BACKGROUND: This study aimed to explore the mechanism of organic zinc and manganese (OZM) influencing eggshell quality of laying hens. A total of 384 21-week-old Jingfen-1 laying hens were randomly divided into four groups (8 replicates per group, 12 hens per replicate): control group (60 mg kg RESULTS: Compared with the control group, OZM_5 and OZM_10 groups significantly increased feed intake and egg weight in laying hens in 28-31 weeks (P <
0.05) while egg weight and shell weight were increased in all the OZM groups (P <
0.05). In addition, the OZM_15 group significantly decreased the mammillary knob width and mammillary layer thickness and increased the palisade layer thickness and total effective thickness (P <
0.05), and had a stronger and more stable Ca signal strength in the mammillary layer and a more uniform and dense distribution of calcium and zinc in the transversal surfaces of eggshell. Furthermore, mRNA levels of Claudin2, Zona Occludens 1, Alkaline phosphatase and Ca CONCLUSION: Dietary supplementation of 10-15 mg kg