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811300.9583Fate of antibiotic resistance genes in mesophilic and thermophilic anaerobic digestion of chemically enhanced primary treatment (CEPT) sludge. Anaerobic digestion (AD) of chemically enhanced primary treatment (CEPT) sludge and non-CEPT (conventional sedimentation) sludge were comparatively operated under mesophilic and thermophilic conditions. The highest methane yield (692.46±0.46mL CH(4)/g VS(removed) in CEPT sludge) was observed in mesophilic AD of CEPT sludge. Meanwhile, thermophilic conditions were more favorable for the removal of total antibiotic resistance genes (ARGs). In this study, no measurable difference in the fates and removal of ARGs and class 1 integrin-integrase gene (intI1) was observed between treated non-CEPT and CEPT sludge. However, redundancy analysis indicated that shifts in bacterial community were primarily accountable for the variations in ARGs and intI1. Network analysis further revealed potential host bacteria for ARGs and intI1.201728797965
811210.9578Fate of antibiotic resistance bacteria and genes during enhanced anaerobic digestion of sewage sludge by microwave pretreatment. The fate of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) were investigated during the sludge anaerobic digestion (AD) with microwave-acid (MW-H), microwave (MW) and microwave-H2O2-alkaline (MW-H2O2) pretreatments. Results showed that combined MW pretreatment especially for the MW-H pretreatment could efficiently reduce the ARB concentration, and most ARG concentrations tended to attenuate during the pretreatment. The subsequent AD showed evident removal of the ARB, but most ARGs were enriched after AD. Only the concentration of tetX kept continuous declination during the whole sludge treatment. The total ARGs concentration showed significant correlation with 16S rRNA during the pretreatment and AD. Compared with unpretreated sludge, the AD of MW and MW-H2O2 pretreated sludge presented slightly better ARB and ARGs reduction efficiency.201626970692
810620.9574Reducing antibiotic resistance genes, integrons, and pathogens in dairy manure by continuous thermophilic composting. This study explored the effects of composting using three temperature regimes, namely, insufficient thermophilic composting (ITC), normal thermophilic composting (NTC), and continuous thermophilic composting (CTC), on antibiotic resistance genes (ARGs), integrons, and human pathogenic bacteria (HPB), as well as the mechanisms involved. The NTC and CTC treatments led to greater decreases in 5/10 ARGs and two integrons than ITC, and the abundances of ARGs (tetC, tetG, and tetQ) and int1 only declined in the NTC and CTC treatments. The abundances of HPB decreased by 82.8%, 76.9%, and 96.9% under ITC, NTC, CTC, respectively. Redundancy analysis showed that both bacterial succession and horizontal gene transfer play important roles in the variation of ARGs, and the changes in different ARGs were due to diverse mechanisms. CTC performed significantly better at reducing ARGs, integrons, and HPB, thus it may be used to manage the public health risks of ARGs in animal manure.201627598571
811530.9569Effects of reductive soil disinfestation on potential pathogens and antibiotic resistance genes in soil. Reductive soil disinfestation (RSD) is commonly employed for soil remediation in greenhouse cultivation. However, its influence on antibiotic resistance genes (ARGs) in soil remains uncertain. This study investigated the dynamic changes in soil communities, potential bacterial pathogens, and ARG profiles under various organic material treatments during RSD, including distillers' grains, potato peel, peanut vine, and peanut vine combined with charcoal. Results revealed that applying diverse organic materials in RSD significantly altered bacterial community composition and diminished the relative abundance of potential bacterial pathogens (P < 0.05). The relative abundance of high-risk ARGs decreased by 10.7%-30.6% after RSD treatments, the main decreased ARG subtypes were AAC(3)_Via, dfrA1, ErmB, lnuB, aadA. Actinobacteria was the primary host of ARGs and was suppressed by RSD. Soil physicochemical properties, such as total nitrogen, soil pH, total carbon, were crucial factors affecting ARG profiles. Our findings demonstrated that RSD treatment inhibited pathogenic bacteria and could be an option for reducing high-risk ARG proliferation in soil.202539306413
810540.9568Refluxing mature compost to replace bulking agents: A low-cost solution for suppressing antibiotic resistance genes rebound in sewage sludge composting. Antibiotic resistance genes (ARGs) rebounding during composting cooling phase is a critical bottleneck in composting technology that increased ARGs dissemination and application risk of compost products. In this study, mature compost (MR) was used as a substitute for rice husk (RH) to mitigate the rebound of ARGs and mobile genetic elements (MGEs) during the cooling phase of sewage sludge composting, and the relationship among ARGs, MGEs, bacterial community and environmental factors was investigated to explore the key factor influencing ARGs rebound. The results showed that aadD, blaCTX-M02, ermF, ermB, tetX and vanHB significantly increased 4.76-32.41 times, and the MGEs rebounded by 38.60% in the cooling phase of RH composting. Conversely, MR reduced aadD, tetM, ermF and ermB concentrations by 59.49-98.58%, and reduced the total abundance of ARGs in the compost product by 49.32% compared to RH, which significantly restrained ARGs rebound. MR promoted secondary high temperature inactivation of potential host bacteria, including Ornithinibacter, Rhizobiales and Caldicoprobacter, which could harbor aadE, blaCTX-M02, and blaVEB. It also reduced the abundance of lignocellulose degrading bacteria of Firmicutes, which were potential hosts of aadD, tetX, ermF and vanHB. Moreover, MR reduced moisture and increased oxidation reduction potential (ORP) that promoted aadE, tetQ, tetW abatement. Furthermore, MR reduced 97.36% of total MGEs including Tn916/1545, IS613, Tp614 and intI3, which alleviated ARGs horizontal transfer. Overall finding proposed mature compost reflux as bulking agent was a simple method to suppress ARGs rebound and horizontal transfer, improve ARGs removal and reduce composting plant cost.202539798649
810050.9566Composting temperature directly affects the removal of antibiotic resistance genes and mobile genetic elements in livestock manure. The high antibiotic resistance gene (ARGs) contents in livestock manure pose a potential risk to environment and human health. The heap composting with an ambient temperature and thermophilic composting are two methods for converting livestock manure into fertilizer. This study investigated the variations in ARGs and mobile genetic elements (MGEs) and revealed potential mechanisms for ARGs removal using the two composting methods. The ARGs abundance were enriched by 44-fold in heap composting, among them, the macrolide-resistance genes increased significantly. On the contrary, the ARGs were removed by 92% in thermophilic composting, among them, tetracycline-resistance genes decreased by 97%. The bacterial hosts of ARGs were associated with the variations of ARGs and MGEs. The tetO was correlated with the most diverse bacteria in heap composting, and Bacteroidetes was the major host bacteria. While tetT was correlated with the most diverse bacteria in thermophilic composting, and Proteobacteria was the major host bacteria. Structural equation models showed that the enrichment of ARGs in heap composting was mainly correlated with bacterial communities, whereas, the removal of ARGs in thermophilic composting was directly affect by MGEs. Composting temperature directly affected the variations in ARGs. Higher and lower temperatures significantly decreased and increased, respectively, ARGs and MGEs abundance levels.202235306090
722160.9565Profiles of bacteria/phage-comediated ARGs in pig farm wastewater treatment plants in China: Association with mobile genetic elements, bacterial communities and environmental factors. In this study, the profiles of bacteria/phage-comediated antibiotic resistance genes (b/pARGs) were monitored in water samples collected from 45 pig farm wastewater treatment plants (WWTPs) in seven different regions of China. We found that 8 major types and 112 subtypes of b/pARGs were detected in all the water samples, and the detected number ranged from 53 to 92. The absolute abundances of bARGs and pARGs in the influent were as high as 10(9) copies/mL and 10(6) copies/mL, respectively. Anaerobic anoxic/oxic (AAO) and anaerobic short-cut nitrification/denitrification (ASND) treatment plants can effectively reduce the absolute abundance and amount of b/pARGs. Anaerobic treatment plants cannot reduce the absolute abundance of pARGs, and even increase the amount of pARGs. Mobile genetic elements (MGEs), bacterial communities and environmental factors were important factors impacting the b/pARG profile. Among these factors, the bacterial community was the major driver that impacted the bARG profile, while bacterial community and MGEs were the major codrivers impacting the pARG profile. This study was the first to investigate the profiles of b/pARGs in pig farm WWTPs in China on such a large scale, providing a reference for the prevention and control of ARG pollution in agricultural environments.202133069996
812370.9564The 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
722080.9564Occurrence and Drivers of Antibiotic Resistance Genes Carried by Bacteriophages in Soils Following Different Fertilization Treatments. Fertilization has an important effect on soil antibiotic resistance. Most recent studies have focused on antibiotic resistance genes (ARGs) harbored by bacteria (bARGs); however, little is known about ARGs carried by soil bacteriophages (pARGs) under different fertilization treatments. Here, 24 pARG subtypes were quantified in soils with long-term application of different fertilizers using droplet digital PCR (ddPCR). The results showed that the detection rates of the target ARGs in bacteriophages were 66.67%, 70.83%, and 75.00% in unfertilized, chemically fertilized, and organically fertilized soils, respectively. The total abundance of pARGs in soils amended with organic fertilizer was significantly higher than that in unfertilized and chemically fertilized soils. The multidrug resistance gene (mexF) exhibited the highest abundance in soils amended with organic fertilizer. A significant positive correlation was observed between bARGs and pARGs, and the detected pARG subtype abundances were one to two orders of magnitude lower than those of the corresponding bARGs. The results of variation partitioning analysis revealed that the interaction between the bacterial community and soil properties drove the variation in soil pARGs. Our findings indicate that bacteriophages are important vectors of ARGs, in addition to bacteria, in agricultural soils, and their contribution to antibiotic resistance should not be overlooked.202540559968
810990.9564The fate of antibiotic resistance genes and their influential factors in swine manure composting with sepiolite as additive. Manures are storages for antibiotic resistance genes (ARGs) entering the environment. This study investigated the effects of adding sepiolite at 0%, 2.5%, 5%, and 7.5% (CK, T1, T2, and T3, respectively) on the fates of ARGs during composting. The relative abundances (RAs) of the total ARGs in CK and T3 decreased by 0.23 and 0.46 logs, respectively, after composting. The RAs of 10/11 ARGs decreased in CK, whereas they all decreased in T3. The reduction in the RA of the total mobile genetic elements (MGEs) was 1.26 times higher in T3 compared with CK after composting. The bacterial community accounted for 47.93% of the variation in the abundances of ARGs. Network analysis indicated that ARGs and MGEs shared potential host bacteria (PHB), and T3 controlled the transmission of ARGs by reducing the abundances of PHB. Composting with 7.5% sepiolite is an effective strategy for reducing the risk of ARGs proliferating.202235063626
8101100.9564Enhanced removal of antibiotic resistance genes and mobile genetic elements during swine manure composting inoculated with mature compost. Livestock manure is a major source of antibiotic resistance genes (ARGs) that enter the environment. This study assessed the effects of inoculation with mature compost (MC) on the fates of ARGs and the bacterial community during swine manure composting. The results showed that MC prolonged the thermophilic period and promoted the decomposition of organic matter, which was due to the rapid growth and reproduction of thermophilic bacteria (Bacillus, Thermobifida, and Thermobacillus). MC significantly reduced the relative abundances of ARGs (1.02 logs) and mobile genetic elements (MGEs) (1.70 logs) after composting, especially sulfanilamide resistance genes. The total ARGs removal rate was 1.11 times higher in MC than the control. Redundancy analysis and structural equation modeling showed that horizontal gene transfer mediated by MGEs (ISCR1 and intI1) was the main direct factor related to the changes in ARGs during composting, whereas the C/N ratio and pH were the two most important indirect factors. Network analysis showed that members of Firmicutes comprising Romboutsia, Clostridisensu_stricto_1, and Terrisporobacter were the main bacterial hosts of ARGs and MGEs. MC reduced the risk of ARGs transmission by decreasing the abundances of bacterial hosts. Thus, MC is a promising strategy for reducing the proliferation risk of ARGs.202133858100
8054110.9563Effects of nanoscale zero-valent iron on the performance and the fate of antibiotic resistance genes during thermophilic and mesophilic anaerobic digestion of food waste. The effects of nanoscale zero-valent iron (nZVI) on the performance of food waste anaerobic digestion and the fate of antibiotic resistance genes (ARGs) were investigated in thermophilic (TR) and mesophilic (MR) reactors. Results showed that nZVI enhanced biogas production and facilitated ARGs reduction. The maximum CH(4) production was 212.00 ± 4.77 ml/gVS with 5 g/L of nZVI in MR. The highest ARGs removal ratio was 86.64 ± 0.72% obtained in TR at nZVI of 2 g/L. nZVI corrosion products and their contribution on AD performance were analyzed. The abundance of tetracycline genes reduced significantly in nZVI amended digesters. Firmicutes, Chloroflexi, Proteobacteria and Spirochaetes showed significant positive correlations with various ARGs (p < 0.05) in MR and TR. Redundancy analysis indicated that microbial community was the main factor that influenced the fate of ARGs. nZVI changed microbial communities, with decreasing the abundance bacteria belonging to Firmicutes and resulting in the reduction of ARGs.201931505392
8120120.9562Insight into the fate of antibiotic resistance genes and bacterial community in co-composting green tea residues with swine manure. Green tea residues (GTRs) are byproducts of tea production and processing, and this type of agricultural waste retains nutritious components. This study investigated the co-composting of GTRs with swine manure, as well as the effects of GTRs on antibiotic resistance genes (ARGs) and the bacterial community during co-composting. The temperature and C/N ratio indicate compost was mature after processing. The addition of GTRs effectively promoted the reduction in the abundances of most targeted ARGs (tet and sul genes), mobile genetic element (MGE; intI1), and metal resistance genes (MRGs; pcoA and tcrB). Redundancy analysis (RDA) showed that GTRs can reduce the abundance of MRGs and ARGs by reducing the bioavailability of heavy metals. Network analysis shows that Firmicutes and Actinobacteria were the main hosts of ARGs and ARGs, MGEs, and MRGs shared the same potential host bacteria. Adding GTRs during composting may reduce ARGs transmission through horizontal gene transfer (HGT). GTRs affected the bacterial community, thereby influencing the variations in the ARG profiles and reducing the potential risk associated with the compost product.202032310121
8117130.9562Composting of oxytetracycline fermentation residue in combination with hydrothermal pretreatment for reducing antibiotic resistance genes enrichment. Hydrothermal pretreatment can efficiently remove the residual antibiotics in oxytetracycline fermentation residue (OFR), but its effect on antibiotic resistance genes (ARGs) during composting remains unclear. This study compared the shifts in bacterial community and evolutions in ARGs and integrons during different composting processes of OFRs with and without hydrothermal pretreatment. The results demonstrated that hydrothermal pretreatment increased the bacterial alpha diversity at the initial phase, and increased the relative abundances of Proteobacteria and Actinobacteria but decreased that of Bacteroidetes at the final phase by inactivating mycelia and removing residual oxytetracycline. Composting process inevitably elevated the abundance and relative abundance of ARGs. However, the increase in ARGs was significantly reduced by hydrothermal pretreatment, because the removal of oxytetracycline decreased their potential host bacteria and inhibited their horizontal gene transfer. The results demonstrated that hydrothermal pretreatment is an efficient strategy to reduce the enrichment of ARGs during the OFR composting.202033099099
6921140.9561Impacts of Chemical and Organic Fertilizers on the Bacterial Communities, Sulfonamides and Sulfonamide Resistance Genes in Paddy Soil Under Rice-Wheat Rotation. The responses of sulfonamides, sulfonamide-resistance genes (sul) and soil bacterial communities to different fertilization regimes were investigated by performing a field experiment using paddy soil with no fertilizer applied, chemical fertilizer applied, organic fertilizer applied, and combination of chemical and organic fertilizer applied. Applying organic fertilizer increased the bacterial community diversity and affected the bacterial community composition. Eutrophic bacteria (Bacteroidetes, Gemmatimonadetes, and Proteobacteria) were significantly enriched by applying organic fertilizer. It was also found organic fertilizer application increased sulfamethazine content and the relative abundances of sul1 and sul2 in the soil. In contrast, applying chemical fertilizer significantly increased the abundance of Nitrospirae, Parcubacteria, and Verrucomicrobia and caused no obvious changes on sul. Correlation analysis indicated that sul enrichment was associated with the increases in sulfamethazine content and potential hosts (e.g., Novosphingobium and Rhodoplanes) population. The potential ecological risks of antibiotics in paddy soil with organic fertilizer applied cannot be ignored.202236547725
7942150.9561Insight into effects of polyethylene microplastics in anaerobic digestion systems of waste activated sludge: Interactions of digestion performance, microbial communities and antibiotic resistance genes. The environmental risks of microplastics (MPs) have raised an increasing concern. However, the effects of MPs in anaerobic digestion (AD) systems of waste activated sludge (WAS), especially on the fate of antibiotic resistance genes (ARGs), have not been clearly understood. Herein, the variation and interaction of digestion performance, microbial communities and ARGs during AD process of WAS in the presence of polyethylene (PE) MPs with two sizes, PE MPs-180μm and PE MPs-1mm, were investigated. The results showed that the presence of PE MPs, especially PE MPs-1mm, led to the increased hydrolysis of soluble polysaccharides and proteins and the accumulation of volatile fatty acids. The methane production decreased by 6.1% and 13.8% in the presence of PE MPs-180μm and PE MPs-1mm, respectively. Together with this process, hydrolytic bacteria and acidogens were enriched, and methanogens participating in acetoclastic methanogenesis were reduced. Meanwhile, ARGs were enriched obviously by the presence of PE MPs, the abundances of which in PE MPs-180μm and PE MPs-1mm groups were 1.2-3.0 times and 1.5-4.0 times higher than that in the control by the end of AD. That was associated with different co-occurrence patterns between ARGs and bacterial taxa and the enrichment of ARG-hosting bacteria caused by the presence of PE MPs. Together these results suggested the adverse effects of PE MPs on performance and ARGs removal during AD process of WAS through inducing the changes of microbial populations.202235944782
8126160.9560Antiallergic drugs drive the alteration of microbial community and antibiotic resistome in surface waters: A metagenomic perspective. Antiallergic drugs (AADs) are emerging contaminants of global concern due to their environmental persistence and potential ecological impacts. This study investigated the effects of seven AADs (chlorpheniramine, diphenhydramine, cetirizine, loratadine, desloratadine, sodium cromoglicate and calcium gluconate) at environmentally relevant concentrations on antibiotic resistome and bacterial community structures in water using microcosm experiments and metagenomic sequencing. The results showed that AADs increased the abundance of antibiotic-resistant bacteria (ARB) by 1.24- to 7.78-fold. Community structure shifts indicated that chlorpheniramine, diphenhydramine, and cetirizine promoted Actinobacteria (e.g., Aurantimicrobium), while the other four AADs favored Proteobacteria (e.g., Limnohabitans). AADs also significantly altered the relative abundance of antibiotic resistance genes (ARGs), with Actinobacteria and Proteobacteria identified as key ARB components and potential hosts of ARGs (e.g., evgS, mtrA, RanA). Host analysis showed ARGs were primarily carried by Actinobacteria (e.g., Aurantimicrobium) under chlorpheniramine, diphenhydramine, and cetirizine exposure, but by Proteobacteria (e.g., Limnohabitans) under the other four AADs. Furthermore, AADs facilitated the horizontal transfer of ARGs (e.g., evgS) within microbial communities, contributing to antibiotic resistance dissemination. This study highlights the ecological risks of AADs in promoting antibiotic resistance spread and provides new insights into their impact on microbial communities and resistome dynamics in aquatic environments.202540570627
6926170.9560Insights into the driving factors of vertical distribution of antibiotic resistance genes in long-term fertilized soils. The prevalence of antibiotic resistance genes (ARGs) in soils has aroused wide attention. However, the influence of long-term fertilization on the distribution of ARGs in different soil layers and its dominant drivers remain largely unknown. In this study, a total of 203 ARGs were analyzed in greenhouse vegetable soils (0-100 cm from a 13-year field experiment applied with different fertilizers (control, chemical fertilizer, organic manure, and mixed fertilizer). Compared with unfertilized and chemically fertilized soils, manure application significantly increased the abundance and alpha diversity of soil ARGs, where the assembly of ARG communities was strongly driven by stochastic processes. The distribution of ARGs was significantly driven by manure application within 60 cm, while it was insignificantly changed in soil below 60 cm under different fertilization regimes. The inter-correlations of ARGs with mobile genetic elements (MGEs) and microbiota were strengthened in manured soil, indicating manure application posed a higher risk for ARGs diffusion in subsurface soil. Bacteria abundance and MGEs directly influenced ARG abundance and composition, whereas soil depth and manure application indirectly influenced ARG abundance and composition by affecting antibiotics. These results strengthen our understanding of the long-term anthropogenic influence on the vertical distribution of soil ARGs and highlight the ecological risk of ARGs in subsurface soil induced by long-term manure application.202337247491
6917180.9560Response characteristics of antibiotic resistance genes and bacterial communities during agricultural waste composting: Focusing on biogas residue combined with biochar amendments. This research investigated biogas residue and biochar addition on antibiotic resistance genes (ARGs), mobile genetic elements (MGEs), and changes in bacterial community during agricultural waste composting. Sequencing technique investigated bacterial community structure and ARGs, MGEs changes. Correlations among physicochemical factors, ARGs, MGEs, and bacterial community structure were determined using redundancy analysis. Results confirmed that biochar and biogas residue amendments effectively lowered the contents of ARGs and MGEs. The main ARGs detected was sul1. Proteobacteria and Firmicutes were the main host bacteria strongly associated with the dissemination of ARGs. The dynamic characteristics of the bacterial community were strongly correlated with pile temperature and pH (P < 0.05). Redundancy and network analysis revealed that nitrate, intI1, and Firmicutes mainly affected the in ARGs changes. Therefore, regulating these key variables would effectively suppress the ARGs spread and risk of compost use.202336657587
8098190.9560Elimination of antibiotic resistance genes and human pathogenic bacteria by earthworms during vermicomposting of dewatered sludge by metagenomic analysis. This study used a metagenomic approach to investigate the effects of earthworms on ARGs and HPB during the vermicomposting of dewatered sludge. Results showed that 139 types of ARGs were found in sludge vermicompost, affiliated to 30 classes. Compared with the control, the total abundance of ARGs in sludge vermicompost decreased by 41.5%. Moreover, the types and sequences of plasmids and integrons were also decreased by vermicomposting. Proteobacteria and Actinobacteria were the most dominant hosts of ARGs in sludge vermicompost. In addition, earthworms reduced the total HPB abundance and modified their diversity, thus leading to higher abundance of Enterobacteriaceae in sludge vermicompost. However, the sludge vermicompost was still ARG and HPB enriched, indicating a remaining environmental risk for agricultural purpose. The observed change of microbial community and the reduction of mobile genetic elements caused by earthworm activity are the main reasons for the alleviation of ARG pollution during vermicomposting.202031787516