Antibiotic Resistance Genes Patterns Research Articles

Effects of nitrogen fertilization on the fate of high-risk antibiotic resistance genes in reclaimed water-irrigated soil and plants

High-risk antibiotic resistance genes (ARGs) in reclaimed water-irrigated soil pose a potential threat to ecosystem and human health. Inorganic fertilization – including with nitrogen, a key ingredient in agricultural production – may affect the ARG profile in soil. However, little is known about nitrogen fertilization's influence on ARGs profiles in the soil–plant system. This study investigated the effects of different nitrogen fertilizer types (CO(NH2)2, NO3–-N (NaNO3) and NH4+-N (NH4HCO3)) and different nitrogen fertilizer application rates (low, medium, high) on the distribution of high-risk ARGs in reclaimed water-irrigated soil and plants using quantitative PCR, high-throughput sequencing and metagenomic sequencing. Soil microcosms results revealed that nitrogen fertilization significantly affected the pattern of high-risk ARGs in soil, and also affected high-risk ARGs abundance and transfer capacity in plants. Compared with nitrogen fertilizer application rate, nitrogen fertilizer types significantly contributed to enhancing the soil resistome, with the order of CO(NH2)2 > NO3–-N ≈ NH4+-N. The medium application of NO3–-N and NH4+-N significantly reduced high-risk ARGs abundance in the leaf endophyte. Bacterial community mainly drove the variation of ARGs in nitrogen-fertilized soil–plant system, and class I integron and metal resistance genes (MRGs) also had direct effects on these high-risk ARGs. A similar high-risk ARGs pattern was also found in field plot experiments, and several dangerous pathogens were observed as the main high-risk ARGs potential hosts in nitrogen-fertilized soil. Based on an economic assessment, application of NH4+-N (NH4HCO3) could reduce costs by $1,312.83 ha−1 compared with NO3–-N (NaNO3). These results showed that the more important role of nitrogen type might be an effective and economical way to control high-risk ARGs spread in soil–plant system under reclaimed water irrigation.

Emergence and spread patterns of antibiotic resistance genes during cultivating aerobic granular sludge under different feeding modes

Aerobic granular sludge (AGS) technology, which is a promising approach for wastewater treatment, encounters challenges associated with antibiotic resistance genes (ARGs) in biological treatment systems. This study investigates the dynamics of ARGs during AGS cultivation under anaerobic plug-flow (APM), anaerobic mixed-flow (AMM), and aerobic feeding mode (AFM). Significant differences (P < 0.05) are observed in the shifts of ARGs among these modes. The abundance of ARGs notably increases under all modes, with AMM exhibiting a lower average relative abundance (0.063 ± 0.025) compared to that of APM (0.148 ± 0.061) and AFM (0.147 ± 0.047). The observed variations in bacterial communities and the co-selection of ARGs among the feeding modes are likely influential factors contributing to the documented variations of ARGs. Furthermore, the study identifies tnpA and intl1 as key contributors to horizontal gene transfer (HGT) during APM and AFM operations. Additionally, a heightened level of HGT is observed in both APM and AFM, with a larger potential host range (12 hosts each) harboring ARGs, in contrast to AMM (4 hosts). The study highlights the pivotal role played by distinct bacterial communities and the co-selection of ARGs in ARG variations across feeding modes. AMM emerges as an effective strategy for mitigating ARG proliferation, thereby enhancing the safety and sustainability of AGS technology in wastewater treatment.

Field-based investigation reveals selective enrichment of companion microbes in vegetables leading to specific accumulation of antibiotic resistance genes

Vegetables capture antibiotic resistance genes (ARGs) from the soil and then pass them on to consumers through the delivery chain and food chain, and are therefore the key node that may increase the risk of human exposure to ARGs. This study investigates the patterns and driving forces behind the transmission of ARGs from soil to vegetables by the commonly planted cash crops in the coastal region of southern China, i.e. broccoli, pumpkin, and broad bean, to investigate. The study used metagenomic data to reveal the microbial and ARGs profiles of various vegetables and the soil they are grown. The results indicate significant differences in the accumulation of ARGs among different vegetables harvested in the same area at the same time frame, and the ARGs accumulation ability of the three vegetables was in the order of broccoli, broad bean, and pumpkin. In addition, broccoli collected the highest number of ARGs in types (n = 14), while pumpkin (n = 13) does not obtain trimethoprim resistance genes and broad beans (n = 10) do not obtain chloramphenicol, fosmidomycin, quinolone, rifamycin, or trimethoprim resistance genes. Host tracking analysis shows a strong positive correlation (|rho| > 0.8, p < 0.05) between enriched ARGs and plant companion microbes. Enrichment analysis of metabolic pathways of companion microbes shows that vegetables exhibit a discernible enrichment of companion microbes, with significant differences among vegetables. This phenomenon is primarily due to the screening of carbohydrate metabolism capabilities among companion microbes and leads varied patterns of ARGs that spread from the soil to vegetables. This offers a novel insight into the intervention of foodborne transmission of ARGs.

Linking bacterial life strategies with the distribution pattern of antibiotic resistance genes in soil aggregates after straw addition

Straw addition markedly affects the soil aggregates and microbial community structure. However, its influence on the profile of antibiotic resistance genes (ARGs), which are likely associated with changes in bacterial life strategies, remains unclear. To clarify this issue, a soil microcosm experiment was incubated under aerobic (WS) or anaerobic (AnWS) conditions after straw addition, and metagenomic sequencing was used to characterise ARGs and bacterial communities in soil aggregates. The results showed that straw addition shifted the bacterial life strategies from K- to r-strategists in all aggregates, and the aerobic and anaerobic conditions stimulated the growth of aerobic and anaerobic r-strategist bacteria, respectively. The WS decreased the relative abundances of dominant ARGs such as QnrS5, whereas the AnWS increased their abundance. After straw addition, the macroaggregates consistently exhibited a higher number of significantly altered bacteria and ARGs than the silt+clay fractions. Network analysis revealed that the WS increased the number of aerobic r-strategist bacterial nodes and fostered more interactions between r-and K-strategist bacteria, thus promoting ARGs prevalence, whereas AnWS exhibited an opposite trend. These findings provide a new perspective for understanding the fate of ARGs and their controlling factors in soil ecosystems after straw addition. Environmental implicationsStraw soil amendment has been recommended to mitigate soil fertility degradation, improve soil structure, and ultimately increase crop yields. However, our findings highlight the importance of the elevated prevalence of ARGs associated with r-strategist bacteria in macroaggregates following the addition of organic matter, particularly fresh substrates. In addition, when assessing the environmental risk posed by ARGs in soil that receives crop straw, it is essential to account for the soil moisture content. This is because the species of r-strategist bacteria that thrive under aerobic and anaerobic conditions play a dominant role in the dissemination and accumulation of ARG.

Co-occurrence patterns of gut microbiome, antibiotic resistome and the perturbation of dietary uptake in captive giant pandas

The microbiome is a key source of antibiotic resistance genes (ARGs), significantly influenced by diet, which highlights the interconnectedness between diet, gut microbiome, and ARGs. Currently, our understanding is limited on the co-occurrence among gut microbiome, antibiotic resistome in the captive giant panda and the perturbation of dietary uptake, especially for the composition and forms in dietary nutrition. Here, a qPCR array with 384 primer sets and 16 S rRNA gene amplicon sequencing were used to characterize the antibiotic resistome and microbiomes in panda feces, dietary bamboo, and soil around the habitat. Diet nutrients containing organic and mineral substances in soluble and insoluble forms were also quantified. Organic and mineral components in water-unextractable fractions were 7.5 to 139 and 637 to 8695 times higher than those in water-extractable portions in bamboo and feces, respectively, while the latter contributed more to the variation (67.5 %) of gut microbiota. Streptococcus, Prevotellaceae, and Bacteroides were the dominant genera in giant pandas. The ARG patterns in panda guts showed higher diversity in old individuals but higher abundance in young ones, driven directly by the bacterial community change and mobile genetic element mediation and indirectly by dietary intervention. Our results suggest that dietary nutrition mainly accounts for the shift of gut microbiota, while bacterial community and mobile genetic elements influenced the variation of gut antibiotic resistome.

Historical trajectories of antibiotics resistance genes assessed through sedimentary DNA analysis of a subtropical eutrophic lake

Investigating the occurrence of antibiotic-resistance genes (ARGs) in sedimentary archives provides opportunities for reconstructing the distribution and dissemination of historical (i.e., non-anthropogenic origin) ARGs. Although ARGs in freshwater environments have attracted great attention, historical variations in the diversity and abundance of ARGs over centuries to millennia remain largely unknown. In this study, we investigated the vertical change patterns of bacterial communities, ARGs and mobile genetic elements (MGEs) found in sediments of Lake Chenghai spanning the past 600 years. Within resistome preserved in sediments, 177 ARGs subtypes were found with aminoglycosides and multidrug resistance being the most abundant. The ARG abundance in the upper sediment layers (equivalent to the post-antibiotic era since the 1940s) was lower than those during the pre-antibiotic era, whereas the ARG diversity was higher during the post-antibiotic era, possibly because human-induced lake eutrophication over the recent decades facilitated the spread and proliferation of drug-resistant bacteria. Statistical analysis suggested that MGEs abundance and the bacterial community structure were significantly correlated with the abundance and diversity of ARGs, suggesting that the occurrence and distribution of ARGs may be transferred between different bacteria by MGEs. Our results provide new perspectives on the natural history of ARGs in freshwater environments and are essential for understanding the temporal dynamics and dissemination of ARGs.

Integrating metagenomic and isolation strategies revealed high contamination of pathogenies and resistome in market shrimps

This study employs a comprehensive approach combining metagenomic analysis and bacterial isolation to elucidate the microbial composition, antibiotic resistance genes (ARGs), and virulence factors (VFGs) present in shrimps from market and supermarket. Metagenomic analysis of shrimps revealed a dominance of Proteobacteria and Bacteroidetes with Firmicutes notably enriched in some samples. On the other hand, the dominant bacteria isolated included Citrobacter portucalensis, Escherichia coli, Salmonella enterica, Vibrio species and Klebsiella pneumonaie. Metagenomic analysis unveiled a diverse spectrum of 23 main types and 380 subtypes of ARGs in shrimp samples including many clinical significant ARGs such as blaKPC, blaNDM, mcr, tet(X4) etc. Genomic analysis of isolated bacterial strains identified 14 ARG types with 109 subtype genes, which complemented the metagenomic data. Genomic analysis also allowed us to identify a rich amount of MDR plasmids, which provided further insights into the dissemination of resistance genes in different species of bacteria in the same samples. Examination of VFGs and mobile genetic elements (MGEs) in both metagenomic and bacterial genomes revealed a complex landscape of factors contributing to bacterial virulence and genetic mobility. Potential co-occurrence patterns of ARGs and VFGs within human pathogenic bacteria underlined the intricate interplay between antibiotic resistance and virulence. In conclusion, this integrated analysis for the first time provides a comprehensive view and sheds new light on the potential hazards associated with shrimp products in the markets. The findings underscore the necessity of ongoing surveillance and intervention strategies to mitigate risks posed by antibiotic-resistant bacteria in the food supply chain using the novel comprehensive approaches.

Impacts of electric field coupled membrane bioreactor on phenol wastewater with high salinity: Performance, membrane fouling and eco-friendly strategy

Electric field coupled membrane bioreactor (EMBR) enhanced the treatment performance of phenol wastewater with high salinity, improved sludge settleability, and alleviated membrane fouling. The sludge volume index (SVI) and fouling rate were positively correlated with the extracellular polymeric substance (EPS) content, indicating that the electric field improved sludge settleability by improving the properties of EPS, successfully reducing the fouling rate by 42.86 %. The phenol-degrading and salt-tolerant genera were enriched, and the EPS secretion genera were reduced in EMBR. Moreover, EMBR effectively resisted salt stress by increasing the relative abundance of betaine synthesis (betA) and transport genes (proV and proW), glutamate synthesis genes (gltB and gltD), salt stress response gene (rpoS), and F-type H+/Na+-transport ATPase genes (atpA and atpD), as well as reducing the relative abundance of genes related to cell membrane permeability. Meanwhile, EMBR decreased the production of intracellular reactive oxygen species (ROS), activated the antioxidant system and inhibited the ROS-mediated Save our Soul (SOS) response, which further improved salt tolerance of microorganisms. Furthermore, the inhibition of the SOS response reduced the dissemination of antibiotic resistance genes (ARGs). The co-occurrence patterns of ARGs and genera indicated that EMBR could also suppress the dissemination of ARGs by reducing the relative abundance of ARGs hosts.

Food waste compost and digestate as novel fertilizers: Impacts on antibiotic resistome and potential risks in a soil-vegetable system

As a novel agricultural practice, the reuse of food waste compost and digestate as fertilizers leads to a circular economy, but inevitably introduces bio-contaminants such as antibiotic resistance genes (ARGs) into the agroecosystem. Moreover, heavy metal and antibiotic contamination in farmland soil may exert selective pressures on the evolution of ARGs, posing threats to human health. This study investigated the fate, influencing mechanisms and potential risks of ARGs in a soil-vegetable system under different food waste fertilization and remediation treatments and soil contamination conditions. Application of food waste fertilizers significantly promoted the pakchoi growth, but resulted in the spread of ARGs from fertilizers to pakchoi. A total of 56, 80, 84, 41, and 73 ARGs, mobile genetic elements (MGEs) and metal resistance genes (MRGs) were detected in the rhizosphere soil (RS), bulk soil (BS), control soil (CS), root endophytes (RE), and leaf endophytes (LE), respectively. Notably, 7 genes were shared in the above five subgroups, indicating a specific soil-root-endophytes transmission pathway. 36 genes were uniquely detected in the LE, which may originate from airborne ARGs. The combined application of biochar and fertilizers reduced the occurrence of ARGs and MGEs to some extent, showing the remediation effect of biochar. The average abundance of ARGs in the RS, BS and CS was 3.15 × 10−2, 1.31 × 10−2 and 2.35 × 10−1, respectively. Rhizosphere effects may reduce the abundance of ARGs in soil. The distribution pattern of ARGs was influenced by the types of soil, endophyte and contaminant. MGEs is the key driver shaping ARGs dynamics. Soil properties and pakchoi growth status may affect the bacterial composition, and consequently regulate ARGs fate, while endophytic ARGs were more impacted by biotic factors. Moreover, the average daily doses of ARGs from pakchoi consumption is 107–109 copies/d/kg, and its potential health risks should be emphasized.

Exploring the effects of zeolite, biochar, and diatomite as additives for enhancing heavy metals passivation and eliminating antibiotic resistance genes in composts during vermicomposting of dewatered sludge and green waste

Vermicomposting is an effective technology for disposing of municipal sewage sludge and green wastes; however, it can contain heavy metals (HMs) and antibiotic resistance genes (ARGs); thus, the application of the final product to land is risky. In this study, zeolite (ZL), bamboo biochar (BB), and diatomite (DM) were individually mixed with dewatered sludge and green waste at a rate of 5% (w/w) to investigate their effects on earthworm activities, physicochemical characteristics of vermicompost, HMs and ARGs patterns, and microbial community structure during vermicomposting. The results showed that ZL, BB, and DM stimulated the growth and fecundity of earthworms, accompanied by higher degradation of organic matter, nutritional value, humification level, and maturity of final products. The distribution rate of OXI-Zn and RES-Pb in vermicomposts could be increased by 3.48%−11.53% and 8.64%−17.53%, respectively, under all additives addition treatments in comparison to the control treatment. Moreover, the addition of ZL, BB, and DM significantly enhanced the reduction rate of intI-1 (2.45%−20.86%), sul1 (8.16%−26.46%), sul2 (18.88%−29.55%), ermB (30.25%−40.08%), ermC (19.25%−28.53%), gryA (22.99%−30.04%), parC (18.75%−20.46%), qnrS (11.58%−28.96%) and tetC (18.99%−21.57%) genes in vermicomposts, as compared to control without additives. ZL, BB, and DM addition optimized the bacterial community by decreasing the abundances of the potential ARGs hosts, such as Steroidobacter and Acidibacter, enhancing important organic-decomposing bacteria, such as cellulomonas and Chryseolinea. This study newly suggests that ZL, BB, and DM addition is beneficial for the passivation of Zn and Pb and decreases the dissemination risk of ARGs in vermicomposting dewatered sludge and green waste.

Occurrence and dissemination of antibiotic resistance genes in the Yellow River basin: focused on family farms.

As an emerging contaminant, antibiotic resistance genes (ARGs) have attracted growing attention, owing to their widespread dissemination and potential risk in the farming environment. However, ARG pollution from family livestock farms in the Yellow River basin, one of the main irrigation water sources in the North China Plain, remains unclear. Herein, we targeted 21 typical family farms to assess the occurrence patterns of ARGs in livestock waste and its influence on ARGs in receiving environment by real-time quantitative PCR (qPCR). Results showed that common ARGs were highly prevalent in family livestock waste, and tet-ARGs and sul-ARGs were the most abundant in these family farms. Most ARG levels in fresh feces of different animals varied, as the trend of chicken farms (broilers > laying hens) > swine farms (piglets > fattening pigs > boars and sows) > cattle farms (dairy cattle > beef cattle). The effect of natural composting on removing ARGs for chicken manure was better than that forcattle manure, while lagoon storage was not effective in removing ARGs from family livestock wastewater. More troublesomely, considerable amounts of ARGs were discharged with manure application, further leading to the ARG increase in farmland soil (up to 58-119 times), which wouldexert adverse impacts on human health and ecological safety.

Linking antibiotic resistance gene patterns with advanced faecal pollution assessment and environmental key parameters along 2300 km of the Danube River

The global spread of antimicrobial resistance (AMR) in the environment is a growing health threat. Large rivers are of particular concern as they are highly impacted by wastewater discharge while being vital lifelines serving various human needs. A comprehensive understanding of occurrence, spread and key drivers of AMR along whole river courses is largely lacking. We provide a holistic approach by studying spatiotemporal patterns and hotspots of antibiotic resistance genes (ARGs) along 2311 km of the navigable Danube River, combining a longitudinal and temporal monitoring campaign. The integration of advanced faecal pollution diagnostics and environmental and chemical key parameters allowed linking ARG concentrations to the major pollution sources and explaining the observed patterns. Nine AMR markers, including genes conferring resistance to five different antibiotic classes of clinical and environmental relevance, and one integrase gene were determined by probe-based qPCR. All AMR targets could be quantified in Danube River water, with intI1 and sul1 being ubiquitously abundant, qnrS, tetM, blaTEM with intermediate abundance and blaOXA-48like, blaCTX−M-1 group, blaCTX−M-9 group and blaKPC genes with rare occurrence. Human faecal pollution from municipal wastewater discharges was the dominant factor shaping ARG patterns along the Danube River. Other significant correlations of specific ARGs were observed with discharge, certain metals and pesticides. In contrast, intI1 was not associated with wastewater but was already established in the water microbiome. Animal contamination was detected only sporadically and was correlated with ARGs only in the temporal sampling set. During temporal monitoring, an extraordinary hotspot was identified emphasizing the variability within natural waters. This study provides the first comprehensive baseline concentrations of ARGs in the Danube River and lays the foundation for monitoring future trends and evaluating potential reduction measures. The applided holistic approach proved to be a valuable methodological contribution towards a better understanding of the environmental occurrence of AMR.

Antibiotic Resistance in a Coastal River in Mississippi, USA – Potential Drivers

Wastewater treatment plants (WWTPs) are major sources of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in water bodies. Most studies on the impact of WWTPs on antibiotic resistance have focused on freshwater systems, with little information on coastal and estuarine waters with variable salinity. This study monitored seasonal levels of ARGs at the effluent and downstream of the Pascagoula— Moss Point WWTP in the lower Pascagoula River, a coastal river in southeastern Mississippi, USA. Surface water samples were collected seasonally at upstream, outflow, and 3 downstream sites from February to November 2016. Bacterial resistance to sulfamethazine, tetracycline, and ciprofloxacin was quantified using selective culture and qPCR. Mixed—effects models were developed to identify potential driving factors of ARG concentrations related to the WWTP and local environmental conditions (salinity, water temperature, and pH). The best model was selected based on the lowest Akaike Information Criterion (AIC) corrected for small sample size. The results show that the genes sul1, sul2, and intI1 were detected, with intI1 having the highest relative concentration. The qPCR analysis suggests a negative relation between ARG levels and temperature and salinity. ARG concentrations peaked immediately downstream of the WWTP and decreased gradually further downstream in some months, but the spatial pattern varied widely between sampling months. The study highlights the complex patterns of environmental ARGs and the importance of accounting for the impact of WWTPs, local environmental factors, and other anthropogenic influences to understand their potential drivers.

Effects of Zinc Thiazole and Oxytetracycline on the Microbial Metabolism, Antibiotic Resistance, and Virulence Factor Genes of Soil, Earthworm Gut, and Phyllosphere.

Pesticides and antibiotics are believed to increase the incidence of antibiotic resistance genes (ARGs) and virulence factor genes (VFGs), constituting a serious threat to global health. However, the impact of this combined pollution on the microbiome and that of the related ARGs and VFGs on soil-plant-animal systems remain unknown. In this study, a 60-day microcosm experiment was conducted to reveal the effects of zinc thiazole (ZT) and oxytetracycline (OTC) on microbial communities, antibiotic resistomes, and virulence factors in soil, earthworm gut, and phyllosphere samples using metagenomics. ZT exposure perturbed microbial communities and nutrient metabolism and increased the abundance of ARGs and VFGs in the gut. Combined exposure changed the profiles of ARGs and VFGs by decreasing microbial diversity in the phyllosphere. Host-tracking analysis identified some genera, such as Citrobacter and Aeromonas, as frequent hosts of ARGs and VFGs in the gut. Notably, some co-occurrence patterns of ARGs and MGEs were observed on the metagenome-assembled contigs. More importantly, ZT markedly increased the abundance of potentially drug-resistant pathogens Acinetobacter soli and Acinetobacter junii in the phyllosphere. Overall, this study expands our current understanding of the spread of ARGs and VFGs in soil-plant-animal systems under pollutant-induced stress and the associated health risks.

Tracking the extracellular and intracellular antibiotic resistance genes across whole year in wastewater of intensive dairy farm

Monitoring the annual variation of antibiotic resistance genes (ARGs) in livestock wastewater is important for determining the high-risk period of transfer and spread of animal-derived antibiotic resistance into the environment. However, the knowledge regarding the variation patterns of ARGs, especially intracellular ARGs (iARGs) and extracellular ARGs (eARGs), over time in livestock wastewater is still unclear. Herein, we conducted a year-round study to trace the profiles of ARGs at a Chinese-intensive dairy farm, focusing on the shifts observed in different months. The results showed significant differences in the composition and variation between iARGs and eARGs. Tetracycline, sulfonamide, and macrolide resistance genes were the major types of iARGs, while cfr was the major type of eARG. The environmental adaptations of the host bacteria determine whether ARGs appear as intracellular or extracellular forms. The total abundance of ARGs was higher from April to September, which can be attributed to the favorable climatic conditions for bacterial colonization and increased antibiotic administration during this period. Integron was found to be highly correlated with most iARGs, potentially playing a role in the presence of these genes within cells and their similar transmission patterns in wastewater. The intracellular and extracellular bacterial communities were significantly different, primarily because of variations in bacterial adaptability to the high salt and anaerobic environment. The intracellular co-occurrence network indicated that some dominant genera in wastewater, such as Turicibacter, Clostridium IV, Cloacibacillus, Subdivision5_genera_incertae_sedis, Saccharibacteria_genera_incertae_sedis and Halomonas, were potential hosts for many ARGs. To the best of our knowledge, this study demonstrates, for the first time, the annual variation of ARGs at critical points in the reuse of dairy farm wastewater. It also offers valuable insights into the prevention and control of ARGs derived from animals.

Impacts of electric field-magnetic powder coupled membrane bioreactor on phenol wastewater treatment: Performance, synergistic mechanism, antibiotic resistance genes, and eco-environmental benefit evaluation

A novel electric field-magnetic powder coupled membrane bioreactor (EM-MBR) was constructed, which was superior on improvement of phenol treatment performance and sludge characteristics, and mitigation of membrane fouling. EM-MBR enhanced the phenol degradation via the improvement activity of phenol degrading enzymes. The EPS contents and SVI of EM-MBR were significantly reduced by 49.3 % and 58.7 % than that of the conventional MBR, respectively. Moreover, EM-MBR successfully reduced fouling rate by 57.0 %, delaying the membrane resistance. The EPS contents were positively correlated with the SVI and fouling rate, implying that the sludge settleability was strengthened by improving the properties of EPS with the assistance of electromagnetic, thus mitigating the membrane fouling. Microbial co-occurrence network demonstrated that EM-MBR enriched phenol-degrading and EPS-degrading genera correlated to Fe redox cycle. Furthermore, the activation of the antioxidant system in the EM-MBR resulted in the suppression of reactive oxygen species (ROS) generation, consequently impeding the dissemination of antibiotic resistance genes (ARGs). Co-occurrence patterns of MGEs and ARGs revealed that intercellular binding facilitated by ist and Integrase may account for the horizontal transfer of ARGs. The reduction of unit capital costs (15.63 %), running costs (53.00 %), and total average carbon emissions (15.18 %) indicated that EM-MBR was environmentally beneficial.

Chicken feedlot revisited: Co-dispersal of antibiotic and metal resistome under banning in-feed veterinary antibiotics

Intensive livestock farming has been implicated as a notorious hotspot for antibiotic resistance genes (ARGs) due to the excessive or inappropriate use of in-feed antibiotics over the past few decades. Since China implemented a ban on the use of antibiotics in animal feed since 2020, the dissemination of ARGs in the vicinity of feedlots has remained unclear. This study presents a case study that aims to investigate the dispersal of antibiotics and ARGs from a chicken feedlot (established in 2020) to the adjacent aquatic and soil environments. Comparing the sample collected from upstream area, the water and sediment samples from midstream and downstream areas showed an increase in total antibiotic residues and metal content (Cu and Zn) by 4.2–5.3 fold and 1.3–22.6 fold, respectively. The downstream water samples exhibited a 2.49–2.93-fold increase in the abundance of ARGs and a 1.48–1.75-fold increase in the abundance of metal resistance genes (MRGs). The results of Pearson correlation and metagenome-assembled genome revealed a tendency for the co-occurrence of ARGs and MRGs. The dissemination of ARGs and MRGs is primarily driven by tetracycline, tylosin, Cu, and, Mn, with mobile genetic elements playing a more significant role than bacterial communities. These findings shed light on the overlooked co-dispersal pattern of ARGs and MRGs in the environment surrounding feedlots, particularly in the context of banning in-feed veterinary antibiotics.

Fates of extracellular and intracellular antibiotic resistance genes in activated sludge and plastisphere under sulfadiazine pressure

Microplastics, antibiotics, and antibiotic resistance genes (ARGs) represent prominent emerging contaminants that can potentially hinder the efficacy of biological wastewater treatment and pose health risks. Plastisphere as a distinct ecological niche for microorganisms, acts as a repository for ARGs and potential pathogenic bacteria. Nonetheless, the spread pattern of extracellular ARGs (eARGs) and intracellular ARGs (iARGs) in plastisphere under antibiotic exposure was not yet known. This study aimed to investigate disparities in extracellular polymeric substances (EPS) production, extracellular and intracellular microbial community structures, as well as the transmission of eARGs and iARGs between activated sludge and plastisphere in an anaerobic/anoxic/oxic system under sulfadiazine (SDZ) exposure. SDZ was found to enhance EPS production in activated sludge and plastisphere. Interestingly, as SDZ removal efficiency increased, EPS content decreased in activated sludge and plastisphere collected from oxic zone, and continued to increase in plastisphere samples collected from anaerobic and anoxic zones. There were significant differences in microbial community structure between activated sludge and plastisphere, and the DNA fragments of potential pathogenic bacteria were detected in extracellular samples. SDZ exhibited a promoting effect on the propagation of eARGs, which were more abundant in the plastisphere than in activated sludge, thus heightening the risk of ARGs dissemination. Extracellular mobile genetic elements played a pivotal role in driving the spread of eARGs, while the microbial community induced the changes of iARGs. Potential pathogenic bacteria emerged as potential hosts for ARGs and mobile genetic elements within activated sludge and plastisphere, leading to more serious environmental threats.

Spatio-temporal distribution and dynamics of antibiotic resistance genes in a water-diversion lake, China

The distribution and dynamics of antibiotic resistance genes (ARGs) in water-diversion lakes are poorly understood. In this study, two comparative in situ investigations of ARG profiles targeting water diversion (DP) and non-diversion periods (NDP) were conducted in Luoma Lake, a vital transfer node for the eastern route of the South-to-North Water Diversion Project in China. The results demonstrated significant spatiotemporal variations in ARG contamination and notable differences in the co-occurrence patterns of ARGs and bacterial communities between DP and NDP. Correlations among ARGs with the 16 S rRNA, and mobile genetic elements indicate that horizontal gene transfer (HGT) and vertical gene transfer (VGT) in NDP, but only HGT in DP, were the primary mechanisms of ARG proliferation and spread, implying that water diversion could be an essential control of the transfer pattern of ARGs in a lake environment. The null model analysis indicated that stochastic processes, with predominant driver of ecological drift in the lake mainly drove the assembly of ARGs. Partial least squares structural equation modeling was developed to analyze the causal effects of the factors in shaping ARG dynamics and identify the major driving forces in the DP and NDP.

Network complexity of bacterial community driving antibiotic resistome in the microbiome of earthworm guts under different land use patterns

Recently, the study of antibiotic resistance in the soil animal microbiome has attracted extensive attention; however, the patterns of antibiotic resistance genes (ARGs) in soil and soil animals related to different land use types remain poorly studied. In the present study, soil and earthworms were collected from four different land-use types (farmland, hospital, park land, and mountain park), and 162 ARGs in the microbiomes of the soil and earthworms were quantified using high-throughput quantitative PCR. Our study showed that the abundance and number of ARGs were higher in soil samples than in earthworm guts, but earthworms as the living organisms created relatively isolated ambient surroundings, which allowed for a more heterogeneous ARGs profile. Meanwhile, land use significantly influenced the abundance, number and co-occurrence pattern of ARGs in the soil and earthworm samples. Furthermore, abiotic and biotic factors had significant effects on the ARGs profile, among which pH had a negative effect on the ARGs profiles of both soil and earthworm microbiomes, and bacterial network complexity had a positive effect on the earthworm ARGs profile. Our study provides new insights into the distribution and dispersal of ARGs in the soil animal gut microbiome under different land use patterns.