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This study attempted to clarify the difference in eggshell pigmentation between blue-shelled ducks (BSD) and white-shelled ducks (WSD). The eggshell pigmentation deposition process is discussed. Ultraviolet spectro-photometer and HPLC were used to determine the biliverdin concentration in the shell gland, uterus liquid and eggshell at 6, 12, 18, 20, 23.5 h post-oviposition. The biliverdin concentration in the eggshell and uterus fluid showed significant differences between BSD and WSD, but not in the shell gland. The heme oxygenase activity in the shell gland of both kinds of ducks remained mostly constant during the ovulatory cycle with no variation. The assay of exogenous biliverdin injection into the shell gland antrum in the WSD indicated that exogenous biliverdin could be deposited continuously into the eggshell until the source was exhausted. A layer-by-layer dissolution assay was used to examine the eggshell pigment deposition process. The biliverdin concentration in the first to sixth layers of the eggshell in the BSD was significantly higher than that in the white-shelled counterpart. The blue pigment concentration increased persistently from the 6 th layer to the 1 st layer. The BSD eggshells did not accumulate a large quantity of biliverdin in the most external layer. They tended to increase the deposition layer by layer. Our results demonstrated that different BSD and WSD eggshell colors were influenced by the amount of biliverdin in the uterus fluid and not determined by the amount of biliverdin in the shell gland. This implies the existence of a mechanism that controls biliverdin transportation from the shell gland into the uterus fluid, thereby playing a key role in regulating duck eggshell color.

Eggshell Pigmentation Study in Blue-shelled and White-shelled Ducks