Multi-omics provide mechanistic insight into the Pb-induced changes in tadpole fitness-related traits and environmental water quality

Abstract

Water pollution from lead/Pb²⁺ poses a significant threat to aquatic ecosystems, and its repercussions on aquatic animals have received considerable attention. Although Pb²⁺ 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 Pb²⁺-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(NO₃)₂ at environmentally relevant concentrations on the Rana omeimontis tadpoles and the water quality around them, using blank and NaNO₃-treated groups as control. Results showed that Pb(NO₃)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 Pb²⁺ and NO₃^-. Tissue metabolomics revealed that Pb(NO₃)₂ exposure influenced animal substrate (i.e., carbohydrate, lipid, and amino acid) and prostaglandin metabolism. Pb(NO₃)₂ produced profound shifts in gut microbiota, with increased Proteobacteria impairing Firmicutes, resulting in higher aerobic and possibly pathogenic bacteria. NaNO₃ also influenced tadpole metabolome and gut microbiome, in a manner different to that of Pb(NO₃)₂. The presence of NO₃^- seemed to counteract some changes caused by Pb²⁺, 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.

Keywords: Amphibian; Gut microbe; Heavy metals; Metabolomics; Toxicological mechanism; Water quality.