Scanning electron microscopy of thin and soft shells induced by feeding calcium-deficient or vitamin D-deficient diets to laying hens.

A scanning electron microscopic study was conducted on shells from eggs laid by four groups of hens maintained on different types of diets: a) control, b) vitamin D3-deficient, c) Ca-deficient, and d) vitamin D3-deficient supplemented with 1,25-(OH)2D3. After 1 week for Ca-deficient hens and after 4 weeks for vitamin D3-deficient hens, the thickness of the shell decreased abruptly and numerous thin-shelled and soft-shelled eggs were laid. The study showed that with both Ca-deficient and vitamin D3-deficient diets, the outer layers of the shell (cuticle and spongy) were reduced or absent but the mammillary layer was present even in the thinnest soft-shelled egg. The order in which layers disappeared as treatment progressed was exactly the reverse of the order in which these layers are formed in normal eggs. No eggs were found without mammillary knobs, which suggests that the hens stop laying before Ca concentrations in blood become too low for the formation of the mammillary knobs. Uncalcified portions of the shell organic matrix were never found, suggesting that Ca deposition and matrix formation were inhibited simultaneously. The relationship between fibers of the shell membrane and mammillary knobs was preserved in all cases. The eggshells from hens on 1,25-(OH)2D3-supplemented diets were ultrastructurally indistinguishable from those of hens on diets adequate in vitamin D3.