Analysis of ultrastructure and microstructure of blackbird (Turdus merula) and song thrush (Turdus philomelos) eggshell by scanning electron microscopy and X-ray computed microtomography.

The unique structure of the egg allows for efficient reproduction on land. Although the functions of the egg are ensured by the concomitant cooperation of all its structures, the eggshell also plays a significant role. Apart from maintaining an aqueous environment within the egg along with controlled gas exchange, the color and pigmentation pattern of eggshell contributes to identification and protection. As a result of all these functions, the structure, shape, and pigmentation of eggshell greatly vary across the class of birds, and understanding these three variability-determining factors may aid in better interpretation of evolutionary mechanisms. In this study, we analyzed for the first time the structure, mineral composition, and characteristics of the pigmentation of blackbird (Turdus merula) and song thrush (Turdus philomelos) eggshells. The shell of blackbird eggs is much thicker compared to the shell of song thrush eggs which is due to a much thicker crystalline and palisade layers. In both species, strongly elongated mammillary knobs are observed, which create a large space between the mineralized shell and the egg membranes. The blackbird egg shell has a higher water vapor conductivity which is due to the larger diameter of the circle and the surface area of individual pores. The primary compound entering the mineral composition of the shell in both species is CaCO however, the thrush egg shells contained more Mg in all layers except the crystalline layer, and S in the crystalline and palisade layers. The two species clearly differ in the size and distribution of pigment spots on the eggshell. We suppose that the differences in shell structure and pigmentation presented in this study may in the future provide a basis for explaining the reasons for the much lower reproductive efficiency of song thrush compared to blackbird.