Can Mercury Influence Carbon Dioxide Levels? Implications for the Implementation of the Minamata Convention on Mercury.

The Paris Agreement and the Minamata Convention on Mercury are two of the most important environmental conventions being implemented concurrently, with a focus on reducing carbon and mercury emissions, respectively. The relation between mercury and carbon influences the interactions and outcomes of these two conventions. This perspective investigates the link between mercury and CO2, assessing the consequences and exploring the policy implications of this link. We present scientific evidence showing that mercury and CO2 levels are negatively correlated under natural conditions. As a result of this negative correlation, the CO2 level under the current mercury reduction scenario is predicted to be 2.4-10.1 ppm higher than the no action scenario by 2050, equivalent to 1.0-4.8 years of CO2 increase due to human activity. The underlying causations of this negative correlation are complex and need further research. Economic analysis indicates that there is a trade-off between the benefits and costs of mercury reduction actions. As reducing mercury emission may inadvertently undermine efforts to achieve climate goals, we advocate for devising a coordinated implementation strategy for carbon and mercury conventions to maximize synergies and reduce trade-offs.

Normalization Approach by a Reference Material to Improve LC-MS-Based Metabolomic Data Comparability of Multibatch Samples.

Large cohorts of samples from multiple batches are usually required for global metabolomic studies to characterize the metabolic state of human disease. As such, it is critical to eliminate systematic variation and truly reveal the biologically associated alterations. In this study, we proposed a reference material-based approach (Ref-M) for data correction by liquid chromatography-mass spectrometry and represented by an analysis of multibatch human serum samples. The reference material was generated by mixing serum from healthy donors and distributed to each extraction batch of subject samples. Pooled quality control samples and isotopic internal standards were then applied in each acquisition batch for data quality control. Finally, each metabolite in subject samples was normalized by its counterpart in the reference serum. We demonstrated that Ref-M significantly enhanced the numbers of efficient features and effectively eliminated the batch variation of 522 serum samples of healthy individuals, benign pulmonary nodules, and lung cancer patients. Twenty differential metabolites were identified to distinguish lung cancer from healthy controls in the training set. The discriminant model was validated in an independent data set with an area under the receiver operating characteristics (ROC) curve (AUC) of 0.853. Another 40 serum samples further tested with Ref-M were achieved an AUC of 0.843 by the established model. Our results showed that the reference material-based approach presents the potential to improve the data comparability and precision for biomarker discovery in large-scale metabolomic studies.

Mercury (Hg) in the "Skin" of the Ocean: Dissolved Gaseous Hg, Total Hg, and Hg Redox Chemistry in Sea Surface Microlayer and Implication for Air-Sea Hg Exchange.

The sea surface microlayer (SML) is the uppermost ∼1000 µm of the surface of the ocean. With distinct physicochemical properties and position relative to the adjacent subsurface waters (SSWs), the ubiquitous distribution and high dynamics of the SML greatly regulate the global air-sea gas exchange and biogeochemistry. Mercury (Hg) redox chemistry in surface seawaters and air-sea exchange of gaseous Hg (mainly Hg(0)) fundamentally control the global oceanic Hg cycle. However, the occurrence and transformation of Hg in the SML have been poorly quantified. Here we optimize the traditional SML sampling system to make it more suitable for dissolved gaseous Hg (DGM, mainly Hg(0)) sampling. We then assess the temporal and spatial variability of DGM, total Hg, dissolved organic carbon (DOC), and Hg redox chemistry in the SML and SSWs of diverse marine environments. Our data suggest a general DGM, total Hg, and DOC enrichment in the SML relative to the SSWs but with complex variability in time and space. The incubation experiments further reveal the complex characteristics of Hg redox chemistry between the SML and SSWs. We discuss important implications of the SML Hg cycle on air-sea Hg exchange and suggest wider investigations of the SML Hg cycle in the global hydrosphere.

Recent progress in analytical strategies of arsenic-binding proteomes in living systems.

Arsenic (As) is one of the most concerning elements due to its high exposure risks to organisms and ecosystems. The interaction between arsenicals and proteins plays a pivotal role in inducing their biological effects on living systems, e.g., arsenicosis. In this review article, the recent advances in analytical techniques and methods of As-binding proteomes were well summarized and discussed, including chromatographic separation and purification, biotin-streptavidin pull-down probes, in situ imaging using novel fluorescent probes, and protein identification. These analytical technologies could provide a growing body of knowledge regarding the composition, level, and distribution of As-binding proteomes in both cells and biological samples, even at the organellar level. The perspectives on analysis of As-binding proteomes are also proposed, e.g., isolation and identification of minor proteins, in vivo targeted protein degradation (TPD) technologies, and spatial As-binding proteomics. The application and development of sensitive, accurate, and high-throughput methodologies of As-binding proteomics would enable us to address the key molecular mechanisms underlying the adverse health effects of arsenicals.

Lead activates neutrophil degranulation to induce early myocardial injury in mice.

Lead (Pb) is a pervasive toxic metal contaminant associated with a high risk of myocardial injury. However, the precise mechanism underlying Pb-induced myocardial injury has yet to be fully elucidated. In this study, a murine model of Pb exposure (0, 1, 5, and 10 mg/kg) was employed to investigate the involvement of neutrophil degranulation in the induction of myocardial injury. Notably, serum levels of cardiac troponin I (cTnI) and creatine kinase-MB (CK-MB) increased significantly in Pb-exposed mice, whereas cTnI levels in cardiomyocytes decreased, suggesting that Pb exposure may cause early myocardial injury. Moreover, Pb exposure was found to promote neutrophil degranulation, as evidenced by elevated myeloperoxidase (MPO) and neutrophil elastase (NE) concentrations in both the serum of Pb-exposed workers and Pb-exposed mice, as well as the extracellular supernatant of neutrophils following exposure. However, we found that serum level of cTnI enhanced by Pb exposure is associated with increased NE levels in the serum, but not with MPO levels. Upon treatment with NE inhibitor (sivelestat), the serum level of cTnI markedly reduced in Pb-exposed mice, we found that early myocardial injury is associated with NE levels in the serum. At the molecular level, western blotting analysis revealed an upregulation of ERK1/2 expression in vitro following Pb exposure, suggesting that the activation of the ERK1/2 signaling pathway may underlie the participation of neutrophil degranulation in Pb-induced myocardial injury. In summary, our findings demonstrate that Pb exposure can initiate early myocardial injury by promoting the neutrophil degranulation process, thereby highlighting the potential role of this process in the pathogenesis of Pb-associated myocardial injury.

Unified Probability Distribution and Dynamics of Lead Contents in Human Erythrocytes Revealed by Single-Cell Analysis.

Understanding the presence and dynamics of chemical pollutants in individual cells is fundamentally important for their trafficking, fate, and toxicity in humans. The presence of molecular components (i.e., proteins and mRNA) in individual cells of higher organisms is considered a stochastic event. The characteristics of chemical pollutants, as extrinsic compounds, in subpopulation of human cells on single-cell basis have not been explored yet. Here, we demonstrated the lead (Pb) content in individual mature erythrocytes (m-erythrocytes) of Pb-intoxicated patients, and healthy subjects exhibited a unified pattern in probability distribution (gamma distribution) and dynamics, despite being highly heterogeneous. The Pb content in individual m-erythrocytes decreased with the lifetime of m-erythrocytes. Meanwhile, the distribution and dynamics were found to be highly related to the Pb content in m-erythrocytes and was independent of patients and their status. This is the first study to analyze the distribution pattern of chemical pollutants at a single-cell level in higher organisms. This study sheds light on the molecular mechanism of Pb trafficking and fate in humans and the search for an efficient strategy to improve Pb excretion during Pb treatment.

Lab-on-Membrane Platform Coupled with Paper Spray Ionization for Analysis of Prostate-Specific Antigen in Clinical Settings.

The analysis of protein antigens as biomarkers in clinical samples is particularly helpful for the early diagnosis of diseases. However, this is difficult to accomplish owing to the presence of the antigens in trace amounts as well as the complexity of the matrixes in clinical samples. In this study, a lab-on-membrane platform that can be combined with paper spray ionization mass spectrometry was developed for the in situ high-throughput sensitive detection of the prostate-specific antigen (PSA). The sensitivity of the proposed platform was enhanced via two strategies: (1) the synthesis of a biotin-streptavidin scaffold caused an increase in the capturing efficiency of PSA by a factor of 5 and (2) the immobilization of a large number of mass tag molecules on the gold nanoparticles allowed for the amplification of the mass spectrometry signals. The limit of detection was approximately 3.0 pg mL-1. The selectivity to PSA was guaranteed by using an antibody-aptamer pairing sandwich immunoassay, and PSA detection was unaffected even when other protein antigens (carcinoembryonic antigen and carbohydrate antigen 125) were present. The modified membranes maintained their performance for at least 30 days when stored at 4 °C. Finally, analysis of human serum samples confirmed that the PSA concentration as determined using the proposed platform was consistent with that determined with a conventional chemiluminescent immunoassay. Thus, this PSA analyzing platform is suitable for prostate cancer diagnosis in clinical settings.

Spatial and seasonal variations of antibiotic resistance genes and antibiotics in the surface waters of Poyang Lake in China.

Antibiotic resistance genes (ARGs) and antibiotics in the aquatic environment raise health concerns particularly on the dispersal and persistence of antibiotic resistance. Large lakes, which serve as catch basins of anthropogenic inputs provide an ideal environment for understanding the occurrence and accumulation of ARGs and antibiotics in freshwater environments. Here, the largest freshwater lake in China, Poyang Lake, located in the developing district of Yangtze valley was used to study the characterization of the spatial and seasonal variation of both ARGs and antibiotics. Results showed that twelve tested ARGs (sul1, sul2, sul3, tetA, tetB, tetC, tetH, tetW, tetO, tetM, qnrS, and qnrB) were detected in the surface waters of Poyang Lake, with a detection frequency ranging from 19.2% to 100%, and sul2 and tetA genes were identified as potential indicators of ARG pollution in this region. Among the 11 analyzed antibiotics, sulfonamides were the predominant antibiotics with a contribution of more than 50% to the total concentrations of tested antibiotics. The total concentrations of both ARGs and antibiotics were higher in the dry season than those in the wet season. Furthermore, ARGs and antibiotics in the surface waters also varied with sampling locations, being consistently at riverine tributaries. Positive correlations were also observed between the concentrations of ARGs and antibiotics, as well as the integron gene (intI1), indicating that antibiotics and intI1 may be playing important roles in the occurrence and dispersal of ARGs in the surface waters. Lastly, our results suggest that intensive anthropogenic activities related to antibiotic usage have substantially contributed to the occurrence and persistence of ARGs and antibiotics in Poyang Lake.

Bacteria and Antibiotic Resistance Genes (ARGs) in PM2.5 from China: Implications for Human Exposure.

Airborne transmission is one of the environmental dissemination pathways of antibiotic resistance genes (ARGs), and has critical implications for human exposure through inhalation. In this study, we focused on three regions of China to reveal some unique spatiotemporal features of airborne bacteria and ARGs in fine aerosols (PM2.5): (1) greater seasonal variations in the abundance of bacteria and ARGs in temperate urban Beijing than in the subtropical urban areas of the Yangtze River Delta (YRD) and Pearl River Delta (PRD) regions, with regional disparities in bacterial communities; (2) geographical fingerprints of ARG profiles independent of seasonal cycles and land-use gradients within each region; (3) region-independent associations between the targeted ARGs and limited bacterial genera; (4) common correlations between ARGs and mobile genetic elements (MGEs) across regions; and (5) PM2.5 at the higher end of ARG enrichment across various environmental and human media. The spatiotemporally differentiated bacterial communities and ARG abundances, and the compositions, mobility, and potential hosts of ARGs in the atmosphere have strong implications for human inhalational exposure over spatiotemporal scales. By comparing other contributing pathways for the intake of ARGs (e.g., drinking water and food ingestion) in China and the U.S.A., we identified the region-specific importance of inhalation in China as well as country-specific exposure scenarios. Our study thus highlights the significance of inhalation as an integral part of the aggregate exposure pathways of environmentally disseminated ARGs, which, in turn, may help in the formulation of adaptive strategies to mitigate the exposure risks in China and beyond.

Prevalence, Biotransformation, and Maternal Transfer of Synthetic Phenolic Antioxidants in Pregnant Women from South China.

Synthetic phenolic antioxidants (SPAs) have been identified as an emerging group of contaminants in recent years. However, there are significant gaps in our knowledge of human prenatal exposure to these synthetic chemicals. In this study, a set of eight SPAs and four major transformation products (TPs) were systematically analyzed in matched samples of maternal plasma, cord plasma, and placenta from a population of pregnant women. Five of the eight target SPAs and all four target TPs were frequently detected in the maternal-placental-fetal unit, indicating prenatal exposure to SPAs and the transfer of SPAs across the placenta. In the three matrices, 2,6-di-tert-butyl-hydroxytoluene (BHT), 2,4-di-tert-butylphenol (DBP), and 2,2'-methylenebis(4-methyl-6-tert-butylphenol) (AO 2246) were identified as the most abundant SPAs, while 2,6-di-tert-butyl-1,4-benzoquinone (BHT-Q) and 2,6-di-tert-butyl-4-hydroxy-4-methyl-2,5-cyclohexadienone (BHT-quinol) were identified as the predominant TPs of BHT. In the maternal plasma, concentrations of both BHT-Q and BHT-quinol were significantly correlated with BHT (p < 0.001), suggesting that the two TPs mainly originated from the biotransformation of BHT itself in pregnant women. The transplacental transfer efficiencies (TTEs) of the SPAs and TPs were structure-dependent and generally less than 1. Significantly higher TTEs for four target TPs than their parent BHT were identified. To our knowledge, this study provides the first evidence that SPAs and TPs transfer across the placenta in pregnant women.

Mercury methylation-related microbes and genes in the sediments of the Pearl River Estuary and the South China Sea.

Methylmercury (MeHg) is a toxicant that mainly originates from in situ microbial methylation of inorganic mercury (Hg) in the environment and poses a severe health risk to the public. However, the characteristics of the Hg-methylating microbial community and its relationship with MeHg production in various environments remain to be understood. In the present study, Hg-methylating microbial communities and genes (hgcAB cluster) in the sediments of the Pearl River (PR), Pearl River Estuary (PRE) and South China Sea (SCS) were investigated at a large spatial scale using high-throughput sequencing-based approaches. The results showed that sulfur-reducing bacteria (SRB) and iron-reducing bacteria (IRB) were consistently the dominant microbial strains responsible for the methylation of inorganic Hg in all three regions investigated. The abundance and diversity of Hg-methylating communities and genes were both found to be higher in the PR sediments compared to that in the PRE and SCS sediments, and in good agreement with the spatial distribution of MeHg. Furthermore, a significant correlation was observed between the MeHg concentration and the abundance of both hgcA and hgcB genes in the sediments of the PR, PRE and SCS regions. Overall, the present study suggested that there was the presence of a close link between MeHg and Hg-methylating communities or genes in the ambient aquatic environment, which could be used to reflect the potential of in situ MeHg production.

Metagenomic characterization of antibiotic resistance genes in Antarctic soils.

Antibiotic resistance genes (ARGs) are considered environmental pollutants. Comprehensive characterization of the ARGs in pristine environments is essential towards understanding the evolution of antibiotic resistance. Here, we analyzed ARGs in soil samples collected from relatively pristine Antarctica using metagenomic approaches. We identified 79 subtypes related to 12 antibiotic classes in Antarctic soils, in which ARGs related to multidrug and polypeptide were dominant. The characteristics of ARGs in Antarctic soils were significantly different from those in active sludge, chicken feces and swine feces, in terms of composition, abundance and potential transferability. ARG subtypes (e.g., bacA, ceoB, dfrE, mdtB, amrB, and acrB) were more abundant than others in Antarctic soils. Approximately 60% of the ARGs conferred antibiotic resistance via an efflux mechanism, and a low fraction of ARGs (∼16%) might be present on plasmids. Culturable bacterial consortiums isolated from Antarctic soils were consistently susceptible to most of the tested antibiotics frequently used in clinical therapies. The amrB and ceoB carried by culturable species did not express the resistance to aminoglycoside and fluoroquinolone at the levels of clinical concern. Our results suggest that the wide use of antibiotics may have contributed to developing higher antibiotic resistance and mobility.

Bacterial resistance to lead: Chemical basis and environmental relevance.

Natural bacterial isolates from heavily contaminated sites may evolve diverse tolerance strategies, including biosorption, efflux mechanism, and intracellular precipitation under the continually increased stress of toxic lead (Pb) from anthropogenic activities. These strategies utilize a large variety of functional groups in biological macromolecules (e.g., exopolysaccharides (EPSs) and metalloproteins) and inorganic ligands, including carboxyl, phosphate and amide groups, for capturing Pb. The amount and type of binding sites carried by biologically originated materials essentially determines their performance and potential for Pb removal and remediation. Many factors, e.g., metal ion radius, electronegativity, the shape of the cell surface sheath, temperature and pH, are thought to exert significant influences on the abovementioned interactions with Pb. Conclusively, understanding the chemical basis of Pb-binding in these bacteria can allow for the development of effective microbial Pb remediation technologies and further elucidation of Pb cycling in the environment.

Natural Porphyrins Accelerating the Phototransformation of Benzo[a]pyrene in Water.

Phototransformation is one of the most important transformation pathways of organic contaminants in the water environment. However, how active compounds enable and accelerate the phototransformation of organic pollutants remains to be elucidated. In this study, the phototransformation of benzo[a]pyrene (BaP, the first class "human carcinogens") by various natural porphyrins under solar irradiation was investigated, including chlorophyll a, sodium copper chlorophyllin, hematin, cobalamin, and pheophorbide a. Transformation efficiency of BaP varied considerably with chemical stabilities of the porphyrins. Porphyrins with a lower stability displayed higher BaP transformation efficiencies. BaP transformation had a significant positive correlation with the production of singlet oxygen. Identical phototransformation products of BaP were observed for all investigated porphyrins, and the main products were identified as BaP-quinones, including BaP-1,6-dione, BaP-3,6-dione, and BaP-6,12-dione. The mechanism of natural porphyrins accelerating the BaP phototransformation in water was proposed to proceed via the photocatalytic generation of singlet oxygen resulting in the transformation of BaP to quinones.

Exploring the differences of antibiotic resistance genes profiles between river surface water and sediments using metagenomic approach.

To better understand the potential genic communication and dissemination of antibiotic resistance genes (ARGs) in different environmental matrices, the differences of ARG profiles between river surface water and sediments were explored. Metagenomic analysis was applied to investigate the comprehensive ARG profiles in water and sediment samples collected from the highly human-impacted catchment of the Beijiang River and its river source. A total of 135 ARG subtypes belonging to 18 ARG types were identified. Generally, ARGs in surface water were more diverse and abundant than those in sediments. ARG profiles in the surface water and sediment samples were distinct from each other, but some ARGs were shared by the surface water and sediments. Results revealed that multidrug and bacitracin resistance genes were the predominant ARGs types in both surface water (0.30, 0.17 copies/cell) and sediments (0.19, 0.15 copies/cell). 73 ARG subtypes were shared by the water and sediment samples and had taken over 90% of the total detected ARG abundance. Most of the shared ARGs are resistant to the clinically relevant antibiotics. Furthermore, significant correlations between the ARGs and 21 shared genera or mobile genetic elements (MGEs) (plasmids and integrons) were found in surface water and sediments, suggesting the important role of genera or MGEs in shaping ARGs profiles, propagation and distribution. These findings provide deeper insight into mitigating the propagation of ARGs and the associated risks to public health.

Characterizing the antibiotic resistance genes in a river catchment: Influence of anthropogenic activities.

Previous studies on environmental antibiotics resistance genes (ARGs) have focused on the pollution sources such as wastewater treatment plants, aquaculture and livestock farms, etc. Few of them had addressed this issue in a regional scale such as river catchment. Hence, the occurrence and abundances of 23 ARGs were investigated in surface water samples collected from 38 sites which located from the river source to estuary of the Beijiang River. Among them, 11 ARGs were frequently detected in this region and 5 ARGs (sulI, sulII, tetB, tetC, and tetW) were selected for their distribution pattern analysis. The abundances of the selected ARGs were higher in the upstream (8.70×106copies/ng DNA) and downstream areas (3.17×106copies/ng DNA) than those in the midstream areas (1.23×106copies/ng DNA), which was positively correlated to the population density and number of pollution sources. Pollution sources of ARGs along the Beijiang River not only had a great impact on the abundances and diversity, but also on the distribution of specific ARGs in the water samples. Both sulI and sulII were likely originated from aquaculture farms and animal farms, tetW gene was possibly associated with the mining/metal melting industry and the electric waste disposal and tetC gene was commonly found in the area with multiple pollution sources. However, the abundance of tetB was not particularly related to anthropogenic impacts. These findings highlight the influence of pollution sources and density of population on the distribution and dissemination of ARGs at a regional scale.

Novosphingobium guangzhouense sp. nov., with the ability to degrade 1-methylphenanthrene.

A novel Gram-stain-negative, flagellated, rod-shaped, yellow-pigmented aerobic bacterium, strain SA925T, that is capable of degrading 1-methylphenanthrene was isolated from oil-polluted soil collected from a refinery located in Guangzhou, China. Phylogenetic analysis based on the 16S rRNA gene sequence demonstrated that strain SA925T belongs to the genus Novosphingobium and is evolutionarily close to the type strains of Novosphingobium gossypii (98.5 % similarity), Novosphingobium panipatense (98.2 %), Novosphingobium mathurense (98.0 %) and Novosphingobium pentaromativorans (96.5 %). The G+C content of the genomic DNA was 60.2 mol%. DNA-DNA hybridization experiments between strain SA925T and the closest strain, Novosphingobium gossypii JM-1396T, revealed a low level of relatedness (35.5 %). Strain SA925T grew at 10-35 °C, at pH 6.0-8.0 and in the presence of 0-4 % (w/v) NaCl. The major fatty acids were C18 : 1ω7c, C16 : 0 and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c). The polar lipid profiles mainly consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidyldimethylethanolamine, phosphatidylethanolamine and sphingoglycolipid (the characteristic polar lipid). The predominant ubiquinone was Q-10. The major polyamine was spermidine. Based on the phylogenetic, phenotypic and physiological characteristics, strain SA925T was considered to represent a novel species of the genus Novosphingobium, for which the name Novosphingobium guangzhouense sp. nov. is proposed. The type strain is SA925T (=DSM 32207T=GDMCC 1.1110T).

Identification of Methylated Dithioarsenicals in the Urine of Rats Fed with Sodium Arsenite.

Biotransformation of inorganic arsenic results in the formation of methylarsenicals of both oxygen and sulfur analogues. Aiming to improve our understanding of metabolism of inorganic arsenic in animals, we conducted an animal feeding study with an emphasis on identifying new arsenic metabolites. Female F344 rats were given 0, 1, 10, 25, 50, and 100 µg/g of arsenite (iAs(III)) in the diet. Arsenic species in rat urine were determined using high performance liquid chromatography (HPLC) separation and inductive coupled plasma mass spectrometry (ICPMS) and electrospray ionization tandem mass spectrometry (ESI MS/MS) detection. Nine arsenic species were detected in the urine of the iAs(III)-dosed rats. Seven of these arsenic species were consistent with previous reports, including iAs(III), arsenate, monomethyarsonic acid, dimethylarsinic acid, trimethylarsine oxide, monomethylmonothioarsonic acid, and dimethylmonothioarsinic acid. Two new methyldithioarsencals, monomethyldithioarsonic acid (MMDTA(V)) and dimethyldithioarsinic acid (DMDTA(V)), were identified for the first time in the urine of rats treated with iAs(III). The concentrations of both MMDTA(V) and DMDTA(V) in rat urine were dependent on the dosage of iAs(III) in diet. The concentration of DMDTA(V) was approximately 5 times higher than that of MMDTA(V). MMDTA(V) has not been identified in any biological samples of animals, and DMDTA(V) has not been reported as a metabolite of inorganic arsenic in the rats. The identification of novel methylated dithioarsenicals as metabolites of inorganic arsenic in the rat urine provided further insights into the understanding of the metabolism of arsenic.

Metagenomic Analysis Revealing Antibiotic Resistance Genes (ARGs) and Their Genetic Compartments in the Tibetan Environment.

Comprehensive profiles of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) in a minimally impacted environment are essential to understanding the evolution and dissemination of modern antibiotic resistance. Chemical analyses of the samples collected from Tibet demonstrated that the region under investigation was almost devoid of anthropogenic antibiotics. The soils, animal wastes, and sediments were different from each other in terms of bacterial community structures, and in the typical profiles of ARGs and MGEs. Diverse ARGs that encoded resistance to common antibiotics (e.g., beta-lactams, fluoroquinolones, etc.) were found mainly via an efflux mechanism completely distinct from modern antibiotic resistome. In addition, a very small fraction of ARGs in the Tibetan environment were carried by MGEs, indicating the low potential of these ARGs to be transferred among bacteria. In comparison to the ARG profiles in relatively pristine Tibet, contemporary ARGs and MGEs in human-impacted environments have evolved substantially since the broad use of anthropogenic antibiotics.