The objective of this study was to evaluate the effects of in ovo injection of histidine on hatch performance and post-hatch development by determining hatchability, hatching time, BW gain, carcass traits, and intestinal morphology in domestic pigeons (Columba livia). A completely randomized design (n = 3) with a L-histidine injection treatment (His group, 0.55mg histidine dissolved in 200 μL 0.75% saline as 1% Conc compared to total histidine in the egg), a 0.75% saline injection treatment (SC group), and non-injection treatment (NC group) was used. Six squabs from each treatment were randomly sampled at day of hatch (DOH), day 7 (D7), and day 14 (D14) post hatch, respectively. Results showed that in ovo injection of histidine solution increased (P < 0.01) the hatchability in comparison with that of other groups. The hatching time of His group was earlier (P = 0.05) than that of the NC group. In ovo injection of histidine had no influence (P > 0.05) on BW gain and carcass traits, but had a significant effect on the organ index of pigeons. The index of pancreas on DOH, the indices of leg, gizzard, proventriculus, small intestine, and pancreas on D7, and the index of head on D14 were significantly increased (P < 0.05) by injection of histidine. Moreover, in ovo injection of histidine had significant effects on the weight indices of different intestinal segments and jejunal morphology in squabs. Ileum weight index on D7 from His group was significantly higher (P < 0.05) than that of other groups. Compared with the NC group, the jejunal crypt depth on DOH and D7 of His group decreased (P < 0.05), while jejunal villus area and villus crypt ratio on D7 of His group increased (P < 0.05). The results of this study indicate that in ovo injection of histidine may have beneficial effects on squabs' hatchability, intestinal development but have negligible effects on their growth performance during early post-hatch period.