The test of the digestive enzyme activity, observation of intestinal tissue and the living imaging technology were applied to investigate the effect of the ferrous chelating peptide on the digestive tract. The test of indexes of biochemical indexes of serum and liver, and observation of the liver were used to investigate the effect of the ferrous chelating peptide on immunological characteristics of Misgurnus anguillicaudatus. The result showed lipase activity was significantly higher than that of the control group when the ferrous chelating peptide was 2 g/kg; amylase activity of pepsin was significantly higher than the normal control group when the ferrous chelating peptide was 1 g/kg; trypsin activity in each experimental group compared with the control group had no significant difference. SOD and lysozyme in serum were significantly higher than those of normal control group when the ferrous chelating peptide was 2 g/kg; compared with the normal control group, CAT at ferrous chelating peptide concentration of 1 g/kg showed significant difference. Compared with control group, the morphological structure of each experimental group had no change. With the application of living imaging technology, complexes of ferrous chelating peptide with FITC marked stayed in M. anguillicaudatus for a long time and fluorescence intensity was mainly enriched in the upper body of M. anguillicaudatus, hence, after absorption, the ferrous chelating peptide was speculated to be distributed in the liver, gallbladder and other organs. The results showed that the hydrolysis of ferrous chelate peptide could improve the digestive enzyme activities of loach and enhance non-specific immunity of loach.