Master's Dissertation
DOI
https://doi.org/10.11606/D.10.2008.tde-18062008-143039
Document
Author
Full name
Rodrigo de Souza Amaral
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Oliveira, Claudio Alvarenga de (President)
Furtado, Priscila Viau
Guimarães, Marcelo Alcindo de Barros Vaz
Title in Portuguese
Uso de diferentes matrizes biológicas na dosagem de andrógenos em peixes-bois da Amazônia machos (Trichechus inunguis) mantidos em cativeiro
Keywords in Portuguese
Peixe-boi Amazônico
Radioimuniensaio
Reprodução
Sirênio
Testosterona
Abstract in Portuguese
O objetivo deste estudo foi verificar a viabilidade da dosagem de andrógenos em amostras de saliva, secreção lacrimal, urina e fezes de peixe-boi da Amazônia realizando um desafio hormonal. Dois peixes-bois amazônicos adultos machos (A-1 e A-2) foram submetidos a um protocolo de experimentação de doze dias (D-1 a D10). No D0 os animais receberam uma injeção intramuscular de GnRH exógeno. Amostras de saliva, secreção lacrimal, urina e fezes foram coletadas diariamente (entre 08h00 e 09h00) e mantidas a -20°C até o ensaio. As amostras de fezes foram liofilizadas, extraídas com metanol 80% e diluídas em tampão antes do radioimunoensaio (RIE). As amostras de urina sofreram hidrólise ácida e foram diluídas em soro bovino depletado. As amostras de saliva e secreção lacrimal foram dosadas sem etapa de extração, porém, o ensaio foi adaptado para aumentar a sensibilidade do teste. Os ensaios hormonais foram realizados utilizando um conjunto comercial de RIE para testosterona total. Um pico de andrógenos (mediana+2DI) somente foi observado nas amostras de saliva, urina e fezes de ambos os animais. Porém, os picos de andrógenos fecais ocorreram depois (cinco dias) dos picos de andrógenos urinários e salivares. Este intervalo está correlacionado com o longo tempo de passagem da digesta pelo trato gastrointestinal na espécie. Os picos salivares e urinários ocorreram muito próximos, provavelmente com poucas horas de intervalo. Esses resultados demonstram que as concentrações de andrógenos em amostras de saliva, urina ou fezes refletem consistentemente os eventos fisiológicos e são ferramentas de grande utilidade no monitoramento reprodutivo de peixes-bois da Amazônia.
Title in English
Use of different biological matrices on androgens measurement in captive male Amazonian manatees (Trichechus inunguis)
Keywords in English
Amazonian manatee
Radioimmunoassay
Reproduction
Sirenian
Testosterone
Abstract in English
The aim of this study was to verify the viability of androgens measurement in saliva, lacrimal secretion, urine and fecal samples of Amazonian manatee by carrying out hormonal challenge. Two adult male manatees (A-1 and A-2) were submitted to an experimentation protocol of twelve days (D-1 to D10). On D0 the animals received an intramuscular injection of GnRH-analogue. Salivary, lacrimal, urinary and fecal samples were collected daily (between 08h00 and 09h00) and frozen at -20°C until assayed. Fecal samples were lyophilized, extracted with 80% methanol and diluted in buffer before the radioimmunoassay (RIA). Urine samples underwent acid hydrolysis and diluted in depleted bovine serum. Salivary and lacrimal samples were assayed without extraction step, but, the assay was adapted to improve the sensibility. Hormonal assays were carried out with a commercial testosterone RIA kit. An androgen peak (>median+2IQR) was observed only in salivary, urinary and fecal samples of both animals. However, the fecal androgens peaks occurred later than urinary and salivary androgens peaks. These intervals are correlated with the long digesta passage time in this species. The salivary and urinary peaks were very close, probably with few hours of interval. These results show that androgens concentrations in saliva, urine or feces samples reflect reliably physiological events and are powerful tool for reproductive monitoring of Amazonian manatees.
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Publishing Date
2008-06-26
WARNING: The material described below relates to works resulting from this thesis or dissertation. The contents of these works are the author's responsibility.
- AMARAL, Rodrigo de Souza, et al. Noninvasive Monitoring of Androgens in Male Amazonian Manatee (Trichechus inunguis) : Biologic Validation [doi:10.1638/2008-0111.1]. Journal of Zoo and Wildlife Medicine [online], 2009, vol. 40, n. 3, p. 458-465.