Copyright 2021 Zeng et al.Farming operation and amino acid profiles of pond-reared Chinese mitten crabs, Eriocheir sinensis (Milne Edwards, 1853), collected from different areas in Jiangsu Province, China were investigated and compared with the aim to elucidate how farming practices affected the nutritional values of three edible tissues (muscle, hepatopancreas and gonad) of crab. The crab pond aquaculture practices including snail input, macrophytes coverage, total commercial feed, the ratio of trash fish to total feed, were much higher in Gaochun and Jintan than that in other sites (having larger pond size), which leads to higher average individual body weight and commercial yields. Further, the mean body weight, muscle weight, carapace length and width, and the ratio of gonad to hepatopancreas were significantly higher in Jintan, Suzhou and Gaochun areas than in other regions. Amino acid assessment showed that all crabs collected delivered high-quality protein (Amino acid score >1 except Valine), the main amino acids being glutamic acid, aspartic acid, and alanine. Significant differences in amino acid profiles were observed between sites, tissues and sexes. Muscles were rich in total amino acids, essential amino acids, and delicious amino acids, followed by gonads and hepatopancreas. The contents of essential amino acids in crab muscles from Gaochun, Jintan, Suzhou and Guannan were significantly higher than those from Suqian, Sihong and Xinghua. All the amino acids except Serine and Glycine were significantly higher in gonads from males than from females. The redundancy analysis revealed that the snail input, trash fish ratio to the total feed, macrophytes coverage and total trash fish supply explained 84.3% of the variation in the amino acid content and structure in crabs from Gaochun, Jintan and Suzhou. Overall, our results show that mitten crabs collected in Jiangsu province had good nutritional quality suitable for human dietary needs, and that farming practices, especially degree of fish-source protein feeding, influence the amino acids composition of crabs.