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Ascorbic acid (AA) is an indispensable nutrient required to sustain optimal poultry health and performance, which is commonly excluded from the diet of broilers. To investigate the synthesis and distribution of AA during broiler growth and clarify its possible turnover, 144 1 d old healthy Arbor Acres broilers with a body weight of approximately 41 g were randomly assigned to eight groups of 18 broilers each. The kidney, liver, ileum, and spleen of one bird from each group were collected every week until 42 d to detect the synthesis capacity, tissue distribution, and transporter gene expression of AA. The results showed that kidney L-gulonolactone oxidase (GLO) activity responded quadratically (p < 0.001), with maximum activity observed at 7 to 21 d old. Hepatic total AA and dehydroascrobate (DHA) concentration increased linearly (p < 0.001) with age, as did splenic total AA (p < 0.001). In the ileum, mRNA expression of sodium vitamin C transporter 1/2 (SVCT1/2) decreased with the growing age of the broilers (p < 0.05). The expression of SVCT1 in the kidney was not influenced by the growing age of the broilers. The progressive buildup of AA in the liver and spleen of broilers as they age implies an amplified demand for this nutrient. The waning synthesis capacity over time, however, raises concerns regarding the possible inadequacy of AA in the latter growth phase of broilers. The addition of AA to the broilers’ diet might have the potential to optimize their performance. However, the effectiveness of such dietary supplementation requires further investigation. Full article