Behavior Of Antibiotic Resistance Genes Research Articles

Vertical migration and leaching behavior of antibiotic resistance genes in soil during rainfall: Impact by long-term fertilization

The vertical migration and leaching behavior of antibiotic resistance genes (ARGs) during rainfall in soils subjected to long-term fertilization remain largely unclear. In this study, ARGs in vertical profiles (0−60 cm) and leachates from three soils (acidic, neutral, and calcareous) in a long-term (13 years) field fertilization experiment were monitored by high-throughput quantitative PCR after each rainfall event throughout an entire year. The results showed that, compared with unfertilized soils, long-term manure fertilization mainly promoted the vertical migration and leaching of aminoglycoside, beta-lactam, and multidrug resistance genes in the soil profiles. As a result, the annual cumulative loads of ARGs in leachates from the three soils with long-term manure fertilization were significantly increased compared to the controls and were in the order of acidic soil > neutral soil > calcareous soil. SourceTracker analyses revealed that manured soil was the predominant source of the ARGs in the soil leachate samples. Pseudomonas, Anaeromyxobacter, IMCC26256, and MND1 were identified as the dominant potential hosts responsible for the vertical migration and leaching of ARGs in the three soils. PiecewiseSEM analysis further showed that long-term manure fertilization affected the vertical migration of ARGs during rainfall mainly by altering soil properties (i.e., pH, soil organic carbon, and sand). Our results suggest that the ARGs in soils with long-term manure fertilization are a significant potential source of ARG pollution in groundwater, and the measures should be taken to mitigate the vertical migration and leaching of ARGs during rainfall.

Earthworm gut digestion drives the transfer behavior of antibiotic resistance genes in layers of extracellular polymeric substances during vermicomposting of dewatered sludge

Gut digestion by earthworms (GDE) is a crucial step in vermicomposting, affecting the fate of antibiotic resistance genes (ARGs) in vermicompost sludge. The extracellular polymeric substance (EPS) matrix of sludge is an important space for ARG transfer. However, the effect of GDE on EPS-associated ARGs remains unclear. Therefore, this study explored the role of GDE in driving the transfer of ARGs within different EPS layers in sludge. For this, the changes in intracellular ARGs and EPS-associated ARGs in sludge were analyzed after 5 days of the GDE process. The results showed that after the GDE process, both nitrate and dissolved organic carbon significantly increased in all EPS layers of sludge, while the proteins and polysaccharides only enhanced in soluble and loosely bound EPS of sludge. In addition, a 7.0% decrease in bacterial diversity was recorded after the GDE process, with a functional bacterial community structure emerging. Moreover, the absolute abundance of total ARGs and mobile genetic elements decreased by 90.71% and 61.83%, respectively, after the GDE process. Intracellular ARGs decreased by 92.1%, while EPS-associated ARGs increased by 4.9%, indicative of intracellular ARG translocation into the EPS during the GDE process. Notably, the ARGs exhibited significant enrichment in both the soluble and loosely bound EPS, whereas they were reduced in the tightly bound EPS. The structural equation modeling revealed that the GDE process effectively mitigated the ARG dissemination risk by modulating both the EPS structure and microenvironment, with the organic structure representing a primary factor influencing ARGs in the EPS.

Transfer, elimination and accumulation of antibiotic resistance genes in decentralized household wastewater treatment facility treating total wastewater from residential complex

The fate and behavior of antibiotic resistance genes (ARGs) in decentralized household wastewater treatment facilities (DHWWTFs) are unclear. In this study, targeting on a representative DHWWTF that receive all wastewater from a residential complex having 150 households, the transfer, elimination and accumulation of tetG, tetM, sul1, sul2 and intl1 were quantitively studied through real-time PCR-based quantification, mass balance evaluation and the existing state analysis based on size fractionation. Significant abundance changes of the genes were observed in involved biological reactions and the sedimentation process due to microbial growth and decomposition as well as the accumulation of the genes to sludge. tetG and sul1 increased in their fluxes against respective input in the influent. Although substantial portions of the increased genes were found in excess sludge compared to the flux of genes in the influent, those remaining in the discharge were still high, with an average about 3.4 × 1014 copies/d. The abundance of all four genes (tetG, tetM, sul1and sul2) in both water and sludge phases showed a general trend of reduction as sludge accumulated gradually in its storage tank within two months after desludging. Classification of ARGs based on particle sizes (>250 μm, 125–250 μm, 75–125 μm, 25–75 μm, 3–25 μm, <3 μm) indicated that while the major part of ARGs were distributed in particles with larger sizes (125–250 μm), ARGs in smaller particles (3–25 μm) and free ARGs (<3 μm) still existed, which may pose a greater threat to water environment due to their poor settleability. The results of this study can benefit the optimization of on-site maintenance and operation of decentralized wastewater treatment facility for elimination of the transfer of ARGs.

Response behavior of antibiotic resistance genes and human pathogens to slope gradient and position: An environmental risk analysis in sloping cultivated land

Soils, especially in farmlands, are key media for the transmission of antibiotic resistance genes (ARGs) and their hosts from the environment to humans. Sloping farmland is an important agricultural resource, but there lack of studies on the fate and risk of ARGs in sloping land. Also, the behavior and drivers of ARGs in response to slope gradient and position are unclear. Here, metagenomics was used to investigate the profiles of ARGs, mobile genetic elements, and microbial communities in soils from lands of five slope gradients (5°, 10°, 15°, 20°, and 25°) with two slope positions (uphill and downhill). Results showed that while the abundance (except 15°) and diversity (except 20°) of ARGs increased as the slope gradient increased, the diversity of ARGs with health risk, especially the high-risk ones, decreased. For slope positions, abundant and diverse ARGs were more likely to accumulate at downhill. Furthermore, 52 bacterial genera and 12 human pathogenic bacteria (HPB) species were identified as the potential hosts for ARGs with high risk, and abundant HPB species were also detected in the soils with low gradients at downhill. Moreover, the structural equation model analysis revealed that the slope gradient and the slope position have both direct and indirect effects on the abundance of ARGs. Further correlation analysis revealed that the slope gradient has a positive effect (p < 0.05) on nitrite nitrogen in the soils. Also, the slope position has a negative effect (p < 0.05) on total phosphorus and microbial nitrogen, while positively affected (p < 0.05) on particulate nitrogen and microbial carbon, which were the key factors driving the behavior of ARGs. Overall, this study provided comprehensive information on ARGs with health risks and their potential pathogenic hosts in sloping farmland. It can be important for controlling antibiotic resistance transmission and be consistent with the One Health framework.

Long-term influence of chloroxylenol on anaerobic microbial community: Performance, microbial interaction, and antibiotic resistance gene behaviors

The use of antibacterial and disinfection products is increasing in recent years. Para-chloro-meta-xylenol (PCMX), a widely used antimicrobial agent, has been detected in various environments. Herein, the impacts of PCMX with long-term exposure on anaerobic sequencing batch reactors were investigated. The high concentration (50 mg/L, GH group) PCMX severely inhibited the nutrient removal process, and the low concentration group (0.5 mg/L, GL group) slightly affected the removal efficiency which was recovered after 120 days of adaptation compared to the control group (0 mg/L, GC group). Cell viability tests indicated that PCMX inactivated the microbes. A significant reduction in bacterial α-diversity was observed in the GH but not the GL group. The microbial communities were shifted upon PCMX exposure, among which Olsenella, Novosphingobium, and Saccharibacteria genera incertae Sedis became the predominant genera in the GH groups. Network analyses showed that PCMX significantly reduced the complexity and interactions of the microbial communities, consistent with the negative impacts on bioreactor performance. Real-time PCR analysis indicated that PCMX affected the behavior of antibiotic resistance genes (ARGs), and the relationship between ARGs and bacterial genera gradually became complicated after long-term exposure. Most detected ARGs decreased on Day 60 but increased on Day 120 especially in the GL group, implying the potential risk of environment-relevant concentration of PCMX in the ecosystems. This study provides new insights into the understanding of the impacts and risks of PCMX on wastewater treatment processes.

Cross-media migration behavior of antibiotic resistance genes (ARGs) from municipal wastewater treatment systems (MWTSs): Fugitive characteristics, sharing mechanisms, and aerosolization behavior

The municipal wastewater treatment systems (MWTSs) are the leading enrichment site of antibiotic resistance genes (ARGs), the occurrence of which in sewage and sludge significantly influences the ARGs burden of aerosols. However, the migration behavior and impact factors of ARGs in gas–liquid–solid phase are still unclear. This study collected gas (aerosol), liquid (sewage), and solid (sludge) samples from three MWTSs to explore the cross-media transport behavior of ARGs. The results showed that the main ARGs detected in the solid–gas-liquid phase were consistent, constituting the central antibiotic resistance system of MWTSs. Multidrug resistance genes dominated cross-media transmission (average relative abundance is 42.01 %). Aminocoumarin, fluoroquinolone, and aminoglycoside resistance genes (aerosolization index of 1.260, 1.329, and 1.609, respectively) were prone to migrating from the liquid to gas phase, resulting in long-distance transmission. Environmental factors (mainly temperature and wind speed), water quality index (mainly COD), and heavy metals may be the key factors affecting the trans-media migration of ARGs between the liquid, gas, and solid phase. Based on partial least squares path modeling (PLS-PM), the migration of ARGs in gas phase is primarily influenced by ARGs' aerosolization potential in liquid and solid phase, while heavy metals indirectly influences almost all categories of ARGs. Impact factors aggravated the migration of ARGs in MWTSs through co-selection pressure. This study clarified the key pathways and impact factors that form the cross-media migration behavior of ARGs, which can more specifically control ARGs pollution from different media.

Effects of micron-scale zero valent iron on behaviors of antibiotic resistance genes and pathogens in thermophilic anaerobic digestion of waste activated sludge

This work investigated the metagenomics-based behavior and risk of antibiotic resistance genes (ARGs), and their potential hosts during thermophilic anaerobic digestion (TAD) of waste activated sludge, enhanced by micron-scale zero valent iron (mZVI). Tests were conducted with 0, 25, 100, and 250 mg mZVI/g total solids (TS). Results showed that up to 7.3% and 4.8% decrease in ARGs’ abundance and diversity, respectively, were achieved with 100 mg mZVI/g TS. At these conditions, ARGs with health risk in abundance and human pathogenic bacteria (HPB) diversity were also decreased by 8.3% and 3.6%, respectively. Additionally, mZVI reduced abundance of 72 potential pathogenic supercarriers for ARGs with high health risk by 2.5%, 5.0%, and 6.1%, as its dosage increased. Overall, mZVI, especially at 100 mg/g TS, can mitigate antibiotic resistance risk in TAD. These findings are important for better understanding risks of ARGs and their pathogenic hosts in ZVI-enhanced TAD of solid wastes.

Effects of particle size and pyrolytic temperature of biochar on the transformation behavior of antibiotic resistance genes

Rampant use of antibiotics has caused a rapid dissemination of antibiotic resistance genes (ARGs) in environment, posing great threats to ecosystems and human health. Applying biochar (BC) in natural systems to combat the spread of ARGs arises as an attention-grabbing solution. Unfortunately, the effectiveness of BC is still unmanageable due to the incomprehensive knowledge over correlations between BC properties and extracellular ARGs transformation. To pinpoint the crucial factors, we primarily explored transformation behaviors of plasmid-mediated ARGs exposed to BC (in suspensions or extraction solutions), adsorption capacities of ARGs on BC, and growth inhibition of E. coli imposed by BC. Specifically, the effects of BC properties including particle size (large-particulate 150 μm and colloidal 0.45–2 μm) and pyrolytic temperature (300, 400, 500, 600, and 700 °C) on the ARGs transformation were emphasized. Results showed that both large-particulate BC and colloidal BC, irrespective of their pyrolytic temperature, would induce significant inhibitory effects on the ARGs transformation, while the BC extraction solutions showed little effect except BC pyrolyzed at 300 °C. Correlation analysis uncovered that the inhibition effect of BC on ARGs transformation was tightly correlated with its adsorption capacity towards plasmid. Accordingly, greater inhibitory effects from those BCs with higher pyrolytic temperatures and smaller particle sizes mainly originated from their greater adsorption capacities. Intriguingly, E. coli was unable to ingest the plasmid adsorbed on BC, which led to ARGs blocked outside the cell membrane, although this inhibitory effect was partially affected by survival inhibition of BC on E. coli. Particularly, significant plasmid aggregation could occur in the extraction solution of large-particulate BC pyrolyzed at 300 °C, leading to a significant inhibition of ARGs transformation. Overall, our findings complete the insufficient understanding over the effects of BC on ARGs transformation behavior, and potentially provide new insights to scientific communities in mitigating ARGs spreading.

The influence of humic substances on environmental behavior of antibiotic resistance genes: A mini-review

The global antibiotic resistance pollution caused by antibiotic resistance genes (ARGs) has led to serious harm to the ecological environment and human beings. As a type of organic matter that commonly exists in the environment, humic substances (HS) may affect the fate of contaminants. Based on the high possibility of ARGs coexisting with HS in the environment, it is necessary to understand the influence of HS on environmental behavior of ARGs. This article reviews the impact of HS on environmental behavior of ARGs, and finds that the topic has received increasing attention by scientometric analysis using CiteSpace software. The environmental behavior of ARGs is not only affected by HS and its by-products, but also frequently implicated through antibiotics, heavy metals and particles in various environments, such as wastewater, activated sludge and soil. The main mechanisms by which HS affects the ARGs evolution include electrostatic interactions, hydrophobic interactions, hydrogen bonding, non-specific van der Waals interactions, and microbial community alterations. Finally, future research directions are proposed to help refine the theoretical basis for the interaction between HS and ARGs, and can contribute to the prevention and control of ARGs pollution in practical applications.

Evolution of antibiotic resistance genes and bacterial community during erythromycin fermentation residue composting

The removal efficiency of antibiotic resistance genes (ARGs) is the biggest challenge for the treatment of erythromycin fermentation residue (EFR). In the current research, 0% (control), 10% (T1), and 30% (T2) spray-dried EFR were composted with bulking materials, consisting of cattle manure and maize straw, for 30 days. Environmental factors and bacterial community on the behaviors of ARGs were further investigated. Apart from the high levels of erythromycin, the electrical conductivities were also increased by 66.7% and 291.7% in the samples of T1 and T2, respectively. After 30 days of composting, total ARGs in the samples of control were decreased by 78.1%–91.2%, but those of T1 and T2 were increased 14.5–16.7- and 38.5–68.7-fold. ARGs related to ribosomal protection (erm) dominated the samples of T1 and T2 at D 13 and 30, especially that ermF accounted for more than 80% of the total ARGs. Furthermore, the results of bacterial community revealed that EFR promoted the growth of Proteobacteria and Bacteroidetes, but inhibited that of Actinobacteria, Verrucomicrobia and Chloroflexi. Network analysis revealed that the enriched ARGs had strong correlation with seven bacterial genera, including Halomonas, Oceanobacillus, and Alcaligenes, most of which are halotolerant. Above all, erythromycin combined with high salinity can have synergistic effect on the enrichment of ARGs and their hosts.

Effects of Chinese medicine herbal residues on antibiotic resistance genes and the bacterial community in chicken manure composting.

The use of livestock manure is an important way for antibiotic resistance genes (ARGs) to enter the environment, and composting is an effective method for removing ARGs from livestock manure. In this study, different volume ratios of Chinese medicinal herbal residues (CMHRs) were added to laboratory-scale chicken manure composting to evaluate their effects, if any, on the behavior of ARGs, mobile genetic elements (MGEs), and the bacterial community. At the end of the composting period, the composition of the microbial community changed. Firmicutes decreased and Bacteroidetes increased. The most striking effect was that the relative abundance of the 21 ARGs and 5 MGEs detected decreased by varying degrees in the different treatments (except for sulI and intI1). The removal rate of the ARGs increased with the increased addition of CMHRs. The correlations between transferase genes (tnpA and tnpA-02) and ARGs were significant (p < 0.05); therefore, transposons play an important role in the horizontal gene transfer of ARGs in chicken manure. The results imply that CMHRs would be an effective bulking agent for the removal of ARGs from chicken manure composting.

Response behavior of antibiotic resistance genes to zinc oxide nanoparticles in cattle manure thermophilic anaerobic digestion process: A metagenomic analysis

This work investigated the metagenomics-based behavior of antibiotic resistance genes (ARGs) during cattle manure anaerobic digestion with zinc oxide nanoparticles (ZnO NPs) that are commonly used as animal feed additives. The 6.6% decrease in total ARGs abundance while remained unchanged ARGs diversity with ZnO NPs (5 mg/g total solid), suggested ZnO NPs may mitigate ARGs risk by abundance. Also, ZnO NPs affected ARGs with mechanisms specifically of antibiotic inactivation and antibiotic target change, and declined potential hosts’ abundance (bacterial genus Ruminiclostridium, Riminococcus, and Paenibacillus) which mainly contributed to the decreased ARGs’ abundance. Besides, microbial chemotaxis decreased by 17% with ZnO NPs compared to that without nanoparticles indicated a depression on potential hosts, who could develop the mechanism to adapt to altered digestion conditions, which probably inhibited the ARGs’ propagation. These findings are important to promote understanding of the potential ARGs risks in treatments of livestock wastes containing animal feed additives.

Effects of Particle Size and Pyrolysis Temperature of Biochar on the Transformation Behavior of Antibiotic Resistance Genes

Effects of Particle Size and Pyrolysis Temperature of Biochar on the Transformation Behavior of Antibiotic Resistance Genes

Behaviors of antibiotic resistance genes (ARGs) and metal resistance genes (MRGs) during the pilot-scale biophysical drying treatment of sewage sludge: Reduction of ARGs and enrichment of MRGs

Biophysical drying (BPD) is one of the best alternatives for reducing the moisture content from sewage sludge by utilizing biological heat from aerobic reactions. However, the fate of emerging pollutants during BPD process is largely unknown. In this study, the fates of antibiotic resistance genes (ARGs) and metal resistance genes (MRGs) were investigated during a pilot-scale BPD treatment of sewage sludge. A total of 20 types (388 subtypes) of ARGs and 16 types (364 subtypes) of MRGs were detected by metagenomic sequencing and annotation. The total abundance of ARGs decreased from 1.78 ± 0.13 copies/16S rRNA to 0.55 ± 0.01 copies/16S rRNA while the total abundance of MRGs increased from 3.81 ± 0.01 copies/16S rRNA to 6.30 ± 0.02 copies/16S rRNA, showing the distinct behaviors of ARGs and MRGs during BPD process. The ARGs were effectively reduced during the mesophilic and thermophilic stages of BPD process and the reduction of ARGs fitted the first-order kinetic model (p < 0.01). Microbial community analysis showed that the abundance of potential pathogens also decreased during BPD process. On the contrary, the abundances of most MRG subtypes (78.3%) were enriched during BPD process with up to 122-fold change, implying the potential threats for the end product of BPD process. These results together indicate that although the ARGs and potential pathogens can be effectively reduced during BPD process, the safety for the end product still needs to be considered due to the enrichment of MRGs.

The behavior of antibiotic-resistance genes and their relationships with the bacterial community and heavy metals during sewage sludge composting

The main contributors to antibiotic resistance genes (ARGs) profiles during the composting process under the addition of biochar (BC) and peat (PT) were not fully explored. This study investigated the influence of BC and PT amendment on ARGs fate, the bacterial community and heavy metals in sewage sludge compost. Compared to control, BC and PT declined the total abundances of ARGs by 17.6% and 43.0% after composting. Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria were the dominant phyla across the composting process, among which, members of Firmicutes (mainly Bacillaceae) were the potential hosts for multiple ARGs. BC and PT addition declined the abundance of bacterial pathogens such as Bacteroides and Pseudomonas. Besides, the concentrations of copper (Cu), zinc (Zn) and lead (Pb) were less in BC and PT treatments than control on day 40, and these metals displayed significant positive correlations to sul1 and intI1. Furthermore, variation partitioning analysis (VPA) revealed that the bacterial community exhibited the most contribution to the ARG patterns, as much as 34.0%, followed by heavy metals (10.8%) and intI1 gene (1.5%). These results suggested that biochar and peat can reduce the risks of ARGs in sewage sludge mainly by sharping the bacterial composition.

Behavior of antibiotic resistance genes in a wastewater treatment plant with different upgrading processes

Wastewater treatment plants (WWTPs) in China have been upgraded or renovated with a variety of emerging processes, but a comprehensive understanding of the behavior of antibiotic resistance genes (ARGs) in these WWTPs is still lacking. Here, the distribution of ARGs and bacterial community were investigated in a wastewater treatment plant with upgrading processes (WWTP-UP). 238 unique ARGs were detected in all samples. During the study period, the average ARGs concentration decreased by 98.4% along the entire treatment process. The removal efficiency of A2/O-membrane bioreactor (MBR) process was significantly higher than that of A2/O-high efficiency flocculent settling/cloth media filter (HEFS/CMF) process (p < 0.05), which corresponded to 3.5 and 2.1 log values on average, respectively. Notably, 35 ARGs and 14 mobile genetic elements (MGEs) were persistent in all samples. Based on the co-occurrence pattern revealed by network analysis, persistent ARGs possibly spread through the transfer of persistent MGEs among persistent bacteria. Using multiple linear regression analysis, we obtained 3 to 5 possible indicators for major ARG types, which might be served to evaluate the general distribution of ARGs or even predict the abundance of different ARG types. Our findings provide new insights into the impacts of upgrading process on ARGs and highlight the need for better strategies to improve ARGs elimination in WWTPs.

Horizontal gene transfer is a key determinant of antibiotic resistance genes profiles during chicken manure composting with the addition of biochar and zeolite

Livestock manure is an important reservoir of antibiotic resistance genes (ARGs). Biochar and zeolite are commonly used to improve the quality of compost, however, little is known about the impacts of these additives on the fate of ARGs during composting and the underlying mechanisms involved. In this study, zeolite (ZL), biochar (BC), or zeolite and biochar (ZB) simultaneously were added to chicken manure compost to evaluate their effects on the ARGs patterns. After composting, the abundance of ARGs reduced by 92.6% in control, while the reductions were 95.9%, 98.7% and 98.2% for ZL, BC, ZB, respectively. Co-occurrence network analysis indicated that the potential hosts for most ARGs were predominantly affiliated to Firmicutes such as Lactobacillus and Fastidiosipila. Furthermore, shifts in ARGs were significantly correlated with class 1 integrase gene (intI1), and structural equation models further revealed that intI1 gene contributed most (standardized total effect 0.92) to the ARGs-removal, which was trigged by horizontal gene transfer. Together these results suggest that the addition of zeolite and biochar mitigate the accumulation and spread of ARGs during composting, and the crucial role of horizontal gene transfer (HGT) on the behaviors of ARGs should pay more attention to in the future.

Response of antibiotic resistance genes in constructed wetlands during treatment of livestock wastewater with different exogenous inducers: Antibiotic and antibiotic-resistant bacteria

This work aimed to study the behavior of antibiotic resistance genes (ARGs) in constructed wetlands with different exogenous inducers additions (oxytetracycline and its resistant bacteria) by high-throughput quantitative polymerase chain reaction. Results indicated that constructed wetlands have the potential to reduce ARGs relative abundances in wastewater, and the total ARGs removal efficiency could exceed 60%. ARGs profile in the effluent differed from that in the influent, and that did not directly reflect the export of dominant ARGs in wetland biofilms. Meanwhile, the highest levels of detected numbers and relative abundances of ARGs were 43 and 3.35 × 10−1 for control system and 44 and 6.40 × 10−1 for treatment system, respectively, which meant that ARGs generation in wetlands were inevitable, and antibiotic and antibiotic-resistant bacteria from wastewater could indeed promote ARGs abundance in the system. Compared to the single roles of inducers, their synergistic role had a more significant influence on ARGs relative abundance.

RETRACTED: The behavior of antibiotic resistance genes and their associations with bacterial community during poultry manure composting

In this paper, the influence of clay amendment on antibiotic resistance genes (ARGs) fate and their relation with bacterial diversity in poultry manure (PM) composting was evaluated. The results confirmed that 4% clay amendment was significantly reduce the relative abundances (RAs) 94% of ARGs, while higher dosage of clay blended treatments (T5 and T6) composts were observed relatively very high percentage of ARGs as compared to other clay blended treatments. Clay addition play potential role on the occurrence and distribution of ARGs during the composting by altering the overall RAs of Firmicutes and Proteobacteria, and there host bacterial species. The heat map correlation figure also indicated that reduction of ARGs was significantly associated with the alteration of microbial dynamics and physicochemical parameters during the composting. Therefore, the addition of 4% clay in to PM composting system could efficiently mitigate the greater concentration of ARGs in PM compost and also environmental risk associated with its application.

Amendment soil with biochar to control antibiotic resistance genes under unconventional water resources irrigation: Proceed with caution

The spread of antibiotic resistance genes (ARGs) has become a cause for serious concern because of its potential risk to public health. The use of unconventional water resources (e.g., reclaimed water or piggery wastewater) in agriculture to relieve groundwater shortages may result in an accumulation of ARGs in soil. Biochar addition has been proven to be a beneficial method to alleviate the pollution of ARGs in manure-amended soil. However, the role of biochar on ARGs in soil-plant systems repeatedly irrigated with unconventional water resources is unknown. Under reclaimed water or piggery wastewater irrigation, rhizobox experiments using maize plants in soil amended with biochar were conducted to investigate the variation of typical ARGs (tet and sul genes) in soil-plant systems during a 60-day cultivation, and ARGs was characterized by high-throughput qPCR with a 48 (assays) × 108 (samples) array. Only piggery wastewater irrigation significantly increased the abundance of ARGs in rhizosphere and bulk soils and root endophytes. Following 30-day cultivation, the abundance of ARGs in soil was significantly lower due to biochar addition. However, by day 60, the abundance of ARGs in soil supplemented with biochar was significantly higher than in the control soils. Antibiotics, bio-available heavy metals, nutrients, bacterial community, and mobile gene elements (MGEs) were detected and analyzed to find factors shaping ARGs dynamics. The behavior of ARGs were associated with antibiotics but not with bio-available heavy metals. The correlation between ARGs and available phosphorus was stronger than that of ARGs with total phosphorus. MGEs had good relationship with ARGs, and MGEs shifts contributed most to ARGs variation in soil and root samples. In summary, this study provides insights into potential options for biochar use in agricultural activities.