >25 % of the viruses detected in downstream habitats originated from glaciers, demonstrating that glaciers could be one of the potential sources of ARGs and viruses in downstream habitats. Increased solar radiation and emission of carbon dioxide mainly influenced the release of the ARGs and viruses from glaciers into downstream habitats. This study provides a systematic insight demonstrating the release of ARGs and viruses from the melting glaciers, potentially increasing ecological pressure.