Non-antibiotic components of feed additives can enter farmland soils via livestock manure and accumulate persistently in agroecosystems, presenting potential environmental risks. We established soil microcosms, integrated metagenomes with viromes, and applied a contig-based horizontal gene transfer (HGT)-resolution pipeline to partition vector-level contributions, to assess how saccharin, copper, and their co-contamination affect soil gene flow and health risk. Results indicate divergent vector responses under additive stress: phage-host associations increased under saccharin (82 pairs vs. control 29 pairs), whereas copper strengthened plasmid-host associations. With saccharin, phage nucleotide diversity rose while synonymous nucleotide diversity declined, consistent with stronger purifying selection atop enhanced mutation supply, whereas copper increased lysogeny. Saccharin significantly elevated HGT frequency (∼50% increase), expanded donor-recipient phylogenetic span (class-level P < 0.05), and raised the phage-mediated share (∼100% increase). Copper primarily modestly increased the plasmid-mediated contribution (Cu 2.7%, HS 1.9%). Two-factor analyses revealed a significant antagonistic interaction between saccharin and copper, reducing overall HGT across taxonomic ranks under co-exposure. Although total ARG abundance did not change significantly, the health-risk index increased under saccharin, driven by enhanced ARG-MGE co-occurrence. Under co-contamination, auxiliary metabolic genes were enriched, suggesting phage-conferred metabolic empowerment that mitigates stress, partly explaining the antagonism. Altogether, our findings reveal that feed additives reshape vector-specific gene mobility and ARG risk, and they underpin a three-tiered risk-assessment framework that progresses from mere abundance to network-structured mobility and finally to mobility drivers incorporating phylogenetic transfer distance, offering a more mechanistic basis for soil-health management.

The environmental accumulation of non-antibiotic pharmaceuticals is an emerging driver of antibiotic resistance. While individual compounds are known to shape the soil resistome, and contaminant diversity also plays a role, the impact of pharmaceutical diversity on the gut resistome of soil invertebrates remains unclear. Here, we combined metagenomics and metaproteomics to examine the collembolan gut and soil resistome across a gradient of pharmaceutical diversity under diurnal warming. Increasing pharmaceutical diversity at a constant total concentration significantly enriched antibiotic resistance genes (ARGs) in the gut microbiome, with no comparable effect in surrounding soils. This enrichment was mainly driven by multidrug resistance associated with efflux activity and biofilm-related processes, accompanied by increases in ARG-carrying taxa such as Gordonia and Ochrobactrum. Notably, Ochrobactrum encoded biofilm-related aryl polyene pathways. In vitro experiments confirmed that biofilm formation promotes resistance through coordinated cellular responses. Metaproteomic data indicated that Ochrobactrum initiates early biofilm formation by recruiting extracellular matrix producers such as Bacillus and Pseudomonas. Diurnal warming modulated these responses, indicating an interaction between chemical diversity and climate stress. These findings identify pharmaceutical diversity as an independent driver of ARG enrichment in host-associated microbiomes and establish chemical complexity as a key factor in assessing the ecological risks of pharmaceutical pollution.

The global spread of antibiotic resistance genes (ARGs) poses critical threats to public health, driven by their ability to persist and migrate across interconnected environmental compartments (e.g. soil and water). While antibiotics can exert strong selective pressure on ARGs, emerging evidence suggests that non-antibiotic pollutants, including artificial sweeteners, may play a significant role in ARG dissemination. However, the effects of chemical co-exposure on ARG dynamics within the soil-water continuum remain poorly understood. In this study, we conducted microcosm experiments to investigate ARG temporal dynamics in the soil-water continuum under co-exposure of antibiotic enrofloxacin and artificial sweetener cyclamate. Co-exposure significantly increased ARG diversity, with a particular enrichment of multidrug resistance genes. Exposure to these chemicals induced notable shifts in bacterial diversity and composition. Enrofloxacin had a greater impact on soil resistomes, while cyclamate primarily affected the resistomes of overlying water. This differential effect was attributed to distinct water solubility profiles of enrofloxacin and cyclamate. Our results demonstrated that frequent migration of ARGs between soil and overlying water, with co-exposure to enrofloxacin and cyclamate enhancing ARG transfer from soil to overlying water. Strong correlations between ARGs, bacterial communities, and chemical concentrations indicated that enrofloxacin and cyclamate are key drivers of ARG temporal dynamics. Metagenomic results further revealed potential mechanisms through which enrofloxacin and cyclamate exposure affect ARGs by affecting bacterial amino acid synthesis and phosphatase metabolism. These findings underscore the role of chemical co-exposure in promoting ARG migration within the soil-water continuum, providing new insights into ARG dissemination in complex multi-media ecosystems.

Global environmental stressors are intimately tied to both planetary and human health. In natural environments, complex chemical stressor systems comprising multiple concurrent stressors are pervasive yet often neglected. A key challenge lies in developing risk assessment methods that do not depend on knowing the exact identity of each individual stressor. Here, an anonymized complex system was constructed through a highly controlled soil microcosm experiment to examine how increasing stressor diversity impairs the life functions of Folsomia candida (F.candida). It shows that increasing stressor diversity notably reduces collembolan survival and reproductive output. This population-level decline corresponds to substantial disruptions in the transcriptome, impacting a wide array of biological pathways. We systematically focus on the high-dimensional, multi-directional shifts in transcriptional profiles and further demonstrate that low-dimensional data analysis can effectively capture this degenerative trend. Crucially, by applying a dual filter of “monotonic change + statistical significance”, a consistent set of differentially expressed genes (DEGs) that function as effective biomarkers were identified under this complex stress regime. Our work not only deepens the mechanistic understanding of how soil fauna responds to chemically complex environments but also offers a practical experimental and analytical process for quantifying anonymous, multi-stressor pollution scenarios. This approach highlights the value of monotonic endpoints in ecological risk assessment of complex chemical mixtures.

The role of bacteria in external niches regulating cadmium (Cd(II)) in plant tissues remains unclear. We explored Cd(II) profiles and identified bacterial contributors among phyllosphere, rhizoplane, and rhizosphere of four Zizania latifolia varieties through integrated metagenomic and chemical analyses. Zizania latifolia accumulated Cd(II) in leaves (0.06-0.77 mg/kg), roots (0.73-1.57 mg/kg), and rhizosphere (0.43-3.15 mg/kg), respectively. The highest enrichment coefficient (leaf-Cd(II)/soil-Cd(II)) was observed in Genotype 3 (0.6). Among top 10 genus-level bacteria, Enterococcus in phyllosphere, Streptomyces and Dechloromonas in rhizoplane, and Bradyrhizobium, Pseudolabrys, Mycobacterium, and Dechloromonas in rhizosphere were significantly related to Cd(II). Enterococcus adsorbed Cd(II) by extracellular polysaccharides and precipitated Cd(II) sulfide. Rhizoplane and rhizosphere bacteria absorbed Cd(II) by cell-surface functional groups, and fixed Cd(II) through synthesizing polyphosphate and driving Fe (II) oxidation. Additionally, 64.4%-80% of bacteria were shared between rhizoplane and rhizosphere, 5.5%-6.9% between rhizoplane and phyllosphere, and 4.4%-6.1% between rhizosphere and phyllosphere. Metagenomic analysis indicated that Cd(II) disturbed bacterial secretion system and amino acid metabolic pathways. These findings provided comprehensive insights into interrelationships between Cd(II) and bacteria in leaves, roots, and rhizosphere of Zizania latifolia, offering valuable foundations for developing targeted strategies to mitigate Cd(II) accumulation in aquatic vegetables.

The escalating dissemination of antibiotic resistance genes (ARGs) in lake ecosystems has drawn substantial attention regarding their potential risks to public health. However, the spatiotemporal patterns and the driving mechanisms of ARGs within lake ecosystems under anthropogenic activities remain incompletely understood. Here, 132 sediment and 132 water samples were collected from the production and living, tourism, and natural areas of Baiyangdian Lake during the dry and wet seasons. The findings showed that the total ARGs abundance in sediments during the dry season was 4.37 to 19.05 times higher than that in the wet season. Conversely, the total ARGs abundance in water was 1.97 to 12.51 times greater in the wet season as compared to the dry season. Notably, the production and living area and the tourism area exhibited significantly higher ARGs abundances in both sediments and water than the natural area. Specifically, 23 and 11 types of potential pathogenic bacteria were identified in sediments and water, respectively, with the abundance of animal-origin pathogenic bacteria reaching up to 4.55%. Network analysis revealed that dominant phyla, including Proteobacteria, Bacteroidota, and Chloroflexi, were potential major hosts of ARGs. Additionally, the intI1 gene significantly correlated with ARGs, indicating its crucial role in the dissemination of ARGs. PLS-PM further demonstrated that biotic factors (intI1 gene, bacterial abundance) and abiotic factors (TN, TP) were crucial for ARG spatiotemporal distribution. Overall, our work provided insights into the impacts of anthropogenic activities on ARGs and pinpointed potential high-risk areas, providing crucial implications for the management of ARGs contamination.

Mollisols are among the world’s most fertile agricultural soils, but long-term intensive management, including widespread pesticide use, has introduced chronic pressures to belowground communities. However, the occurrence of multi-class pesticide residues and their impacts on multi-trophic biodiversity in black-soil croplands remain poorly understood. Here, we surveyed 111 topsoil samples from Northeast China’s Mollisol croplands to quantify 22 pesticides, assess ecological risks, and characterize multi-trophic biodiversity (bacteria, fungi, protists, and metazoa) using DNA metabarcoding. Pesticide residues were widely detected, with high frequencies observed for herbicides such as nicosulfuron (79.3%), atrazine (52.3%), and acetochlor (49.5%); insecticides dominated by neonicotinoids (e.g., imidacloprid 70.3%, clothianidin 64.9%) and pymetrozine (49.5%); and fungicides including chloroneb (53.2%). Risk quotient screening identified atrazine, clothianidin, and difenoconazole as compounds of elevated potential concern. At the mixture level, 85.6% of samples exceeded the cumulative risk threshold, although cumulative risk was generally dominated by one or a few compounds, with multi-compound contributions observed in only 20.7% of samples. Biodiversity associations were strongly trophic-level specific. Pesticide residues showed the strongest negative association with metazoan α-diversity, accounting for approximately 29% of the independently explained model variance, whereas bacterial diversity was more closely associated with soil properties and fungal diversity with cropping systems. However, protistan diversity showed weak explanatory patterns within the current framework. Overall, pesticide residues were widespread in Northeast China’s Mollisol croplands, with potential ecological concern largely attributable to a few high-risk compounds. The pronounced association with metazoan diversity supports incorporating higher-trophic soil fauna and compound-specific mixture risk into future soil-health assessments.

The global spread of antibiotic resistance genes (ARGs) has been a hotspot of international concern. Livestock manure application and wastewater irrigation are crucial pathways through which ARGs are dispersed into the receiving environments. However, the current understanding of the vertical migration of ARGs in soils amended with manure and irrigated with wastewater remains inadequate. This study explored the vertical distribution of ARGs across 0-100 cm soil profile under swine manure amendment and swine wastewater irrigation, based on a five-year field experiment including swine manure application and swine wastewater irrigation treatment (SM + SW), swine wastewater irrigation (SW) treatment and deep groundwater irrigation treatment (Control). Compared with the Control treatment, the abundance of ARGs in the SM + SW treatment exhibited a 2.06 to 31.36-fold increase, while in the SW treatment, the increase was 0.99 to 5.86-fold. The abundance of ARGs displayed a decreasing trend with the increase in soil depth. ARGs were predominantly distributed in the 0-25 cm soil layers. Notably, the abundance of ARGs in the 75-100 cm still reached 105 copies & sdot;g- 1 (dry soil), suggesting the potential risk of ARGs leaching into deeper soil strata. High-throughput sequencing results illustrated that soil bacterial diversity and community assembly mechanisms were mainly affected by soil depth. Furthermore, a partial least squares path model indicated that soil depth and soil properties predominantly influence the vertical distribution of ARGs through indirect effect, whereas bacterial abundance and intI1 gene mainly act through direct effects. This study holds crucial importance for evaluating environmental health risks and optimizing agricultural management strategies regarding ARGs pollution.

BACKGROUND:Antimicrobial resistance (AMR) emerges primarily through antibiotic exposure and the resulting selection pressure, but climate change is likely to accelerate the dissemination of AMR, particularly for zoonotic diseases, such as those caused by Salmonella. However, the link between climatic factors and antimicrobial resistance genes (ARGs) carried by Salmonella remains poorly characterised. This longitudinal ecological study aimed to link climate change to ARGs using multiple regression models. METHODS:We analysed a comprehensive dataset of 488 232 Salmonella genomes and multiple potential predictors from 139 countries or regions over the period 1940-2023. Robustness was verified via Tobit and generalised additive models. Climate-related changes of average ARG abundance in Salmonella were quantified through counterfactual scenarios. Future ARG trends were projected to 2100 using integrated Shared Socioeconomic Pathways (SSPs) with Representative Concentration Pathways scenarios (SSP1-1.9, SSP1-2.6, SSP2-4.5, SSP3-7.0, and SSP5-8.5). FINDINGS:The global average ARG abundance in Salmonella has increased by 38% (0·50 copies per cell) in the time period considered. Multiple regression models revealed that variability in ARGs follows a non-linear quadratic response to temperature and precipitation. Climate change is associated with a 10% (95% CI 5·4-13·3) global rise in the abundance of Salmonella ARGs, with increases observed in 82 (82%) of 100 countries. By 2100, the emergence of ARGs is projected to be further intensified by warming; however, achieving low-emission (SSP1-2.6) targets alongside strengthened antibiotic stewardship programmes could reduce Salmonella ARGs by 24% (95% CI 21-29) as compared with high-emission scenarios (SSP5-8.5). INTERPRETATION:This study provides global evidence linking climate change to ARG dynamics in Salmonella. Warming and shifting precipitation patterns are associated with rising ARG abundance and are projected to further exacerbate AMR risks under high-emission scenarios (SSP2-4.5, SSP3-7.0, and SSP5-8.5). These findings highlight the need to integrate climate considerations into AMR surveillance and stewardship, providing a quantitative basis for climate-informed strategies to restrict future resistance escalation. FUNDING:National Key Research and Development Program of China, the National Natural Science Foundation of China, Zhejiang Provincial Natural Science Foundation of China, and Beijing Municipal Sci-Tech Project on Ecology and Environment.

Antimicrobial resistance is an escalating global threat, with soils serving as reservoirs and conduits for the dissemination of antibiotic resistance genes (ARGs). Pesticide use in agriculture contributes to ARG proliferation, and ~60% of agricultural soils contain multiple pesticide residues. However, how pesticide diversity influences ARG dynamics in active microbial populations (active ARGs) remains unclear. Here, we evaluate the effects of pesticide diversity on active soil ARGs through a long-term field experiment integrating bioorthogonal non-canonical amino acid tagging (BONCAT), fluorescence-activated cell sorting (FACS), and metagenomics. We show that both low and high pesticide diversity significantly increase active ARG abundance relative to untreated control, whereas total ARG levels remain largely unchanged. The underlying mechanisms differ with pesticide diversity. At low diversity, active ARG co-selection via efflux pumps in Acinetobacter baumannii is a prominent mechanism. At high diversity, elevated reactive oxygen species and SOS responses promote horizontal gene transfer of active ARGs, as validated by culture experiments. These findings demonstrate that increasing pesticide diversity accelerates the emergence and dissemination of active ARGs, highlighting the need for integrated pesticide management strategies that consider both application intensity and diversity to mitigate resistance risks under the One Health framework.

Straw incorporation and nitrogen amendment in agricultural soils have been shown to increase the diversity of bacterial communities and antibiotic resistance genes (ARGs). However, the effects of straw types and nitrogen amendment levels on ARG dissemination potential lack genetic evidence. Here, we conducted a metagenomic analysis of 24 agricultural soils amended with wheat or maize straw under a nitrogen fertilization gradient (0, 200, 400, and 600 kya). Our results showed that the incorporation of wheat straw in soils significantly increased the abundance of ARGs and mobile genetic elements (MGEs) compared with maize straw. Moreover, genetic evidence of the coexistence of ARGs and MGEs (distance < 5 000 bp) demonstrated that the dissemination potential of ARGs was significantly greater in wheat than in maize straw-returning soils. Glycopeptide, fluoroquinolone and diaminopyrimidine resistance were the dominant ARGs and were assigned to Pseudomonadota, Actinobacteria and Firmicutes, which were also the predominant bacteria harboring ARG-MGE. Compared with the absence of nitrogen amendment or at 600 kya, nitrogen amendment at 200 and 400 kya increased the ARG dissemination potential in wheat straw-returning soils. The different correlation patterns between the dominant ARGs and the carbon and nitrogen metabolism genes implied that bacteria involved in degrading organic substrates and nitrogen metabolism may have antibiotic resistance ability. This study suggested that wheat straw incorporation and nitrogen fertilization contribute to the spread of ARGs in agricultural soils and should not be neglected.

Large-scale mining of ion-adsorption rare earth elements (REEs) generates acidic mine drainage (AMD) laden with REEs and heavy metals (HMs), yet its cascading impacts on microbial community assembly and antibiotic resistance genes (ARGs) dissemination across multi-matrices remain poorly characterized. By integrating high-throughput sequencing, co-occurrence network analysis, and partial least squares path modeling (PLS-PM), we unraveled that mining-induced geochemical divergence between mining-impacted areas and adjacent watersheds acts as a dominant environmental filter, reshaping the microbial community assembly and ARGs endowment. Key findings were in three areas: (1) Pollution-driven microbial adaptation. Significant shifts in microbial composition fostered "alternative steady states" without altering richness and induced network polarization-simplification in mining water (degree/density reduced by about 50 %) versus complexification in mining soil (degree/connections increased by about 3-fold), with enhanced mutualistic interactions and a 3-fold reduction in keystone species complexity. (2) ARGs risk-abundance decoupling. Mining areas exhibited 2.6-fold higher ARGs health risks than watersheds (p = 0.019), despite comparable abundance levels (p > 0.05), necessitating a paradigm shift from quantitative surveillance to health risks monitoring. (3) Dual ARGs regulation. While acidic REEs co-contamination directly promoted ARGs proliferation via co-selection (pathway coefficient = 0.254), it concurrently mitigated overall ecological risks through host community restructuring and potential horizontal gene transfer suppression (total effects = -0.186). These findings elucidate the ecological trade-offs between microbial adaptive resilience and ARGs dissemination in mining-impacted ecosystems, while establishing a mechanistic framework for optimizing targeted remediation strategies and sustainable resource extraction protocols.

Pathogenic Enterobacteriaceae (PE) pose significant public health threats, particularly with the emergence of antibiotic-resistant isolates that frequently cause foodborne infection outbreaks through the consumption of contaminated leafy vegetables. However, there is currently a lack of comprehensive genomic analysis for PE in leafy vegetables, hindering insights into the distribution patterns and resistance capacity of these PE in diverse agricultural settings around the world. This study addresses this gap by performing a genomic analysis of PE related to leafy vegetables, encompassing 458 publicly available genomes and 49 newly sequenced genomes from our study. The newly sequenced genomes correspond to PE isolates from organic fertilizer, fertilizer-applied soils, and various parts of Brassica rapa grown in fertilizer-applied soils (31 from B. rapa plants, 18 from organic fertilizer and soils), for which we also performed antibiotic susceptibility testing as a supplement. Results showed extensive multidrug resistance among PE isolates (>95 %), and a significant progressive increase of high-risk antibiotic resistance genes (ARGs) from roots to leaves. We established a genome dataset comprising 489 PE isolates (458 public isolates plus 31 B. rapa isolates from our study) from leafy vegetables worldwide and identified Escherichia coli as harboring substantially higher numbers of both high-risk ARGs (mean 9.71 per genome) and virulence factor genes (mean 72.45 per genome) compared to other PE taxa. This study provides a valuable genomic resource for investigating PE contamination in leafy vegetables and underscores the urgent need for enhanced surveillance targeting antibiotic-resistant PE, particularly E. coli, in leafy vegetables to mitigate emerging public health threats.

As widespread toxicants that cause cancer and affect the endocrine system, persistent organic pollutants, such as polycyclic aromatic hydrocarbons, pesticides, and chlorinated solvents, are harmful to the environment and human health. This review aims to highlight biochar-supported microbial systems as a transformative solution for remediating these contaminants, with a particular focus on current challenges and future perspectives. Conventional pollutant-remediation techniques based on physicochemical treatments are expensive and inefficient. Bioremediation technology faces challenges, such as low microbial survival and environmental sensitivity. Biochar-supported microbial systems have become attractive because of their strong adsorption characteristics and microbial degradation. Biochar-supported microbial systems offer promising solutions that combine the superior adsorption capacity of biochar with its microbial degradation capabilities. Biochar produced from pyrolyzed biomass has a porous structure and functional groups that immobilize pollutants and support microbial growth. Recent research demonstrates that integrating nutrient-enriched biochar with symbiotic microbial communities extends their remediation potential to a wider range of pollutants, including persistent organic pollutants. However, challenges, such as long-term microbial viability, biochar aging, and field-scale economic feasibility, remain unresolved. Further research is required to optimize these systems for real-world applications. By addressing these gaps, biochar-microbial remediation can become a sustainable and scalable strategy for environmental rehabilitation, supporting circular economic goals.

Soils are critical reservoirs of antibiotic-resistance genes (ARGs) and antibiotic-resistant bacteria (ARB), serving as interfaces among human, animal, plant, and environmental microbiomes. While many studies have profiled soil ARGs, most rely on DNA-based methods that cannot distinguish total from metabolically active ARB, limiting risk assessment and mitigation. This review outlines soil ARG sources, their mobility, and potential transmission to plants and the food chain. We highlight advances in community- and single-cell-level approaches for characterizing active ARB and explore emerging mitigation strategies such as advanced waste treatment and bioremediation. This review aims to bridge the gap between ARG pollution and its risk mitigation, contributing to a comprehensive framework for tackling active ARB in soils.

Manure applications can facilitate the transmission of antibiotic resistance genes (ARGs) from the soil to the plant microbiome, with detrimental effects on human health through the food chain. Interventions to mitigate the spread of ARGs from animal waste to plants are essential for food safety. We previously demonstrated that converting composted manure into biochar can effectively mitigate the spread of ARGs into soil. However, it remains unclear whether compost-derived biochar can decrease the spread of ARGs in the vegetable endosphere. In this study, a pot experiment involving pakchoi (Brassica chinensis) was conducted to investigate the effects of compost-derived biochar on endophytic ARGs and the pakchoi microbiome. A total of 99 ARGs and 7 mobile genetic elements (MGEs) were identified in endophytes via high-throughput quantitative PCR. Compared with the compost amendment, the application of biochar produced from composted pig manure significantly decreased the diversity of ARGs and MGEs in vegetable endophytes. The abundance of ARGs, MGEs, and bacteria closely related to known pathogens in roots treated with biochar was much lower than that in roots treated with compost, but was similar to that in the control treatment. Fertilizer treatments influenced endophytic bacterial community assembly, while bacterial communities played an important role in shaping ARG profiles. Overall, the transmission of ARGs from animal waste to endophytic microbiomes in vegetables can be effectively mitigated using biochar derived from manure.

The growing accumulation of plastic waste in the environment has created novel habitats known as the “plastisphere”, where microorganisms can thrive. Concerns are rising about the potential for pathogenic microorganisms to proliferate in the plastisphere, posing risks to human health. However, our knowledge regarding the virulence and pathogenic potential of these microorganisms in the plastisphere remains limited. This study quantified the abundance of virulence factor genes (VFGs) in the plastisphere and its surrounding environments (water and soil) to better assess pathogenic risks. Our findings revealed a selective enrichment of VFGs in the plastisphere, which were attributed to the specific microbial community assembled. The presence of arsenic and ciprofloxacin in the plastisphere exerted additional co-selective pressures, intensifying the enrichment of VFGs. Notably, VFGs that encoded multiple functions or enhanced the survival of host microorganisms (e.g., encoding adherence functions) tended to accumulate in the plastisphere. These versatile and environmentally adaptable VFGs are more likely to be favored by bacteria in the environment, warranting increased attention in future investigations due to their potential for widespread dissemination. In terms of virulence and pathogenicity, this research offers new insights into evaluating pathogen-related risks in the plastisphere.

The impact of microplastics on antibiotic resistance has attracted widespread attention. However, previous studies primarily focused on the effects of individual microplastics. In reality, diverse microplastic types accumulate in soil, and it remains less well studied whether microplastic diversity (i.e., variations in color, shape or polymer type) can be an important driver of increased antibiotic resistance gene (ARG) abundance. Here, we employed microcosm studies to investigate the effects of microplastic diversity on soil ARG dynamics through metagenomic analysis. Additionally, we evaluated the associated potential health risks by profiling virulence factor genes (VFGs) and mobile genetic elements (MGEs). Our findings reveal that as microplastic diversity increases, there is a corresponding rise in the abundance of soil ARGs, VFGs and MGEs. We further identified microbial adaptive strategies involving genes (changed genetic diversity), community (increased specific microbes), and functions (enriched metabolic pathways) that correlate with increased ARG abundance and may thus contribute to ARG dissemination. Additional global change factors, including fungicide application and plant diversity reduction, also contributed to elevated ARG abundance. Our findings suggest that, in addition to considering contamination levels, it is crucial to monitor microplastic diversity in ecosystems due to their potential role in driving the dissemination of antibiotic resistance through multiple pathways. The effects of microplastics (MPs) on soil microbial communities and antimicrobial resistance genes are not well understood. Here, the authors used microcosm studies to show that MP diversity, and partially fungicide treatment and reduced plant diversity, correlate with higher levels of ARG and related genetic factors.