Evaluating the reproductive status of the male budgerigar (Melopsittacus undulatus).

Limited knowledge about male psittacine reproduction reduces the success of breeding programmes. Within the scope of fecundity assessment, classification of male sexual status is essential for effective conservation of the species. The aim of the present study was to investigate the testes of male budgerigars (Melopsittacus undulatus), as psittaciform model species to verify their reproductive status by morphological and immunocytochemical examination. Using light microscopy, gonadal samples were categorized resulting in three reproductive states (active, intermediate, non-active). Calculation of testes weights plus measurement of tubular and interstitial dimensions displayed significant (p ≤ 0.05) differences between all three reproductive stages. Lipids in the testicular tubules, analysed by Sudan black staining and fluorescence microscopy (DAPI(2) mode) were highly present in non-active status. Immunocytochemistry involved two different hydroxysteroid dehydrogenases (HSD), 3ß-HSD and 17ß-HSD-2, as markers for steroidogenesis, as well as steroid receptors for androgens (AR), oestrogen (ER) and progesterone (PR). Both HSDs and AR declined in non-active gonads compared to active and intermediate stages, with a positive signal in germ and somatic cells of testis and epididymis. ER and PR were detected in testicular and epididymal cells, similarly expressed in all three stages. The proliferation rate of germ cells in the testicular tubules, obtained by Ki67, differed significantly in active (38.67%), intermediate (32.40%) and non-active (6.01%) status. According to this morphological study, we have been able to establish markers for the reproductive staging of psittacine testes. This knowledge will be useful to deepen reproductive biology in budgerigars.

Antibody selection for immunocytochemical characterization of the male reproductive system in Psittaciformes.

The success of breeding programs is limited by the sparse knowledge about endocrine regulation and biochemical reactions in the psittacine male tract. The immunocytochemical analysis of parrots' testicular tissues provides an insight into their reproductive system but is often hampered by the lack of reliable antibodies. In the present study, we tested a large panel of antibodies raised against steroid receptors, steroidogenic enzymes, relaxin peptides including their receptors, and proliferation markers on paraffin sections of testicular tissue from eight psittacine genera representing three continents. All investigated species displayed the tested markers in somatic and germ cells of testis and epididymis, even though cell distribution was partly heterogenous and in species-specific patterns. The 17ß-hydroxysteroid-dehydrogenase-2, 3ß-hydroxysteroid-dehydrogenase, and smooth muscle actin allowed the cross-species differentiation between active and nonactive gonads. The remaining steroidogenic enzymes, steroid receptors, relaxin peptides, and Ki67 proved to be suitable to define reproductive activity depending on the parrot species. Adapting immunocytochemical methods to different psittacines was successful, though various cellular expression patterns do not allow the transfer of results among different parrot species. However, the availability of a reliable repertory of sexual markers is important to examine reproductive biology of psittacine birds.