Physiological validation of enzyme immunoassays for analysis of cortisol and its metabolites in neotropical deer

• Grant number: 14/23549-0
• Support Opportunities: Scholarships abroad - Research Internship - Scientific Initiation
• Start date: March 23, 2015
• End date: July 22, 2015
• Field of knowledge: Agronomical Sciences - Veterinary Medicine
• Principal Investigator: José Maurício Barbanti Duarte
• Grantee: Cláudia Maria Herédias Ribas
• Supervisor: Rupert Palme
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil
• Institution abroad: University of Veterinary Medicine, Vienna, Austria
• Associated to the scholarship: 14/18954-3 - Physiological validation of enzyme immunoassays for cortisol and its fecal metabolites in neotropical deer, BP.IC

Abstract

In response to a stressor stimulus, animals present a physiological response to stimulation of the Hypothalamic-Pituitary-Adrenal glands, with release of hormones such as cortisol. After their immediate release into the bloodstream, they are metabolized and excreted via bile, and released in the feces after 12 to 24 hours. Deer are known to be susceptible to stress, making it extremely important to determine noninvasive methods for determination of stress-related hormones. Thus, this project proposes 3 different tests for analysis of glucocorticoids in the feces of seven species of Neotropical deer, through physiological validation. To this end, animals (Blastocerus dichotomus, Ozotoceros bezoarticus, Odocoileus virginianus, Mazama americana, M. gouazoubira, M. nana, M. nemorivaga) will be anesthetized (Ketamine/Xylazine/Isoflurane) and given an IV application of ACTH, after which their blood will be collected every 15 minutes for the next 2 hours. There will also be a previous collection before ACTH application. Feces will be collected every 2 hours for 24 hours before and 72 hours after application of ACTH. Cortisol analysis will be performed in serum, to demonstrate the effect of ACTH. Analysis of glucocorticoid will be done in fecal extracts, using antibodies to cortisol (R4866) and corticosterone (CJM006), which will be analyzed at the Endocrinology Laboratory at NUPECCE/UNESP/Brazil, and to 11.17-dioxoandrostanes (11-oxoaetiocholanolone-3-HS: BSA), which will be analyzed at the University of Veterinary Medicine Vienna, Austria, in collaboration with Dr. Rupert Palme. Thus, it is expected that we will be able to determine which tests better reflect the peak of blood cortisol in each of the studied species, giving us basis for future research on stress evaluation in Neotropical Deer. (AU)