Silver nanoparticles regulate antibiotic resistance genes by shifting bacterial community and generating anti-silver genes in estuarine biofilms. - Related Documents




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795601.0000Silver nanoparticles regulate antibiotic resistance genes by shifting bacterial community and generating anti-silver genes in estuarine biofilms. Biofilms are thought to be sinks for antibiotic resistance genes (ARGs) and nanoparticles (NPs), however, studies on the interactions between NPs and ARGs in biofilms are limited. This study focused on the occurrence and regulatory mechanisms of ARGs during the formation of biofilms with continuous treatment of zero-valent silver nanoparticles (Ag(0)-NPs) and Ag ions at an environmental concentration of 10 µg/L in the Yangtze Estuary. The biofilms could enrich large amounts of Ag, with the highest concentration of 97.60 mg/kg and 111.08 mg/kg in the Ag(0)-NPs and Ag ions group at 28 days. Compared to the blank at 28 days, the abundance of ARGs was reduced 2.2 times in the Ag(0)-NPs group, whereas it increased 1.3 times in the Ag ion group. Ag(0)-NPs and Ag ions induced the production of silver resistance genes (SRGs) or selected bacteria with SRGs in biofilms. Based on machine learning, the bacterial community, SRGs, and Ag concentration were the top three dominant regulators of ARGs, with 27.74 %, 25.57 %, and 17.93 % contributions, respectively. Structural equation modeling revealed that Ag could indirectly regulate ARGs by regulating the bacterial community in the Ag(0)-NPs group. Metagenomic sequencing further showed that most of the decreased ARGs were hosted by Betaproteobacteria in the Ag(0)-NPs groups. According to the KEGG pathway database, the possible molecular mechanism of Ag(0)-NPs/Ag ions regulating ARGs may be through the two-component system (arlS/silS-arlR) and beta-lactam resistance system (mexI-mexV-oprM/oprZ/smeF). Overall, this study provides new insights into the effects of Ag(0)-NPs at environmental concentrations on the ecological environment, especially regarding the mechanism of regulating ARGs in estuarine biofilms.202439489934
758510.9998Impacts of engineered nanoparticles and antibiotics on denitrification: Element cycling functional genes and antibiotic resistance genes. The wide presence of antibiotics and minerals warrants their combined effects on the denitrification in natural aquatic environment. Herein, we investigated the effects of two antibiotics, sulfamethazine (SMZ) and chlortetracycline (CTC), on the reduction of NO(3)(-)-N and accumulation of NO(2)(-)-N in the absence and presence of engineered nanoparticles (NPs) (Al(2)O(3), SiO(2), and geothite) using 16 S rRNA sequencing and high-throughput quantitative PCR. The results showed that the addition of antibiotics inhibited the reduction of NO(3)(-)-N by changing the bacterial community structure and reducing the abundance of denitrification genes, while engineered NPs promoted the denitrification by increasing the abundance of denitrification genes. In the binary systems, engineered NPs alleviated the inhibitory effect of antibiotics through enriching the denitrification genes and adsorbing antibiotics. Antibiotics and its combination with engineered NPs changed the composition of functional genes related to C, N, P, S metabolisms (p < 0.01). The addition of antibiotics and/or engineered NPs altered the bacterial community structure, which is dominated by the genera of Enterobacter (40.7-90.5%), Bacillus (4.9-58.5%), and Pseudomonas (0.21-12.7%). The significant relationship between denitrification, carbon metabolism genes, and antibiotic resistance genes revealed that the heterotrophic denitrifying bacteria may host the antibiotic resistance genes and denitrification genes simultaneously. The findings underscore the significance of engineered NPs in the toxicity assessment of pollutants, and provide a more realistic insight into the toxicity of antibiotics in the natural aquatic environment.202235738104
763020.9998Coexistence of silver ion and tetracycline at environmentally relevant concentrations greatly enhanced antibiotic resistance gene development in activated sludge bioreactor. Antibiotic resistance has become a global public health problem. Recently, various environmental pollutants have been reported to induce the proliferation of antibiotic resistance. However, the impact of multiple pollutants (e.g., heavy metals and antibiotics), which more frequently occur in practical environments, is poorly understood. Herein, one widely distributed heavy metal (Ag(+)) and one frequently detected antibiotic (tetracycline) were chosen to investigate their coexisting effect on the proliferation of antibiotic resistance in the activated sludge system. Results show that the co-occurrence of Ag(+) and tetracycline at environmentally relevant concentrations exhibited no distinct inhibition in reactor performances. However, they inhibited the respiratory activity by 42%, destroyed the membrane structure by 218%, and increased membrane permeability by 29% compared with the blank control bioreactor. Moreover, the relative abundances of target antibiotic resistance genes (ARGs) (e.g., tetA, bla(TEM-1), and sulII) in effluent after exposure of coexisting Ag(+) and tetracycline were increased by 92-1983% compared with those in control reactor, which were 1.1-4.3 folds higher than the sum of the sole ones. These were possibly attributed to the enrichments of antibiotic-resistant bacteria. The results would illumine the coexisting effect of heavy metals and antibiotics on the dissemination of ARGs in activated sludge system.202234482077
795930.9997Evolution of microbial community and antibiotic resistance genes in anammox process stressed by oxytetracycline and copper. The individual and combined impacts of copper ion (Cu(2+)) and oxytetracycline (OTC) on anaerobic ammonium oxidation (anammox) performance and its self-recovery process were examined. Experimental results showed that the anammox performance and activity of anammox bacteria were inhibited by 1.0 mg L(-1) OTC, Cu(2+) and OTC + Cu(2+), and both single and combined inhibitions were reversible. The abundance of functional genes and parts of antibiotic resistance genes (ARGs) were positively related to the dominant bacterium Ca. Kuenenia, implying that the recovery of the performance was associated with the progressive induction of potentially resistant species after inhibition. The above outcomes illustrated that anammox bacteria were stressed by metals and antibiotics, but they still could remove nitrogen at a rate higher than 20.6 ± 0.8 kg N m(-3) d(-1), providing guidance for engineering applications of anammox processes.202132949830
759840.9997Aspartame affects methane yield and enhances transmission of antibiotic resistance genes during anaerobic digestion of sludge. Aspartame (ASP) is a widely used artificial sweetener, yet recent studies have shown that ASP have potential toxic effect. ASP is also detected in sludge, however, the influence of ASP on the performance of sludge anaerobic digestion and the fate of antibiotic resistance genes (ARGs) have not been thoroughly investigated. Under stress of 0, 0.5, 5 and 50 mg/L ASP, cumulative methane production was 181.7, 167.0, 154.0 and 140.8 mlCH(4)/g VSS, respectively. ASP inhibited the dissolution and conversion of organic matter in sludge. Sequencing data revealed a decline in the abundance of functional microorganisms compared to control, such as hydrolytic-acidifying bacteria and methanogens, potentially attributed to increased intracellular reactive oxygen species and damaged cell membranes caused by ASP addition. Specifically, 50 mg/L ASP reduced the total abundance of methanogens by 59.40 % compared to control. Concurrently, alterations in microbial communities along with an increase in Tn916 and intI1 were observed, increasing the abundance of ARGs. The total abundance of five ARGs peaked at 1.43E+ 12 copies/g at 5 mg/L ASP, representing 139 % of the control. This research contributes valuable insights into the alterations in anaerobic digestion caused by ASP, emphasizing the potential risks in the overall environmental system.202539616849
808050.9997Fates of antibiotic resistance genes and bacterial/archaeal communities of activated sludge under stress of copper: Gradient increasing/decreasing exposure modes. Effect of copper (Cu) on antibiotic resistance genes (ARGs) and bacterial/archaeal community of activated sludge under gradient increasing (0.5-10 mg/L) or decreasing exposure (10-0.5 mg/L) modes was explored. Here, 29 genes were detected among 48 selected ARGs and mobile gene elements (MGEs). Two exposure modes showed dissimilar effects on ARGs and distribution was more affected by environmental concentrations of Cu, which promoted transmission of ARGs (multiple drug resistance and sulfonamide). Cellular protection was main resistance mechanism, which was less inhibited than efflux pumps. The tnpA-02, as main MGE, interacted closely with ARGs (sul2, floR, etc.). Gradient increasing exposure mode had more effects on bacterial/archaeal structure and composition. Bacteria were main hosts for specific ARGs and tnpA-02, while archaea carried multiple ARGs (cmx(A), adeA, etc.), and bacteria (24.24 %) contributed more to changes of ARGs than archaea (19.29 %). This study clarified the impacts of Cu on the proliferation and transmission of ARGs.202236096328
796060.9997Diversity evolution of functional bacteria and resistance genes (CzcA) in aerobic activated sludge under Cd(II) stress. An activated sludge sequencing batch reactor (SBR) was used to treat divalent cadmium (Cd(II)) wastewater for 60 d to investigate the overall treatment performance, evolution of the bacterial community, and abundance of the Cd(II) resistance gene CzcA and shifts in its potential host bacteria. During stable operation with a Cd(II) concentration of 20 mg/L, the average removal efficiencies of Cd(II) and chemical oxygen demand (COD) were more than 85% and that of total phosphorus was greater than 70%, while the total nitrogen (TN) was only about 45%. The protein (PN) content in the extracellular polymeric substances (EPS) increased significantly after Cd(II) addition, while polysaccharides displayed a decreasing trend (p < 0.05), indicating that EPS prefer to release PN to adsorb Cd(II) and protect bacteria from damage. Three-dimensional fluorescence spectral analysis showed that fulvic acid-like substances were the most abundant chemical components of EPS. The addition of Cd(II) adversely affected most denitrifying bacteria (p < 0.05), which is consistent with the low TN removal. In addition, quantitative polymerase chain reaction analysis revealed that CzcA gene abundance decreased as the Cd(II) concentration increased, possibly because expression of the CzcA gene was inhibited by Cd(II) stress. The majority of CzcA gene sequences were carried by Pseudomonas, making it the dominant genus among Cd(II)-resistant bacteria.201931514000
796670.9997How heavy metal stress promotes dissemination of antibiotic resistance genes in the activated sludge process. Heavy metals have been recently revealed as promoters to antibiotic resistance gene (ARG) dissemination in water environment, but their influence on ARG transfer in the activated sludge process has not been clear. In this study, a set of sequencing batch reactors (SBRs) and micro-scale microfluidic chips were established to quantify the impacts of heavy metals (0.5 mM of Pb, 0.1 mM of As, and 0.005 mM of Hg) on the ARG spreading in the activated sludge consortium. Under heavy metal stress, transfer frequencies were 1.7-3.6 folds increase compared to the control. Gram-negative bacteria increased significantly after heavy metal added, which were more prone to receiving resistant plasmid from donors. Meanwhile, the relative expression of genes related to conjugation changed in activated sludge, especially the expression of outer membrane protein and oxidative stress regulatory genes increased by 2.9-7.4 folds and 7.8-13.1 folds, respectively. Furthermore, using microfluidic chips, the dynamics of ARG transfer was observed at single cell level under heavy metal pressure. Heavy metals firstly promoted conjugation and then vertical gene transfer played an important part for ARG spreading. The results provided in-depth understanding of the influence of heavy metals on ARG behavior in the environment.202235724617
758480.9997Responses of microbial community and antibiotic resistance genes to co-existence of chloramphenicol and salinity. In recent years, the risk from environmental pollution caused by chloramphenicol (CAP) has emerged as a serious concern worldwide, especially for the co-selection of antibiotic resistance microorganisms simultaneously exposed to CAP and salts. In this study, the multistage contact oxidation reactor (MCOR) was employed for the first time to treat the CAP wastewater under the co-existence of CAP (10-80 mg/L) and salinity (0-30 g/L NaCl). The CAP removal efficiency reached 91.7% under the co-existence of 30 mg/L CAP and 10 g/L NaCl in the influent, but it fluctuated around 60% with the increase of CAP concentration and salinity. Trichococcus and Lactococcus were the major contributors to the CAP and salinity shock loads. Furthermore, the elevated CAP and salinity selection pressures inhibited the spread of CAP efflux pump genes, including cmlA, tetC, and floR, and significantly affected the composition and abundance of antibiotic resistance genes (ARGs). As the potential hosts of CAP resistance genes, Acinetobacter, Enterococcus, and unclassified_d_Bacteria developed resistance against high osmotic pressure and antibiotic environment using the efflux pump mechanism. The results also revealed that shifting of potential host bacteria significantly contributed to the change in ARGs. Overall, the co-existence of CAP and salinity promoted the enrichment of core genera Trichococcus and Lactococcus; however, they inhibited the proliferation of ARGs. KEY POINTS: • Trichococcus and Lactococcus were the core bacteria related to CAP biodegradation • Co-existence of CAP and salinity inhibited proliferation of cmlA, tetC, and floR • The microorganism resisted the CAP using the efflux pump mechanism.202236205764
757190.9997Ancient Oriental Wisdom still Works: Removing ARGs in Drinking Water by Boiling as compared to Chlorination. Antibiotic resistance genes (ARGs) in municipal drinking water may not be effectively removed during centralized treatment. To reduce potential health risks, water disinfection at the point-of-use scale is warranted. This study investigated the performance of boiling, a prevalent household water disinfection means, in response to ARGs contamination. We found that boiling was more efficient in inactivating both Escherichia coli and environmental bacteria compared to chlorination and pasteurization. Boiling of environmental bacteria suspension removed a much broader spectrum of ARGs and mobile genetic elements (up to 141 genes) than chlorination (up to 13 genes), such better performance was largely attributed to a stronger inactivation of chlorine-tolerant bacteria including Acinetobacter and Bacillus. Accumulation of extracellular ARGs was found during low-temperature heating (≤ 80°C) and in the initial stage of chlorination (first 3 min when initial chlorine was 5 mg/L and first 12 min when initial chlorine was 1 mg/L). These extracellular ARGs as well as the intracellular ARGs got removed as the heating temperature increased or the chlorination time prolonged. Under the same treatment time (30 min), high-temperature heating (≥ 90.1°C) damaged the DNA structure more thoroughly than chlorination (5 mg/L). Taking into account the low transferability of ARGs after DNA melting, boiling may provide an effective point-of-use approach to attenuating bacterial ARGs in drinking water and is still worth promoting in the future.202234910990
7843100.9997Inactivation of chlorine-resistant bacteria (CRB) via various disinfection methods: Resistance mechanism and relation with carbon source metabolism. With the widespread use of chlorine disinfection, chlorine-resistant bacteria (CRB) in water treatment systems have gained public attention. Bacterial chlorine resistance has been found positively correlated with extracellular polymeric substance (EPS) secretion. In this study, we selected the most suitable CRB controlling method against eight bacterial strains with different chlorine resistance among chloramine, ozone, and ultraviolet (UV) disinfection, analyzed the resistance mechanisms, clarified the contribution of EPS to disinfection resistance, and explored the role of carbon source metabolism capacity. Among all the disinfectants, UV disinfection showed the highest disinfection capacity by achieving the highest average and median log inactivation rates for the tested strains. For Bacillus cereus CR19, the strain with the highest chlorine resistance, 40 mJ/cm(2) UV showed a 1.90 log inactivation, which was much higher than that of 2 mg-Cl(2)/L chlorine (0.67 log), 2 mg-Cl(2)/L chloramine (1.68 log), and 2 mg/L ozone (0.19 log). Meanwhile, the UV resistance of the bacteria did not correlate with EPS secretion. These characteristics render UV irradiation the best CRB controlling disinfection method. Chloramine was found to have a generally high inactivation efficiency for bacteria with high chlorine-resistance, but a low inactivation efficiency for low chlorine-resistant ones. Although EPS consumed up to 56.7% of chloramine which an intact bacterial cell consumed, EPS secretion could not explain chloramine resistance. Thus, chloramine is an acceptable CRB control method. Similar to chlorine, ozone generally selected high EPS-secreting bacteria, with EPS consuming up to 100% ozone. Therefore, ozone is not an appropriate method for controlling CRB with high EPS secretion. EPS played an important role in all types of disinfection resistance, and can be considered the main mechanism for bacterial chlorine and ozone disinfection resistance. However, as EPS was not the main resistance mechanism in UV and chloramine disinfection, CRB with high EPS secretion were inactivated more effectively. Furthermore, carbon source metabolism was found related to the multiple resistance of bacteria. Those with low carbon source metabolism capacity tended to have higher multiple resistance, especially to chlorine, ozone, and UV light. Distinctively, among the tested gram-negative bacteria, in contrast to other disinfectants, chloramine resistance was negatively correlated with EPS secretion and positively correlated with carbon source metabolism capacity, suggesting a special disinfection mechanism.202337659185
8082110.9997Deciphering the toxic effects of antibiotics on denitrification: Process performance, microbial community and antibiotic resistance genes. The extensive application of antibiotics, and the occurrence and spread of antibiotic resistance genes (ARGs) shade health risks to human and animal. The long-term effects of sulfamethoxazole (SMX) and tetracycline (TC) on denitrification process were evaluated in this study, with the focus on nitrogen removal performance, microbial community and ARGs. Results showed that low-concentration SMX and TC (<0.2 mg L(-1)) initially caused a deterioration in nitrogen removal performance, while higher concentrations (0.4-20 mg L(-1)) of both antibiotics had no further inhibitory influences. The abundances of ARGs in both systems generally increased during the whole period, and most of them had significant correlations with intI1, especially efflux-pump genes. Castellaniella, which was the dominant genus under antibiotic pressure, might be potential resistant bacteria. These findings provide an insight into the toxic effects of different antibiotics on denitrification process, and guides future efforts to control antibiotics pollution in ecosystems.202032250829
7572120.9997Stormwater runoff treatment through electrocoagulation: antibiotic resistant bacteria removal and its transmission risks. Recently, increasing attention has been paid to antibiotic resistant bacteria (ARB) in stormwater runoff. However, there were little data on ARB removal through electrocoagulation (EC) treatment. In this study, batch experiments were conducted to investigate key designs for ARB removal, role of SS, effects of water matrix, and potential risks after EC treatment under the pre-determined conditions. EC treatment with 5 mA/cm(2) of current density and 4 cm of inter-electrode distance was optimal with the highest ARB removal (3.04 log reduction for 30 min). The presence of SS significantly improved ARB removal during EC treatment, where ARB removal increased with the increase of SS levels when SS less than 300 mg/L. Large ARB removal was found under particles with size lower than 150 μm with low contribution (less than 10%) of the settlement without EC treatment, implying that the enhancement of ARB adsorption onto small particles could be one of the reasonable approaches for ARB removal through EC treatment. ARB removal increased firstly and then decreased with the increase of pH, while had proportional relationship with conductivity. After the optimal condition, there were weak conjugation transfer but high transformation frequency (5.5 × 10(-2) for bla(TEM)) for target antibiotic resistance genes (ARGs), indicating that there could be still a risk of antibiotic resistance transformation after EC treatment. These suggested that the combination of EC and other technologies (like electrochemical disinfection) should be potential ways to control antibiotic resistance transmission through stormwater runoff.202436848218
7844130.9997Insight into using a novel ultraviolet/peracetic acid combination disinfection process to simultaneously remove antibiotics and antibiotic resistance genes in wastewater: Mechanism and comparison with conventional processes. In this study, the simultaneous removal mechanism of antibiotics and antibiotic resistance genes (ARGs) was investigated using the novel ultraviolet/peracetic acid (UV/PAA) combination disinfection process and conventional disinfection processes were also applied for comparison. The results showed that UV/PAA disinfection with a high UV dosage (UV/PAA-H) was most effective for the removal of tetracyclines, quinolones, macrolides and β-lactams; their average removal efficiencies ranged from 25.7% to 100%, while NaClO disinfection was effective for the removal of sulfonamides (∼81.6%). The majority of ARGs were well removed after the UV/PAA-H disinfection, while specific genes including tetB, tetC, ermA and bla(TEM) significantly increased after NaClO disinfection. In addition, β-lactam resistance genes (-35.9%) and macrolides resistance genes (-12.0%) remarkably augmented after UV/NaClO disinfection. The highly reactive oxidation species generated from UV/PAA process including hydroxyl radicals (•OH) and carbon-centered organic radicals (R-C•), were responsible for the elimination of antibiotics and ARGs. Correlation analysis showed that tetracycline, sulfonamide and macrolide antibiotics removal showed a positive correlation with the corresponding ARGs, and a low dose of antibiotic residues played an important role in the distribution of ARGs. Metagenomic sequencing analysis showed that UV/PAA disinfection could not only greatly decrease the abundance of resistant bacteria but also downregulate the expression of key functional genes involved in ARGs propagation and inhibit the signal transduction of the host bacteria, underlying that its removal mechanism was quite different from that of NaClO-based disinfection processes. Our study provides valuable information for understanding the simultaneous removal mechanism of antibiotics and ARGs in wastewater during the disinfection processes, especially for the novel UV/PAA combination process.202234982977
7568140.9997Self-rescue of nitrogen-cycling bacteria under β-lactam antibiotics stress during managed aquifer recharge (MAR): Microbial collaboration and anti-resistance. Storing reclaimed water via managed aquifer recharge (MAR) is an effective strategy for alleviating groundwater overdraft and achieving water resource recycling simultaneously. However, β-lactam antibiotics in the reclaimed water can induce stress on aquifer system, reshape microbial community, and affect the emergence and prevalence of antibiotic resistance genes (ARGs). In this study, three sandy soil columns (H 1.5 m, ID 14 cm) were employed to simulate MAR, and synthetic reclaimed water containing either amoxicillin (AMO), ampicillin (AMP) or oxacillin (OXA) was continuously recharged for 120 d The temporal and spatial attenuation of β-lactams and nitrogen was studied, and microbial collaboration and the resistance mechanism were elaborated. Biodegradation is the main pathway for β-lactams elimination, AMO and AMP were eliminated when migrating 30 cm, while the attenuation of OXA experienced in the whole column with final removal efficiency of 82%. Moreover, refractory OXA induced more ARGs production, and approximately 10% and 13% higher than that of AMO and AMP columns. Efflux pump and antibiotics inactivation were the two major resistance mechanisms. NO(3)(-)-N gradually decreased (by 26%, 38%, and 49% for AMO, AMP, and OXA, respectively) along the recharge direction. Microbial co-occurrence network revealed that nitrogen-cycling bacteria were the keystone species in aquifer community, and ammonation provided NH(4)(+)-N for the nitrification process of ammonia-oxidizing archaea (AOA), promoting the further denitrification for nitrogen removal in MAR process. Nitrogen-cycling bacteria were the key and active ARG hosts, which could keep nitrogen transformation activity under antibiotics stress. In sum, nitrogen-cycling bacteria exhibited intimate collaboration and elastic resistance in response to the malnutrition environment and β-lactams exposure during MAR.202336689880
6748150.9997Time-dependent effects of ZnO nanoparticles on bacteria in an estuarine aquatic environment. Many studies have examined the acute toxicity of nanoparticles (NPs) towards model bacteria. In this study, we report the time-dependent effects of ZnO NPs on native, selected Zn-resistant and dominant bacteria in estuarine waters. An initial inhibition of bacterial growth followed by a recovery at 24 h was observed, and this rebound phenomenon was particularly notable when the raw water samples were treated with relatively high ZnO NP concentrations (1 and 10 mg/L).By comparing the groups treated with Zn(2+), Zn(2+) was shown to largely explain the acute cytotoxic effect of ZnO NPs on bacteria in raw waters. Furthermore, similar to the native bacteria, especially the dominant bacteria, the viability of Escherichia coli (E. coli) decreased with the increasing treatments time and the concentrations of ZnO NPs in water with different salinities. Moreover, the expression of Zn-resistance genes including zntA and zntR in E. coli suggested that the Zn-resistance system in E. coli can be activated to defend against the stress of Zn(2+) released from ZnO NPs, and salinity may promote this process in estuarine aquatic systems. Thus, the effect of ZnO NPs on bacteria in estuarine water bodies is likely determined by the synergistic effect of environmental salinity and dissolved Zn ions. As such, our findings are of high relevance and importance for understanding the ecological disturbances caused by anthropogenic NPs in estuarine environments.202031505343
7845160.9997Mechanism and potential risk of antibiotic resistant bacteria carrying last resort antibiotic resistance genes under electrochemical treatment. The significant rise in the number of antibiotic resistance genes (ARGs) that resulted from our abuse of antibiotics could do severe harm to public health as well as to the environment. We investigated removal efficiency and removal mechanism of electrochemical (EC) treatment based on 6 different bacteria isolated from hospital wastewater carrying 3 last resort ARGs including NDM-1, mcr-1 and tetX respectively. We found that the removal efficiency of ARGs increased with the increase of both voltage and electrolysis time while the maximum removal efficiency can reach 90%. The optimal treatment voltage and treatment time were 3 V and 120 min, respectively. Temperature, pH and other factors had little influence on the EC treatment process. The mechanism of EC treatment was explored from the macroscopic and microscopic levels by scanning electron microscopy (SEM) and flow cytometry. Our results showed that EC treatment significantly changed the permeability of cell membrane and caused cells successively experience early cell apoptosis, late cell apoptosis and cell necrosis. Moreover, compared with traditional disinfection methods, EC treatment had less potential risks. The conjugative transfer frequencies of cells were significantly reduced after treatment. Less than 1% of bacteria entered the viable but nonculturable (VBNC) state and less than 5% of intracellular ARGs (iARGs) turned into extracellular ARGs (eARGs). Our findings provide new insights into as well as important reference for future electrochemical treatment in removing ARB from hospital wastewater.202235085630
7193170.9997Plasmid-mediated transfer of antibiotic resistance genes and biofilm formation in a simulated drinking water distribution system under chlorine pressure. The effects of disinfectants and plasmid-based antibiotic resistance genes (ARGs) on the growth of microorganisms and the plasmid-mediated transfer of ARGs in the water and biofilm of the drinking water distribution system under simulated conditions were explored. The heterotrophic plate count of the water in reactors with 0.1 mg/L NaClO and NH(2)Cl was higher than in the control groups. There was no similar phenomenon in biofilm. In the water of reactors containing NaClO, the aphA and bla genes were lower than in the antibiotic resistant bacteria group, while both genes were higher in the water of reactors with NH(2)Cl than in the control group. Chloramine may promote the transfer of ARGs in the water phase. Both genes in the biofilm of the reactors containing chlorine were lower than the control group. Correlation analysis between ARGs and water quality parameters revealed that the copy numbers of the aphA gene were significantly positively correlated with the copy numbers of the bla gene in water and significantly negatively correlated in biofilm (p < 0.05). The results of the sequencing assay showed that bacteria in the biofilm, in the presence of disinfectant, were primarily Gram-negative. 1.0 mg/L chlorine decreased the diversity of the community in the biofilm. The relative abundance of some bacteria that may undergo transfer increased in the biofilm of the reactor containing 0.1 mg/L chlorine.202539617560
7583180.9997Insights into the combined effect of ofloxacin and humic substances on sewage sludge anaerobic digestion. Humic substances (HS) and antibiotics are present simultaneously in various environments. However, the influence path and consequences of HS on antibiotics behaviors in complex anaerobic microbial systems are rarely known, hindering the understanding and control of antibiotics risks. This study for the first time investigated the combined effects of ofloxacin (OFL) and HS in sewage sludge anaerobic digestion system. Experimental results showed that OFL alone reduced the cumulative methane production and the maximum methane production rate by 14.6 % and 33.5 %, respectively. The methane production curves showed step by step adaption, which might be related with the increase of antibiotics resistance genes and their potential hosts. The coexistence of low concentration (6 % of sludge volatile solid) HS could alleviate the inhibition of OFL on hydrolysis-related bacteria and genes to a certain extent, thereby enhanced the methane production by 4.8 %. However, the coexistence of high concentration (12-24 % of sludge volatile solid) HS intensified the inhibition on hydrolysis-related bacteria and genes, and had more potential to combine with organic matters to prevent sludge solubilization, macromolecular organics hydrolysis and OFL degradation, thereby further decreasing the methane production by 7.6-15.9 %. Besides, the coexistence of OFL and high concentration HS increased the antimicrobial resistance and pathogenicity risks of digested sludge, by enhancing the residual level of verified pathogens, antibiotics resistance genes and virulence factor genes. This study provides new insights into the environmental risks of combined antibiotics and HS pollution, and offers a basis for strengthening the safe treatment and disposal of sewage sludge.202540752562
7590190.9997Characteristics, Performance and Microbial Response of Aerobic Granular Sludge for Treating Tetracycline Hypersaline Pharmaceutical Wastewater. Salt-tolerant aerobic granular sludge(AGS) was successfully cultivated under the dual stress of tetracycline and 2.5% salinity, resulting in an average particle size of 435.0 ± 0.5 and exhibiting a chemical oxygen demand(COD) removal rate exceeding 80%, as well as excellent sedimentation performance. The analysis of metagenomics technology revealed a significant pattern of succession in the development of AGS. The proportion of Oleiagrimonas, a type of salt-tolerant bacteria, exhibited a gradual increase and reached 38.07% after 42 days, which indicated that an AGS system based on moderate halophilic bacteria was successfully constructed. The expression levels of targeted genes were found to be reduced across the entire AGS process and formation, as evidenced by qPCR analysis. The presence of int1 (7.67 log10 gene copies g(-1) in 0 d sludge sample) enabled microbes to horizontally transfer ARGs genes along the AGS formation under the double pressure of TC and 2.5% salinity. These findings will enhance our understanding of ARG profiles and the development in AGS under tetracycline pressure, providing a foundation for guiding the use of AGS to treat hypersaline pharmaceutical wastewater.202438930555