Dominant Antibiotic Resistance Genes Research Articles

Antibiotic resistome alteration along a full-scale drinking water supply system deciphered by metagenome assembly: Regulated by seasonality, mobile gene elements and antibiotic resistant gene hosts

Both drinking water treatment processes and distribution can lead to antibiotic resistome variation, yet the variation of antibiotic resistome in the whole drinking water supply system (DWSS) combined with seasonality remains unknown. In this study, microbial community, antibiotic resistome, mobile genetic elements (MGEs) co-existing with antibiotic resistance genes (ARGs) and ARG hosts would be explored along a DWSS for four seasons with metagenome assembly. Multidrug and bacitracin ARGs were dominant ARGs in DWSS. Integrase, plasmids, recombinase and transposase were major MGEs co-existing with ARGs. Filtration and disinfection treatments could alter the ARG relative abundance, mainly via changing the abundance of ARG hosts (Limnohabitans and Polynucleobacter), which was influenced by water total organic carbon (TOC) content. When TOC was relatively high, filtration could proliferate ARGs via promoting antibiotic resistance bacteria (ARB) but chlorine dioxide could decrease ARGs via killing ARB. Filtration played an important role in controlling ARGs by reducing ARB when TOC was relatively low. The stimulation effect of disinfection on ARGs existed in more oligotrophic environment. Distribution could enrich ARGs in higher temperature by increasing MGEs co-occurring with ARGs and diversifying ARG hosts. MGEs co-occurring with ARGs became more abundant and diverse in disinfected water in warmer seasons. Microbial community was the most important factor determining the antibiotic resistome along a DWSS. These findings extend the knowledge about how and why water treatment processes and pipe distribution shape drinking water antibiotic resistome in different seasons.

The global distribution and environmental drivers of the soil antibiotic resistome

BackgroundLittle is known about the global distribution and environmental drivers of key microbial functional traits such as antibiotic resistance genes (ARGs). Soils are one of Earth’s largest reservoirs of ARGs, which are integral for soil microbial competition, and have potential implications for plant and human health. Yet, their diversity and global patterns remain poorly described. Here, we analyzed 285 ARGs in soils from 1012 sites across all continents and created the first global atlas with the distributions of topsoil ARGs.ResultsWe show that ARGs peaked in high latitude cold and boreal forests. Climatic seasonality and mobile genetic elements, associated with the transmission of antibiotic resistance, were also key drivers of their global distribution. Dominant ARGs were mainly related to multidrug resistance genes and efflux pump machineries. We further pinpointed the global hotspots of the diversity and proportions of soil ARGs.ConclusionsTogether, our work provides the foundation for a better understanding of the ecology and global distribution of the environmental soil antibiotic resistome.CYsYwEfC-LPouz_MuhvMM4Video

Data-driven discoveries on widespread contamination of freshwater reservoirs by dominant antibiotic resistance genes

The propagation of antibiotic resistance genes (ARGs) in freshwater reservoirs threatens ecosystem security and human health, and has attracted increasing attention. A series of recent research articles on ARGs provides a unique opportunity for data-driven discoveries in this emerging field. Here, we mined data from a total of 290 samples from 60 reservoirs worldwide with a data-driven framework (DD) developed to discover geographical distribution, influencing factors and pollution hotspots of ARGs in freshwater reservoirs. Most data came from Asia and Europe where nine classes of ARGs were most frequently detected in reservoirs with multi-drug resistance and sulfonamide resistance genes prevailing. Factors driving distribution of reservoir ARGs differed between reservoir waters and sediments, and interactions among these factors had linear or nonlinear enhancement effects on the explanatory power of ARG distribution. During the cold season, small-sized reservoir waters rich in organic carbon, mobile genetic elements (MGEs) and antibiotics had a higher pollution potential of ARGs; during the spring drought, sediments in large reservoirs located in densely populated areas were more conducive to dissemination of ARGs due to their richness in antibiotics and MGEs. Thus, distribution pattern of ARG pollution hotspots in reservoir waters and sediments varies greatly depending on the differences of internal and external factors. From the “One Health” perspective, this widespread contamination of freshwater reservoirs by ARGs we discovered through the DD framework should be a push to promote integrated research across regions and disciplines. Especially the human - food-chain - ecosystem interface needs an improved understanding of ARG contamination mechanisms and targeted monitoring and evaluation systems should be developed to maintain all ecosystem services in freshwater reservoirs as well as to safeguard human health.

Tracking antibiotic resistance genes in microplastic-contaminated soil

Agricultural soils and microplastics (MPs) are hotspots for antibiotic resistance genes (ARGs). Plastic mulch is the most important source of MPs in agricultural soil. ARGs, mobile genetic elements (MGEs), and their host profiles in long-term mulch MP-exposed soils remain unclear. In the present study, metagenomics was used to investigate the distribution patterns of ARGs and MGEs in eight Chinese provinces with a long history of plastic mulch use. A total of 204 subtypes of ARGs and thousands of MGEs (14 integrons, 28 insertions, and 2993 plasmids) were identified. A similar diversity of ARGs was found among MPs film-contaminated sites. The types of ARGs with a high abundance were more concentrated, and multidrug resistance genes were the dominant ARGs. Soils from regions with a longer history of plastic film use (such as Xinjiang province) had a higher abundance of ARGs and MGEs. The distribution of ARGs and MGEs exhibited a modular network distribution pattern. A total of 27 ARG subtypes and 29 MGEs showed co-occurrence network relationships. More than 10 common hosts of ARGs and MGEs, such as Pseudomonas, were found, and their abundances were highest in three provinces, including Xinjiang. This study may help elucidate the impact mechanism of long-term MP residues on the occurrence and spread of ARGs in soil.

Study on microbes and antibiotic resistance genes in karst primitive mountain marshes – A case study of Niangniang Mountain in Guizhou, China

Previous research on antibiotic resistance genes and microorganisms centered on those in urban sewage treatment plants, breeding farms, hospitals and others with serious antibiotic pollution. However, at present, there are evident proofs that antibiotic resistance genes (ARGs) indeed exist in a primitive environment hardly without any human’s footprints. Accordingly, an original karst mountain swamp ecosystem in Niangniang Mountain, Guizhou, China, including herbaceous swamp, shrub swamp, sphagnum bog and forest swamp, was selected to analyze the physical and chemical parameters of sediments. Moreover, microbial compositions, functions, as well as their connections with ARGs were assayed and analyzed using metagenomic technology. The results showed that there was no significant difference in the dominant microorganisms and ARGs in the four marshes, in which the dominant bacteria phyla were Proteobacteria (37.82 %), Acidobacteriota (22.17 %) and Actinobacteriota (20.64 %); the dominant archaea Euryarchaeota. (1.00 %); and the dominant eukaryotes Ascomycota (0.07 %), with metabolism as their major functions. Based on the ARDB database, the number of ARGs annotated reached 209 including 30 subtypes, and the dominant ARGs were all Bacitracin resistance genes (bacA, 84.77 %). In terms of the diversity of microorganisms and ARGs, the herbaceous swamp ranked the top, and the shrub swamp were at the bottom. Correlation analysis between microorganisms and resistance genes showed that, apart from aac2ic, macB, smeE, tetQ, and tetL, other ARGs were positively correlated with microorganisms. Among them, baca coexisted with microorganisms. Pearson correlation analysis results showed that contrary to ARGs, microorganisms were more affected by environmental factors.

The Sources and Potential Hosts Identification of Antibiotic Resistance Genes in the Yellow River, Revealed by Metagenomic Analysis.

The fate of antibiotic resistance genes (ARGs) has been revealed in various environmental media in recent years. Namely, the emergence of genes that resist colistin and carbapenems has attracted wide attention. However, the pollution condition of ARGs and sources in the Yellow River is still little understood, despite the river being the second longest in China. The present study determined the levels of ARG pollution in the Henan section of the Yellow River and evaluated the role of the aquaculture industry in the spread of ARGs. As revealed by the results, a total of 9 types of ARGs were detected in the sediments of the Yellow River, and the total ARG content in the Yellow River ranges from 7.27 to 245.45 RPKM. Sul1 and sul2 are the dominant ARGs, and the huge usage of sulfonamides, horizontal gene transfer, and wide bacteria host contribute to the prevalence of these two genes. The results of Spearman correlation analysis indicate that the breeding industry has little influence on ARGs in the Yellow River. Network analysis reveals that the opportunistic pathogen Pseudomonas is the potential host of sul1, tetG, and ANT(3′′)-IIa, which can pose a risk to human health.

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

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

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

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

Metagenomic insights into the antibiotic resistome in freshwater and seawater from an Antarctic ice-free area

The comprehensive profiles of antibiotic resistance genes (ARGs) in the Antarctic water environments and their potential health risks are not well understood. The present study characterized the bacterial community compositions and ARG profiles of freshwater (11 samples) and seawater (28 samples) around the Fildes Region (an ice-free area in Antarctica) using a shotgun metagenomic sequencing approach for the first time. There were significant differences in the compositions of the bacterial community and ARG profiles between freshwater and seawater. In the 39 water samples, 114 ARG subtypes belonging to 15 ARG types were detectable. In freshwater, the dominant ARGs were related to multidrug and rifamycin resistance. In seawater, the dominant ARGs were related to peptide, multidrug, and beta-lactam resistance. Both the bacterial community compositions and ARG profiles were significantly related to certain physicochemical properties (e.g., pH, salinity, NO3−). Procrustes analysis revealed a significant correlation between the bacterial community compositions and ARG profiles of freshwater and seawater samples. A total of 31 metagenome-assembled genomes (MAGs) carrying 35 ARG subtypes were obtained and identified. The results will contribute to a better evaluation of the ARG contamination in relation to human health in the Antarctic aquatic environments.

Reduction of antibiotic resistance genes (ARGs) in swine manure-fertilized soil via fermentation broth from fruit and vegetable waste

The issue of growing increase of antibiotic resistance genes (ARGs) in manure-fertilized soil needs urgently addressing. In this study, fermentation broth from fruit and vegetable waste was prepared to reduce ARG abundance in swine manure-fertilized soils. With a six-month field experiment, we found that swine manure-fertilized soil had significantly higher ARG abundance than soil applied with chemical fertilizer. As expected, the homemade fermentation broth significantly reduced ARG abundance in swine manure-fertilized soil, possibly through the decrease of abundance of Actinomyces, in which there was a 48.0%, 51.9%, and 66.7% decrease in the abundance of Nocardioides, Streptomyces, and Nonomuraea, respectively. With the bacteriostatic experiment, we observed that fermentation broth (5 mL/L) significantly inhibited the growth and metabolism in Actinomycetes spp. and Nocardioides sp., in terms of ATPase and PDH activity. These findings confirmed that the inhibition of Actinobacteria, some of the most dominant ARG hosts, was one of the main mechanisms responsible for the decrease in ARG abundance in fermentation broth-treated soil. This study provides field-scale evidence of a feasible strategy for controlling farmland ARG pollution, which is of utmost importance for soil health in the context of sustainable agriculture.

Metagenomic analysis of the abundances, diversity, and distribution of antibiotic resistance genes and their potential bacterial hosts in two types of shrimp-rearing farms in South China

Antibiotic resistance genes (ARGs) are attracting increasing concern worldwide. Many previous studies have investigated the occurrence and concentrations of ARGs in aquaculture. However, the sources of ARGs and the links with their potential bacterial hosts have not yet been explored. This study investigated the abundances and diversity of ARGs in two types of shrimp farms in South China through metagenomic sequencing. In total, 14 ARG types were detected. Tetracycline was the dominant ARG type. The abundances of ARGs in samples decreased in the order of duck feces > water source > sediment > shrimp gut > pond water. The samples from the duck–shrimp integrated farm contained 1.29–3.81-fold more abundant ARGs than those from the shrimp monoculture farm (p < 0.05). Several ARGs, that were most predominant in the duck feces samples, were also the most predominant in the shrimp gut samples from the duck–shrimp integrated farm. Redundancy analysis indicated that the abundances and distribution of ARGs formed three clusters: duck feces, water samples, and sediment and shrimp gut samples. The dominant genera in duck feces known as human pathogenic bacteria were potential hosts of ARGs, and were also dominant in the shrimp gut samples in the duck–shrimp integrated farm. Additionally, the abundances of dominant genera in the shrimp gut samples of the duck–shrimp integrated farm were 1.74–35.07-fold higher than those in the shrimp monoculture farm (p < 0.01). The duck–shrimp integrated farm had 3.36-fold and 4.94-fold higher abundances of ARGs and mobile genetic elements in the shrimp gut samples than those from the shrimp monoculture farm, respectively (p < 0.05). The results indicate that duck feces may be a crucial source of diverse and abundant ARGs spreading to reared shrimps in duck–shrimp integrated farms, posing a severe risk to public health.

Higher spreading risk of antibacterial biocide and heavy metal resistance genes than antibiotic resistance genes in aerobic granular sludge

Metagenomic approach was applied to simultaneously reveal the antibiotic resistance genes (ARGs) and antibacterial biocide & metal resistance genes (BMRGs), and the corresponding microbial hosts with high mobility during aerobic granular sludge (AGS) formation process. The results showed that the relative abundance of BMRGs was 88–123 times that of ARGs. AGS process was easier to enrich BMRGs, leading to a greater risk of drug resistance caused by BMRGs than that by ARGs. The enrichments of ARGs and BMRGs in AGS were closely related to several enhanced microbial metabolisms (i.e., cell motility, transposase and ATP-binding cassette transporters) and their corresponding regulatory genes. Several enhanced KEGG Orthologs (KO) functions, such as K01995, K01996, K01997 and K02002, might cause a positive impact on the spread of ARGs and BMRGs, and the main contributors were the largely enriched glycogens accumulating organisms. The first dominant ARGs (adeF) was carried by lots of microbial hosts, which might be enriched and propagated mainly through horizontal gene transfer. Candidatus Competibacter denitrificans simultaneously harbored ARG (cmx) and Cu related RGs (corR). Many enriched bacteria contained simultaneously multiple BMRGs (copR and corR) and mobile genetic elements (integrons and plasmids), granting them high mobility capabilities and contributing to the spread of BMRGs. This study might provide deeper understandings of the proliferation and mobility of ARGs and BMRGs, importantly, highlighted the status of BMRGs, which laid the foundation for the controlling widespread of resistance genes in AGS.

Metagenomic investigation of the seasonal distribution of bacterial community and antibiotic-resistant genes in Day River Downstream, Ninh Binh, Vietnam

In this study, we use high-throughput sequencing-based metagenomic methods to investigate the differences in seasonal structures of the bacterial community and the abundance and diversity of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) in both shrimp ponds and river water samples downstream of the Day River, Ninh Binh, Vietnam. The structure of the central bacterial community, ARGs, and MGEs was found to be regardless of the seasons and locations. The predominant phyla found in all samples was Proteobacteria, Bacteroidetes, and Actinobacteria. Multi-drug resistance (MDR) genes and transposases are the most dominant ARG types and MGEs, respectively. Our data showed a higher abundance of bacterial communities, ARGs, and MGEs in the river water during the rainy season. There is a significant correlation between the abundance of ARGs, MGEs, and environmental factors. Our results indicate that water environments containing ARGs/MGEs carrying bacteria pose a risk to shrimp and human health, especially during the rainfall-polluted water season.

Metagenomic analysis of microbial community structure and distribution of resistance genes in Daihai Lake, China

The emergence of resistance genes is a global phenomenon that poses a significant threat to both animals and humans. Lakes are important reservoirs of genes that confer resistant to antibiotics and metals. In this study, we investigated the distribution and diversity of antibiotic resistance genes (ARGs) and metal resistance genes (MRGs) in the sediment of Daihai Lake using high-throughput sequencing and metagenomic analysis. The results indicated that all sampling sites had similar bacterial community structures, with Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes being the most abundant. A total of 16 ARG types containing 111 ARG subtypes were deposited in the sediment. Among the resistance genes to bacitracin, multidrug, macrolide-lincosamide-streptogramin (MLS), tetracycline, beta-lactam, and sulfonamide were the dominant ARG types, accounting for 89.9–94.3% of the total ARGs. Additionally, 15 MRG types consisting of 146 MRG subtypes were identified. In all samples, MRGs of the same type presented resistance to Pb, Ni, Hg, W, Zn, Ag, Cr, Fe, As, Cu, and multimetals. Overall, the distribution and diversity of antibiotic and metal resistance genes showed no significant differences in the samples. Plasmids (91.03–91.82%) were the most dominant mobile genetic elements in the sediments of Daihai Lake. Network analysis indicated that the target ARGs and MRGs were significantly positively correlated with the microorganisms. Potential hosts for various ARGs and MRGs include Proteobacteria, Euryarchaeota, Actinobacteria, Chloroflexi, and Bacteroidetes.

Pollution Characteristics and Driving Factors of Antibiotic Resistance Genes in Dexing Copper Mine

Environmental antibiotic resistance genes (ARGs) are a type of emerging pollutant that has been widely concerning. However, investigations into the contamination of ARGs in mining areas have been scarce. Here, the types, abundances, and influencing factors of ARGs and mobile genetic elements (MGEs) were investigated in soil/sediment of the Dexing copper mine area in June 2019 by using high-throughput quantitative polymerase chain reaction (HT-qPCR). Furthermore, the influence of heavy metals and MGEs factors on ARGs was studied using the multivariate statistical analysis method. The results showed that there were a variety of ARGs in the Dexing copper mining area, and the maximum detected number of ARGs was 70. At the relative abundance level, the relative abundance of individual sites reached 0.085. In the Dexing copper mine, multidrug, MLSB, β-lactamases, tetracycline, and aminoglycoside resistance genes were the dominant ARG classes based on their numbers. The efflux pump was the most dominant resistance mechanism, followed by antibiotic deactivation and cellular protection. There was a significant positive correlation between the abundance of ARGs and MGEs (P<0.05), and TnpA04 and Inti1 were the most important MEGs in Dexing copper mine samples, indicating that horizontal gene transfer might be an important mechanism for the spread of environmental ARGs. The results of Pearson correlation analysis and RDA analysis showed that the content of Cu was significantly positively correlated with the detected numbers and abundance of ARGs (P<0.05), suggesting that the high content of Cu in the Dexing copper mining area might be an important driving factor for the formation of ARGs.

Antibiotic resistomes in water supply reservoirs sediments of central China: main biotic drivers and distribution pattern.

Water supply reservoirs form one of the critical drinking water resources. Their water quality directly affects human health. However, reservoir sediments have not received adequate attention in antibiotic resistance genes (ARGs) dissemination, though they reflect long-term ARGs contamination of water supply reservoirs. Moreover, the physicochemical parameters in water supply reservoir sediments are generally better than those in the other media. Thus, the main ARGs biotic drivers of the media would demonstrate their unique characteristics. In this study, sediment samples were collected from 10 water supply reservoirs in central China, and the antibiotic resistomes were determined with the metagenomic method. As revealed from the results, 174 ARGs (18 ARG types) were detected in the reservoir sediment. Besides, multidrug-, sulfonamide-, and vancomycin-ARGs were the dominant ARGs in the sediment samples. The macrolide-resistant Microcystis was prevalent (100% detection frequency with 0.35% average percentage) in reservoir sediments and posed potential risks to human health. Furthermore, the results of the Mantel test and VPA demonstrated that mobile genetic elements (MGEs) were the more essential biotic drivers in ARG contents of reservoir sediments rather than the bacteria community.

Intervention of antimicrobial peptide usage on antimicrobial resistance in aquaculture

Antibiotic resistance genes (ARGs) widely exist in diverse matrices to pose potential risks and attract global attention. This study performed tilapia-cultivation experiment to discuss the intervention of antimicrobial peptide usage on antimicrobial resistance in aquaculture by analyzing 29 target ARGs in water, residue, and fish muscle samples. The sul1 gene was the dominant ARG in most of samples while absolute abundances of total ARGs were in the range of 1.5 × 102-2.3 × 107 copies/mL in water samples and 2.1 × 108-3.9 × 109 copies/g in the residue and fish muscle samples. Compared with antibiotic treatment, the cecropin treatment could significantly reduce the absolute abundance of ARGs and intI1 in all samples. Proteobacteria were the dominant phylum for most of samples while Aeromonas/Cetobacterium served as the dominant genus for samples with antibiotic/cecropin treatment. Network analysis showed that cecropin treatment could significantly reduce occurrence of multi-drug resistant bacteria and the correlation between genera and ARGs. Results of partial least squares structural equation modeling which was used to discuss the influential factors of ARGs exhibited that antibiotics could increase ARGs, further proving that cecropin peptide could effectively reduce ARGs in aquaculture. This study will provide important information on controlling ARGs in aquaculture in terms of antimicrobial peptide application.

Characteristics and removal of antibiotics and antibiotic resistance genes in a constructed wetland from a drinking water source in the Yangtze River Delta

Safe drinking water is crucial to public health. However, approximately one-third of the world's population lacks access to clean drinking water. The presence of antibiotics and antibiotic resistance genes (ARGs) in drinking water sources has become a severe problem worldwide due to its potential threat to human health. We monitored the occurrence and variations of 23 antibiotics and 9 ARGs in different treatment processes in a constructed wetland serving as drinking water source in the Yangtze River Delta, China. The studied wetland is consisted of four treatment processes: pretreatment area, pump station lifting, root-channel ecological purification area and deep purification area. Except for sulfapyridine and roxithromycin, 21 antibiotics were detected at concentrations ranging from 0.15 to 59.52 ng/L. The concentration of macrolides was the highest in this wetland, especially tylosin (42.86–59.52 ng/L). TetG, tetX and sul2 were the dominant ARGs in both water (2.41 × 10−4-1.87 × 10−2) and sediment (6.65 × 10−5-4.92 × 10−3). In addition, a strong correlation between ARGs in water and ARGs in sediment (Pearson, R2 > 0.9, p < 0.05) indicated an exchange between the two phases. Moreover, the significantly positive correlation of ARGs between the inlet and outlet of each subsystem illustrated that upstream pollution was the primary source for downstream processes. In general, the wetland system could efficiently eliminate antibiotics (9.0–53.8%) and ARGs (14.5–94.1%), with the deep purification area having the highest removal efficiency. Overall, our results provide important insights into the occurrence, abundance and removal of antibiotics and ARGs in the constructed wetland serving as drinking water sources.

How Does Vegetable Waste Decomposition Influence the Antibiotic Resistome and the Human Bacterial Pathogen Structure in Leachates?

Understanding how antibiotic-resistance genes (ARGs) and pathogens in household waste leachates contribute to acquiring antibiotic resistance is profoundly significant. Leachates can impair water quality and harm the environment and human health. This study explored co-occurrence and accumulation of pathogens and ARGs in leachates during decomposition of vegetable wastes. Watercress leachates had more average ARG numbers and abundance. ARG abundances were 2.17 × 107 copies/mL for pepper, 1.27 × 107 copies/mL for watercress, and 4.93 × 105 copies/mL for onion. Vegetable decomposition enriched ARGs first and then decreased. Genes conferring resistance to multidrug, aminoglycosides, and β-lactam were the dominant ARGs; the main mechanism was antibiotic deactivation. Preferential co-occurrence patterns were observed across ARG types, indicating coeffects of antibiotic selection pressure and coresistance when shaping the antibiotic resistome. The pathogenic community varied significantly during vegetable decomposition. ARG levels positively correlated with the dominant Enterococcus but negatively correlated with Bartonella and Bacteroides. Functional genes shared by all waste leachates were mainly linked to carbohydrate (9.2%) and amino acid transport and metabolism (8.2%). ARGs were positively associated with defensive virulence factors and autoinducers. The quorum sensing system regulated ARG levels by balancing signal molecule synthesis and degradation gene expression. These findings provide insights into the mechanism underlining vegetable waste decomposition regarding occurrence and spread of ARGs in leachates.

Bacterial Concentrations and Water Turbulence Influence the Importance of Conjugation Versus Phage-Mediated Antibiotic Resistance Gene Transfer in Suspended Growth Systems.

Despite the abundance of phage-borne antibiotic resistance genes (ARGs) in the environment, the frequency of ARG propagation via phage-mediated transduction (relative to via conjugation) is poorly understood. We investigated the influence of bacterial concentration and water turbulence level [quantified as Reynold's number (Re)] in suspended growth systems on the frequency of ARG transfer by two mechanisms: delivery by a lysogenic phage (phage λ carrying gentamycin-resistance gene, genR) and conjugation mediated by the self-transmissible plasmid RP4. Using Escherichia coli (E. coli) as the recipient, phage delivery had a comparable frequency (1.2 ± 0.9 × 10-6) to that of conjugation (1.1 ± 0.9 × 10-6) in suspensions with low cell concentration (104 CFU/mL) and moderate turbulence (Re = 5 × 104). Turbulence affected cell (or phage)-to-cell contact rates and detachment (due to shear force), and thus, it affected the relative importance of conjugation versus phage delivery. At 107 CFU/mL, no significant difference was observed between the frequencies of ARG transfer by the two mechanisms under quiescent water conditions (2.8 ± 0.3 × 10-5 for conjugation vs 2.2 ± 0.5 × 10-5 for phage delivery, p = 0.19) or when Re reached 5 × 105 (3.4 ± 1.5 × 10-5 for conjugation vs 2.9 ± 1.0 × 10-5 for phage delivery, p = 0.52). Transcriptomic analysis of genes related to conjugation and phage delivery and simulation of cell (or phage)-to-cell collisions at different Re values corroborate that the importance of phage delivery relative to conjugation increases under either quiescent or turbulent conditions. This finding challenges the prevailing view that conjugation is the dominant ARG transfer mechanism and underscores the need to consider and mitigate potential ARG dissemination via transduction.