Resistance Genes In Soil Research Articles

Effects of glyphosate on antibiotic resistance in soil bacteria and its potential significance: A review.

The evolution and spread of antibiotic resistance are problems with important consequences for bacterial disease treatment. Antibiotic use in animal production and the subsequent export of antibiotic resistance elements in animal manure to soil is a concern. Recent reports suggest that exposure of pathogenic bacteria to glyphosate increases antibiotic resistance. We review these reports and identify soil processes likely to affect the persistence of glyphosate, antibiotic resistance elements, and their interactions. The herbicide molecular target of glyphosate is not shared by antibiotics, indicating that target-site cross-resistance cannot account for increased antibiotic resistance. The mechanisms of bacterial resistance to glyphosate and antibiotics differ, and bacterial tolerance or resistance to glyphosate does not coincide with increased resistance to antibiotics. Glyphosate in the presence of antibiotics can increase the activity of efflux pumps, which confer tolerance to glyphosate, allowing for an increased frequency of mutation for antibiotic resistance. Such effects are not unique to glyphosate, as other herbicides and chemical pollutants can have the same effect, although glyphosate is used in much larger quantities on agricultural soils than most other chemicals. Most evidence indicates that glyphosate is not mutagenic in bacteria. Some studies suggest that glyphosate enhances genetic exchange of antibiotic-resistance elements through effects on membrane permeability. Glyphosate and antibiotics are often present together in manure-treated soil for at least part of the crop-growing season, and initial studies indicate that glyphosate may increase abundance of antibiotic resistance genes in soil, but longer term investigations under realistic field conditions are needed. Although there are demonstratable interactions among glyphosate, bacteria, and antibiotic resistance, there is limited evidence that normal use of glyphosate poses a substantial risk for increased occurrence of antibiotic-resistant, bacterial pathogens. Longer term field studies using environmentally relevant concentrations of glyphosate and antibiotics are needed.

Microplastic diversity increases the abundance of antibiotic resistance genes in soil

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.

Temporal dynamic of soil microbial communities and antibiotic resistance markers exposed to increasing concentrations of sulfamethoxazole

The reuse of treated wastewater (TWW) for irrigation is widely applied to alleviate pressure on freshwater resources. However, TWW contains antibiotics that once in soils, can exert selective pressure, promoting the emergence and spread of antimicrobial resistance (AMR) in the environment. Current environmental risk assessments for antibiotic residues rely on indicators such as Predicted No Effect Concentrations (PNECs), usually determined in liquid media. These PNECs aim to predict antibiotic concentrations that may promote resistance in the environment. Given the complexity of soil matrices, few studies have established PNEC values for soil, which likely differ significantly from aquatic environments.To address this gap, we developed a simplified experimental model using soil microcosms irrigated with TWW and the antibiotic sulfamethoxazole (SMX) to estimate threshold concentrations favouring resistance transfer or/and emergence within the soil microbiome. We identified SMX concentrations between 0.01 and 0.1 mg/kgdry soil that likely increased the abundance of sulfonamide resistance genes in soil. A time window of 1 to 7 days post-exposure showed a temporary rise in sul1 and intl1 gene abundance (over 1 log/soil 16S rDNA), the appearance of SMX transformation products, and an increase in some Rhodocyclaceae. After 1.5 months of incubation and complete SMX transformation, the relative abundance of sul1 and intl1 remained about 0.5 log higher than in SMX-free controls and soils with SMX levels below 0.1 mg/kg dry soil. A persistent transformation product, 4-N-glucuronide-SMX, was also observed.Here, the estimated PNEC for SMX in soil, between 0.01 and 0.1 mg/kg, exceeds typical SMX concentrations found in soils exposed to TWW. This may suggest low impact on resistance selection for this compound in the context of TWW exposure. However further studies on other soils, water, and antibiotics need to be conducted to expand our knowledge on soil PNECs.

Culture-based and molecular investigation of antibiotic and metal resistance in a semi-arid agricultural soil repeatedly amended with urban sewage sludge

Unsustainable agricultural intensification and climate change effects have caused chronic soil depletion in most arid and semi-arid croplands. As such, the land application of urban sewage sludge (USS) has been regulated in several countries as an alternative soil conditioner with recycling benefits. However, the risks of multi-contamination have made its agricultural reuse debatable. Accordingly, this study explored the long-term the impact of repetitive USS applications with increasing rates (0, 40, 80, and 120 t ha−1 year−1) on a sandy soil properties. A special focus was on the spread of antibiotic-resistant bacteria, metal-resistant bacteria and corresponding resistance genes in soil (ARB, MRB, ARGs and MRGs, respectively). The outcomes showed a dose-dependent variation of different soil parameters including the increase of heavy metal content and total heterotrophic bacteria (THB) up to the highest sludge application rate. Besides, the two last sludge lots applied in fall 2019 and 2020 contained cultivable ARB for all addressed antibiotics at much higher counts than in corresponding treated soils. Interestingly, the average index of antibiotic resistance (ARB/THB) increased in the USS used in fall 2020 compared to 2019 (from 6.2% to 9.4%). This indicates that factors such as fluctuations in wastewater quality, treatments operations, and extensive antibiotic use following the outbreak of the COVID-19 pandemic in early 2020 could have caused this variation. The molecular assessment of bacterial resistance resulted in the identification of three ARGs (mefA, sul1 and sul2), one MRG (czcA) and one integron (intI1). This might have implications on resistance co-selection, which can pose a threat to human health via contaminated crops.

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.

Nanoscale zero-valent iron alleviated horizontal transfer of antibiotic resistance genes in soil: The important role of extracellular polymeric substances

Extracellular polymeric substances (EPS) are tightly related to the horizontal gene transfer (HGT) of antibiotic resistance genes (ARGs), but often neglected in soil. In this study, nanoscale zero-valent iron (nZVI) was utilized for attenuation of ARGs in contaminated soil, with an emphasis on its effects on EPS secretion and HGT. Results showed during soil microbe cultivation exposed to tetracycline, more EPS was secreted and significant increase of tet was observed due to facilitated HGT. Notably, copies of EPS-tet accounted for 71.39 % of the total tet, implying vital effects of EPS on ARGs proliferation. When co-exposed to nZVI, EPS secretion was decreased by 38.36–71.46 %, for that nZVI could alleviate the microbial oxidative stress exerted by tetracycline resulting in downregulation of genes expression related to the c-di-GMP signaling system. Meanwhile, the abundance of EPS-tet was obviously reduced from 7.04 to 5.12–6.47 log unit, directly causing decrease of total tet from 7.19 to 5.68–6.69 log unit. For the reduced tet, it was mainly due to decreased EPS secretion induced by nZVI resulting in inhibition of HGT especially transformation of the EPS-tet. This work gives an inspiration for attenuation of ARGs dissemination in soil through an EPS regulation strategy.

Antibiotic Susceptibility of Bacteria Isolated from Livestock-exposed and Unexposed Areas in Perlite-rich Soil in Thailand

Antibiotic resistant bacteria are present in various environments and are concerning for public health. Antibiotic resistance genes in soil may proliferate throughout different ecosystems and be acquired by pathogens that pose health risks to humans and animals. This study investigated the antibiotic resistance of cultivable bacteria isolated from livestock-exposed and unexposed areas in a former perlite mining area in the Fa-La-Mee Mountain range, Lopburi, Thailand. Soil samples were collected in five different locations. Soil bacteria were isolated and tested for susceptibility against seven antibiotics, followed by 16S rRNA gene-based identification. A total of 56 bacterial isolates were isolated from the perlite-rich soil samples, including 34 isolates from the livestock-exposed areas and 22 isolates from the unexposed areas. Most of the isolates were resistant to cefepime (26.5% in exposed areas and 22.7% in unexposed areas) and ampicillin (23.5% in exposed areas and 22.7% in unexposed areas). 16S rRNA gene sequences revealed that most of the resistant bacteria isolated from perlite-rich soil were Bacillus spp. A multidrug-resistant isolate of Bacillus cereus resistant to ampicillin, trimethoprim/sulfamethoxazole, and cefepime was found in a livestock-unexposed area. The majority of the culturable antibiotic-resistant bacteria isolated from perlite-rich soil were cefepime-resistant Bacillus. Interestingly, the pattern of antibiotic resistance in exposed and unexposed areas was not different, which indicated that antibiotic resistance in perlite-rich soil is likely caused by other factors, such as physicochemical characteristics and/or the microbial population of the soil rather than by antibiotics used in livestock husbandry.

Presence of Antimicrobial-Resistant Bacteria and Resistance Genes in Soil Exposed to Wastewater Treatment Plant Effluent

Presence of Antimicrobial-Resistant Bacteria and Resistance Genes in Soil Exposed to Wastewater Treatment Plant Effluent

Fertilizing drug resistance: Dissemination of antibiotic resistance genes in soil and plant bacteria under bovine and swine slurry fertilization

The increasing global demand for food production emphasizes the use of organic animal fertilizers, such as manure and slurry, to support sustainable agricultural practices. However, recent studies highlight concerns about antibiotic resistance determinants in animal excrements, posing a potential risk of spreading antibiotic resistance genes (ARGs) in agricultural soil and, consequently, in food products.This study examines the dissemination of ARGs within the soil and plant-associated microbiomes in cherry radish following the application of swine and bovine slurry. In a 45-day pot experiment, slurry-amended soil, rhizospheric bacteria, and endophytic bacteria in radish roots and leaves were sampled and analyzed for 21 ARGs belonging to 7 Antibiotic Resistance Phenotypes (ARPs). The study also assessed slurry's impact on soil microbiome functional diversity, enzymatic activity, physicochemical soil parameters, and the concentration of 22 selected antimicrobials in soil and plant tissues.Tetracyclines and β-lactams were the most frequently identified ARGs in bovine and swine slurry, aligning with similar studies worldwide. Swine slurry showed a higher prevalence of ARGs in soil and plant-associated bacteria, particularly TET genes, reflecting pig antibiotic treatments. The persistent dominance of TET genes across slurry, soil, and plant microbiomes highlights significant influence of slurry application on gene occurrence in plant bacteria. The presence of ARGs in edible plant parts underscores health risks associated with raw vegetable consumption. Time-dependent dynamics of ARG occurrence highlighted their persistent presence throughout the experiment duration, influenced by the environmental factors and antibiotic residuals. Notably, ciprofloxacin, which was the only one antimicrobial detected in fertilized soil, significantly impacted bovine-amended variants. Soil salinity modifications induced by slurry application correlated with changes in ARG occurrence. Overall, the research underscores the complex relationships between agricultural practices, microbial activity, and antibiotic resistance dissemination, emphasizing the need for a more sustainable and health-conscious farming approaches.

Body size: A hidden trait of the organisms that influences the distribution of antibiotic resistance genes in soil

Body size is a key life-history trait of organisms, which has important ecological functions. However, the relationship between soil antibiotic resistance gene (ARG) distribution and organisms' body size has not been systematically reported so far. Herein, the impact of organic fertilizer on the soil ARGs and organisms (bacteria, fungi, and nematode) at the aggregate level was analyzed. The results showed that the smaller the soil aggregate size, the greater the abundance of ARGs, and the larger the body size of bacteria and nematodes. Further analysis revealed significant positive correlations of ARG abundance with the body sizes of bacteria, fungi, and nematodes, respectively. Additionally, the structural equation model demonstrated that changes in soil fertility mainly regulate the ARG abundance by affecting bacterial body size. The random forest model revealed that total phosphorus was the primary soil fertility factor influencing the body size of organisms. Therefore, these findings proposed that excessive application of phosphate fertilizers could increase the risk of soil ARG transmission by increasing the body size of soil organisms. This study highlights the significance of organisms' body size in determining the distribution of soil ARGs and proposes a new disadvantage of excessive fertilization from the perspective of ARGs.

Assessment of antibiotic resistance genes in soils polluted by chemical and technogenic ways with poly-aromatic hydrocarbons and heavy metals

Anthropogenic activities are leaving lots of chemical footprints on the soil. It alters the physiochemical characteristics of the soil thereby modifying the natural soil microbiome. The prevalence of antimicrobial-resistance microbes in polluted soil has gained attention due to its obvious public health risks. This study focused on assessing the prevalence and distribution of antibiotic-resistance genes in polluted soil ecosystems impacted by industrial enterprises in southern Russia. Metagenomic analysis was conducted on soil samples collected from polluted sites using various approaches, and the prevalence of antibiotic-resistance genes was investigated. The results revealed that efflux-encoding pump sequences were the most widely represented group of genes, while genes whose products replaced antibiotic targets were less represented. The level of soil contamination increased, and there was an increase in the total number of antibiotic-resistance genes in proteobacteria, but a decrease in actinobacteria. The study proposed an optimal mechanism for processing metagenomic data in polluted soil ecosystems, which involves mapping raw reads by the KMA method, followed by a detailed study of specific genes. The study's conclusions provide valuable insights into the prevalence and distribution of antibiotic-resistance genes in polluted soils and have been illustrated in heat maps.

Effects of reductive soil disinfestation on potential pathogens and antibiotic resistance genes in soil

Reductive soil disinfestation (RSD) is commonly employed for soil remediation in greenhouse cultivation. However, its influence on antibiotic resistance genes (ARGs) in soil remains uncertain. This study investigated the dynamic changes in soil communities, potential bacterial pathogens, and ARG profiles under various organic material treatments during RSD, including distillers’ grains, potato peel, peanut vine, and peanut vine combined with charcoal. Results revealed that applying diverse organic materials in RSD significantly altered bacterial community composition and diminished the relative abundance of potential bacterial pathogens (P < 0.05). The relative abundance of high-risk ARGs decreased by 10.7%-30.6% after RSD treatments, the main decreased ARG subtypes were AAC(3)_Via, dfrA1, ErmB, lnuB, aadA. Actinobacteria was the primary host of ARGs and was suppressed by RSD. Soil physicochemical properties, such as total nitrogen, soil pH, total carbon, were crucial factors affecting ARG profiles. Our findings demonstrated that RSD treatment inhibited pathogenic bacteria and could be an option for reducing high-risk ARG proliferation in soil.

Effect of wheat crops on the persistence and attenuation of antibiotic resistance genes in soil after swine wastewater application

Swine wastewater (SW) application introduces antibiotic resistance genes (ARGs) into farmland soils. However, ARG attenuation in SW-fertigated soils, especially those influenced by staple crops and soil type, remains unclear. This study investigated twelve soil ARGs and one mobile genetic element (MGE) in sandy loam, loam, and silt loam soils before and after SW application in wheat-planted and unplanted soils. The results revealed an immediate increase in the abundance of ARGs in soil by two orders of magnitude above background levels following SW application. After SW application, the soil total ARG abundance was attenuated, reaching background levels at 54 days; However, more individual ARGs were detected above the detection limit than pre-application. Among the 13 genes, acc(6′)-lb, tetM, and tetO tended to persist in the soil during wheat harvest. ARG half-lives were up to four times longer in wheat-planted soils than in bare soils. Wheat planting decreased the persistence of acc(6′)-lb, ermB, ermF, and intI2 but increased the persistence of others such as sul1 and sul2. Soil type had no significant impact on ARG and MGE fates. Our findings emphasize the need for strategic SW application and the consideration of crop cultivation effects to mitigate ARG accumulation in farmland soils.

Pentachlorophenol affects doxycycline and tetracycline resistance genes in soil by altering microbial structure

Tetracycline antibiotics play a vital role in animal husbandry, primarily employed to uphold the health of livestock and poultry. Consequently, when manure is reintegrated into farmland, tetracycline antibiotics can persist in the soil. Simultaneously, to ensure optimal crop production, organochlorine pesticides (OCPs) are frequently applied to farmland. The coexistence of tetracycline antibiotics and OCPs in soil may lead to an increased risk of transmission of tetracycline resistance genes (TRGs). Nevertheless, the precise mechanism underlying the effects of OCPs on tetracycline antibiotics and TRGs remains elusive. In this study, we aimed to investigate the effects of OCPs on soil tetracycline antibiotics and TRGs using different concentrations of doxycycline (DOX) and pentachlorophenol (PCP). The findings indicate that PCP and DOX mutually impede their degradation in soil. Furthermore, our investigation identifies Sphingomonas and Bacillus as potential pivotal microorganisms influencing the reciprocal inhibition of PCP and DOX. Additionally, it is observed that the concurrent presence of PCP and DOX could impede each other's degradation by elevating soil conductivity. Furthermore, we observed that a high concentration of PCP (10.7 mg/kg) reduced the content of efflux pump tetA, ribosome protective protein tetM, tetQ, and passivating enzyme tetX. In contrast, a low PCP concentration (6.4 mg/kg) only reduced the content of ribosome protective protein tetQ. This suggests that PCP may reduce the relative abundance of TRGs by altering the soil microbial community structure and inhibiting the potential host bacteria of TRGs. These findings have significant implications in understanding the combined pollution of veterinary antibiotics and OCPs. By shedding light on the interactions between these compounds and their impact on microbial communities, this study provides a theoretical basis for developing strategies to manage and mitigate their environmental impact, and may give some information regarding the sustainable use of antibiotics and pesticides to ensure the long-term health and productivity of agricultural systems.

Response characteristics and functional predictions of soil microorganisms to heavy metals, antibiotics, and their resistance genes originating from different animal farms amended with Herbaspirillum huttiense

Current understanding is limited regarding technologies that use biochar and microorganisms to simultaneously treat soils contaminated with both veterinary antibiotics (VAs) and heavy metals (HMs) from different animal farms. The contributions of the keystone taxa and their similarities from different animal farms under VA and HM stresses before and after soil remediation should be further investigated as well. An innovative treatment of Herbaspirillum huttiense (HHS1) inoculated waste fungus chaff-based (WFCB) biochar was designed for immobilization of copper (Cu) and zinc (Zn), and the removal of oxytetracycline (OTC), enrofloxacin (ENR), and a subsequent reduction in their resistance genes in soils from pig, cow, and chicken farms. Roles of indigenous microorganisms which can treat soils contaminated with VAs and HMs were summarized. Results showed that available Cu and Zn were reduced by 19.5% and 28.1%, respectively, while 49.8% of OTC and 85.1% of ENR were removed by WFCB-HHS1. The decrease in ENR improved overall microbial community diversity, and the increases in genera HHS1, Pedobacter, Flavobacterium and Aequorivita, along with the decreases of genera Bacillus, Methylobacter, and Fermentimonas were indirectly favorable to treat HMs and VAs in soils from different animal farms. Bacterial communities in different animal farm soils were predominantly influenced by stochastic processes. The regulations of functional genes associated with metabolism and environmental information processing, which contribute to HM and VA defense, were altered when using WFCB-HHS1. Furthermore, the spread of their antibiotic resistance genes was restricted.

Effects of anaerobic soil disinfestation on antibiotics, human pathogenic bacteria, and their associated antibiotic resistance genes in soil

Despite the widespread use of anaerobic soil disinfestation (ASD) to improve soil health, its impact on soil antibiotics, human pathogenic bacteria (HPB) and antibiotic resistance genes (ARGs) remains unclear. To address this knowledge gap, we conducted an incubation study in manure-amended soil with three treatments: no treatment (CK), crop straw only (CS), and CS combined with water flooding and plastic covering (ASD). The results showed that ASD increased the concentrations of tetracycline by 27 % and β-lactam by 22 %, while depleting macrolide by 51 %. Furthermore, ASD increased the numbers of potential HPB species and genes by 4.1 % and 23 %, respectively. Co-occurrence network analysis demonstrated intensified cooperation and competition among HPB and HNPB (human non-pathogenic bacteria) species under ASD, favoring the prevalence of HPB species such as Escherichia coli, Streptococcus equi, Clostridium tetani, and Bacillus anthracis. Potential HPB species in Proteobacteria exhibited negative correlations with macrolide and β-lactam antibiotics, whereas those in Firmicutes showed positive correlations with tetracycline antibiotics, suggesting that specific bacterial taxa contributed to the production and degradation of antibiotics in response to ASD. Interestingly, CS mitigated the presence of ARGs in soil, while ASD further amplified this reduction, almost eliminating nalC, acrB, OXA-60, VIM-7, tetL, and tetQ. The variation in ARG profiles (91.7 %) could be explained by antibiotics, HPB, and HNPB species, indicating their collective contribution to reducing soil ARGs under ASD. Moreover, host prediction and network analysis revealed that, compared with CK, where potential HPB species were the main carriers of ARGs, soil ARGs were less frequently carried by potential HPB species after ASD, indicating a reduction in the collaborative dissemination of HPB and ARGs in manured soil. In conclusion, ASD provides a promising strategy for mitigation of soil biologic contamination, which can have profound significance for environmental safety and agricultural sustainability.

Effects of chicken manure‐modified biochar on the adsorption capacity of tetracycline and abundance of antibiotic resistance genes in soil

AbstractLarge quantities of livestock manure not only occupy farmland but also serve as the primary source of antibiotics and antibiotic resistance genes (ARGs) in agricultural soil. To enhance the resource utilization of animal manure and mitigate ecological risks associated with antibiotics in agroecosystems, chicken manure was used as a raw material for biochar production. The manure derived biochar (MB) was modified with an alkali–acid two‐step modification process. The results showed that the total pore volume and surface area of alkali–acid modified MB (AMMB) was significantly increased about three and five times, respectively after modification. The SEM images showed that the surface of AMMB had a more regular and clearer structure with fewer impurities. The improvement of surface properties of AMMB significantly increased its adsorbing capacity of tetracycline in the soil. The adsorption kinetics of tetracycline by AMMB were better fitted by the pseudo‐second‐order kinetic model, suggesting that the adsorption of tetracycline on biochar followed the process of chemisorption. The adsorption mechanisms mainly involved the pore filling, hydrophobic effect, and π–π EDA interaction. Moreover, after a 60 days amendment of AMMB, the total relative abundances (RAs) of tested tetracycline resistance genes (TRGs) in soil decreased significantly, which suggested that AMMB can be a potential material to reduce risks of antibiotics and its resistance genes.

Diversity of antibiotic resistance genes in soils with four different fertilization treatments.

Although the enrichment of resistance genes in soil has been explored in recent years, there are still some key questions to be addressed regarding the variation of ARG composition in soil with different fertilization treatments, such as the core ARGs in soil after different fertilization treatments, the correlation between ARGs and bacterial taxa, etc. For soils after different fertilization treatments, the distribution and combination of ARG in three typical fertilization methods (organic fertilizer alone, chemical fertilizer alone, and conventional fertilizer) and non-fertilized soils were investigated in this study using high-throughput fluorescence quantitative PCR (HT-qPCR) technique. The application of organic fertilizers significantly increased the abundance and quantity of ARGs and their subtypes in the soil compared to the non-fertilized soil, where sul1 was the ARGs specific to organic fertilizers alone and in higher abundance. The conventional fertilizer application also showed significant enrichment of ARGs, which indicated that manure addition often had a more decisive effect on ARGs in soil than chemical fertilizers, and three bacteria, Pseudonocardia, Irregularibacter, and Castllaniella, were the key bacteria affecting ARG changes in soil after fertilization. In addition, nutrient factors and heavy metals also affect the distribution of ARGs in soil and are positively correlated. This paper reveals the possible reasons for the increase in the number of total soil ARGs and their relative abundance under different fertilization treatments, which has positive implications for controlling the transmission of ARGs through the soil-human pathway.

Accumulation of Sulfonamides and Macrolides Antibiotic Resistance Genes in Soils with Different Utilization Patterns in China

To explore the variation in the absolute abundance of antibiotic resistance genes (ARGs) in different regions of China under different land use modes and different planting years, the qualitative and quantitative study of sulfonamide (sul1,sul2) and macrolide (ermB,mefA) ARGs and an integron gene (intl1) were conducted using ordinary PCR and the fluorescence quantitative technique. The results revealed that the frequencies of sulfonamides (sul1,sul2) and intl1 were all 100% in different soils, and the detection frequencies of macrolides (ermB,mefA) were 100% in facilities vegetable fields; however, in open vegetable fields, and open grain fields, the frequencies of ermB were 94%, and those of mefA were 92% and 90%, respectively. The absolute abundance of sulfonamide and macrolide ARGs was the highest in 15 years facilities vegetable soil in Heilongjiang Province. The absolute abundance of intl1 was the highest in 15 years facilities vegetable soil in Neimenggu Province. The absolute abundances of ARGs and intl1 in facilities vegetable soil of 3, 7, and 15 years were significantly higher than that in open grain fields and open vegetable fields of the same years. The absolute abundance of ARGs and intl1 in facilities vegetable fields for 7 years and 15 years were significantly higher than that for 3 years. There was no significant linear relationship between the gene accumulation and planting life in open vegetable fields and open grain fields, except for sul1 andsul2, whereas there was a significantly positive correlation in facilities vegetable soil. Correlation analysis demonstrated that there was a significant positive correlation between the abundance of ARGs and the abundance of intl1 in different soils. This reveals the accumulation of ARGs and intl1 in soils with different utilization patterns, thereby providing reference and support for secure agricultural production.

Uncovering the prevalence and drivers of antibiotic resistance genes in soils across different land-use types

The emergence and spread of antibiotic resistance genes (ARGs) in soil due to animal excreta and organic waste is a major threat to human health and ecosystems, and global efforts are required to tackle the issue. However, there is limited knowledge of the variation in ARG prevalence and diversity resulting from different land-use patterns and underlying driving factors in soils. This study aimed to comprehensively characterize the profile of ARGs and mobile genetic elements and their drivers in soil samples collected from 11 provinces across China, representing three different land-use types, using high-throughput quantitative polymerase chain reaction and 16S rRNA amplicon sequencing. Our results showed that agricultural soil had the highest abundance and diversity of ARGs, followed by tea plantation and forest land. A total of 124 unique ARGs were detected in all samples, with shared subtypes among different land-use patterns indicating a common origin or high transmission frequency. Moreover, significant differences in ARG distribution were observed among different geographical regions, with the greatest enrichment of ARGs found in southern China. Biotic and abiotic factors, including soil properties, climatic factors, and bacterial diversity, were identified as the primary drivers associated with ARG abundance, explaining 71.8% of total ARG variation. The findings of our study demonstrate that different land-use patterns are associated with variations in ARG abundance in soil, with agricultural practices posing the greatest risk to human health and ecosystems regarding ARGs. Our identification of biotic and abiotic drivers of ARG abundance provides valuable insights into strategies for mitigating the spread of these genes. This study emphasizes the need for coordinated and integrated approaches to address the global antimicrobial resistance crisis.