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To understand the composition and dynamic changes of metabolites during the formation of goose fatty liver, the metabolite profiles of goose liver at three overfeeding stages were analyzed using widely targeted metabolomics. Three 70-day-old Landes geese with similar body conditions from the same batch were selected randomly for slaughter at the early (day 7), middle (day 16) and late (day 25) overfeeding stages, separately. The tip of the larger liver lobe was collected for widely targeted metabolomic analysis. The results showed that: (1) a total of 1 153 metabolites belonging to 19 classes including amino acids, organic acids, nucleotides and lipids were detected in the liver of geese at the three overfeeding stages; (2) principal component analysis (PCA) showed significant differences in the metabolic profiles of goose liver at the three stages, and identified 142 and 92 differential metabolites at the early versus middle stage, and the middle versus late stage, respectively, the major ones being amino acids and their derivatives, as well as organic acids and their derivatives; and (3) Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway analysis indicated that the pathways involved in fatty acid biosynthesis, vitamin B6 metabolism, linoleic acid metabolism, lysine degradation, arginine biosynthesis, arachidonic acid metabolism and amino acid biosynthesis changed significantly during the formation of goose fatty liver. This study found that most of the differential metabolites were involved in fatty acid synthesis during goose fatty liver formation. Moreover, the contents of transport-related metabolites showed a continuous increasing trend. Findings in this study will not only enrich the theoretical knowledge of poultry liver metabolism, but also provide a theoretical basis for the precise nutritional regulation and efficient production of high-quality goose fatty liver.

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