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

  • Liu, H.C. (Ilan Branch, Livestock Research Institute) ;
  • Hsiao, M.C. (Ilan Branch, Livestock Research Institute) ;
  • Hu, Y.H. (Ilan Branch, Livestock Research Institute) ;
  • Lee, S.R. (Ilan Branch, Livestock Research Institute) ;
  • Cheng, W.T.K. (Department of Animal Science and Technology, National Taiwan University)
  • Received : 2009.04.24
  • Accepted : 2009.10.10
  • Published : 2010.02.01


This study attempted to clarify the difference in eggshell pigmentation between blue-shelled ducks (BSD) and whiteshelled 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 6th 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.


  1. Baird, T., S. E. Solomon and D. R. Tedstone. 1975. Localisation and characterisation of egg shell porphyrin in several avian species. Br. Poult. Sci. 16:201-208
  2. Barnett, G. M., M. F. Hudson and K. M. Smith. 1975. Concerning of meso-tetraphenylporphyrin purification. J. Chem. Soc. 14:1041-1043
  3. Chen, D. T., S. R. Lee, Y. H. Hu, C. C. Huang, Y. S. Cheng, C. Tai, J. P. Poivey and R. Rouvier. 2003. Genetic trends for laying traits in the Brown Tsaiya (Anas platyrynchos) selected with restricted genetic selection index. Asian-Aust. J. Anim. Sci. 16(12):1705-1710
  4. Cornelius, C. E. 1991. Bile pigments in fishes: a review. Vet. Clin. Pathol. 20:106-115
  5. Johnson, A. L. 2000. Reproduction in the female. In: Sturkie’s avian physiology (Ed. G. C. Whittow). Academic press, USA. pp. 569-596
  6. Gosler, A. G., J. P. Higham and S. J. Reynolds. 2005. Why are birds' eggs speckled? Ecol. Lett. 8(10):1105-1113
  7. Kennedy, G. Y. and H. G. Ververs. 1973. Eggshell pigments of the Arauano fowl. Comp. Biochem. Physiol. 44B:11-35
  8. Kennedy, G. Y. and H. G. Ververs. 1975. A survey of avian eggshell pigments. Comp. Biochem. Physiol. 55B:117-123
  9. Lang, M. R. and J. W. Wells. 1987. A review of eggshell pigmentation. World's Poult. Sci. J. 43(3):238-246
  10. Liu, S. C., J. F. Huang, T. J. Sun, S. R. Lee and C. T. Wang. 1998. The inheritance of blue eggshell in Brown Tsaiya. J. Taiwan Lives. Res. 31(4):373-382
  11. Ma, R. C. S. 1968. The time of release of the luteinizing hormone from the adenohypophysis of laying domestic ducks. Poult. Sci. 47:404-410
  12. Mik$\check{s}$$\acute{i}$k, I., V. Hol$\acute{a}$$\check{n}$ and Z. Deyl. 1994. Quantification and variability of eggshell pigment content. Comp. Biochem. Physiol. 109A:769-772
  13. Moreno, J. and J. L. Osorno. 2003. Avian egg colour and sexual selection: does eggshell pigmentation reflect female condition and genetic quality? Ecol. Lett. 6(9):803-806
  14. Nakano, T., N. I. Ikawa and L. Ozimek. 2003. Chemical composition of chicken eggshell and shell membranes. Poult. Sci. 82:510-514
  15. Polin, D. 1957. Formation of porphyrin from delta-aminolevulenic acid by uterine and liver tissue from laying hens. Proc. Soc. Exp. Biol. Med. 94:276-279
  16. Poole, H. K. 1965. Spectrophotometric identification of eggshell pigments and timing of supercial pigment deposition in the Japanese quail. Proc. Soc. Exp. Biol. Med. 119:547-551
  17. Ryter, S. W., E. Kvam and R. W. Tyrrell. 2000. Heme oxygenase activity. Methods Mol. Biol. 99:369-391
  18. SAS Institute Inc. 2002. SAS Qualification Tools User's Guide, version 9.1.3 SAS Institute Inc., Cary, North Carolina
  19. Schwartz, S. W., A. Raux, B. A. Schacter, B. D. Stephenson and R. N. Shoffner. 1980. Loss of hereditary uterine protoporphyria through chromosomal rearrangement in mutant Rhode Island Red hens. Int. J. Biochem. 12:935-940
  20. Solomon, S. E. 1991. Eggshell colour. In: Egg and eggshell quality. Wolfe publishing Ltd. England. pp. 123-129
  21. Solomon, S. E. 2002. The oviduct in chaos. World's Poult. Sci. J. 58:41-48
  22. Tai, C., R. Rouvier and J. P. Poivey. 1989. Genetic parameters of some growth and egg production traits in laying Brown Tsaiya. Genet. Sel. Evol. 21:377-384
  23. Wang, X. T., X. M. Deng, C. J. Zhao, J. Y. Li, G. Y. Xu, L. S. Lian and C. X. Wu. 2007. Study of the deposition process of eggshell pigments using an improved dissolution method. Poult. Sci. 86:2236-2238
  24. Warren, D. C. and R. M. Conrad. 1942. Time of pigment deposition in brown shelled hen eggs and in turkey eggs. Poult. Sci. 21:515-520
  25. Wicke, W. 1858. Ueber das pigment in den Eischalen der V$\ddot{o}$gel. Naumannia. 8:393-397
  26. Zhao, R., G. Y. Xu, Z. Z. Liu, X. Y. Li and N. Yang. 2006. A study on eggshell pigmentation: Biliverdin in blue-shelled chickens. Poult. Sci. 85:546-549
  27. Zhao, R., Z. Z. Liu, G. Y. Xu and N. Yang. 2007. Analysis of SNP markers for chicken blue-shelled gene using PCR-SSCP. Chin. J. Agric. Biotech. 4(1):53-56

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