Metagenomic analysis of antibiotic resistance genes (ARGs) during refuse decomposition. - Related Documents




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857001.0000Metagenomic analysis of antibiotic resistance genes (ARGs) during refuse decomposition. Landfill is important reservoirs of residual antibiotics and antibiotic resistance genes (ARGs), but the mechanism of landfill application influence on antibiotic resistance remains unclear. Although refuse decomposition plays a crucial role in landfill stabilization, its impact on the antibiotic resistance has not been well characterized. To better understand the impact, we studied the dynamics of ARGs and the bacterial community composition during refuse decomposition in a bench-scale bioreactor after long term operation (265d) based on metagenomics analysis. The total abundances of ARGs increased from 431.0ppm in the initial aerobic phase (AP) to 643.9ppm in the later methanogenic phase (MP) during refuse decomposition, suggesting that application of landfill for municipal solid waste (MSW) treatment may elevate the level of ARGs. A shift from drug-specific (bacitracin, tetracycline and sulfonamide) resistance to multidrug resistance was observed during the refuse decomposition and was driven by a shift of potential bacteria hosts. The elevated abundance of Pseudomonas mainly contributed to the increasing abundance of multidrug ARGs (mexF and mexW). Accordingly, the percentage of ARGs encoding an efflux pump increased during refuse decomposition, suggesting that potential bacteria hosts developed this mechanism to adapt to the carbon and energy shortage when biodegradable substances were depleted. Overall, our findings indicate that the use of landfill for MSW treatment increased antibiotic resistance, and demonstrate the need for a comprehensive investigation of antibiotic resistance in landfill.201829660875
857810.9996Impact of earthworms on suppressing dissemination of antibiotic resistance genes during vermicomposting treatment of excess sludge. Earthworms play a crucial role in suppressing the dissemination of antibiotic resistance genes (ARGs) during vermicomposting. However, there is still a lack of how earthworms influence the spread of ARGs. To address this gap, a microcosm experiment was conducted, incorporating earthworms and utilizing metagenomics and quantitative PCR to assess the impact of earthworms on microbial interactions and the removal of plasmid-induced ARGs. The findings revealed that vermicomposting led to a reduction in the relative abundance of ARGs by altering microbial communities and interactions. Significantly, vermicomposting demonstrated an impressive capability, reducing 92% of ARGs donor bacteria and impeding the transmission of 94% of the RP4 plasmid. Furthermore, through structural equation model analysis, it was determined that mobile genetic elements and environmental variables were the primary influencers of ARG reduction. Overall, this study offers a fresh perspective on the effects of vermicomposting and its potential to mitigate the spread of ARGs.202438885722
810320.9996Anaerobic/aerobic conditions determine antibiotic resistance genes removal patterns from leachate by affecting bacteria taxa-genes co-occurrence modules. Landfill treatment of municipal solid waste treatment produces a large amount of leachate, which has been an important hotspot of ARGs. This study aimed to investigate the ARGs removal potential, kinetics and mechanism from leachate in aerobic and anaerobic conditions. Simulated landfill reactors showed the efficacy in reducing ARGs, and the removal efficiencies depended on ARGs types and aerobic/anaerobic conditions. The ARGs tetQ and bla(CTX-M) were more likely to attenuate with the log-removal efficiencies of 1.50-3 order of magnitude. The ARGs removal kinetic was well fitted by modified Collins-Selleck model, and aerobic condition showed better removal capacities and kinetics than anaerobic condition. Among the ARGs with great removal performance, sul2, aadA1and bla(CTX-M) were eliminated from leachate and refuse simultaneously, but tetM, ermB, and mefA were removed from leachate but enriched in refuse. Aerobic/anaerobic states might drive the bacterial community shift of leachate and refuse, and topology property comparison of co-occurrence networks suggested that refuse had a closer non-random host relationship between ARGs and microbial taxa than leachate. Further module analyses revealed that ARGs removal efficiencies depended on the taxonomy of host bacteria in leachate, while the refuse taxa-ARGs correlation determined ARGs removal patterns. By selecting distinct bacteria cluster in different conditions, aerobic treatment benefited ARGs reduction in leachate and refuse, while anaerobic treatment enhanced the enrichment of ARGs in refuse. These findings can potentially foster the understanding of ARGs removal mechanism in biological treatment processes.201930763913
691630.9996Mobile genetic elements in potential host microorganisms are the key hindrance for the removal of antibiotic resistance genes in industrial-scale composting with municipal solid waste. During the municipal solid waste (MSW) composting, antibiotic resistance genes (ARGs) could be one of the concerns to hinder the application of MSW composting. However, the understanding of enrichment and dissemination of ARGs during the industrial-scale composting is still not clear. Hence, this study aimed to investigate the ARG distributions at different stages in an industrial-scale MSW composting plant. Seven target ARGs and four target mobile genetic elements (MGEs) and bacterial communities were investigated. The abundances of ARGs and MGEs increased during two aerobic thermophilic stages, but they decreased in most ARGs and MGEs after composting. Network analysis showed that potential host bacteria of ARGs were mainly Firmicutes and Actinobacteria. The reduction of potential host bacteria was important to remove ARGs. MGEs were an important factor hindering ARG removal. Water-extractable S and pH were two main physicochemical factors in the changes of microbial community and the abundance of ARGs.202031962245
698340.9996Deciphering Potential Roles of Earthworms in Mitigation of Antibiotic Resistance in the Soils from Diverse Ecosystems. Earthworms are capable of redistributing bacteria and antibiotic resistance genes (ARGs) through soil profiles. However, our understanding of the earthworm gut microbiome and its interaction with the antibiotic resistome is still lacking. Here, we characterized the earthworm gut and soil microbiome and antibiotic resistome in natural and agricultural ecosystems at a national scale, and microcosm studies and field experiments were also employed to test the potential role of earthworms in dynamics of soil ARGs. The diversity and structure of bacterial communities were different between the earthworm gut and soil. A significant correlation between bacterial community dissimilarity and spatial distance between sites was identified in the earthworm gut. The earthworm gut consistently had lower ARGs than the surrounding soil. A significant reduction in the relative abundance of mobile genetic elements and dominant bacterial phylotypes that are the likely hosts of ARGs was observed in the earthworm gut compared to the surrounding soil, which might contribute to the decrease of ARGs in the earthworm gut. The microcosm studies and field experiments further confirmed that the presence of earthworms significantly reduced the number and abundance of ARGs in soils. Our study implies that earthworm-based bioremediation may be a method to reduce risks associated with the presence of ARGs in soils.202133977709
698150.9996Decline in the Relative Abundance of Antibiotic Resistance Genes in Long-Term Fertilized Soil and Its Driving Factors. The changes in antibiotic resistance genes (ARGs) in long-term fertilized soil remain controversial. We aimed to analyze the variation characteristics of ARGs in long-term fertilized soil using metagenomic sequencing. The relative abundance of ARGs did not increase significantly after 7 years of fertilization. However, a clear decline in the relative abundance of ARGs was observed compared to the data from the 4th year. Microbial adaptation strategies in response to changes in the ARG abundance were associated with shifts in microbiome composition and function. Among these, bacterial abundance was the primary driving factor. Additionally, total heavy metal content might serve as the most significant co-selective pressure influencing ARG number. We believe that increasing the selective pressure from heavy metals and antibiotics might result in the loss of certain microbial species and a decrease in ARG abundance. This study provides novel insights into the variations of soil resistance genes under long-term fertilization.202540785530
701360.9996Bacterial Community Shift Drives Antibiotic Resistance Promotion during Drinking Water Chlorination. For comprehensive insights into the effects of chlorination, a widely used disinfection technology, on bacterial community and antibiotic resistome in drinking water, this study applied high-throughput sequencing and metagenomic approaches to investigate the changing patterns of antibiotic resistance genes (ARGs) and bacterial community in a drinking water treatment and distribution system. At genus level, chlorination could effectively remove Methylophilus, Methylotenera, Limnobacter, and Polynucleobacter, while increase the relative abundance of Pseudomonas, Acidovorax, Sphingomonas, Pleomonas, and Undibacterium in the drinking water. A total of 151 ARGs within 15 types were detectable in the drinking water, and chlorination evidently increased their total relative abundance while reduced their diversity in the opportunistic bacteria (p < 0.05). Residual chlorine was identified as the key contributing factor driving the bacterial community shift and resistome alteration. As the dominant persistent ARGs in the treatment and distribution system, multidrug resistance genes (mainly encoding resistance-nodulation-cell division transportation system) and bacitracin resistance gene bacA were mainly carried by chlorine-resistant bacteria Pseudomonas and Acidovorax, which mainly contributed to the ARGs abundance increase. The strong correlation between bacterial community shift and antibiotic resistome alteration observed in this study may shed new light on the mechanism behind the chlorination effects on antibiotic resistance.201526397118
691870.9996Variations in antibiotic resistance genes and removal mechanisms induced by C/N ratio of substrate during composting. For a comprehensive insight into the potential mechanism of the removal of antibiotic resistance genes (ARGs) removal induced by initial substrates during composting, we tracked the dynamics of physicochemical properties, bacterial community composition, fungal community composition, the relative abundance of ARGs and mobile genetic genes (MGEs) during reed straw and cow manure composting with different carbon to nitrogen ratio. The results showed that the successive bacterial communities were mainly characterized by the dynamic balance between Firmicutes and Actinobacteria, while the fungal communities were composed of Ascomycota. During composting, the interactions between bacteria and fungi were mainly negative. After composting, the removal efficiency of ARGs in compost treatment with C/N ≈ 26 (LL) was higher than that in compost treatment with C/N ≈ 35 (HL), while MGEs were completely degraded in HL and enriched by 2.3% in LL. The large reduction in the relative abundance of ARGs was possibly due to a decrease in the potential host bacterial genera, such as Advenella, Tepidimicrobium, Proteiniphilum, Acinetobacter, Pseudomonas, Flavobacteria and Arcbacter. Partial least-squares path modeling (PLS-PM) revealed that the succession of bacterial communities played a more important role than MGEs in ARGs removal, while indirect factors of the fungal communities altered the profile of ARGs by affecting the bacterial communities. Both direct and indirect factors were affected by composting treatments. This study provides insights into the role of fungal communities in affecting ARGs and highlights the role of different composting treatments with different carbon to nitrogen ration on the underlying mechanism of ARGs removal.202134375241
858980.9996Impacts of microplastic type on the fate of antibiotic resistance genes and horizontal gene transfer mechanism during anaerobic digestion. Microplastics (MPs) and antibiotic resistance genes (ARGs) are important pollutants in waste activated sludge (WAS), but their interactions during anaerobic digestion (AD) still need to be further explored. This study investigated variations in ARGs, mobile genetic elements (MGEs), and host bacteria during AD under the pressure of polyamide (PA), polyethylene (PE), and polypropylene (PP). The results showed that the MPs increased methane production by 11.7-35.5%, and decreased ARG abundance by 5.6-24.6%. Correlation analysis showed that the decrease of MGEs (plasmid, prophage, etc.) promoted the decrease of the abundance of multidrug, aminoglycoside and tetracycline resistance genes. Metagenomic annotation revealed that the reduction of key host bacteria (Arenimonas, Lautropia, etc.) reduced the abundance of major ARGs (rsmA, rpoB2, etc.). Moreover, PP MPs contributed to a reduction in the abundance of functional genes related to the production of reactive oxygen species, ATP synthesis, and cell membrane permeability, which was conducive to reducing the potential for horizontal gene transfer of ARGs. These findings provide insights into the treatment of organic waste containing MPs.202438772228
856590.9996Deciphering the transfers of antibiotic resistance genes under antibiotic exposure conditions: Driven by functional modules and bacterial community. Antibiotics can exert selective pressures on sludge as well as affect the emergence and spread of antibiotic resistance genes (ARGs). However, the underlying mechanisms of ARGs transfers are still controversial and not fully understood in sludge system. In present study, two anaerobic sequence batch reactors (ASBR) were constructed to investigate the development of ARGs exposed to two sulfonamide antibiotics (SMs, sulfadiazine SDZ and sulfamethoxazole SMX) with increasing concentrations. The abundance of corresponding ARGs and total ARGs obviously increased with presence of SMs. Functional analyses indicated that oxidative stress response, signal transduction and type IV secretion systems were triggered by SMs, which would promote ARGs transfers. Network analysis revealed 18 genera were possible hosts of ARGs, and their abundances increased with SMs. Partial least-squares path modeling suggested functional modules directly influenced mobile genetic elements (MGEs) as well as the ARGs might be driven by both functional modules and bacteria community, while bacteria community composition played a more key role. Sludge with refractory antibiotics (SDZ) may stimulate the relevant functions and shift the microbial composition to a greater extent, causing more ARGs to emerge and spread. The mechanisms of ARGs transfers are revealed from the perspective of functional modules and bacterial community in sludge system for the first time, and it could provide beneficial directions, such as oxidative stress reduction, cellular communication control, bacterial composition directional regulation, for ARGs spread controlling in the future.202134563930
6982100.9996Viral Communities Suppress the Earthworm Gut Antibiotic Resistome by Lysing Bacteria on a National Scale. Earthworms are critical in regulating soil processes and act as filters for antibiotic resistance genes (ARGs). Yet, the geographic patterns and main drivers of earthworm gut ARGs remain largely unknown. We collected 52 earthworm and soil samples from arable and forest ecosystems along a 3000 km transect across China, analyzing the diversity and abundance of ARGs using shotgun metagenomics. Earthworm guts harbored a lower diversity and abundance of ARGs compared to soil, resulting in a stronger distance-decay rate of ARGs in the gut. Greater deterministic assembly processes of ARGs were found in the gut than in soil. The earthworm gut had a lower frequency of co-occurrence patterns between ARGs and mobile genetic elements (MGEs) in forest than in arable systems. Viral diversity was higher in the gut compared to soil and was negatively correlated with bacterial diversity. Bacteria such as Streptomyces and Pseudomonas were potential hosts of both viruses and ARGs. Viruses had negative effects on the diversity and abundance of ARGs, likely due to the lysis on ARG-bearing bacteria. These findings provide new insights into the variations of ARGs in the earthworm gut and highlight the vital role of viruses in the regulation of ARGs in the soil ecosystem.202439037720
6929110.9996Root exudates regulate soil antibiotic resistance genes via rhizosphere microbes under long-term fertilization. Organic fertilizer application promotes the prevalence of antibiotic resistance genes (ARGs), yet the factors driving temporal differences in ARG abundance under long-term organic fertilizer application remain unclear. This study investigated the temporal dynamics of ARG diversity and abundance in both bulk and rhizosphere soils over 17 years (2003-2019), and explored microbial evolution strategies, ARG hosts succession and the influence of root exudates on ARGs regulation. The results showed that the ARGs abundance in rhizosphere soil was lower than that in bulk soil under long-term fertilization, and ARGs abundance exhibited a decrease and then remained stable in rhizosphere soil over time. There was a strong association between host bacteria and dominant ARGs (p < 0.05). Structural equations demonstrated that bacterial community had a most pronounced influence on ARGs (p < 0.05), and metabolites exhibited an important mediation effect on bacterial community (p < 0.05), thereby impacting ARGs. The metabolome analysis evidenced that significant correlations were found between defensive root exudates and most ARGs abundance (p < 0.05), like, luteolin-7-glucoside was negatively correlated with tetA(58). These findings provide deeper insights into the dynamics of soil ARGs under long-term fertilization, and identify critical factors that influence ARGs colonization in soils, providing support for controlling the spread of ARGs in agriculture soils.202539700687
7560120.9996The effect of bacterial functional characteristics on the spread of antibiotic resistance genes in Expanded Granular Sludge Bed reactor treating the antibiotic wastewater. To explore the fate and spreading mechanism of antibiotics resistance genes (ARGs) in antibiotics wastewater system, a laboratory-scale (1.47 L) Expanded Granular Sludge Bed (EGSB) bioreactor was implemented. The operating parameters temperature (T) and hydraulic retention time (HRT) were mainly considered. This result showed the removal of ARGs and COD was asynchronous, and the recovery speed of ARGs removal was slower than that COD removal. The decreasing T was attributed to the high growth rate of ARGs host bacteria, while the shortened HRT could promote the horizontal and vertical gene transfer of ARGs in the sludge. The analysis result of potential bacterial host showed more than half of the potential host bacteria carried 2 or more ARGs and suggested an indirect mechanism of co-selection of multiple ARGs. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) was used to investigate the functional characteristics of bacterial community. This result showed the bacterial functional genes contributed 40.41% to the abundance change of ARGs in the sludge, which was higher that of bacterial community. And the function genes of "aromatic hydrocarbon degradation", "Replication, recombination and repair proteins" and "Flagellar assembly" were mainly correlated with the transfer of ARGs in the sludge. This study further revealed the mechanism of ARGs spread in the EGSB system, which would provide new ideas for the development of ARGs reduction technology.202134488144
7015130.9996Comparison of bacterial communities and antibiotic resistance genes in oxidation ditches and membrane bioreactors. Oxidation ditches (ODs) and membrane bioreactors (MBRs) are widely used in wastewater treatment plants (WWTPs) with bacteria and antibiotic resistance genes (ARGs) running through the whole system. In this study, metagenomic sequencing was used to compare the bacterial communities and ARGs in the OD and MBR systems, which received the same influent in a WWTP located in Xinjiang, China. The results showed that the removal efficiency of pollutants by the MBR process was better than that by the OD process. The composition and the relative abundance of bacteria in activated sludge were similar at the phylum and genus levels and were not affected by process type. Multidrug, fluoroquinolones and peptides were the main ARG types for the two processes, with macB being the main ARG subtype, and the relative abundance of ARG subtypes in MBR effluent was much higher than that in the OD effluent. The mobile genetic elements (MGEs) in the activated sludge were mainly transposons (tnpA) and insertion sequences (ISs; IS91). These results provide a theoretical basis for process selection and controlling the spread of ARGs.202133903636
7038140.9996Interactions between fungi and bacteria hosts carrying MGEs is dominant for ARGs fate during manure mesophilic composting. The mycelial networks of fungi promote the interaction between the originally isolated bacteria, thereby potentially enhancing the exchange of nutrients and the horizontal transfer of genetic materials. However, the driving effect of fungi on antibiotic resistance genes (ARGs) during mesophilic facultative composting is still unclear. This study aims to elucidate the changes in ARGs and underlying mechanisms during the mesophilic composting of manure. Results indicated that reduction rates of ARGs in sheep and pig manure over a 90-day composting period were 34.68% and 60.10%, respectively. The sul1, sul2 and tetX were identified as recalcitrant ARGs in both composting treatments, with the additional unique recalcitrant gene addA observed in sheep manure. Fungal communities appeared to have a more significant influence on the cooperation between bacteria and fungi. Massive fungi interacted intensively with bacterial hosts carrying both ARGs and mobile genetic elements (MGEs). In sheep and pig manure, there were 53 and 38 potential bacterial hosts (genus level) carrying both ARGs and MGEs, associated close interactions with fungi. Structural equation modeling revealed that compost properties influence ARGs by affecting the abundance of core fungi and the hosts carrying MGEs, and that core fungi could also impact ARGs by influencing the bacterial hosts carrying MGEs. Core fungi have the potential to facilitate the horizontal transfer of ARGs by enhancing bacterial network interactions.202539764902
7041150.9995Antibiotic and heavy metal resistance genes in sewage sludge survive during aerobic composting. Municipal sewage sludge has been generated in increasing amounts with the acceleration of urbanization and economic development. The nutrient rich sewage sludge can be recycled by composting that has a great potential to produce stabilized organic fertilizer and substrate for plant cultivation. However, little is known about the metals, pathogens and antibiotic resistance transfer risks involved in applying the composted sludge in agriculture. We studied changes in and relationships between heavy metal contents, microbial communities, and antibiotic resistance genes (ARGs), heavy metal resistance genes (HMRGs) and mobile genetic elements (MGEs) in aerobic composting of sewage sludge. The contents of most of the analyzed heavy metals were not lower after composting. The bacterial α-diversity was lower, and the community composition was different after composting. Firmicutes were enriched, and Proteobacteria and potential pathogens in the genera Arcobacter and Acinetobacter were depleted in the composted sludge. The differences in bacteria were possibly due to the high temperature phase during the composting which was likely to affect temperature-sensitive bacteria. The number of detected ARGs, HMRGs and MGEs was lower, and the relative abundances of several resistance genes were lower after composting. However, the abundance of seven ARGs and six HMRGs remained on the same level after composting. Co-occurrence analysis of bacterial taxa and the genes suggested that the ARGs may spread via horizontal gene transfer during composting. In summary, even though aerobic composting is effective for managing sewage sludge and to decrease the relative abundance of potential pathogens, ARGs and HMRGs, it might include a potential risk for the dissemination of ARGs in the environment.202336608829
7040160.9995The maturity period is the main stage of antibiotic resistance genes reduction in aerobic composting process of swine manure in sub-scale farms. This study was focused on the changes of antibiotic resistance genes (ARGs) and their potential host bacteria during the swine manure composting on sub-scale farms. Eight target ARGs increased 427% on average, with a trend of increase at early stage and decrease at later stage, and the main reduction stage appeared in maturity stage. The abundance of ARGs was mainly affected by the community succession of potential host bacteria. Composting could reduce the abundance of potential host bacteria of ARGs as well as pathogens such as Pseudomonas, and reduce the environmental risks of swine manure. N/C and S levels had a positive effect on the potential host of most ARGs. Prolonging the maturity period would inhibit the growth of potential host bacteria of ARGs during composting, therefore inhibiting the transmission of ARGs.202132971337
7011170.9995Dynamics of bacterial composition and the fate of antibiotic resistance genes and mobile genetic elements during the co-composting with gentamicin fermentation residue and lovastatin fermentation residue. Dynamics in bacterial community composition, along with 13 antibiotic resistance genes (ARGs) and eight mobile genetic elements (MGEs), were assessed during co-composting with gentamicin and lovastatin fermentation residue (GFR and LFR, respectively). Using next generation sequencing, the key bacterial taxa associated with the different stages of composting were identified. Most importantly, Bacillus, belonging to Phylum Firmicutes, was associated with enhanced degradation of gentamicin, decomposition of organic matter (OM) and dissolved organic carbon (DOC), and also extension of the thermophilic phase of the composting cycle. During the course of composting, the patterns of different ARGs/MGEs varied. However, the total and the normalized (to bacterial numbers) copies both remained high. The abundance of various ARGs was related to bacterial abundance and community composition, and the changing pattern of individual ARGs was influenced by the selectivity of MGEs and bacteria.201829673993
7563180.9995Polystyrene microplastics alleviate the effects of sulfamethazine on soil microbial communities at different CO(2) concentrations. Microplastics were reported to adsorb antibiotics and may modify their effects on soil systems. But there has been little research investigating how microplastics may affect the toxicities of antibiotics to microbes under future climate conditions. Here, we used a free-air CO(2) enrichment system to investigate the responses of soil microbes to sulfamethazine (SMZ, 1 mg kg(-1)) in the presence of polystyrene microplastics (PS, 5 mg kg(-1)) at different CO(2) concentrations (ambient at 380 ppm and elevated at 580 ppm). SMZ alone decreased bacterial diversity, negatively affected the bacterial structure and inter-relationships, and enriched the sulfonamide-resistance genes (sul1 and sul2) and class 1 integron (intl1). PS, at both CO(2) conditions, showed little effect on soil bacteria but markedly alleviated SMZ's adverse effects on bacterial diversity, composition and structure, and inhibited sul1 transmission by decreasing the intl1 abundance. Elevated CO(2) had limited modification in SMZ's disadvantages to microbial communities but markedly decreased the sul1 and sul2 abundance. Results indicated that increasing CO(2) concentration or the presence of PS affected the responses of soil microbes to SMZ, providing new insights into the risk prediction of antibiotics under future climate conditions.202133592488
7552190.9995Effect of temperature on removal of antibiotic resistance genes by anaerobic digestion of activated sludge revealed by metagenomic approach. As antibiotic resistance continues to spread globally, there is growing interest in the potential to limit the spread of antibiotic resistance genes (ARGs) from wastewater sources. In particular, operational conditions during sludge digestion may serve to discourage selection of resistant bacteria, reduce horizontal transfer of ARGs, and aid in hydrolysis of DNA. This study applied metagenomic analysis to examine the removal efficiency of ARGs through thermophilic and mesophilic anaerobic digestion using bench-scale reactors. Although the relative abundance of various ARGs shifted from influent to effluent sludge, there was no measureable change in the abundance of total ARGs or their diversity in either the thermophilic or mesophilic treatment. Among the 35 major ARG subtypes detected in feed sludge, substantial reductions (removal efficiency >90%) of 8 and 13 ARGs were achieved by thermophilic and mesophilic digestion, respectively. However, resistance genes of aadA, macB, and sul1 were enriched during the thermophilic anaerobic digestion, while resistance genes of erythromycin esterase type I, sul1, and tetM were enriched during the mesophilic anaerobic digestion. Efflux pump remained to be the major antibiotic resistance mechanism in sludge samples, but the portion of ARGs encoding resistance via target modification increased in the anaerobically digested sludge relative to the feed. Metagenomic analysis provided insight into the potential for anaerobic digestion to mitigate a broad array of ARGs.201525994259