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854500.9886Role of anaerobic sludge digestion in handling antibiotic resistant bacteria and antibiotic resistance genes - A review. Currently, anaerobic sludge digestion (ASD) is considered not only for treating residual sewage sludge and energy recovery but also for the reduction of antibiotic resistance genes (ARGs). The current review highlights the reasons why antibiotic resistant bacteria (ARB) and ARGs exist in ASD and how ASD performs in the reduction of ARB and ARGs. ARGs and ARB have been detected in ASD with some reports indicating some of the ARGs can be completely removed during the ASD process, while other studies reported the enrichment of ARB and ARGs after ASD. This paper reviews the performance of ASD based on operational parameters as well as environmental chemistry. More studies are needed to improve the performance of ASD in reducing ARGs that are difficult to handle and also differentiate between extracellular (eARGs) and intracellular ARGs (iARGs) to achieve more accurate quantification of the ARGs.202133735726
793510.9882Removal of antibiotic resistance genes by Cl(2)-UV process: Direct UV damage outweighs free radicals in effectiveness. Antibiotic resistance genes (ARGs) pose significant environmental health problems and have become a major global concern. This study investigated the efficacy and mechanism of the Cl(2)-UV process (chlorine followed by UV irradiation) for removing ARGs in various forms. The Cl(2)-UV process caused irreversible damage to nearly all ARB at typical disinfectant dosages. In solutions containing only extracellular ARGs (eARGs), the Cl₂-UV process achieved over 99.0 % degradation of eARGs. When both eARGs and intracellular ARGs (iARGs) were present, the process reached a 97.2 % removal rate for iARGs. While the abundance of eARGs initially increased due to the release of iARGs from lysed cells during pre-chlorination, subsequent UV irradiation rapidly degraded the released eARGs, restoring their abundance to near-initial levels by the end of the Cl₂-UV process. Analysis of the roles in degrading eARGs and iARGs during the Cl(2)-UV process revealed that UV, rather than free radicals, was the dominant factor causing ARG damage. Pre-chlorination enhanced direct UV damage to eARGs and iARGs by altering plasmid conformation and promoting efficient damage to high UV-absorbing cellular components. Furthermore, no further natural transformation of residual ARGs occurred following the Cl(2)-UV treatment. This study demonstrated strong evidence for the effectiveness of the Cl(2)-UV process in controlling antibiotic resistance.202540048777
855220.9881Sustainable material platforms for multi-log removal of antibiotic-resistant bacteria and genes from wastewater: A review. Antibiotic-resistant bacteria (ARB) and the associated resistance genes (ARGs) are now recognized as emerging contaminants that can disseminate via wastewater streams, posing significant risks to both human and ecosystem health. Conventional physicochemical treatment approaches (e.g., chlorination, ozonation, advanced oxidation processes) typically suppress these contaminants but may also result in the formation of hazardous by-products. This critical review comprehensibly evaluates bio-based and other sustainable materials designed for the removal of ARB and ARGs from aqueous environments. The materials are systematically categorized into (i) biopolymers and their composites (chitosan, alginate, cellulose), (ii) carbon-rich adsorbents and (photo-)catalysts (biochar, activated carbon, graphene), (iii) metal- and semiconductor-based nanomaterials, and (iv) nature-based treatment solutions (constructed wetlands, soil-aquifer treatment, clay sorbents). Observed log-reduction value range from 2 to 7 for ARB with platforms such as zinc oxide/activated-carbon alginate beads, Fe/N-doped biochars, and graphene-supramolecular-porphyrin hybrids demonstrating high multifunctional efficacy. Mechanistic studies reveal that removal involves synergistic adsorption, photodynamic or Fenton-like oxidation, cell-membrane disruption, and inhibition of horizontal gene transfer. This review emphasizes the advancing potential of sustainable material solutions for mitigating antibiotic resistance and highlights the urgent need to develop scalable, environmentally sustainable treatment methods for protecting water resources and public health.202540763861
793430.9878Mitigated membrane fouling and enhanced removal of extracellular antibiotic resistance genes from wastewater effluent via an integrated pre-coagulation and microfiltration process. Antibiotic resistance genes (ARGs) have been regarded as an emerging pollutant in municipal wastewater treatment plant (WWTP) effluents due to their potential risk to human health and ecological safety when reused for landscape and irrigation. Conventional wastewater treatment processes generally fail to effectively reduce ARGs, especially extracellular ARGs (eARGs), which are persistent in the environment and play an important role in horizontal gene transfer via transformation. Herein, an integrated process of pre-coagulation and microfiltration was developed for removal of ARGs, especially eARGs, from wastewater effluent. Results show that the integrated process could effectively reduce the absolute abundances of total ARGs (tARGs) (>2.9 logs) and eARGs (>5.2 logs) from the effluent. The excellent performance could be mainly attributed to the capture of antibiotic resistant bacteria (ARB) and eARGs by pre-coagulation and co-rejection during subsequent microfiltration. Moreover, the integrated process exhibited a good performance on removing common pollutants (e.g., dissolved organic carbon and phosphate) from the effluent to improve water quality. Besides, the integrated process also greatly reduced membrane fouling compared with microfiltration. These findings suggest that the integrated process of pre-coagulation and microfiltration is a promising advanced wastewater treatment technology for ARGs (especially eARGs) removal from WWTP effluents to ensure water reuse security.201931085389
854640.9878A review of emerging organic contaminants (EOCs), antibiotic resistant bacteria (ARB), and antibiotic resistance genes (ARGs) in the environment: Increasing removal with wetlands and reducing environmental impacts. Emerging organic contaminants (EOCs) include a diverse group of chemical compounds, such as pharmaceuticals and personal care products (PPCPs), pesticides, hormones, surfactants, flame retardants and plasticizers. Many of these compounds are not significantly removed in conventional wastewater treatment plants and are discharged to the environment, presenting an increasing threat to both humans and natural ecosystems. Recently, antibiotics have received considerable attention due to growing microbial antibiotic-resistance in the environment. Constructed wetlands (CWs) have proven effective in removing many EOCs, including different antibiotics, before discharge of treated wastewater into the environment. Wastewater treatment systems that couple conventional treatment plants with constructed and natural wetlands offer a strategy to remove EOCs and reduce antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) far more efficiently than conventional treatment alone. This review presents as overview of the current knowledge on the efficiency of different wetland systems in reducing EOCs and antibiotic resistance.202032247686
855350.9877Unveiling the power of nanotechnology: a novel approach to eliminating antibiotic-resistant bacteria and genes from municipal effluent. The increasing global population and declining freshwater resources have heightened the urgency of ensuring safe and accessible water supplies.Query The persistence of antibiotic-resistant bacteria (ARB) and antibiotic-resistant genes (ARGs) in municipal effluents poses a significant public health threat, exacerbated by the widespread use of antibiotics and the inadequate removal of contaminants in wastewater treatment facilities. Conventional treatment methods often fail to eliminate these emerging pollutants, facilitating their entry into agricultural systems and natural water bodies, thereby accelerating the spread of antimicrobial resistance. To address these challenges, interdisciplinary strategies in water treatment are essential. Nanotechnology has emerged as a promising approach due to its unique physicochemical properties, biocompatibility, and high efficiency in detecting and removing biological and chemical contaminants. Various nanomaterials, including graphene-based structures, Carbon nanotubes (CNTs), noble metal nanoparticles (gold (Au) and silver (Ag)), silicon and chitosan-based nanomaterials, as well as titanium and Zinc oxide (ZnO) nanomaterials, demonstrate potent antimicrobial effects. Moreover, nanosensors and photocatalysts utilizing these nanomaterials enable precise detection and effective degradation of ARB and ARGs in wastewater. This review examines the mechanisms by which nanotechnology-based materials can mitigate the risks associated with antibiotic resistance in urban effluents, focusing on their applications in pathogen detection, pollutant removal, and wastewater treatment. By integrating nanotechnology into existing treatment frameworks, we can significantly enhance the efficiency of water purification processes, ultimately contributing to global water security and the protection of public health.202540512401
854960.9877Current perspectives on microalgae and extracellular polymers for reducing antibiotic resistance genes in livestock wastewater. Antibiotic resistance genes (ARGs) in livestock wastewater resulting from excessive antibiotics used in animal farming pose significant environmental and public health risks. Conventional treatment methods are often costly, inefficient, and may inadvertently promote ARG transmission. Microalgae, with their long genetic distance from bacteria and strong ability to utilize wastewater nutrients, offer a sustainable solution for ARG mitigation. This review studied the abundance and characterization of ARGs in livestock wastewater, highlighted microalgal-based removal mechanisms of ARGs, including phagocytosis, competition, and absorption by extracellular polymeric substances (EPS), and explored factors influencing their efficacy. Notably, the microalgae-EPS system reduced ARGs by 0.62-3.00 log, demonstrating significant potential in wastewater treatment. Key challenges, such as optimizing algal species, understanding EPS-ARG interactions, targeted reduction of host bacteria, and scaling technologies, were discussed. This work provides critical insights for advancing microalgal-based strategies for ARG removal, promoting environmentally friendly and efficient wastewater management.202540324729
785470.9874Removal of antibiotic resistant bacteria and plasmid-encoded antibiotic resistance genes in water by ozonation and electro-peroxone process. The electro-peroxone (EP) process is an electricity-based oxidation process enabled by electrochemically generating hydrogen peroxide (H(2)O(2)) from cathodic oxygen (O(2)) reduction during ozonation. In this study, the removal of antibiotic resistant bacteria (ARB) and plasmid-encoded antibiotic resistance genes (ARGs) during groundwater treatment by ozonation alone and the EP process was compared. Owing to the H(2)O(2)-promoted ozone (O(3)) conversion to hydroxyl radicals (•OH), higher •OH exposures, but lower O(3) exposures were obtained during the EP process than ozonation alone. This opposite change of O(3) and •OH exposures decreases the efficiency of ARB inactivation and ARG degradation moderately during the EP process compared with ozonation alone. These results suggest that regarding ARB inactivation and ARG degradation, the reduction of O(3) exposures may not be fully counterbalanced by the rise of •OH exposures when changing ozonation to the EP process. However, due to the rise of •OH exposure, plasmid DNA was more effectively cleaved to shorter fragments during the EP process than ozonation alone, which may decrease the risks of natural transformation of ARGs. These findings highlight that the influence of the EP process on ARB and ARG inactivation needs to be considered when implementing this process in water treatment.202336738938
639580.9874Risk control of antibiotics, antibiotic resistance genes (ARGs) and antibiotic resistant bacteria (ARB) during sewage sludge treatment and disposal: A review. Sewage sludge is an important reservoir of antibiotics, antibiotic resistance genes (ARGs), and antibiotic resistant bacteria (ARB) in wastewater treatment plants (WWTPs), and the reclamation of sewage sludge potentially threats human health and environmental safety. Sludge treatment and disposal are expected to control these risks, and this review summarizes the fate and controlling efficiency of antibiotics, ARGs, and ARB in sludge involved in different processes, i.e., disintegration, anaerobic digestion, aerobic composting, drying, pyrolysis, constructed wetland, and land application. Additionally, the analysis and characterization methods of antibiotics, ARGs, and ARB in complicate sludge are reviewed, and the quantitative risk assessment approaches involved in land application are comprehensively discussed. This review benefits process optimization of sludge treatment and disposal, with regard to environmental risks control of antibiotics, ARGs, and ARB in sludge. Furthermore, current research limitations and gaps, e.g., the antibiotic resistance risk assessment in sludge-amended soil, are proposed to advance the future studies.202336933744
784890.9873Simultaneous Removal of Antibiotic Resistant Bacteria, Antibiotic Resistance Genes, and Micropollutants by FeS(2)@GO-Based Heterogeneous Photo-Fenton Process. The co-occurrence of various chemical and biological contaminants of emerging concerns has hindered the application of water recycling. This study aims to develop a heterogeneous photo-Fenton treatment by fabricating nano pyrite (FeS(2)) on graphene oxide (FeS(2)@GO) to simultaneously remove antibiotic resistant bacteria (ARB), antibiotic resistance genes (ARGs), and micropollutants (MPs). A facile and solvothermal process was used to synthesize new pyrite-based composites. The GO coated layer forms a strong chemical bond with nano pyrite, which enables to prevent the oxidation and photocorrosion of pyrite and promote the transfer of charge carriers. Low reagent doses of FeS(2)@GO catalyst (0.25 mg/L) and H(2)O(2) (1.0 mM) were found to be efficient for removing 6-log of ARB and 7-log of extracellular ARG (e-ARG) after 30 and 7.5 min treatment, respectively, in synthetic wastewater. Bacterial regrowth was not observed even after a two-day incubation. Moreover, four recalcitrant MPs (sulfamethoxazole, carbamazepine, diclofenac, and mecoprop at an environmentally relevant concentration of 10 μg/L each) were completely removed after 10 min of treatment. The stable and recyclable composite generated more reactive species, including hydroxyl radicals (HO(•)), superoxide radicals (O(2)(• -)), singlet oxygen ((1)O(2)). These findings highlight that the synthesized FeS(2)@GO catalyst is a promising heterogeneous photo-Fenton catalyst for the removal of emerging contaminants.202235759741
7932100.9873How multi-walled carbon nanotubes in wastewater influence the fate of coexisting antibiotic resistant genes in the subsequent disinfection process. Wastewater treatment plants (WWTPs) are important hubs for the spread of antibiotic resistance genes (ARGs). Engineered nanoparticles, which was inevitably released to WWTPs, could change environmentally sensitive of antibiotic resistant bacteria (ARB). This would influence the fate of ARGs in subsequent disinfection process and consequent health risk. In this study, the ARGs fate of the effluent in conventional sodium hypochlorite (NaClO) disinfection process was investigated as multi-walled carbon nanotubes (MWCNTs) existed in sequencing batch reactor (SBR). The results showed the existence of MWCNTs in SBR could enhance the removal efficiency of intracellular 16S rRNA gene and intI1, extracellular intI1, sul2 and tetX in the effluent by NaClO. This is mainly due to the variation of bacterial physiological status, bacterial population structure and the activation of NaClO under the role of MWCNTs. MWCNTs in SBR could increase in membrane permeability of bacterial cells, which would be conducive to the penetration of chlorination to cytoplasm. MWCNTs in SBR also could change the bacterial population structure and induce the chlorine-sensitive bacteria; thus the potential hosts of ARGs in the effluent would be more easily inactivated by NaClO. Moreover, the residual MWCNTs in the effluent could activate NaClO to generate various free radical, which would enhance the oxidizing capacity of chlorination.202235500623
7853110.9872Natural pyrite and ascorbic acid co-enhance periodate activation for inactivation of antibiotic resistant bacteria and inhibition of resistance genes transmission: A green disinfection process dominated by singlet oxygen. The transmission of antibiotic resistance genes (ARGs) and the propagation of antibiotic resistant bacteria (ARB) threaten public health security and human health, and greener and more efficient disinfection technologies are expected to be discovered for wastewater treatment. In this study, natural pyrite and ascorbic acid (AA) were proposed as environmental-friendly activator and reductant for periodate (PI) activation to inactivate ARB. The disinfection treatment of PI/pyrite/AA system could inactivate 5.62 log ARB within 30 min, and the lower pH and higher PI and natural pyrite dosage could further boost the disinfection efficiency. The (1)O(2) and SO(4)(•-) were demonstrated to be crucial for the inactivation of ARB in PI/pyrite/AA system. The disinfection process destroyed the morphological structure of ARB, inducing oxidative stress and stimulating the antioxidant system. The PI/pyrite/AA system effectively reduced the intracellular and extracellular DNA concentration and ARGs abundance, inhibiting the propagation of ARGs. The presence of AA facilitated the activation of PI with natural pyrite and significantly increased the concentration of Fe(2+) in solution. The reusability of natural pyrite, the safety of the disinfection by-products and the inhibition of ARB regeneration indicated the application potential of PI/pyrite/AA system in wastewater disinfection.202439038380
8580120.9872Mitigation of microplastic-associated emerging pollutants by chlorination using field-collected microplastic: Antimicrobial-resistant genes and pathogens. The ubiquity of microplastics (MPs) in aquatic environments has raised significant concerns regarding their roles as vectors for antibiotic-resistance genes (ARGs) and antibiotic-resistant pathogens (ARPs). This study investigated the mitigation of ARGs and ARPs associated with field-collected MPs through chlorination using free available chlorine (FAC) at varying concentrations. FAC effectively reduced the absolute abundance of ARGs on MPs by up to 99.69 %, although the relative abundance of certain ARGs persisted or increased after treatments. Results revealed that the three-dimensional structure of biofilms on MPs significantly influenced FAC efficacy, with interior biofilm bacteria demonstrating greater resistance than outer biofilm. Additionally, FAC induced fragmentation of MPs, particularly increasing the proportion of particles smaller than 100 μm. Notably, ARGs such as sul1 and ermB showed substantial reductions in absolute abundance, whereas ermC and sul2 exhibited less reduction, highlighting the complexity of disinfection in MP-associated biofilms. These findings underscore the need for optimizing disinfection strategies to mitigate ARG dissemination and address environmental risks posed by MPs in wastewater effluents.202540436100
6502130.9872A critical review on the occurrence of resistomes in the environment and their removal from wastewater using apposite treatment technologies: Limitations, successes and future improvement. Recent reports are pointing towards the potential increasing risks of resistomes in human host. With no permissible limit in sight, resistomes are continually multiplying at an alarming rate in the ecosystem, with a disturbing level in drinking water source. The morphology and chemical constituent of resistomes afford them to resist degradation, elude membrane and counter ionic charge, thereby, rendering both conventional and advanced water and wastewater treatment inefficient. Water and wastewater matrix may govern the propagation of individual resistomes sub-type, co-selection and specific interaction towards precise condition may have enhanced the current challenge. This review covers recent reports (2011-2019) on the occurrence of ARB/ARGs and ease of spread of resistance genes in the aquatic ecosystem. The contributions of water matrix to the spread and mitigation, treatment options, via bulk removal or capture, and intracellular and extracellular DNA lysis were discussed. A complete summary of recent occurrences of ARB/ARGs, fate after disinfection and optimum conditions of individual treatment technology or in tandem, including process limitations, with a brief assessment of removal or degradation mechanism were highlighted.202032224385
8554140.9872Nanomaterial-Enhanced Hybrid Disinfection: A Solution to Combat Multidrug-Resistant Bacteria and Antibiotic Resistance Genes in Wastewater. This review explores the potential of nanomaterial-enhanced hybrid disinfection methods as effective strategies for addressing the growing challenge of multidrug-resistant (MDR) bacteria and antibiotic resistance genes (ARGs) in wastewater treatment. By integrating hybrid nanocomposites and nanomaterials, natural biocides such as terpenes, and ultrasonication, this approach significantly enhances disinfection efficiency compared to conventional methods. The review highlights the mechanisms through which hybrid nanocomposites and nanomaterials generate reactive oxygen species (ROS) under blue LED irradiation, effectively disrupting MDR bacteria while improving the efficacy of natural biocides through synergistic interactions. Additionally, the review examines critical operational parameters-such as light intensity, catalyst dosage, and ultrasonication power-that optimize treatment outcomes and ensure the reusability of hybrid nanocomposites and other nanomaterials without significant loss of photocatalytic activity. Furthermore, this hybrid method shows promise in degrading ARGs, thereby addressing both microbial and genetic pollution. Overall, this review underscores the need for innovative wastewater treatment solutions that are efficient, sustainable, and scalable, contributing to the global fight against antimicrobial resistance.202439591087
8551150.9871Promising bioprocesses for the efficient removal of antibiotics and antibiotic-resistance genes from urban and hospital wastewaters: Potentialities of aerobic granular systems. The use, overuse, and improper use of antibiotics have resulted in higher levels of antibiotic-resistant bacteria (ARB) and antibiotic-resistance genes (ARGs), which have profoundly disturbed the equilibrium of the environment. Furthermore, once antibiotic agents are excreted in urine and feces, these substances often can reach wastewater treatment plants (WWTPs), in which improper treatments have been highlighted as the main reason for stronger dissemination of antibiotics, ARB, and ARGs to the receiving bodies. Hence, achieving better antibiotic removal capacities in WWTPs is proposed as an adequate approach to limit the spread of antibiotics, ARB, and ARGs into the environment. In this review, we highlight hospital wastewater (WW) as a critical hotspot for the dissemination of antibiotic resistance due to its high level of antibiotics and pathogens. Hence, monitoring the composition and structure of the bacterial communities related to hospital WW is a key factor in controlling the spread of ARGs. In addition, we discuss the advantages and drawbacks of the current biological WW treatments regarding the antibiotic-resistance phenomenon. Widely used conventional activated sludge technology has proved to be ineffective in mitigating the dissemination of ARB and ARGs to the environment. However, aerobic granular sludge (AGS) technology is a promising technology-with broad adaptability and excellent performance-that could successfully reduce antibiotics, ARB, and ARGs in the generated effluents. We also outline the main operational parameters involved in mitigating antibiotics, ARB, and ARGs in WWTPs. In this regard, WW operation under long hydraulic and solid retention times allows better removal of antibiotics, ARB, and ARGs independently of the WW technology employed. Finally, we address the current knowledge of the adsorption and degradation of antibiotics and their importance in removing ARB and ARGs. Notably, AGS can enhance the removal of antibiotics, ARB, and ARGs due to the complex microbial metabolism within the granular biomass.202438086508
7621160.9871Pre-chlorination in source water endows ARB with resistance to chlorine disinfection in drinking water treatment. Chlorine disinfection is widely used to ensure biosafety of drinking water. However, antibiotic resistance bacteria (ARB) and antibiotic resistance genes (ARGs) are often detected in treated drinking water. The impact of chlorine disinfection on the abundance of ARGs in drinking water is currently contradictory. Some studies suggested that chlorine disinfection could reduce the abundance of ARGs, while others had found that chlorine disinfection increased the abundance of ARGs. Pre-chlorination is widely applied in raw water to kill the algae cells in source water Pump Station. Different distances between the source water Pump Station and the drinking water treatment plants (DWTPs) resulted in different degrees of residual chlorine decay in the incoming raw water. This study found that the abundance of ARGs in drinking water would be increased during chlorine disinfection when the chlorine concentration in raw water was higher (> 0.2 mg/L). On the contrary, chlorine disinfection would decrease the abundance of ARGs in drinking water when the chlorine concentration in raw water was lower (< 0.09 mg/L). Pre-chlorination in source water with sub-lethal concentration could allow ARB to adapt to the chlorine environment in advance, endowing ARB with chlorine resistance, which resulted in ineffective removal of ARB and increased ARGs abundance during subsequent chlorine disinfection. High abundance of chlorine and antibiotics co-resistance bacteria in raw water was the main reason for the increase in ARGs abundance in chlorine treated drinking water. It should be noticed that, pre-chlorination treatment in source water would increase the difficulty of removing ARGs in subsequent chlorine disinfection process.202540398032
6407170.9871Urban wastewater treatment plants as hotspots for antibiotic resistant bacteria and genes spread into the environment: a review. Urban wastewater treatment plants (UWTPs) are among the main sources of antibiotics' release into the environment. The occurrence of antibiotics may promote the selection of antibiotic resistance genes (ARGs) and antibiotic resistant bacteria (ARB), which shade health risks to humans and animals. In this paper the fate of ARB and ARGs in UWTPs, focusing on different processes/technologies (i.e., biological processes, advanced treatment technologies and disinfection), was critically reviewed. The mechanisms by which biological processes influence the development/selection of ARB and ARGs transfer are still poorly understood. Advanced treatment technologies and disinfection process are regarded as a major tool to control the spread of ARB into the environment. In spite of intense efforts made over the last years to bring solutions to control antibiotic resistance spread in the environment, there are still important gaps to fill in. In particular, it is important to: (i) improve risk assessment studies in order to allow accurate estimates about the maximal abundance of ARB in UWTPs effluents that would not pose risks for human and environmental health; (ii) understand the factors and mechanisms that drive antibiotic resistance maintenance and selection in wastewater habitats. The final objective is to implement wastewater treatment technologies capable of assuring the production of UWTPs effluents with an acceptable level of ARB.201323396083
6417180.9870Fate of environmental pollutants: A review. A review of the literature published in 2019 on topics associated with the fate of environmental pollutants is presented. Environmental pollutants covered include pharmaceuticals, antibiotic-resistant bacteria and genes, pesticides and veterinary medicines, personal care products and emerging pollutants, PFAS, microplastics, nanomaterials, heavy metals and radionuclides, nutrients, pathogens and indicator organisms, and oil and hydrocarbons. For each pollutant, the occurrence in the environment and/or their fate in engineered as well as natural systems in matrices including water, soil, wastewater, stormwater, runoff, and/or manure is presented based on the published literature. The review includes current developments in understanding pollutants in natural and engineered systems, and relevant physico-chemical processes, as well as biological processes.202032671926
8123190.9870The effect of bulk-biochar and nano-biochar amendment on the removal of antibiotic resistance genes in microplastic contaminated soil. Biochar amendment has significant benefits in removing antibiotic resistance genes (ARGs) in the soil. Nevertheless, there is little information on ARGs removal in microplastic contaminated soil. Herein, a 42-day soil microcosm experiment were carried out to study how two coconut shell biochars (bulk- and nano-size) eliminate soil ARGs with/without microplastic presence. The results showed that microplastic increased significantly the numbers and abundances of ARGs in soil at 14d of cultivation. And, two biochars amendment effectively inhibited soil ARGs spread whether or not microplastic was present, especially for nano-biochar which had more effective removal compared to bulk-biochar. However, microplastic weakened soil ARGs removal after applying same biochar. Two biochars removed ARGs through decreasing horizontal gene transfer (HGT) of ARGs, potential host-bacteria abundances, some bacteria crowding the eco-niche of hosts and promoting soil properties. The adverse effect of microplastic on ARGs removal was mainly caused by weakening mobile genetic elements (MGEs) removal, and by changing soil properties. Structural equation modeling (SEM) analysis indicated that biochar's effect on ARGs profile was changed by its size and microplastic presence through altering MGEs abundances. These results highlight that biochar amendment is still an effective method for ARGs removal in microplastic contaminated soil.202437907163