Relative Abundance Of Antibiotic Resistance Genes Research Articles

Bacterial Community Structure and Antibiotic Resistance Gene Changes in IFAS+Magnetic Coagulation Process Wastewater Treatment Plant in Cold Regions

The main objective of this study was to explore the changes in bacterial communities and antibiotic resistance genes (ARGs) in an integrated fixed-film activated sludge (IFAS)+magnetic coagulation process wastewater treatment plant (WWTP) in Xinjiang. The bacterial communities and ARGs in the influent, suspended activated sludge, attached biofilm, and effluent were studied using 16S rRNA gene sequencing and metagenomic sequencing. The results showed that the average relative abundances of Chloroflexi and Nitrospirae in activated sludge were 3.50% and 0.03%, respectively, and their relative abundances in biofilm reached 10.02% and 2.12%, respectively. The average removal rates of NH4+-N and TN increased from 91.89% and 66.76% to 97.71% and 91.90% after the reformation of this wastewater treatment plant, respectively, indicating that IFAS enhanced the biological nitrogen removal capacity of wastewater treatment plants in cold regions. The average relative abundances of Ferruginibacter and Rhodoferax related to iron redox in the biological treatment section were 5.24% and 3.72%, respectively, and the relative abundance of Rhodoferax in effluent reached 9.48%, indicating that the magnetic powder had an impact on the bacterial community. The IFAS wastewater treatment plant had an obvious removal effect on ARGs, and the relative abundance of ARGs decreased from 191.08×10-3‰ in the influent to 32.58×10-3‰ in the effluent. The relative abundance of ARGs in activated sludge was 63.25×10-3‰-72.38×10-3‰, which was significantly higher than 41.31×10-3‰ in biofilm. However, the relative abundances of dominant subtypes of ARGs such as sul2, floR, and rpoB2 in biofilm were 5.77×10-3‰, 2.52×10-3‰, and 2.03×10-3‰, respectively, which were higher than the 3.15×10-3‰-3.57×10-3‰, 1.73×10-3‰-2.24×10-3‰, and 1.28×10-3‰-1.76×10-3‰ in activated sludge. The network analysis indicated that Caldilineaceae_norank and Trichococcus were respectively positively correlated with sul2 and floR. These results can provide theoretical reference for the optimal operation and ARGs control of WWTPs in cold regions.

Application of spray-dried erythromycin fermentation residue as a soil amendment: antibiotic resistance genes, nitrogen cycling, and microbial community structure.

Erythromycin fermentation residue (EFR) after spray drying could be reused as a soil amendment. However, the effects of spray-dried EFR on antibiotic resistance genes (ARGs), nitrogen cycling, and microbial community structure in soil are rarely reported. In this study, a pot experiment was conducted by adding spray-dried EFR to soil. For the application of 1.0% spray-dried EFR, the residual erythromycin (ERY) could be rapidly removed with the half-life of 22.2 d; the total relative abundance of ARGs increased at first, but decreased to the initial level of the control group in the end; genes related to ammonium assimilation (glnA, gltB, gltD), ammonification (gdhA, gudB, cynT, cynS, ncd2), denitrification (narI, narG, narH), assimilatory nitrate reduction (nirA, nasA), and dissimilatory nitrate reduction (nirD) were enriched; soil microbial community structure presented temporary variation. Network analysis showed significant negative correlations between ARGs and nitrogen cycling genes. The addition of 6.0% spray-dried EFR resulted in the amplification of ARGs and inhibition of nitrogen cycling. This work provides new insights into the effects of spray-dried EFR on ARGs, nitrogen cycling, and microbial community structure within the fertilized soil.

City-scale distribution of airborne antibiotic resistance genes

Concerns around urban air quality have been increasing worldwide due to large-scale urbanization. A large volume of work has been focused on the chemical pollutants in the air and their impacts on human health. However, the profile of airborne microbial contaminants, especially antibiotic resistance genes (ARGs), is largely understudied. Here, high-throughput quantitative PCR (HT-qPCR) was employed to explore the temporal and spatial distribution of airborne ARGs from 11 sites with various functional zones and different urbanization levels within Xiamen, China. A total of 104 unique ARGs and 23 mobile genetic elements (MGEs) were detected across all samples. Temporal shift was observed in the distribution of ARG profiles, with significantly higher relative abundance of ARGs detected in summer than that in spring. Temperature is the key predictor of the total relative abundance of ARGs and MGEs in summer, while PM2.5 and PM10 were the two most important factors affecting the abundance in spring. Our findings suggest that urban aerosols accommodate rich and dynamic ARGs and MGEs, and emphasize the role of temperature, air quality and anthropogenic activities in shaping the profile of ARGs.

Dual character of methane production improvement and antibiotic resistance genes reduction by nano-Fe2O3 addition during anaerobic digestion of swine manure

In this study, the dual character of the methane production and fate of antibiotic resistance genes (ARGs) were evaluated after adding nano-Fe2O3 (NF) during anaerobic digestion (AD) of swine manure, concerning the different exposure times (ET). NF has a decreasing effect on methane production, and the inhibitory effect is proportional to the dosage. NF350 decreased the accumulative methane production by about 7.8% when the ET was 30 days due to the disruption of cellular integrity and the damage to the bacterial communities. Nonetheless, NF could improve the maximum specific methane production rate by up to 24.5% when the ET was shorter than 12 days. And the changes in the relative abundance of ARGs were proportional to the daily methane production at this stage. The addition of NF in a short time is helpful for methanogenic activities, which might be correlated with direct interspecies electron transfer (DIET). NF specifically enhanced the reduction of tetracycline resistance genes with antibiotic target protection (40.7%) and aminoglycoside resistance genes with antibiotic inactivation (34.0%) when the ET was 30 days. It was speculated that the specifical reduction was mainly due to that higher NF could produce the ROS (reactive oxygen species), down regulate key enzyme actions in phosphorylation, acetylization, and nucelotidylalation with antibiotic inactivation, and reduces the reduction of selective pressure for tetracycline. Environmental variables and microbial community were the major factors causing the changes of the ARGs by Structural equation models (SEM) and Procrustes analysis. These results illustrated that resonable NF addition could help to improve the accumulative methane production and reduce ARGs during AD of swine manure.

Industrial-scale aerobic composting of livestock manures with the addition of biochar: Variation of bacterial community and antibiotic resistance genes caused by various composting stages

The presence of large amounts of antibiotic resistance genes (ARGs) in livestock manures poses an impending, tough safety risk to ecosystems. To investigate more comprehensively the mechanisms of ARGs removal from industrial-scale composting of livestock manure based on biochar addition, we tracked the dynamics of bacterial community and ARGs at various stages of aerobic composting of livestock manures with 10% biochar. There were no significant effects of biochar on the bacterial community and the profiles of ARGs. During aerobic composting, the relative abundance of ARGs and mobile genetic elements (MGEs) showed overall trends of decreasing and then increasing. The key factor driving the dynamics of ARGs was bacterial community composition, and the potential hosts of ARGs were Caldicoprobacter, Tepidimicrobium, Ignatzschineria, Pseudogracilibacillus, Actinomadura, Flavobacterium and Planifilum. The retention of the thermophilic bacteria and the repopulation of the initial bacteria were the dominant reasons for the increase in ARGs at maturation stage. Additionally, among the MGEs, the relative abundance of transposon gene was substantially removed, while the integron genes remained at high relative abundance. Our results highlighted that the suitability of biochar addition to industrial-scale aerobic composting needs to be further explored and that effective measures are needed to prevent the increase of ARGs content on maturation stage.

Effects of g-C3N4 on bacterial community and tetracycline resistance genes in two typical sediments in tetracycline pollution remediation.

Photocatalysis, as a novel technique, has been widely used for antibiotic pollution remediation in wastewater. In the processes of degradation and removal of antibiotics, the impact of photocatalysts on microenvironment is very important but remains poorly understood. In the present study, the effect of typical photocatalyst g-C3N4 (Graphitic carbon nitride) on microbial community was investigated in two sediment types (riverbed sediment and pig-farm sediment) polluted by tetracycline (TC) in central southern China. The riverbed sediment and pig farm sediment samples were respectively exposed to g-C3N4 (25, 75, 125 mg⋅kg–1) and TC (60, 120, 180 mg⋅L–1) treatments alone or combination for 30 days, respectively. The bacterial community and antibiotic resistance genes (ARGs) of the treated sediments were analyzed by Illumina sequencing and metagenomic sequencing. Studies had shown that: TC, g-C3N4, and TC/g-C3N4 have significant effects on the changes of microbial communities and components in riverbed sediment, but they do not exist in pig farm sediment. The most alterations of microbial taxa were Acidobacteriota, Actinobacteriota, and Desulfobacterota in riverbed sediment, and Elusimicrobiota in the pig farm sediment under various treatments. Through network analysis, it was found that the distribution of microorganisms in the pig farm sediment is more complex and more stable. The addition of g-C3N4 reduced the absolute abundance of ARGs in the two examined sediments, but not significantly changed their relative abundance of ARGs. The g-C3N4 application was beneficial to the removal of TC residues and to the prevention of the generation and transmission of ARGs in sediments. Our results suggested that g-C3N4 was a suitable photocatalyst with excellent application prospect for the removal of TC residues and the control of ARGs in environment.

Occurrence and driving mechanism of antibiotic resistance genes in marine recreational water around Qinhuangdao, China

Rapid urbanization and other human activities have exacerbated the global human health risks associated with antibiotic resistance genes (ARGs). Swimming in contaminated waters is one of important pathways by which humans can be exposed to ARGs. However, little is currently known about the overall levels of ARGs in recreational waters and the forces driving their occurrence. In this study, we analyzed the ARGs and bacterial communities in water samples taken from five recreational coastal waters (RCWs) and their adjacent estuaries in the Qinhuangdao area of Bohai Bay, China, using high-throughput quantitative PCR and 16S rRNA gene amplicon sequencing technique. The relative abundances of ARGs in RCWs varied greatly, and occurred at levels close to, or even higher than, those in the neighboring estuaries, ranging from 9.4×10-2 to 2.5 copies/16S rRNA gene. A total of 159 different ARGs able to express resistance to almost all antibiotics were detected in all of the water samples. β-lactamase resistance genes (blaTEM gene) were the most abundance of all ARGs. Patterns of ARG occurrence were significantly correlated with those of bacterial communities (Mantel test, P < 0.05). A partial redundancy analysis showed that differences in bacterial communities accounted for 50.3% of the total ARG variations, which was much higher than the sum of the contribution of mobile genetic elements (18.1%) and the other selected environmental factors (8.3%). Principal coordinate analysis based on Bray-Curtis distance revealed the similarities between ARG profiles in the RCWs and in their adjacent estuaries. To our knowledge, this is the first study to report the ARG profiles in RCWs using high throughput qPCR technology. These results implied that it is necessary to develop proper regulations and monitoring strategies of RCWs for reducing risks of ARGs on human health.

Watershed urbanization enhances the enrichment of pathogenic bacteria and antibiotic resistance genes on microplastics in the water environment

Microplastics (MPs) serve as vectors for microorganisms and antibiotic resistance genes (ARGs) and contribute to the spread of pathogenic bacteria and ARGs across various environments. Patterns of microbial communities and ARGs in the biofilm on the surface of MPs, also termed as plastisphere, have become an issue of global concern. Although antibiotic resistome in the plastisphere has been detected, how watershed urbanization affects patterns of potential pathogens and ARGs in the microplastic biofilms is still unclear. Here, we compared the bacterial communities, the interaction between bacterial taxa, pathogenic bacteria, and ARGs between the plastisphere and their surrounding water, and revealed the extensive influence of urbanization on them. Our results showed that bacterial communities and interactions in the plastisphere differed from those in their surrounding water. Microplastics selectively enriched Bacteroidetes from water. In non-urbanized area, the abundance of Oxyphotobacteria was significantly (p < 0.05) higher in plastisphere than that in water, while α-Proteobacteria was significantly (p < 0.05) higher in plastisphere than those in water of urbanized area. Pathogenic bacteria, ARGs, and mobile genetic elements (MGEs) were significantly (p < 0.05) higher in the urbanized area than those in non-urbanized area. MPs selectively enriched ARG-carrying potential pathogens, i.e., Klebsiella pneumoniae and Enterobacter cloacae, and exhibited a distinct effect on the relative abundance of ARG and pathogens in water with different urbanization levels. We further found ARGs were significantly correlated to MGEs and pathogenic bacteria. These results suggested that MPs would promote the dissemination of ARGs among microbes including pathogenic bacteria, and urbanization would affect the impact of MPs on microbes, pathogens, and ARGs in water. A high level of urbanization could enhance the enrichment of pathogens and ARGs by MPs in aquatic systems and increase microbial risk in aquatic environments. Our findings highlighted the necessity of controlling the spread of ARGs among pathogens and the usage of plastic products in ecosystems of urban areas.

Spread and driving factors of antibiotic resistance genes in soil-plant system in long-term manured greenhouse under lead (Pb) stress

Livestock manure is often used as fertilizer in greenhouses, resulting in simultaneous enrichment of heavy metals and antibiotic resistance genes (ARGs) in soils. The soil-plant system is a non-negligible way to spread ARGs; however, the effects of lead (Pb) on the spread of ARGs and their driving factors in the greenhouse soil-plant system remain unclear. In this present study, the occurrence of ARGs in greenhouse soils and their spread into plants under Pb stress were studied. Overall, Pb promoted the accumulation of ARGs in root endophytes at 10, 50, and 100 mg/kg as well as in soils at 10 and 200 mg/kg, but reduced the total relative abundance of ARGs in leaf endophytes. Particularly, Pb increased the mobile genetic elements (MGEs) relative abundance and endophytic bacterial community diversity in roots, consistent with the change in the total relative abundance of ARGs. Network analysis revealed that bacterial community and MGEs may jointly affect the migration of ARGs in the soil-plant system of greenhouses. Overall, this study extended our knowledge of how Pb can promote the transmission of ARGs to plant roots from greenhouse soils receiving long-term manure applications, which must be considered when assessing the risk of ARGs to public health.

ARGs distribution and high-risk ARGs identification based on continuous application of manure in purple soil

Tetracyclines (TCs) are one of the most widely used antibiotics in livestock and poultry industry. Sichuan province and Chongqing city are the provinces and cities with the most concentrated purple soil distribution in China, as well as the largest livestock and poultry breeding scale. Purple soils with low organic carbon content and poor structure, and manure fertilizers were heavily applied to improve soil quality. However, the research on antibiotic resistance genes (ARGs) dispersal in purple soils is limited. Overall, 234 ARGs subtypes belonging to 15 types were detected in all soils. Long-term application of livestock manures significantly changed the characteristics of ARGs, mobile genetic elements (MGEs), pathogen species and bacterial communities in the soil, and increased by 103.2 %, 13.1 %, 188.6 % and 43.7 % in chicken manure treatments compared with the control, respectively. Partial equation PLS-PM analysis further reveals that the main driving factor of ARGs is bacterial abundance, while bacterial diversity has a negative effect on ARGs abundance. Through ANOVA and network analysis, it was found that 30 ARGs were significantly affected by manure, its relative abundance is 0.8–1.4 times that of the control treatment. The qPCR results also proved that the relative abundance of ARGs including sul2, etc. increased with the increase of manure application and these resistance genes in chicken manure treatments were higher than in pig manure treatments. These resistance genes pose a high risk to public health, and chicken manure application posed a higher risk than pig manure. The TC content in the 40–60 cm soil layer was higher in medium- and high-volume pig manure, which was at risk of being transported to groundwater. Our research results deepen the understanding of ARGs transmission in purple soil under agricultural activities and emphasize the species and distribution of antibiotic resistance genes may differ across soil and manure types.

Metagenomic assembly reveals hosts and mobility of common antibiotic resistome in animal manure and commercial compost

BackgroundAntibiotics and antibiotic resistance genes (ARGs) used in intensive animal farming threaten human health worldwide; however, the common resistome, ARG mobility, and ARG host composition in different animal manures and mixed manure composts remain unclear. In the present study, metagenomic assembly and cross-sample mapping were used to comprehensively decipher the common resistome and its potential mobility and hosts in animal manure and composts.ResultsIn total, 201 ARGs were shared among different animal (layer, broiler, swine, beef cow, and dairy cow) manures and accounted for 86–99% of total relative abundance of ARGs. Except for multidrug, sulfonamide, and trimethoprim resistance genes, the relative abundance of most ARGs in composts was significantly lower than that in animal manure. Procrustes analysis indicated that antibiotic residues positively correlated with ARG composition in manure but not in composts. More than 75% ARG subtypes were shared between plasmids and chromosomes in our samples. Transposases could play a pivotal role in mediating the transfer of ARGs between different phyla in animal manure and composting. Cross-sample mapping to contigs carrying ARGs showed that the hosts of common resistome in manure had preference on animal species, and the dominant genus of ARG host shifted from Enterococcus in manure to Pseudomonas in composts. The broad host range and linking with diverse mobile genetic elements (MGEs) were two key factors for ARGs, such as sul1 and aadA, which could survive during composting. The multidrug resistance genes represented the dominant ARGs in pathogenic antibiotic-resistant bacteria in manure but could be effectively controlled by composting.ConclusionsOur experiments revealed the common resistome in animal manure, classified and relative quantified the ARG hosts, and assessed the mobility of ARGs. Composting can mitigate ARGs in animal manure by altering the bacterial hosts; however, persistent ARGs can escape from the removal because of diverse host range and MGEs. Our findings provide an overall background for source tracking, risk assessment, and control of livestock ARGs.

Distribution Characteristics of Antibiotics and Antibiotic Resistance Genes in Manure and Surrounding Soil of Poultry Farm in Ningxia

Antibiotics and antibiotic resistance genes (ARGs) in livestock and poultry manure pose potential ecological risks. In order to understand the distribution characteristic of antibiotics and ARGs in manure and surrounding soils of poultry farms in Ningxia, the poultry manure and relative soil samples were collected from 12 layers of different poultry breeding farms. The compositions of antibiotics and ARGs in the samples were analyzed using UPLC-MS/MS and HT-qPCR. The results showed that:① tetracycline, aminoglycoside, and sulfonamide were the dominant antibiotics in poultry manure. The types and contents of antibiotics in poultry manure were different in different breeding periods. There were more types of antibiotics in the brooding period, the average content was high, and the initial stage showed the opposite trend. ② A small amount of antibiotics was detected in the surrounding soil only 20 m away from the poultry farm, and the poultry farm had little effect on the distribution of antibiotics in the surrounding soil. The content of quinolone in the soils with poultry manure application was significantly higher than that in the control and surrounding soil. ③ We detected 132-168 ARGs in poultry manure, and the number of aminoglycosides and tetracycline was higher. The relative abundance of ARGs in the rearing period was highest, and the initial stage showed the opposite trend. The total relative abundance of ARGs in the brooding period was highest, but the terminal period showed the opposite. There were 110 ARGs in poultry manure during all breeding periods. ④ There were 23-105 ARGs in the soils, and the number of aminoglycoside was highest, followed by multidrug ARGs. The poultry farm had a great effect on the number and relative abundance of ARGs in the surrounding soil. For example, the number and relative abundance of ARGs in the surrounding soil of poultry farms gradually decreased with the increase in the distance from the poultry farms. The number and relative abundance of ARGs in the soil with applied poultry manure were significantly increased; however, these values were lower than those in the soil 20 m away from the poultry farm. ⑤ β-lactamases, aminoglycosides, and macrolide lincosamide-streptogramin B (MLSB) ARGs were all at risk of horizontal movement in manure, and chloramphenicol ARGs were at risk of horizontal movement in soil. Correlation analysis showed that the relative abundance of aminoglycoside, tetracycline, sulfonamide, β-lactamase, and MLSB were not significantly correlated with their contents. ⑥ Different types of ARGs had related co-occurrence phenomena, such as the positive correlation between the relative abundance of ARGs in poultry manure, and aminoglycoside and β-lactamases, MGEs, multidrugs and vancomycins. The relative abundances of ARGs in soil, aminoglycoside and tetracyclines, vancomycins, sulfonamides, and MLSBs; tetracyclines and MLSBs; etc., all showed a significant positive correlation. In short, the co-occurrence among the relative abundance of ARGs in soil was significantly stronger than that in poultry manure. These results could provide the theoretical basis for the site selection of poultry farms, the selection of antibiotic types and dosages for large-scale breeding of laying hens, and the application of poultry manure.

Distribution and driving factors of antibiotic resistance genes in treated wastewater from different types of livestock farms

Treated wastewater from livestock farms is an important reservoir for antibiotic resistance genes (ARGs), and is a main source of ARGs in the environment. However, the distribution and driving factors of ARGs in treated wastewater from different types of livestock farms are rarely reported. In this study, treated wastewater from 69 large-scale livestock farms of different types, including broiler, layer, and pig farms, was collected, and 11 subtypes of ARGs, 2 mobile genetic elements (MGEs) and bacterial community structure were analyzed. The results revealed detection rates of NDM-1 and mcr-1 of 90 % and 43 %, respectively, and the detection rates of other ARGs were 100 %. The relative abundance of ARGs, such as tetA, tetX and strB, in broiler farms was significantly higher than that in layer farms, but the bacterial α diversity was significantly lower than that in other farm types. Furthermore, although the treatment process had a greater impact on the physicochemical properties of the treated wastewater than the livestock type, livestock type was the main factor affecting the bacterial community in the treated wastewater. The analysis of potential host bacteria of ARGs revealed significant differences in the host bacteria of ARGs in treated wastewater from different types of livestock farms. The host bacteria of ARGs in broiler farms mainly belonged to Actinobacteria, layer farms mainly belonged to Proteobacteria, and pig farms mainly belonged to Firmicutes. Additionally, redundancy analysis showed that the distribution of ARGs may have resulted from the combination of multiple driving factors in different types of livestock farms, among which tnpA and NH4+-N were the main influencing factors. This study revealed multiple driving factors for the distribution of typical ARGs in treated wastewater from different types of livestock farms, providing basic data for the prevention and control of ARG pollution in agricultural environments.

Anaerobic electrochemical membrane bioreactor effectively mitigates antibiotic resistance genes proliferation under high antibiotic selection pressure.

The spread of antibiotics and antibiotic resistance genes (ARGs) in environments has posed potential threats to public health. Unfortunately, conventional biological wastewater treatment technologies generally show insufficient removal of antibiotics and ARGs. Bioelectrochemical systems, which can effectively degrade refractory organic pollutants via enhancing microbial metabolisms through electrochemical redox reaction, may provide an alternative for the control of antibiotics and ARGs. Herein, an anaerobic electrochemical membrane bioreactor (AnEMBR) was conducted by combining bioelectrochemical system and anaerobic membrane bioreactor to treat antibiotic-containing wastewater. The AnEMBR at open circuit showed stable CH4 production and high removal of COD and chlortetracycline (CTC) in treating 2.5-15mg/L CTC. However, increasing CTC to 45mg/L completely inhibited the methanogenesis of AnEMBR at open circuit. After applying external voltage in AnEMBR, the performances of AnEMBR were significantly improved (e.g., increased CH4 production and CTC removal). Moreover, CTC exposure significantly increased the relative abundances of ARGs in sludge, supernatant, and effluent in AnEMBR at open circuit. Applying voltage greatly attenuated the total relative abundances of ARGs in the supernatant and effluent of AnEMBR compared to those at open circuit. This could be attributed to the enrichment of tetracycline degradation gene tetX, which greatly enhanced the removal of CTC by the AnEMBR and thus reduced the selective pressure of CTC on the microorganisms in supernatant and effluent for ARGs proliferation. These results would provide an effective wastewater treatment technology for treating high-level antibiotic-containing wastewater to mitigate the potential risk of ARGs and antibiotics spread in receiving water body.

Persistent versus transient, and conventional plastic versus biodegradable plastic? —Two key questions about microplastic-water exchange of antibiotic resistance genes

The ubiquitous microplastics (MPs) in water environment play an important role in the dissemination of antibiotic resistance genes (ARGs) due to their exchange between floating MPs and receiving waters. However, whether the ARG exchange is persistent or transient and what are the differences in ARG exchange between conventional plastics and biodegradable plastics are the two key issues to be addressed. In this study, biodegradable PBAT and non-biodegradable PET MPs were chosen to explore the MP-water ARG exchange after the MPs floated to the receiving waters. The results demonstrated that the active exchange of ARGs between MPs and receiving waters occurred, which, however, were transient for most of ARGs. The relative abundance of ARGs both on the MPs and in the waters rapidly decreased to the initial or lower levels within 4 weeks. Approximately 25–50% (ARG subtype number ratio) of studied ARG subtypes were introduced into the receiving waters by MPs, and 35–65% of studied ARG subtypes went through fluctuation in terms of abundance on MPs and in the receiving water. ARGs tended to converge between MPs and the receiving waters with time. Furthermore, the ARG exchange between MPs and waters facilitated horizontal gene transfer (HGT). IntI1 and tnpA05 played the crucial roles in HGT, which was indicated by their correlated change with most ARGs; in contrast, tnpA04 showed the obvious lagging responses. The biodegradable MP of PBAT generally accumulated higher levels of most ARGs including multidrug resistant genes than the non-biodegradable MP of PET. The transient exchange of most ARGs between MPs and water implies that the on-off hitchhiking of ARGs on MPs in aquatic environment may not exert significant influence on ARG transmission. However, compared with the conventional plastics, the biodegradable MPs might pose much higher ARG dissemination risks due to the higher enrichment of ARGs particularly with people's ever-increasingly usage. Enough attention must be paid to this emerging issue.

Effects of thermal hydrolysis on anaerobic digestion and abundance of antibiotic resistance genes during recuperative thickening digestate treatment of sewage sludge

The effects of recuperative thickening (RT) and thermal hydrolysis during anaerobic digestion (AD) on AD performance, microbial communities and reduction of antibiotic resistance genes (ARGs) were investigated. Compared to the results in conventional AD (Phase I), biogas production increased slightly (10%) through RT (Phase II) but increased (63%) via RT with a digestate treatment system (DTS, thermal hydrolysis of thickened digestate) during AD (Phase III). Meanwhile, the three different operating conditions further influenced microbial community structures as well as efficiencies in reducing ARGs: The abundance of hydrolytic bacteria in Phase II and the abundance of acidogenic bacteria in Phase III increased, and 64.3% and 47.4% of the total sum of the relative abundance of ARGs in the substrate were removed in Phases II and III, respectively. Thus, RT-AD with DTS could represent a promising process for improved AD performance, but requires additional refinements to further reduce concentrations of residual ARGs in digestate.

Bioavailability of potentially toxic elements influences antibiotic resistance gene and mobile genetic element abundances in urban and rural soils

Antibiotic resistance genes (ARGs) that can encode resistance traits in bacteria are found across the environment. While it is often difficult to discern their origin, their prevalence and diversity depends on many factors, one of which is their exposure to potentially toxic elements (PTE, i.e., metals and metalloids) in soils. Here, we investigated how ambient ARGs and mobile genetic elements (MGEs) relate to the relative bioavailability of different PTEs (total versus exchangeable and carbonate-bound PTE) in rural and urban soils in northeast England. The average relative abundances of ARGs in rural sites varied over a 3-log range (7.24 × 10−7 to 1.0 × 10−4 genes/16S rRNA), and relative ARG abundances in urban sites varied by four orders of magnitude (1.75 × 10−6 to 2.85 × 10−2 genes/16S rRNA). While beta-lactam and aminoglycoside resistance genes dominated rural and urban sites, respectively, non-specific ARGs, also called multidrug-resistance genes, were significantly more abundant in urban sites (p < 0.05). Urban sites also had higher concentrations of total and exchangeable forms of PTE than rural sites, whereas rural sites were higher in carbonate-bound forms. Significant positive Spearman correlations between PTEs, ARGs and MGEs were apparent, especially with bioavailable PTE fractions and at urban sites. This study found significant positive correlations between ARGs and beryllium (Be), which has not previously been reported. Overall, our results show that PTE bioavailability is important in explaining the relative selection of ARGs in soil settings and must be considered in future co-selection and ARG exposure studies.

Occurrence and distribution of antibiotics and antibiotic resistance genes in water of Liaohe River Basin, China

Surface water environment is an important repository of antibiotics and antibiotic resistance genes (ARGs). It is of great significance to study the occurrence and distribution of antibiotics and ARGs in surface water environment. In this study, the Liaohe River Basin, China was taken as the study area, and the concentrations of antibiotics and ARGs in water were investigated by HPLC-MS/MS and HT-qPCR. The results showed that a total of 53 antibiotics of 6 types were detected in water, and the pollution level was at ND~331.64 ng/L, where PCG was the highest. Totally 164 ARGs and 10 mobile genetic elements (MGEs) were detected in the water, and the absolute abundances were at 2.18 × 104~3.95 × 107 copies/L and 1.82 × 105–3.78 × 107 copies/L, respectively. The multidrug and aminoglycoside were the dominant ARG types. Amoxicillin, erythromycin, anhydroerythromycin and ofloxacin posed certain ecological risks for sensitive aquatic organisms. In spatial distribution, the pollution of antibiotics and ARGs in the Daliao River system was higher than that in the Liaohe River system. There was a significant positive correlation between total concentrations of antibiotics and total relative abundance of ARGs (r = 0.66, p < 0.05). The co-occurrence of multiple antibiotics promoted the pollution and spread of ARGs. In addition, the total relative abundance of MGEs and ARGs showed a significant positive correlation (r = 0.946, p < 0.01), and MGEs played an important role in the occurrence and evolution of ARGs in water.

Subsurface Manure Injection Reduces Surface Transport of Antibiotic Resistance Genes but May Create Antibiotic Resistance Hotspots in Soils.

Compared to surface application, manure subsurface injection reduces surface runoff of nutrients, antibiotic resistant microorganisms, and emerging contaminants. Less is known regarding the impact of both manure application methods on surface transport of antibiotic resistance genes (ARGs) in manure-amended fields. We applied liquid dairy manure to field plots by surface application and subsurface injection and simulated rainfall on the first or seventh day following application. The ARG richness, relative abundance (normalized to 16s rRNA), and ARG profiles in soil and surface runoff were monitored using shotgun metagenomic sequencing. Within 1 day of manure application, compared to unamended soils, soils treated with manure had 32.5-70.5% greater ARG richness and higher relative abundances of sulfonamide (6.5-129%) and tetracycline (752-3766%) resistance genes (p ≤ 0.05). On day 7, soil ARG profiles in the surface-applied plots were similar to, whereas subsurface injection profiles were different from, that of the unamended soils. Forty-six days after manure application, the soil ARG profiles in manure injection slits were 37% more diverse than that of the unamended plots. The abundance of manure-associated ARGs were lower in surface runoff from manure subsurface injected plots and carried a lower resistome risk score in comparison to surface-applied plots. This study demonstrated, for the first time, that although manure subsurface injection reduces ARGs in the runoff, it can create potential long-term hotspots for elevated ARGs within injection slits.

Tracking the influence of antibiotics, antibiotic resistomes, and salinity gradient in modulating microbial community assemblage of surface water and the ecological consequences

The ecological impacts of antibiotics and antibiotic resistance genes (ARGs) on water ecology remain elusive in natural environments. We investigated the influence of antibiotics, ARGs and salinity gradient on the surface water ecosystem. Cefquinome (104.2 ± 43.6 ng/L) and cefminox (16.2 ± 7.50 ng/L) cephalosporins were predominant in all sites. Antibiotic contamination was increased in the estuary ecosystems compared to the freshwater ecosystems by 6%. Bacterial diversity could resist changes in salinity, but the relative abundance of some bacterial genera; Pseudoalteromonas, Glaciecola, norank_f__Arcobacteraceae, and Pseudohongiella was increased in the estuary zone (salinity>0.2%). The eukaryotic composition was increased in the subsaline environments (<0.2%), but the higher salinity in the saline zone inhibited the eukaryotic diversity. The relative abundance of ARGs was significantly higher in the estuary than in freshwater ecosystems, and ARGs interactions and mobile elements (aac(6′)-Ib(aka_aacA4)-01, tetR-02, aacC, intI1, intI-1(clinic), qacEdelta1-01, and strB) were the predominant factors responsible for the ARGs propagation. Antibiotics associated with corresponding and non-corresponding ARGs and potentially created an adverse environment that increased the predation and pathogenicity of the aquatic food web and inhibited the metabolic functions. Surface water are first-line-ecosystems receiving antibiotics and ARGs hence our findings provided vital insights into understanding their ecological consequences on surface water ecosystems.