Antibiotic Resistance Genes In Wastewater Research Articles

Comparison of qPCR and metagenomic sequencing methods for quantifying antibiotic resistance genes in wastewater.

Surveillance methods of circulating antibiotic resistance genes (ARGs) are of utmost importance in order to tackle what has been described as one of the greatest threats to humanity in the 21st century. In order to be effective, these methods have to be accurate, quickly deployable, and scalable. In this study, we compare metagenomic shotgun sequencing (TruSeq DNA sequencing) of wastewater samples with a state-of-the-art PCR-based method (Resistomap HT-qPCR) on four wastewater samples that were taken from hospital, industrial, urban and rural areas. ARGs that confer resistance to 11 antibiotic classes have been identified in these wastewater samples using both methods, with the most abundant observed classes of ARGs conferring resistance to aminoglycoside, multidrug-resistance (MDR), macrolide-lincosamide-streptogramin B (MLSB), tetracycline and beta-lactams. In comparing the methods, we observed a strong correlation of relative abundance of ARGs obtained by the two tested methods for the majority of antibiotic classes. Finally, we investigated the source of discrepancies in the results obtained by the two methods. This analysis revealed that false negatives were more likely to occur in qPCR due to mutated primer target sites, whereas ARGs with incomplete or low coverage were not detected by the sequencing method due to the parameters set in the bioinformatics pipeline. Indeed, despite the good correlation between the methods, each has its advantages and disadvantages which are also discussed here. By using both methods together, a more robust ARG surveillance program can be established. Overall, the work described here can aid wastewater treatment plants that plan on implementing an ARG surveillance program.

Sensitivity and consistency of long- and short-read metagenomics and epicPCR for the detection of antibiotic resistance genes and their bacterial hosts in wastewater

Wastewater surveillance is a powerful tool to assess the risks associated with antibiotic resistance in communities. One challenge is selecting which analytical tool to deploy to measure risk indicators, such as antibiotic resistance genes (ARGs) and their respective bacterial hosts. Although metagenomics is frequently used for analyzing ARGs, few studies have compared the performance of long-read and short-read metagenomics in identifying which bacteria harbor ARGs in wastewater. Furthermore, for ARG host detection, untargeted metagenomics has not been compared to targeted methods such as epicPCR. Here, we 1) evaluated long-read and short-read metagenomics as well as epicPCR for detecting ARG hosts in wastewater, and 2) investigated the host range of ARGs across the wastewater treatment plant (WWTP) to evaluate host proliferation. Results highlighted long-read revealed a wider range of ARG hosts compared to short-read metagenomics. Nonetheless, the ARG host range detected by long-read metagenomics only represented a subset of the hosts detected by epicPCR. The ARG-host linkages across the influent and effluent of the WWTP were characterized. Results showed the ARG-host phylum linkages were relatively consistent across the WWTP, whereas new ARG-host species linkages appeared in the WWTP effluent. The ARG-host linkages of several clinically relevant species found in the effluent were identified.

The fate of antibiotic resistance genes in wastewater containing microalgae treated by chlorination, ultra-violet, and Fenton reaction

Antibiotic resistance genes (ARGs) and bacteria (ARBs) in the effluent of wastewater treatment plants (WWTPs) are of utmost importance for the dissemination of ARGs in natural aquatic environments. Therefore, there is an urgent need for effective technologies to eliminate WWTP ARGs/ARBs and mitigate the associated risks posed by the discharged ARG in aquatic environments. To test the effective technology for eliminating ARGs/ARBs, we compared the removal of ARGs and ARBs by three different tertiary treatments, namely ultra-violet (UV) disinfection, chlorination disinfection, and Fenton oxidation. Then, the treated wastewater was co-cultured with Chlorella vulgaris (representative of aquatic biota) to investigate the fate of discharged ARGs into the aquatic environment. The results demonstrated that chlorination (at a chlorine concentration of 15 mg/L) and Fenton (at pH 2.73, with 0.005 mol/L Fe2+ and 0.0025 mol/L H2O2) treatment showed higher efficacy in ARG removal (1.8 - 4.17 logs) than UV treatment (15 min) (1.29 - 3.87 logs). Moreover, chlorine at 15 mg/L and Fenton treatment effectively suppressed ARB regeneration while UV treatment for 15 min could not. Regardless of treatments tested in this study, the input of treated wastewater to the Chlorella system increased the number of ARGs and mobile genetic elements (MGEs), indicating the potential risk of ARG dissemination associated with WWTP discharge. Among the wastewater-Chlorella co-culture systems, chlorination resulted in less of an increase in the number of ARGs and MGEs compared to Fenton and UV treatment. When comparing the wastewater systems to the co-culture systems, it was observed that Chlorella vulgaris reduced the number of ARGs and MGEs in chlorination and UV-treated wastewater; however, Chlorella vulgaris promoted ARG survival in Fenton-treated water, suggesting that aquatic microalgae might act as a barrier to ARG dissemination. Overall, chlorination treatment not only effectively removes ARGs and inhibits ARB regeneration but also shows a lower risk of ARG dissemination. Therefore, chlorination is recommended for practical application in controlling the spread of discharged ARGs from WWTP effluent in natural aquatic environments.

Wastewater surveillance for antibiotics and resistance genes in a river catchment: Spatiotemporal variations and the main drivers

Wastewater-based epidemiology (WBE) is used for mining information about public health such as antibiotics resistance. This study investigated the distribution profiles of six types of antibiotic resistance genes (ARGs) in wastewater and rivers in Wuhu City, China. The levels of ARGs found in the Qingyijiang River were significantly higher than other rivers, and were comparable to effluent levels. Among the ARGs, sulfonamides ARGs and intI1 were the predominant in both wastewaters and rivers. Additionally, the concentrations of ARGs were higher on weekends as opposed to weekdays. Their distribution patterns remained consistent inter-week and inter-season using linear regression analysis (p < 0.001). Interestingly, the occurrence levels of ARGs in wastewaters during spring were significantly higher than in autumn, although insignificant in rivers. The apparent removal rate of ARGs in domestic wastewater sources ranged from 61.52–99.29%, except for qepA (-1.91% to 81.09%), whereas the removal rates in mixed domestic and industrial wastewaters showed a marked decrease (-92.94% to 76.67%). A correlation network analysis revealed that azithromycin and erythromycin were key antibiotics, while blaNDM-1, tetM, tetB, and ermB were identified as key ARGs. Sulfonamide and fluoroquinolone antibiotics, and tetracycline and macrolide ARGs were the primary contributors. Linear mixed models demonstrated that socio-economic variables positively impacted the occurrence levels of ARGs, whereas wastewater flow and river runoff were the negative drivers for their concentrations in wastewaters and surface waters, respectively. Overall, this WBE study contributes to the understanding of spatiotemporal profiles and main drivers of the occurrence of ARGs in wastewater and receiving water.

Ozone micron bubble pretreatment for antibiotic resistance genes reduction in hospital wastewater treatment

Ozone micron bubble (OMB) treatment offers a promising approach to effectively eliminate Antibiotic Resistance Genes (ARGs) from infectious medical wastewater and mitigate the threat of drug resistance transmission. This study evaluated the effectiveness of OMB treatment for reducing ARGs from infectious medical wastewater in laboratory and on-site pilot treatment setups. In part, the presence of antibiotic residues in a hospital wastewater treatment plant (WWTP) and the impact of hospital wastewater on the distribution of ARGs in a wastewater collection system were also investigated. The results of wastewater collection system survey revealed a high prevalence of ARGs in the system, particularly mcr-1, largely originating from medical wastewater discharges. Furthermore, analysis of antibiotic residues in the hospital wastewater treatment system showed significant accumulation, particularly of quinolone antibiotics, in the biomass of the biological oxidation tank, suggesting a potential risk of ARG proliferation within the system. Comparison of wastewater samples from domestic and hospital WWTPs revealed a relatively higher abundance of ARGs in the latter, with differences ranging from 2.2 to sixfold between corresponding locations in the treatment plants. Notably, the biological oxidation unit of both WWTPs exhibited a greater proportion of ARGs among all sampled points, indicating the potential proliferation of ARGs within the biomass of the treatment units. ARG degradation experiments showed that OMB treatment resulted in a significantly lower CT value (9.3 mg O3 L−1 min) compared to ozone coarse bubble treatment (102 mg O3 L−1 min) under identical test conditions. Moreover, the use of OMB on site significantly reduced the accumulation of ARGs in hospital wastewater, underscoring its potential as an effective solution for mitigating ARG spread.

Occurrence and removal prediction of pharmaceuticals positively correlated with antibiotic resistance genes in wastewater treatment processes

Wastewater treatment plants (WWTPs) are important source of pharmaceuticals in the environment, identifying their relationship with antibiotic resistance genes (ARGs) and removal behavior is important for risk control. In this study, the correlations of common pharmaceuticals (including antibiotics and non-antibiotics) and ARGs both in wastewater and sludge were investigated, and machine learning was applied to predict the removal efficiencies of typical pharmaceuticals positively correlated with ARGs. Among the 23 kinds of target pharmaceuticals, both antibiotics and non-antibiotics were found to be positive with typical ARGs in wastewater (e.g. correlation coefficients between caffeine and blaTEM, aadD and qnrS were 0.738, 0.609, 0.936, p < 0.01) but negative in sludge (e.g. correlation coefficients between ofloxacin and tetO, tetW and qnrS were −0.922, −0.933 and −0.902, p < 0.01), indicating the high risk of pharmaceuticals in wastewater promoting ARG spread. Furthermore, based on pharmaceutical removal efficiencies in different wastewater treatment processes in this and previous studies, the removal efficiencies of typical pharmaceuticals highly correlated with ARGs were well predicted through process operation parameters and wastewater characteristics by Random Forest model. HRT and temperature were identified as the most important explanatory variables. This study could provide comprehensive references for controlling pharmaceuticals in WWTPs.

Research Progress on Removing Antibiotic Resistance Genes in Constructed Wetlands

Recently, the issue of environmental pollution and emerging antibiotic resistance genes (ARGs) has gradually gained attention. Antibiotic resistant bacteria (ARB) can be effectively reduced via the conventional water treatment processes. Although the ARB are completely inactivated during the disinfection process, the free ARGs can be incorporated into other microorganisms through transformation or transduction, allowing the ARGs to spread and propagate. Therefore, ARGs in wastewater must be handled by a specific process. It has been demonstrated in several studies that treatment using constructed wetland is an effective, economical, and environmentally friendly method of removing antibiotics and resistance genes from wastewater. Here, the research progress on the removal effect of constructed wetland water treatment systems on ARGs at home and abroad was reviewed. The results revealed that the removal efficiency of ARGs in subsurface flow constructed wetland was higher than that in surface flow-constructed wetlands. The composite-constructed wetland had significantly improved removal efficiency of ARGs compared to that in the ordinarily constructed wetland; however, the parameter setting of the composite process still requires further research. Several studies have reported that the removal efficiency of ARGs using constructed wetlands varies depending on the type of constructed wetland enhancement, plant, temperature, pH, and other factors. The results of the current study revealed that cross-mixing was the best way to combine plants, whereas the selection of plant species has not yet shown a clear dominant species. Temperature and pH affected the removal of ARGs by altering the microbial community in constructed wetlands. Although longer hydraulic residence time could increase the removal efficiency of ARGs, it also increased the enrichment risk of ARGs. The selection of constructed wetland substrate type should focus on fillers with a high specific surface area; the flow direction of the up-flow type was generally more efficient than the down-flow type in removing ARGs. In conclusion, the various factors (such as, the constructed wetland type, substrate type, hydraulic retention time, ambient temperature, and plant species) need to be integrated into the design of the constructed wetland system parameters to achieve the most effective treatment effect. The application of constructed wetlands in removing ARGs from the environment has broad prospects but also faces challenges.

Comparison of concentration and extraction workflows for qPCR quantification of intI1 and vanA in untreated wastewater

Quantitative polymerase chain reaction (qPCR) measurement of antibiotic resistance genes (ARGs) in untreated municipal wastewater may prove useful in combating the antimicrobial resistance crisis. However, harmonizing and optimizing qPCR-based workflows is essential to facilitate comparisons across studies, and includes achieving highly-effective ARG capture through efficient concentration and extraction procedures. In the current study, combinations of sample volume, membrane types and DNA extraction kits within filtration and centrifugation-based workflows were used to quantify 16S ribosomal RNA (16S rRNA), class 1 integron-integrase gene (intI1) and an ARG encoding resistance to vancomycin (vanA) in untreated wastewater sampled from three wastewater treatment plants (WWTPs). Highly abundant 16S rRNA and intI1 were detected in 100 % of samples from all three WWTPs using both 2 and 20 mL sample volumes, while lower prevalence vanA was only detected when using the 20 mL volume. When filtering 2 mL of wastewater, workflows with 0.20-/0.40-μm polycarbonate (PC) membranes generally yielded greater concentrations of the three targets than workflows with 0.22-/0.45-μm mixed cellulose ester (MCE) membranes. The improved performance was diminished when the sample volume was increased to 20 mL. Consistently greater concentrations of 16S rRNA, intI1 and vanA were yielded by filtration-based workflows using PC membranes combined with a DNeasy PowerWater (DPW) Kit, regardless of the sample volume used, and centrifugation-based workflows with DNeasy Blood & Tissue Kit for 2-mL wastewater extractions. Within the filtration-based workflows, the DPW kit yielded more detection and quantifiable results for less abundant vanA than the DNeasy PowerSoil Pro Kit and FastDNA™ SPIN Kit for Soil. These findings indicate that the performance of qPCR-based workflows for surveillance of ARGs in wastewater varies across targets, sample volumes, concentration methods and extraction kits. Workflows must be carefully considered and validated considering the target ARGs to be monitored.

Prevalence of antibiotic-resistant Gram-negative bacteria having extended-spectrum β-lactamase phenotypes in polluted irrigation-purpose wastewaters from Indian agro-ecosystems.

Antibiotic resistance in bacteria has emerged as a serious public health threat worldwide. Aquatic environments including irrigation-purpose wastewaters facilitate the emergence and transmission of antibiotic-resistant bacteria and antibiotic resistance genes leading to detrimental effects on human health and environment sustainability. Considering the paramount threat of ever-increasing antibiotic resistance to human health, there is an urgent need for continuous environmental monitoring of antibiotic-resistant bacteria and antibiotic resistance genes in wastewater being used for irrigation in Indian agro-ecosystems. In this study, the prevalence of antibiotic resistance in Gram-negative bacteria isolated from irrigation-purpose wastewater samples from Sirmaur and Solan districts of Himachal Pradesh was determined. Bacterial isolates of genera Escherichia, Enterobacter, Hafnia, Shigella, Citrobacter, and Klebsiella obtained from 11 different geographical locations were found to exhibit resistance against ampicillin, amoxyclav, cefotaxime, co-trimoxazole, tobramycin, cefpodoxime and ceftazidime. However, all the isolates were sensitive to aminoglycoside antibiotic gentamicin. Enterobacter spp. and Escherichia coli showed predominance among all the isolates. Multidrug-resistance phenotype was observed with isolate AUK-06 (Enterobacter sp.) which exhibited resistant to five antibiotics. Isolate AUK-02 and AUK-09, both E. coli strains showed resistant phenotypes to four antibiotics each. Phenotypic detection revealed that six isolates were positive for extended-spectrum β-lactamases which includes two isolates from Enterobacter spp. and E. coli each and one each from Shigella sp. and Citrobacter sp. Overall, the findings revealed the occurrence of antibiotic resistant and ESBL-positive bacterial isolates in wastewaters utilized for irrigation purpose in the study area and necessitate continuous monitoring and precautionary interventions. The outcomes of the study would be of significant clinical, epidemiological, and agro-environmental importance in designing effective wastewater management and environmental pollution control strategies.

Enhanced removal of antibiotic and antibiotic resistance genes by coupling biofilm electrode reactor and manganese ore substrate up-flow microbial fuel cell constructed wetland system

Manganese ore substrate up-flow microbial fuel cell constructed wetland (UCW-MFC(Mn)) as an innovative wastewater treatment technology for purifying antibiotics and electricity generation with few antibiotic resistance genes (ARGs) generation has attracted attention. However, antibiotic purifying effects should be further enhanced. In this study, a biofilm electrode reactor (BER) that needs direct current driving was powered by a Mn ore anode (UCW-MFC(Mn)) to form a coupled system without requiring direct-current source. Removal efficiencies of sulfadiazine (SDZ), ciprofloxacin (CIP) and the corresponding ARGs in the coupled system were compared with composite (BER was powered by direct-current source) and anaerobic systems (both of BER and UCW-MFC were in open circuit mode). The result showed that higher antibiotic removal efficiency (94% for SDZ and 99.1% for CIP) in the coupled system was achieved than the anaerobic system (88.5% for SDZ and 98.2% for CIP). Moreover, electrical stimulation reduced antibiotic selective pressure and horizontal gene transfer potential in BER, and UCW-MFC further reduced ARG abundances by strengthening the electro-adsorption of ARG hosts determined by Network analysis. Bacterial community diversity continuously decreased in BER while it increased in UCW-MFC, indicating that BER mitigated the toxicity of antibiotic. Degree of modularity, some functional bacteria (antibiotic degrading bacteria, fermentative bacteria and EAB), and P450 enzyme related to antibiotic and xenobiotics biodegradation genes were enriched in electric field existing UCW-MFC, accounting for the higher degradation efficiency. In conclusion, this study provided an effective strategy for removing antibiotics and ARGs in wastewater by operating a BER-UCW-MFC coupled system.

The effect of synthetic silver nanoparticles on the antibiotic resistome and the removal efficiency of antibiotic resistance genes in a hybrid filter system treating municipal wastewater

Engineered nanoparticles, including silver nanoparticles (AgNPs), are released into the environment mainly through wastewater treatment systems. Knowledge of the impact of AgNPs on the abundance and removal efficiency of antibiotic resistance genes (ARGs) in wastewater treatment facilities, including constructed wetlands (CWs), is essential in the context of public health. This study evaluated the effect of increased (100-fold) collargol (protein-coated AgNPs) and ionic Ag+ in municipal wastewater on the structure, abundance, and removal efficiency of the antibiotic resistome, integron-integrase genes, and pathogens in a hybrid CW using quantitative PCR and metagenomic approaches. The abundance of ARGs in wastewater and the removal efficiency of ARGs in the hybrid system were significantly affected by higher Ag concentrations, especially with collargol treatment, resulting in an elevated ARG discharge of system effluent into the environment. The accumulated Ag in the filters had a more profound effect on the absolute and relative abundance of ARGs in the treated water than the Ag content in the water. This study recorded significantly enhanced relative abundance values for tetracycline (tetA, tetC, tetQ), sulfonamide (sul1, sul2), and aminoglycoside (aadA) resistance genes, which are frequently found on mobile genetic elements in collargol- and, to a lesser extent, AgNO3-treated subsystems. Elevated plasmid and integron-integrase gene levels, especially intI1, in response to collargol presence indicated the substantial role of AgNPs in promoting horizontal gene transfer in the treatment system. The pathogenic segment of the prokaryotic community was similar to a typical sewage community, and strong correlations between pathogen and ARG proportions were recorded in vertical subsurface flow filters. Furthermore, the proportion of Salmonella enterica was positively related to the Ag content in these filter effluents. The effect of AgNPs on the nature and characteristics of prominent resistance genes carried by mobile genetic elements in CWs requires further investigation.

Simultaneous removal of antibiotics and antibiotic resistance genes in wastewater by a novel nonthermal plasma/peracetic acid combination system: Synergistic performance and mechanism

In this study, a novel and green method combining plasma with peracetic acid (plasma/PAA) was developed to simultaneously remove antibiotics and antibiotic resistance genes (ARGs) in wastewater, which achieves significant synergistic effects in the removal efficiencies and energy yield. At a plasma current of 2.6 A and PAA dosage of 10 mg/L, the removal efficiencies of most detected antibiotics in real wastewater exceeded 90 % in 2 min, with the ARG removal efficiencies ranging from 6.3 % to 75.2 %. The synergistic effects of plasma and PAA could be associated with the motivated production of reactive species (including •OH, •CH3, 1O2, ONOO-, •O2- and NO•), which decomposed antibiotics, killed host bacteria, and inhibited ARG conjugative transfer. In addition, plasma/PAA also changed the contributions and abundances of ARG host bacteria and downregulated the corresponding genes of two-component regulatory systems, thus reducing ARG propagation. Moreover, the weak correlations between the removal of antibiotics and ARGs highlights the commendable performance of plasma/PAA in the simultaneous removal of antibiotics and ARGs. Therefore, this study affords an innovative and effective avenue to remove antibiotics and ARGs, which relies on the synergistic mechanisms of plasma and PAA and the simultaneous removal mechanisms of antibiotics and ARGs in wastewater.

Surveillance on ESBL-Escherichia coli and Indicator ARG in Wastewater and Reclaimed Water of Four Regions of Spain: Impact of Different Disinfection Treatments

In the present study, the occurrence of indicator antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) both in the influent and the effluent of four Spanish wastewater treatment plants (WWTPs) was monitored for 12 months, and the susceptibility profiles of 89 recovered extended spectrum β-lactamase (ESBL)-producing Escherichia coli isolates were obtained against a wide range of antimicrobials. The aim of the study was to better understand whether the current wastewater treatment practices allow us to obtain safe reclaimed water mitigating the spread of ARB and ARGs to the environment. Results showed high concentrations of ESBL-producing E. coli as well as a high prevalence of a range of ARGs in the influent samples. The reclamation treatments implemented in the WWTPs were effective in reducing both the occurrence of ESBL E. coli and ARGs, although significant differences were observed among WWTPs. Despite these reductions in occurrence observed upon wastewater treatment, our findings suggest that WWTP effluents may represent an important source of ARGs, which could be transferred among environmental bacteria and disseminate antimicrobial resistance through the food chain. Remarkably, no major differences were observed in the susceptibility profiles of the ESBL E. coli isolated from influent and effluent waters, indicating that water treatments do not give rise to the emergence of new resistance phenotypes.

HT-ARGfinder: A Comprehensive Pipeline for Identifying Horizontally Transferred Antibiotic Resistance Genes and Directionality in Metagenomic Sequencing Data

Antibiotic resistance is a continually rising threat to global health. A primary driver of the evolution of new strains of resistant pathogens is the horizontal gene transfer (HGT) of antibiotic resistance genes (ARGs). However, identifying and quantifying ARGs subject to HGT remains a significant challenge. Here, we introduce HT-ARGfinder (horizontally transferred ARG finder), a pipeline that detects and enumerates horizontally transferred ARGs in metagenomic data while also estimating the directionality of transfer. To demonstrate the pipeline, we applied it to an array of publicly-available wastewater metagenomes, including hospital sewage. We compare the horizontally transferred ARGs detected across various sample types and estimate their directionality of transfer among donors and recipients. This study introduces a comprehensive tool to track mobile ARGs in wastewater and other aquatic environments.

Metagenomic Analysis Reveals Changes in Bacterial Communities and Antibiotic Resistance Genes in an Eye Specialty Hospital and a General Hospital Before and After Wastewater Treatment.

The spread of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in hospital wastewater poses a great threat to public health, and wastewater treatment plants (WWTPs) play an important role in reducing the levels of ARB and ARGs. In this study, high-throughput metagenomic sequencing was used to analyze the bacterial community composition and ARGs in two hospitals exposed to different antibiotic use conditions (an eye specialty hospital and a general hospital) before and after wastewater treatment. The results showed that there were various potential pathogenic bacteria in the hospital wastewater, and the abundance and diversity of the influent ARGs in the general hospital were higher than those in the eye hospital. The influent of the eye hospital was mainly composed of Thauera and Pseudomonas, and sul1 (sulfonamide) was the most abundant ARG. The influent of the general hospital contained mainly Aeromonas and Acinetobacter, and tet39 (tetracycline) was the most abundant ARG. Furthermore, co-occurrence network analysis showed that the main bacteria carrying ARGs in hospital wastewater varied with hospital type; the same bacteria in wastewater from different hospitals could carry different ARGs, and the same ARG could also be carried by different bacteria. The changes in the bacterial community and ARG abundance in the effluent from the two hospitals showed that the activated sludge treatment and the direct chlorination disinfection can effectively remove some bacteria and ARGs in wastewater but have limitations. The species diversity increased significantly after the activated sludge treatment, while the direct chlorination disinfection did not increase the diversity. The activated sludge treatment has a better effect on the elimination of ARGs than the direct chlorination disinfection. In summary, we investigated the differences in bacterial communities and ARGs in wastewater from two hospitals exposed to different antibiotic usage conditions, evaluated the effects of different wastewater treatment methods on the bacterial communities and ARGs in hospital wastewater, and recommended appropriate methods for certain clinical environments.

Removal of Sulfonamide Resistance Genes in Fishery Reclamation Mining Subsidence Area by Zeolite.

A majority of subsidence lakes were reclaimed as fish ponds, but the widespread use of antibiotics has caused the pollution of antibiotic resistance genes (ARGs). This paper uses zeolite as a filter material to construct a horizontal submersible wastewater treatment device and explores its effect on the removal of conventional pollutants and sulfonamide ARGs in wastewater. The results showed that the removal of total nitrogen and ammonia nitrogen by the zeolite filter media were 59.0% and 63.8%, respectively, which were higher than the removal of total phosphorus and COD. The absolute abundances of sul1 and sul2 in wastewater were 2.81 × 104 copies·L−1 and 2.42 × 103 copies·L−1. On average, 60.62% of sul1 and 75.84% of sul2 can be removed, and more than 90% of sul1 and sul2 can be removed. Experiments showed that the residence time of wastewater in the treatment device had a significant impact on removal. The microbial community structure of aquaculture wastewater was quite different before and after wastewater treatment. The abundance changes of Saccharimonadales and Mycobacterium affect the removal of sulfonamide ARGs.

The prevalence and removal of antibiotic resistance genes in full-scale wastewater treatment plants: Bacterial host, influencing factors and correlation with nitrogen metabolic pathway

This study investigated the prevalence of antibiotic resistance genes (ARGs) in the influent, effluent, and waste activated sludge (WAS) of eight full-scale municipal wastewater treatment plants (WWTPs) in Shanghai, China. A comprehensive understanding of the correlation between various influencing factors (characteristics of wastewater and WAS, antibiotics, metals, mobile genetic elements) and ARGs was explored. Among the eight full-scale WWTPs, the Unitank process was inefficient in removing typical ARGs compared with continuous-flow anaerobic-anoxic-aerobic and oxidation ditch processes. Antibiotic was identified as the most influential factor affecting the occurrence of ARGs in wastewater, followed by flow rate and nutrients. Positive correlations were observed between antibiotics and their corresponding ARGs in the influent, while this correlation disappeared in the WAS. Class I integron, wastewater characteristics (nitrogen and flow rate), antibiotics (ofloxacin, sulfamethazine, and erythromycin), metals (Mg, Al, Fe, and Mn) were identified as crucial factors comprehensively affecting the distribution of ARGs in WAS. Dissimilatory nitrate reduction profoundly influenced the fate of ARGs during wastewater treatment processes, and K04561 (norB), K02567 (napA), K00262 (gdhA), K00284 (gltS) were identified as the most significant genes in the nitrogen metabolism pathway (ko00910). This study provides a new perspective for comprehensively understanding the occurrence and dissemination of ARGs in WWTPs.

Circulation of Antibiotic Resistance Genes in Wastewater in Mediterranean Basin Countries: A Review

Circulation of Antibiotic Resistance Genes in Wastewater in Mediterranean Basin Countries: A Review

Insight 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 blaTEM 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.

Antibiotic resistance genes of emerging concern in municipal and hospital wastewater from a major Swedish city

The spread of antibiotic resistance among bacterial pathogens is to a large extent mediated by mobile antibiotic resistance genes (ARGs). The prevalence and geographic distribution of several newly discovered ARGs, as well as some clinically important ARGs conferring resistance to last resort antibiotics, are largely unknown. Targeted analysis of wastewater samples could allow estimations of carriage in the population connected to the sewers as well as release to the environment. Here we quantified ARGs conferring resistance to linezolid (optrA and cfr(A)) and colistin (mcr-1, -2, -3, -4 and -5) and the recently discovered gar (aminoglycoside ARG) and sul4 (sulphonamide ARG) in raw hospital and municipal wastewater as well as treated municipal wastewater during five years in a low antibiotic resistance prevalence setting (Gothenburg, Sweden). Additionally, variations in bacterial composition of the wastewaters characterized by 16S rRNA sequencing were related to the variations of the ARGs in an attempt to reveal if the presence of known or suspected bacterial host taxa could explain the presence of the ARGs in wastewater. The mcr-1, mcr-3, mcr-4, mcr-5, sul4 and gar genes were detected regularly in all types of wastewater samples while optrA and cfr(A) were detected only in hospital wastewater. The most abundant genes were mcr-3 and mcr-5, especially in municipal wastewater. The detection of optrA was restricted to a peak during one year. Most of the ARGs correlated with taxa previously described as bacterial hosts and associated with humans. Although some of the tentative hosts may include bacteria also thriving in wastewater environments, detection of the ARGs in the wastewaters could reflect their presence in the gut flora of the contributing populations. If so, they could already today or in the near future hinder treatment of bacterial infections in a setting where they currently are rarely targeted/detected during clinical surveillance.