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Water pollution from lead/Pb 2+ poses a significant threat to aquatic ecosystems, and its repercussions on aquatic animals have received considerable attention. Although Pb 2+ has been found to affect numerous aspects of animals, including individual fitness, metabolic status, and symbiotic microbiota, few studies have focused on the associations between Pb 2+ -induced variations in fitness, metabolome, symbiotic microbiome, and environmental parameters in the same system, limiting a comprehensive understanding of ecotoxicological mechanisms from a holistic perspective. Moreover, most ecotoxicological studies neglected the potential contributions of anions to the consequences generated by inorganic lead compounds. We investigated the effects of Pb(NO 3 ) 2 at environmentally relevant concentrations on the Rana omeimontis tadpoles and the water quality around them, using blank and NaNO 3 -treated groups as control. Results showed that Pb(NO 3 ) 2 not only induced a rise in water nitrite level, but exposure to this chemical also impaired tadpole fitness-related traits (e.g., growth and development). The impacts on tadpoles were most likely a combination of Pb 2+ and NO 3 - . Tissue metabolomics revealed that Pb(NO 3 ) 2 exposure influenced animal substrate (i.e., carbohydrate, lipid, and amino acid) and prostaglandin metabolism. Pb(NO 3 produced profound shifts in gut microbiota, with increased Proteobacteria impairing Firmicutes, resulting in higher aerobic and possibly pathogenic bacteria. NaNO 2 produced profound shifts in gut microbiota, with increased Proteobacteria impairing Firmicutes, resulting in higher aerobic and possibly pathogenic bacteria. NaNO 3 also influenced tadpole metabolome and gut microbiome, in a manner different to that of Pb(NO 3 ) 2 . The presence of NO 3 , particularly on the microbiota. Piecewise structural equation model and correlation analyses demonstrated connections between tissue metabolome and gut microbiome, and the variations in tadpole phenotypic traits and water quality were linked to changes in tissue metabolome and gut microbiome. These findings emphasized the important roles of gut microbiome in mediating the effects of toxin on aquatic ecosystem. Moreover, it is suggested to consider the influences of anions in the risk assessment of heavy metal pollutions.- seemed to counteract some changes caused by Pb 2+ , particularly on the microbiota. Piecewise structural equation model and correlation analyses demonstrated connections between tissue metabolome and gut microbiome, and the variations in tadpole phenotypic traits and water quality were linked to changes in tissue metabolome and gut microbiome. These findings emphasized the important roles of gut microbiome in mediating the effects of toxin on aquatic ecosystem. Moreover, it is suggested to consider the influences of anions in the risk assessment of heavy metal pollutions., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Antibiotic resistance is a growing problem for ecosystem health and public healthcare. Hence, the transmission of antibiotic resistance from human and animal origins to natural environments requires careful investigation. In this study, nine antibiotic resistance genes (ARGs), three mobile genetic elements (MGEs), and their relations with antibiotics, heavy metals, and microbiota were investigated in 16 sample sites (Xinxiang, China). Fluoroquinolones (0.13-14.22 μg/L) were most abundant in hospital effluent and oxytetracycline (251.86-5817.47 μg/kg) in animal manure. Animal manure showed the highest levels of zinc (80.79-2597.14 mg/kg) and copper (32.47-85.22 mg/kg), possibly affecting the prevalence of intI1 and aac(6')-Ib genes. Aminoglycoside and sulfonamide resistance genes (aac(6')-Ib, aadA, and sul1) were the main ARGs in this area. In addition, the detected ARGs and MGEs were higher in animal manure than in hospital effluent, except for the sul1 gene. On the other hand, the incomplete removal of antibiotics (29.76-100%), heavy metals (31.25-100%), and ARGs (1-3 orders of magnitude) in MWWTPs resulted in the accumulation of these contaminants in the receiving river. Network analysis suggested that the potential hosts (Jeotgalibaca, Atopostipes, Corynebacterium_1, etc.) of ARGs were more predominant in animal manure rather than hospital effluent, indicating a higher ARG transfer potential in animal manure compared with hospital sources. These results provide useful insights into the different migration and dissemination routes of antibiotics, heavy metals, ARGs, and microbiota from anthropogenic and animal origins to their receiving environments via MWWTP discharge and manure fertilization., (Copyright © 2021 Elsevier Ltd. All rights reserved.)