A Comparative Methodical Study of the Faecal Steroid Analysis on Birds: Looking for A Valid Method of Testosterone Determination


In a comparative study, a relatively simple and high sensitivity method was developed for analysis of testosterone-equivalent(s) in the faeces of different bird species. To determine the recovery of extractions and purifications, tritium-labelled testosterone was added to the wet samples. Then the samples were treated with sodium dodecil sulphate (SDS), an emulsificator to “open-up” the complex, lipid-coated particles of faecal samples. This emulsification resulted in the decrease of the quantity of interfering substances after diethyl-ether extraction and the linearity of the measured testosterone equivalents from aliquots in the range of 2 and 10 mg of faeces. In the RIA, we applied a group specific polyclonal testosterone antibody which cross-reacted with reduced metabolites and at a certain level with sulphate conjugates as well. The use of Helix enzymes did not modified significantly the results of the analysis relating to a low level of conjugated androgens in the faecal extracts. The biological validity of the method was tested on domestic cockerels, where between the plasma and faecal testosterone values a four hours phase shift was observed, with a correlation of 0.6355. This method is suitable for “non invasive”, behavioural-ethological studies.


  1. 1.

    al-Alousi, M. L., Anderson, R. A. (2002) A. relatively simple and rapid multi-component method for analysis of steroid profiles in blood, fecal and liver samples. Steroids 67, 269–275.

    CAS  Article  Google Scholar 

  2. 2.

    Bishop, C. M., Hall, M. R. (1991) Non-invasing monitoring of avian reproduction by simplified fecal steroid analysis. J. Zool. London 224, 649–668.

    Article  Google Scholar 

  3. 3.

    Cockrem, J. E., Rounce, J. R. (1994) Fecal measurements of oestradiol and testosterone allowed non-invasive estimation of plasma steroid concentrations in domestic fowl. Brit. Poultry Sci. 35, 433–143.

    CAS  Article  Google Scholar 

  4. 4.

    Hirschenhauser, K., Möstl, E., Kotrschal, K. (1998) The relationship between social stress and dom-inance is seasonal in Greylag geese (Anser anser). Anim. Behav. 55, 171–176.

    Article  Google Scholar 

  5. 5.

    Hirschenhauser, K., Möstl, E., Kotrschal, K. (1999a) Seasonal patterns of sex steroids determined from feces in different social categories of Greylag geese. Gen. Comp. Endocr. 114, 67–79.

    CAS  Article  Google Scholar 

  6. 6.

    Hirschenhauser, K., Möstl, E., Kotrschal, K. (1999b) Within-pair testosterone covariation and repro-ductive output in Greylag geese, Anser anser. Ibis 141, 577–586.

    Article  Google Scholar 

  7. 7.

    Hirschenhauser, K., Möstl, E., Wallner, B., Dittami, J., Kotrschal, K. (2000a) Endocrine and behav-ioural responses of male Greylag geese (Anser anser) to pairbound challenges during the reproduc-tive season. Ethology 106, 63–77.

    Article  Google Scholar 

  8. 8.

    Hirschenhauser, K., Möstl, E., Péczely, R., Wallner, B., Kotrschal, K. (2000b) Seasonal relationships between plasma and fecal testosterone in response to GnRH in domestic ganders. Gen. Comp. Endocr. 118, 262–272.

    CAS  Article  Google Scholar 

  9. 9.

    Ishii, S., Wada, M., Wakabyashi, S., Sakai, H., Kubudera, Y., Yamagochi, N., Kikuchi, M. (1994) Endocrinological studies for artificial breeding of the Japanese ibis, Nipponia nippon, an endangered avian species in Asia. J. Biosci. 19, 491–502.

    CAS  Article  Google Scholar 

  10. 10.

    Kikuchi, M., Yamaguchi, N., Sato, F., Ishii, S. (1994) Extraction methods for fecal hormone analy-sis in birds. J. Ornithol. 135, 64.

  11. 11.

    Krawny, M. (1996) Die Entwicklung einer nicht invasiven Methode zum Nachweis von Steroidhormonmetaboliten im Kot von Gansen. Dissertation, Inst. of Biochemie, Vet. Univ., Wien.

    Google Scholar 

  12. 12.

    Miller, W. M., Hobbs, N. T., Sousa, M. C. (1991) Detectin stress responses in Rocky Mountain Bighorn sheep (Ovis canadensis canadensis) reliability of cortisol concentrations in urine and feces. Can.J.Zool. 69, 15–24.

    CAS  Article  Google Scholar 

  13. 13.

    Palme, R., Fischer, R., Schildorfer, H., Ismail, M. N. (1996) Excretion of infused 14C steroid hor-mones via faeces and urine in domestic livestock. Anim. Reprod. Sci. 43, 43–63.

    CAS  Article  Google Scholar 

  14. 14.

    Péczely, R., El Halawani, M. E., Hargitai, Cs., Mézes, M., Forgó, V., Janosi, Sz. (1993) The photore-fractoriness in domestic geese: effect of gonads and thyroid ont he development of postbreeding pro-lactinemia. Acta Biol. Hung. 44, 329–352.

    PubMed  Google Scholar 

  15. 15.

    Schwarzenberger, F., Möstl, E., Palme, R., Bamberg, E. (1996) Faecal steroid analysis for non-inva-sive monitoring of reproductive status in farm, wild and Zoo animals. Anim. Reprod. Sci. 42, 513–526.

    Article  Google Scholar 

  16. 16.

    Sturkie, R. D. (1976) Avian Physiology, 3rd ed., Springer Verlag, New York - Heidelberg - Berlin.

  17. 17.

    Walpach, R. (1998) Corticosterone Ausscheidung beim Haushuhn (Gallusgallus). Dissertation, Inst. of Biochemie, Vet. Univ., Wien.

    Google Scholar 

  18. 18.

    Wasser, S. K., Thomas, R., Nair, R. P., Guidry, C., Southers, J., Lucas, J., Wildt, D. E., Monfort, S. L. (1993) effects of dietary fibers on faecal steroid measurements in baboons (Papio cynocephalus). J. Reprod. Fert. 97, 569–574.

    CAS  Article  Google Scholar 

  19. 19.

    Wasser, S. K., Bevis, K., King, G., Hanson, E. (1997) Noninvasive physiological measures of dis-turbance in the northern spotted owl. Conserv. Biol. 11, 1019–1022.

    Article  Google Scholar 

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This work was supported by a grant from OTKA, No. T030769. We are grateful to Mrs Katalin Bara for her excellent technical assistance and to Mr. Szilamér Ferenczi for his consultative assistance.

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Correspondence to K. Kelemen.

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Kelemen, K., Péczely, P., Szőke, Z. et al. A Comparative Methodical Study of the Faecal Steroid Analysis on Birds: Looking for A Valid Method of Testosterone Determination. BIOLOGIA FUTURA 54, 285–298 (2003). https://doi.org/10.1556/ABiol.54.2003.3-4.7

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  • Steroids
  • testosterone
  • faeces
  • birds
  • methods
  • biochemistry