Baggott, Glenn K.

Beckett, M.

Graeme-Cook, K.

English

Birkbeck Institutional Research Online

  Birkbeck ePrints: an open access repository of the research output of Birkbeck College  http://eprints.bbk.ac.uk  Beckett, M., Baggott, G. K. and Graeme-Cook, K. (2003) Bacterial degradation of the eggshell cuticle of the mandarin duck (Aix galericulata). Avian and Poultry Biology Reviews 14 pp 196-197   This is an author-produced version of an article published in Avian and Poultry Biology Reviews. This version has been peer-reviewed but does not include the final publisher proof corrections, journal layout or journal pagination.  All articles available through Birkbeck ePrints are protected by intellectual property law, including copyright law. Any use made of the contents should comply with the relevant law.   Citation for this version: Beckett, M., Baggott, G. K. and Graeme-Cook, K. (2003) Bacterial degradation of the eggshell cuticle of the mandarin duck (Aix galericulata). London: Birkbeck ePrints. Available at: http://eprints.bbk.ac.uk/archive/00000222  Citation for the publisher’s version: Beckett, M., Baggott, G. K. and Graeme-Cook, K. (2003) Bacterial degradation of the eggshell cuticle of the mandarin duck (Aix galericulata). Avian and Poultry Biology Reviews 14 pp 196-197    http://eprints.bbk.ac.ukContact Birkbeck ePrints at lib-eprints@bbk.ac.uk  1Beckett, M., Baggott, G.K. & Graeme-Cook, K. University of Hertfordshire & Birkbeck, University of London  Bacterial degradation of eggshell cuticle of the Mandarin Duck (Aix galericulata)  The Mandarin Duck is a cavity nester in the UK (Davis & Baggott, 1989). Females incubate clutches of up to 30 eggs for about 33 days. During natural incubation GH2O can increase by about 70% in the first 7 days of incubation (Baggott & Graeme-Cook, 1997). The initial GH2O is low, for egg weight, due to cuticle completely covering pore apertures. During the first week of natural incubation counts of surface bacteria capable of culture at incubation temperature decrease but the proportion of Bacillus licheniformis increases (Baggott & Graeme-Cook, 2002). As this species has the capability to digest duck cuticle (Baggott & Graeme-Cook, op. cit.), it is hypothesised that it may be responsible for the increase in GH2O in the first week of incubation. The objective was to identify what cuticle proteins, if any, were capable of degradation by B. licheniformis cultured from the Mandarin eggshell surface.  Using SDS-polyacrylamide gel electrophoresis (PAGE), one major protein was identified in the cuticle of unincubated eggs of Mandarin Duck that was not detectable in the shell matrix. This protein had a molecular weight of 30 kDa as estimated using molecular weight standards and did not react with Schiff’s reagent indicating that it was not a glycoprotein.  Eight additional protein bands, found in the cuticular sample were also present in the shell matrix fraction. These had approximate molecular weights of >80 kDa (x2), 79kDa, 71kDa, 60kDa, 40kDa, 15kDa,12kDa  and all stained with Schiff’s reagent suggesting that they contain carbohydrate moieties. In the Pekin duck, matrix proteins of 15k (lysozyme), 17k, 32k, 66k and 78kDa (ovotransferrin) have been identified (Panheleux, et al., 1999); as in this species, a 45kDa matrix protein (ovalbumin) was absent from mandarin profiles. Analysis of proteins in the cuticle of commercial duck consumption eggs also showed the presence of a protein exclusive to the cuticle but in this case it had an apparent molecular weight of 40kDa.    SDS-PAGE electrophoresis of cuticular and shell fractions of Mandarin duck eggs obtained from nests after natural incubation showed the loss of all detectable cuticle proteins with the exception of the 30 kDa protein. In vitro at natural incubation temperatures, shell samples incubated with a strain of Bacillus licheniformis, originally isolated from the surface of incubated eggs, degraded all proteins of the cuticle.    2In summary, only one protein exclusive to the cuticle fraction was identified. Other proteins found on the surface of the egg were also located within the shell matrix. Previously, the only protein reported to be localised in the cuticle, that of the hen, is a 32kDa protein, ovocalyxin (Gautron et al., 2001). Natural incubation of eggs, and incubation in vitro, with a species of bacteria, Bacillus licheniformis, led to the loss of proteins within the cuticular layer. This suggests to us that the bacteria may be responsible for increasing the GH2O off the egg during incubation by removing the cuticle layer over the pores.  Baggott, G. K., & Graeme-Cook, K. (1997). Variable shell conductance during natural incubation. Poultry and Avian Biology Reviews, 8, 158. Baggott, G.K. & Graeme-Cook, K. (2002). Microbiology of natural incubation, in Avian Incubation, Behaviour, Environment and Evolution, (ed. Deeming, D.C.) pp.179-191. Oxford, OUP. Davies, A. K. and Baggott, G. K. (1989). Egg-laying, incubation and intraspecific nest parasitism by the Mandarin Duck Aix galericulata. Bird Study, 36, 115-22. Gautron J., Hincke, M.T., Mann, K., Panheleux, M., Bain, M., McKee, M.D., Solomon, S.E. & Nys, Y (2001) Ovocalyxin-32, a novel chicken eggshell matrix protein - Isolation, amino acid sequencing, cloning, and immunocytochemical localization. Journal of Biological Chemistry, 276 (42): 39243-39252. Panheleux, M., Bain, M., Fernandez, M.S., Morales, Gautron, J., Arias, J.L., Solomon, S.E., Hincke, M. & Nys, Y.  (1999) Organic matrix composition and ultrastructure of eggshell: a comparative study. British Poultry Science (1999) 40: 240–252.    

Bacterial degradation of the eggshell cuticle of the mandarin duck (Aix galericulata)

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