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Psittacine sex determination by radioimunoassay (RIA) of sex steroids and by polimerase chain reaction (PCR) using fecal samples

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Author(s):
Eduardo Antunes Dias
Total Authors: 1
Document type: Master's Dissertation
Press: São Paulo.
Institution: Universidade de São Paulo (USP). Faculdade de Medicina Veterinária e Zootecnia (FMVZ/SBD)
Defense date:
Examining board members:
Claudio Alvarenga de Oliveira; Marcelo Alcindo de Barros Vaz Guimarães; Alda Maria Backx Noronha Madeira
Advisor: Claudio Alvarenga de Oliveira
Abstract

For the current study, it were used fecal samples from 65 birds of Psittacine Family, previously sexed by PCR from blood cells. The fecal androgens and estrogens metabolites were extracted with PBS (Phosfate Buffer Saline) or PBS :Ethil Alchool (4:1) and measured by comercial radioimunoassay kits at the Hormonal Measurement Laboratory of the Department of Animal Reproduction of the Faculty of Veterinary Medicine and Zootechny of the University of São Paulo. The sex determination of the birds were performed using the confidence interval (95%) for the mean of transformed values of estrogens/androgen rate or transformed androgens values. The fecal DNA extraction of some samples were performed using comercial kits or by resines at the Molecular Biology Laboratory of the Pathology Department of the Faculty of Veterinary Medicine and Zootechny of the University of São Paulo. The DNA were amplified by PCR using specific primers for the target region of bird gene that express sex. Evaluation of the effect of blood cells adding on samples also had been done. 70% of birds had the sex confirmed by radioimunoassay. The amplification of PCR products occured only in samples added with blood cells and extracted by a specific comercial kit. The results showed that further studies for sex determination on monomorphic birds by non-invasive techniques are necessary. Two comercial kits were validated for the measurement of fecal steroid metabolites by radioimunoassay. (AU)