Antibiotic Resistance Genes Attenuation Research Articles

Effect of wheat crops on the persistence and attenuation of antibiotic resistance genes in soil after swine wastewater application

Swine wastewater (SW) application introduces antibiotic resistance genes (ARGs) into farmland soils. However, ARG attenuation in SW-fertigated soils, especially those influenced by staple crops and soil type, remains unclear. This study investigated twelve soil ARGs and one mobile genetic element (MGE) in sandy loam, loam, and silt loam soils before and after SW application in wheat-planted and unplanted soils. The results revealed an immediate increase in the abundance of ARGs in soil by two orders of magnitude above background levels following SW application. After SW application, the soil total ARG abundance was attenuated, reaching background levels at 54 days; However, more individual ARGs were detected above the detection limit than pre-application. Among the 13 genes, acc(6′)-lb, tetM, and tetO tended to persist in the soil during wheat harvest. ARG half-lives were up to four times longer in wheat-planted soils than in bare soils. Wheat planting decreased the persistence of acc(6′)-lb, ermB, ermF, and intI2 but increased the persistence of others such as sul1 and sul2. Soil type had no significant impact on ARG and MGE fates. Our findings emphasize the need for strategic SW application and the consideration of crop cultivation effects to mitigate ARG accumulation in farmland soils.

Attenuation effects of ZVI/PDS pretreatment on propagation of antibiotic resistance genes in bioreactors: Driven by antibiotic residues and sulfate assimilation

Sulfate radical-based advanced oxidation processes (AOPs) combined biological system was a promising technology for treating antibiotic wastewater. However, how pretreatment influence antibiotic resistance genes (ARGs) propagation remains largely elusive, especially the produced by-products (antibiotic residues and sulfate) are often ignored. Herein, we investigated the effects of zero valent iron/persulfate pretreatment on ARGs in bioreactors treating sulfadiazine wastewater. Results showed absolute and relative abundance of ARGs reduced by 59.8%− 81.9% and 9.1%− 52.9% after pretreatments. The effect of 90-min pretreatment was better than that of the 30-min. The ARGs reduction was due to decreased antibiotic residues and stimulated sulfate assimilation. Reduced antibiotic residues was a major factor in ARGs attenuation, which could suppress oxidative stress, inhibit mobile genetic elements emergence and resistant strains proliferation. The presence of sulfate in influent supplemented microbial sulfur sources and facilitated the in-situ synthesis of antioxidant cysteine through sulfate assimilation, which drove ARGs attenuation by alleviating oxidative stress. This is the first detailed analysis about the regulatory mechanism of how sulfate radical-based AOPs mediate in ARGs attenuation, which is expected to provide theoretical basis for solving concerns about by-products and developing practical methods to hinder ARGs propagation.

Mitigation of antibiotic resistome in swine manure by black soldier fly larval conversion combined with composting

The increasing prevalence of antibiotic resistance genes (ARGs) in animal manure has attracted considerable attention because of their potential contribution to the development of multidrug resistance worldwide. Insect technology may be a promising alternative for the rapid attenuation of ARGs in manure; however, the underlying mechanism remains unclear. This study aimed to evaluate the effects of black soldier fly (BSF, Hermetia illucens [L.]) larvae conversion combined with composting on ARGs dynamics in swine manure and to uncover the mechanisms through metagenomic analysis. Compared to natural composting (i.e. without BSF), BSFL conversion combined with composting reduced the absolute abundance of ARGs by 93.2 % within 28 days. The rapid degradation of antibiotics and nutrient reformulation during BSFL conversion combined with composting indirectly altered manure bacterial communities, resulting in a lower abundance and richness of ARGs. The number of main antibiotic-resistant bacteria (e.g., Prevotella, Ruminococcus) decreased by 74.9 %, while their potential antagonistic bacteria (e.g., Bacillus, Pseudomonas) increased by 128.7 %. The number of antibiotic-resistant pathogenic bacteria (e.g., Selenomonas, Paenalcaligenes) decreased by 88.3 %, and the average number of ARGs carried by each human pathogenic bacterial genus declined by 55.8 %. BSF larvae gut microbiota (e.g., Clostridium butyricum, C. bornimense) could help reduce the risk of multidrug-resistant pathogens. These results provide insight into a novel approach to mitigate multidrug resistance from the animal industry in the environment by using insect technology combined with composting, in particular in light of the global “One Health” requirements.

Hazard reduction and persistence of risk of antibiotic resistance during thermophilic composting of animal waste

Animal production contributes substantially to the dissemination of antibiotic resistance genes (ARGs) in the environment. Thermophilic composting is cost-effective for reducing hazards in animal wastes. However, ARGs can still persist in composts and the mechanism remains largely unclear. In the present study, metagenomics and real-time quantitative PCR were employed to assess the dynamics of ARGs, especially those of high risk to human health, their host species and pathogenicity of the bacterial community during thermophilic composting of chicken manure. The results showed that thermophilic composting was effective at reducing the majority of the spreading risk of antibiotic resistance by 1) eliminating more than half of allARGs that accounted for 80% of the total abundance, 2) reducing the number and abundance of high-risk ARGs significantly, and 3) removing the ARG hosts and the pathogenicity of the bacterial community substantially. The attenuation of ARGs was primarily driven by the bacterial community succession. However, the abundance of some ARGs, especially those associated with plasmids, rebounded during the cooling phase, likely caused by horizontal gene transfer. Our research reveals that minimising the horizontal transfer of plasmid-associated ARGs during the cooling phase of thermophilic composting is crucial for a more effective reduction of the hazards associated with the spread of antibiotic resistance.

A review of the impact of conductive materials on antibiotic resistance genes during the anaerobic digestion of sewage sludge and animal manure

The urgent need to reduce the environmental burden of antibiotic resistance genes (ARGs) has become even more apparent as concerted efforts are made globally to tackle the dissemination of antimicrobial resistance. Concerning levels of ARGs abound in sewage sludge and animal manure, and their inadequate attenuation during conventional anaerobic digestion (AD) compromises the safety of the digestate, a nutrient-rich by-product of AD commonly recycled to agricultural land for improvement of soil quality. Exogenous ARGs introduced into the natural environment via the land application of digestate can be transferred from innocuous environmental bacteria to clinically relevant bacteria by horizontal gene transfer (HGT) and may eventually reach humans through food, water, and air. This review, therefore, discusses the prospects of using carbon- and iron-based conductive materials (CMs) as additives to mitigate the proliferation of ARGs during the AD of sewage sludge and animal manure. The review spotlights the core mechanisms underpinning the influence of CMs on the resistome profile, the steps to maximize ARG attenuation using CMs, and the current knowledge gaps. Data and information gathered indicate that CMs can profoundly reduce the abundance of ARGs in the digestate by easing selective pressure on ARGs, altering microbial community structure, and diminishing HGT.

Alteration of Manure Antibiotic Resistance Genes via Soil Fauna Is Associated with the Intestinal Microbiome.

ABSTRACTLivestock wastes contain high levels of antibiotic resistance genes (ARGs) and a variety of human-related pathogens. Bioconversion of livestock manure using larvae of the beetle Protaetia brevitarsis is an effective technique for waste reduction and value creation; however, the fate of manure ARGs during gut passage and interaction with the gut microbiome of P. brevitarsis remains unclear. To investigate this, we fed P. brevitarsis with dry chicken manure for 6 days and measured bacterial community dynamics and ARG abundance and diversity along the P. brevitarsis gut tract using high-throughput quantitative PCR and metagenomics approaches. The diversity of ARGs was significantly lower in larval midgut, hindgut, and frass than in raw chicken manure, and around 80% of pathogenicity-related genes (PRGs) exhibited reduced abundance. Network analysis demonstrated that Bacteroidetes and Firmicutes were the key bacterial phyla associated with ARG reduction. Metagenomic analysis further indicated that ARGs, mobile genetic elements (MGEs), and PRGs were simultaneously attenuated in the hindgut, implicating a decreased likelihood for horizontal gene transfer (HGT) of ARGs among bacteria and pathogens during manure bioconversion. Our findings demonstrated that the attenuation of ARGs is strongly associated with the variation of the gut microbiome of P. brevitarsis, providing insights into mechanisms of risk mitigation of ARG dissemination during manure bioconversion.IMPORTANCE Saprophagous fauna like the oriental edible beetle (P. brevitarsis) plays a fundamental role in converting organic wastes into biofertilizer. Accumulating evidence has shown that soil fauna can reduce the abundance of ARGs, although the underlying mechanism of ARG reduction is still unclear. In our previous research, we found a large reduction of ARGs in vegetable roots and leaves from frass compared with raw manure, providing a promising biofertilizer for soil-vegetable systems. Therefore, in this study, temporal dynamic changes in the microbiomes of the donor (chicken manure) and host (P. brevitarsis) were investigated, and we found a close association between the gut microbiome and the alteration of ARGs. These results shed new light on how the insect gut microbiome can mitigate manure-borne ARGs and provide insights into the bioconversion process via a typical member of the saprophagous fauna, P. brevitarsis.

Microalgae simultaneously promote antibiotic removal and antibiotic resistance genes/bacteria attenuation in algal-bacterial granular sludge system

This study investigated the effects of microalgae growth on antibiotic removal and the attenuation of antibiotic resistance genes (ARGs)/ARGs host bacteria in algal-bacterial granular sludge (ABGS) system. In the presence of tetracycline (TC) and sulfadiazine (SDZ) mixture (2–4 mg/L), microalgae could grow on bacterial granular sludge (BGS) to form ABGS, with a chlorophyll-a content of 7.68–8.13 mg/g-VSS being achieved. The removal efficiencies of TC and SDZ by ABGS were as high as 79.0 % and 94.0 %, which were 4.3–5.0 % higher than those by BGS. Metagenomic analysis indicated that the relative abundances of TC/SDZ- related ARGs and mobile genetic elements (MGEs) in BGS were 56.1 % and 22.1 % higher than those in ABGS. A total of 26 ARGs were detected from the granules, and they were identified to associate with 46 host bacteria. 13 out of 26 ARGs and 13 out of 46 hosts were shared ARGs and hosts, respectively. The total relative abundance of host bacteria in BGS was 30.8 % higher than that in ABGS. Scenedesmus and Chlorella were the dominant microalgae that may reduce the diversity of ARGs hosts. Overall, ABGS is a promising biotechnology for antibiotic-containing wastewater treatment.

Characteristics of antibiotic resistance genes in full-scale anaerobic digesters of food waste and the effects of application of biogas slurry on soil antibiotic resistance genes.

The fate of antibiotic resistance genes (ARGs) in full-scale anaerobic digestion (AD) of food waste (FW) and in the soil applied with biogas slurry has not been fully understood. In this study, 12 targeted ARGs and intI1 in FW, intermediate product, and biogas slurry from three full-scale ADwere analyzed. The results showed that subcritical water pretreatment was aneffective method for ARG attenuation, by which the absolute abundance of total targeted ARGs was removed by 99.69%. The predominant ARGs (ermB, tetM, and tetW) in FW were removed more than 99% after subcritical water pretreatment. The result of field experiments with biogas slurry as fertilizer showed that the absolute abundance of several ARGs (sul2, tetM, blaOXA-1, blaTEM) and intI1 accumulated significantly compared to the control group (CK) during three consecutive growth stages of the rice. The detected abundance of ARGs in paddy field soil increased from 190.50 (CK) to 8.87 × 104 copies/g (wet weight) (soil) during tillering stage, and increased from 4102.65 (CK) to 4.38 × 104 copies/g (wet weight) (soil) during heading time. Biogas slurry improved the soil nutrients (TN, AN, TP, and AP); meanwhile, the concentrations of total salt and Cl- increased. Network analysis indicated that 28 genera were the possible hosts of ARGs; variation partitioning analysis (VPA) indicated that microbial communities (contribution 59.30%) were the main factors that affected the fate of ARGs and intI1.

Attenuation of antibiotic resistance genes in livestock manure through vermicomposting via Protaetia brevitarsis and its fate in a soil-vegetable system

Scarab larvae (Protaetia brevitarsis) could transform large quantities of agricultural waste into compost, providing a promising bio-fertilizer for soil management. There is an urgent need to assess the risk of antibiotic resistance genes (ARGs) in soil-vegetable system with application of compost derived from P. brevitarsis larvae. We conducted a pot experiment to compare the changes of ARGs in the soil and lettuce by adding four types of manure, livestock manure (chicken and swine manure) and the corresponding larval frass. Significantly low numbers of ARGs and mobile genetic elements (MGEs) were detected in both larval frass compared with the corresponding livestock manure. Pot experiment showed that the detected numbers of ARGs and MGEs in bulk soil, rhizosphere soil, and root endophytes were significantly lower in the frass-amended treatments than the raw manure-amended treatments. Furthermore, the relative abundance of ARGs and MGEs with application of chicken-frass was significant lower in rhizosphere soil and leaf endophyte. Using non-metric multidimensional scaling analysis, the patterns of soil ARGs and MGEs with chicken-frass application were more close to those from the bulk soil in the control. Structural equation models indicated that livestock manure addition was the main driver shaping soil ARGs with raw manure application, while MGEs were the key drivers in frass-amended treatments. These findings demonstrated that application of livestock manure vermicomposting via scarab larvae (P. brevitarsis) may be at low risk in spreading manure-borne ARGs through soil-plant system, providing an alternative technique for reducing ARGs in organic waste.

When dewatered swine manure-derived biochar meets swine wastewater in anaerobic digestion: A win-win scenario towards highly efficient energy recovery and antibiotic resistance genes attenuation for swine manure management

This work explored the feasibility of dewatered swine manure-derived biochar (DSMB) as an additive to facilitate anaerobic digestion (AD) of swine wastewater for energy recovery and antibiotic resistance genes (ARG) attenuation enhancements. With 20 g/L DSMB assistance, the methanogenic lag time of swine wastewater was shortened by 17.4-21.1%, and the maximum CH4 production rate increased from 40.8 mL/d to 48.3-50.5 mL/d, among which DSMB prepared under 300 °C exhibited a better performance than that prepared under 500 °C and 700 °C. Integrated analysis of DSMB electrochemical properties, microbial electron transfer system activity, and microbial community succession revealed the potential of DSMB-300 to act as redox-active electron transfer mediators between syntrophic microbes to accelerate syntrophic methanogenesis via potential direct interspecies electron transfer. Meanwhile, DSMB preparation by pyrolysis dramatically reduced ARG abundance by almost 4 logs. Adding DSMB into AD not only strengthened the attenuation efficiency of ARG in the original swine wastewater, but also effectively controlled the potential risk of horizontal gene transfer by mitigating 74.8% of the mobile gene elements abundance. Accordingly, we proposed a win-win scenario for bio-waste management in swine farms, highlighting the more advanced energy recovery and ARG attenuation compared to the current status.

Mechanisms of metabolic performance enhancement and ARGs attenuation during nZVI-assisted anaerobic chloramphenicol wastewater treatment

Anaerobic wastewater treatment is a promising technology for refractory pollutant treatment. The nano zero-valent iron (nZVI) assisted anaerobic system could enhance contaminant removal. In this work, we added nZVI into an anaerobic system to investigate the effects on system performances and metabolic mechanism for chloramphenicol (CAP) wastewater treatment. As nZVI concentrations increased from 0 to 1 g/L, the CAP removal efficiency was appreciably improved from 46.5% to 99.2%, while the CH4 production enhanced more than 20 times. The enhanced CAP removal resulted from the enrichments of dechlorination-related bacteria (Hyphomicrobium) and other functional bacteria (e.g., Zoogloea, Syntrophorhabdus) associated with refractory contaminants degradation. The improved CH4 production was ascribed to the increases in fermentative-related bacteria (Smithella and Acetobacteroides), homoacetogen (Treponema), and methanogens. The increased abundances of anaerobic functional genes further verified the mechanism of CH4 production. Furthermore, the abundances of potential hosts of antibiotic resistance genes (ARGs) were reduced under high nZVI concentration (1 g/L), contributing to ARGs attenuation. This study provides a comprehensive analysis of the mechanism in metabolic performance enhancement and ARGs attenuation during nZVI-assisted anaerobic CAP wastewater treatment.

Attenuation effects of iron on dissemination of antibiotic resistance genes in anaerobic bioreactor: Evolution of quorum sensing, quorum quenching and dynamics of community composition

Zero valent iron (ZVI) coupled with bioreactors is arising as a promising technology for antibiotic resistance genes (ARGs) mitigation, whereas the succession and behaviors of microbes caused by ZVI in relieving ARGs propagation remain unclear. Herein, the effects of ZVI on microbial quorum sensing (QS), quorum quenching (QQ) system and community dynamics were examined in anaerobic bioreactor fed with oxytetracycline (tet), to illustrate the roles of evolutive microbial communication and community composition in ARGs attenuation. With the addition of 5 g/L ZVI, the total absolute abundance of tet ARGs was retarded by approximate 95% and 72% in sludge and effluent after 25 days operation. The abundance of mobile genetic elements and the heredity of antibiotic resistant bacteria revealed the declined horizontal and vertical transfer of ARGs, which directly led to the reduced ARGs propagation. Potential mechanisms are that the positive effects of ZVI on QQ activity via the functional bacteria enrichment inhibited QS system and thus ARGs transfer. Partial least--squares path modeling further demonstrated that ARGs abundance was strongly limited by the dynamics of bacterial composition and thereby less frequent microbial communication. These results provide new insights into the mechanisms of antibiotic resistome remission in anaerobic bioreactor modified by ZVI.

Antibiotic resistance genes attenuation in anaerobic microorganisms during iron uptake from zero valent iron: An iron-dependent form of homeostasis and roles as regulators

Zero valent iron (ZVI) has been previously documented to attenuate the propagation of antibiotic resistance genes (ARGs) in microbes, while how ZVI affects the evolution of ARGs remains unclear. Herein, we investigated the influences of ZVI on ARGs dissemination in anaerobic bioreactor treating oxytetracycline (tet) containing wastewater, by deciphering the roles of iron homeostasis and regulatory effects. A net reduction of tet gene targets ranging from 0.75 to 1.88 and 0.67 to 2.08 log unit in intracellular and extracellular DNA was achieved at the optimal dosage of 5 g/L ZVI, whereas 20 g/L ZVI made no effects on ARGs reduction. The reduced ARGs abundance by ZVI was directly related to the inhibited horizontal transfer of ARGs and decreased proliferation of resistant strains (mainly Paludibacter and WCHB1–32). The potential mechanisms included the increased antioxidant capacity, the depressed efflux pump system and the weakened energy driving force by Fur regulon in microbes (especially for Cloacibacterium and Dechloromonas). The negligible influence of 20 g/L ZVI on ARGs reduction was ascribed to the iron-catalyzed oxidative damage and reduced physiological activity. This study firstly illustrated the potential relationships among activation of iron uptake regulator leading to protection against oxidative stress, alternation of physiological metabolisms and reduction of ARGs dissemination. This work extents our understanding about the priority of ZVI in mitigating ARGs proliferation and sheds light on its potential application in wastewater treatment plants.

Fitness reduction of antibiotic resistome by an extra carbon source during swine manure composting.

This study employed high-throughput quantitative polymerase chain reaction to evaluate the effects of specific co-substrate and additive on the fitness of antibiotic resistome during swine manure composting. The results showed that corncob particle as a co-substrate significantly reduced the relative abundances of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) simultaneously. The diversity of ARGs was also reduced more effectively by corncob addition. Brick granule as an additive reduced the concentrations of bioavailable Cu and Zn. However, the relative abundances of ARGs and MGEs were not reduced by the addition of brick granule subsequently. Redundancy analysis indicated a negative effect of the C content and positive effects of class I integrase gene (intI) and bioavailable metals on the variation of the relative abundance of ARGs (p<0.01). The Procrustes test showed a higher goodness-of-fit between the relative abundance of ARGs and 16S rRNA genes (r=0.8166; p<0.0001). Our results suggests that the effect of corncob particle on the relative abundance of ARGs was achieved by driving the changes in physicochemical properties and microbial communities. This study confirmed the hypothesis of fitness cost and demonstrated the contribution of extra C source to ARG attenuation during composting.

Distinct effects of hypochlorite types on the reduction of antibiotic resistance genes during waste activated sludge fermentation: Insights of bacterial community, cellular activity, and genetic expression

The effectiveness of hypochlorites (NaClO and Ca(ClO)2) on the reduction of antibiotic resistance genes (ARGs) during waste activated sludge (WAS) fermentation was determined by the quantitative PCR. NaClO and Ca(ClO)2 exhibited distinct effects on ARGs fates. Ca(ClO)2 was effective in removing all investigated ARGs, and the efficiency was highly dose-dependent. Unexpectedly, the NaClO treatment attenuated ARGs with lower efficiency and even caused the propagation of certain ARGs (i.e., aadA1 and tetQ) at higher doses. The extracellular polymeric substances dissolution and membrane integrity suggested that unstable NaClO had acute effects on bacteria initially, while it was ineffective to further attenuate ARGs released from hosts due to the rapid consumption of oxidative ClO-. Without lasting and strong oxidative stress, the microbial activities of tolerant ARGs hosts will partially recover and then contribute to the ARGs dissemination across genera. In contrast, solid-state Ca(ClO)2 was slowly released and exhibited prolonged effects on bacteria by disrupting cell membranes and removing the susceptible ARGs released from hosts. Furthermore, bacterial taxa-ARG network analysis indicated that Ca(ClO)2 reduced the abundance of potential hosts, and the metabolic pathway and gene expression related to ARGs propagation were significantly downregulated by Ca(ClO)2, which contributed to efficient ARGs attenuation.

Deciphering the role of calcium peroxide on the fate of antibiotic resistance genes and mobile genetic elements during bioelectrochemically-assisted anaerobic composting of excess dewatered sludge

The integration of bioelectrochemical system (BES) into conventional anaerobic composting process can accelerate the degradation of excess dewatered sludge (ES). However, no reports have been made concerning antibiotic resistance genes (ARGs) profile and abundance in BESs fuelled with ES, and the effect of CaO2 on the fate of ARGs within BESs treating excess sludge has rarely been investigated. The aim of this study was to investigate the effect of CaO2 and the bioelectrochemical process on the abundances of ARGs and MGEs and to unravel the mechanism of ARGs attenuation. Results showed that CaO2 addition could enhance the reduction of ARG levels in ES within AnCBE. The mean ARG abundances in the AnCBE samples with high CaO2 doses (0.4 g CaO2/g VSS and 0.5 g CaO2/g VSS) were significantly lower than the values in samples with other CaO2 doses. The abundances of lincosamide nucleotidyltransferase, macB, macrolide transporter ATP-binding proteins, and macrolide-efflux proteins significantly decreased with the increase in the CaO2 dose. Both CaO2 addition and bioelectrochemical assistance played important roles in shaping the ARG composition during the AnCBE process. The variation in the microbial community composition is the most important contributor to the variation in the ARGs composition. The abundances of Actinobacteria and Firmicutes explained 52.8% of the total ARG variance. Of the MGEs, the abundances of plasmids, insertion sequences, and integrons were all reduced in the sludge metagenomes. There was a significantly positive correlation between the abundances of sulI, sulII, tetG, and blaTEM and those of metal resistance genes (MRGs), which decreased significantly after the AnCBE process. This study revealed the potential of the combination of CaO2 and bioelectrogenesis for ARG attenuation.

Evolutions of antibiotic resistance genes (ARGs), class 1 integron-integrase (intI1) and potential hosts of ARGs during sludge anaerobic digestion with the iron nanoparticles addition

In this work, we investigated the impact of iron nanoparticle, including magnetite nanoparticles (Fe3O4 NPs) and nanoscale zero-valent iron (nZVI), on the anaerobic digestion (AD) performance. Moreover, the evolutions of antibiotic resistance genes (ARGs), class 1 integrons-integrase (intI1) and potential hosts of ARGs were also investigated. The optimal addition of Fe3O4 NPs and nZVI to promote methane production was 0.5 g/L and 1 g/L, which led to 22.07% and 23.02% increase in methane yield, respectively. The degradation rate of organic matter was also enhanced with the addition of Fe3O4 NPs or nZVI. The results of high-throughput sequencing showed that the reactors with iron NPs exhibited significant differences in microbial community structure, compared to the reactors with the non‑iron NPs. Iron NPs have caused the relative abundance of the dominant bacteria (Proteobacteria, Firmicutes and Actinobacteria) generally decreased, while the dominant archaea (Euryarchaeota) increased in AD sludge. Quantitative PCR results revealed that iron NPs accelerated the reductions in total absolute abundance of ARGs, especially a beta-lactamase resistance encoded gene (blaOXA). Network analysis displayed that the attenuation of ARGs was mainly attributed to the decline of potential hosts (Proteobacteria, Firmicutes and Actinobacteria). Meanwhile, environmental factors (such as pH, soluble chemical oxygen demand and heavy metals) were also strongly correlated with ARGs.

Lateral flow sand filters are effective for removal of antibiotic resistance genes from domestic wastewater

The ability of lateral flow sand filters, used as on-site wastewater treatment systems (OWTS), to remove antibiotic resistance genes (ARGs), antibiotic resistant bacteria (ARB), and other relevant genetic markers (HF183, 16S rRNA, and int1) was assessed. Municipal wastewater was settled in a septic tank prior to loading into six pilot-scale lateral flow sand filters comprised of three different sand media types, at 5 and 30% slopes. The sand filters were sampled bi-weekly for: 9 ARGs and 3 other complimentary gene markers (sul1, sul2, qnrS, tetO, ermB, blaTEM, blaCTX-M, mecA, vanA, int1, HF183, 16S rRNA), and conventional microbial and water quality indicators, from July to November in 2017, and four times in the summer of 2018. The sand filters were observed to attenuate 7 of the ARGs to mostly below 2 log gene copies per mL. Log reductions ranging from 2.9 to 5.4 log were observed for the removal of absolute abundances of ARGs from septic tank effluent in 5 of the 6 sand filters. The fine-grained filter on the 5% slope did not perform as well for ARG attenuation due to hydraulic failure. The apportionment of cell-associated versus cell-free DNA was determined for the gene markers and this indicated that the genes were primarily carried intracellularly. Average log reductions of ARB with resistance to either sulfamethoxazole, erythromycin, or tetracycline were approximately 2.3 log CFU per mL within the filters compared to the septic tank effluent. This field study provides in-depth insights into the attenuation of ARB, ARGs, and their genetic compartmentalization in variably saturated sand OWTS. Overall, this type of OWTS was found to pose little risk of antimicrobial resistance contamination spread into surrounding environments when proper hydraulic function was maintained.

Metallic nanoparticles induced antibiotic resistance genes attenuation of leachate culturable microbiota: The combined roles of growth inhibition, ion dissolution and oxidative stress

The dissemination and propagation of antibiotic resistance genes (ARGs) is an emerging global health concern, and the potential effects of nanomaterials on ARGs fates have drawn much attention recently. In the current study, the effects of metallic nanoparticles on ARGs occurrence of leachate culturable microbiota were investigated by four typical metal and metal oxide nanoparticles (Cu, Zn, CuO, and ZnO). The ARGs diversity was remarkably decreased during the cultivation and enrichment of leachate microbiota, and their abundances decreased for 1.4–3.2 orders of magnitude. The presence of nanoparticles facilitated the ARGs attenuation, and the magnitude of effects depended on types of nanoparticles and ARGs. Metal oxide nanoparticles caused more remarkable effects than metal nanoparticles. Mechanism analysis indicated that bacterial growth was inhibited, and the dissolved metal ions from nanoparticles partially contributed to nanoparticles decreasing ARGs. Flow cytometry experiments further confirmed that nanoparticles could enter bacterial cells, and then induce excessive reactive oxygen species (ROS) generation and increase membrane permeability. Finally, the possible mechanisms were put forward, and the structural equation models (SEM) differentiated the contribution of different factors shaping ARGs. The dissolved metal ions and growth inhibition caused by nanoparticles decreased ARGs transfer frequencies via exerting excessive metal stress and lowering population density. On the other hand, nanoparticles were incorporated into the cells, and then induced the generation of ROS, which might facilitate ARGs horizontal transfer via increasing membrane permeability, or decrease ARGs via the damage of genomic and plasmid DNA. Therefore, nanoparticles could affect ARGs fates via several ways, and combined effects finally determined the ARGs variations.

Antibiotic resistance genes attenuated with salt accumulation in saline soil.

Salt accumulation on the surface of the soil layer driven by the strong evaporation is a natural phenomenon that usually happens in the dry season, particularly on the coastal lands reclaimed from tidal flats. However, the influence of salt accumulation on the distribution profile of antibiotic resistance genes (ARGs) and mobile gene elements (MGEs) remains unclear. In this study, we sampled a wild saline soil where the salt accumulation was frequently observed to investigate the vertical distribution profiles of ARGs and MGEs. The results showed that an increasing gradient of ARGs and MGEs was observed from the top to deep layer with the decreasing of electrical conductivity (EC1:5 values) indicating the salt-influenced attenuation of ARGs in the saline soil. The competing test suggested that the attenuation of ARGs in response to salinity gradient was attributable to the elimination of the ARG-harboring plasmids, due to the reduction of the relative fitness of plasmid-harboring strains. Additionally, the network analyses showed that the attenuation of ARGs might be associated with decreased abundance of Actinobacteria. Overall, this study identifies that salinity as an abiotic stress could re-shape the distribution of ARGs, which may influence the dissemination of ARGs in the environment.