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