Ocean acidification (OA) and trace metal pollutants coexist to exert combined effects on the functions and services of marine ecosystems. Increasing atmospheric carbon dioxide has caused a decrease in the pH of the ocean, affecting the bioavailability and speciation of trace metals and consequently altering metal toxicity in marine organisms. As an important trace metal functioned in hemocyanin, the richness of Copper (Cu) in octopuses is remarkable. Therefore, the biomagnification and bioaccumulation capacities of Cu in octopuses may be a non-negligible risk of contamination. Here, Amphioctopus fangsiao was continuously exposed to acidified seawater (pH 7.8) and copper (50 μg/L) to investigate the combined effect of ocean acidification and Cu exposure on marine mollusks. Our results showed that A. fangsiao could adapt well to ocean acidification after 21 days of the rearing experiment. However, the accumulation of Cu in A. fangsiao intestine increased significantly in acidified seawater under high levels of Cu stress. In addition, Cu exposure can influence the physiological function of A. fangsiao, including growth and feeding. This study also demonstrated that Cu exposure disturbed glucolipid metabolism and induced oxidative damage to intestine tissue, and ocean acidification further exacerbated these toxic effects. The obvious histological damage and microbiota alterations were also caused by Cu stress and its combined effect with ocean acidification. At the transcription level, we found numerous differentially expressed genes (DEGs) and significantly enriched KEGG pathways, involving glycolipid metabolism, transmembrane transport, glucolipid metabolism, oxidative stress, mitochondrial, protein and DNA damage, all revealing the strong toxicological synergetic effect of Cu and OA exposure and the molecular adaptation mechanism of A. fangsiao. Collectively, this study demonstrated that octopuses may withstand future ocean acidification conditions, however, the complex interactions of future OA and trace metal pollution need to be emphasized. OA can influence the toxicity of trace metals, inducing a potential threat to marine organism safety.
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