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Validation of an Enzyme Immunoassay for Measuring Fecal Cortisol Metabolites of Bearded (Sapajus libidinosus) and Black (Sapajus nigritus) Capuchins

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Abstract

Fecal steroid analysis is a powerful noninvasive tool for behavioral endocrinology, but enzyme immunoassays (EIAs) require experimental validation before they are applied. We conducted a physiological validation of an in-house EIA measuring fecal cortisol metabolites (FCMs) by performing an adrenocorticotrophic hormone (ACTH) challenge, dexamethasone suppression, and control saline solution injection experiment with six male and five female bearded capuchins (Sapajus libidinosus). We also took advantage of a presumably stressful incident to perform a biological validation for females. In addition, we conducted high-performance liquid chromatography (HPLC) immunograms to characterize the FCMs measured in both sexes of bearded capuchin, and in a closely related species (S. nigritus, black capuchin monkeys). Male and female S. libidinosus showed FCM peaks after ACTH injections, and females also showed FCM peaks in the biological validation. Three of four individuals (two males and one female) had an FCM peak shortly after injection of dexamethasone and both sexes then showed prolonged low FCM levels after dexamethasone injection. We observed no effects of saline solution injections. The time to peak FCM excretion after ACTH injection or a stressful incident varied 1.5–8.5 h. HPLC results revealed no differences in FCM profiles between sexes or species and suggest that the EIA can also be used in male and female S. nigritus. Our results validate an in-house EIA for both sexes of S. libidinosus but show large individual variation in FCM excretion, which highlights the need for carefully planned feces collection in endocrinologial research.

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Acknowledgments

The authors thank Victor Shiramizu, Edilma Chagas, and Eliane Morgado for help with samples in the LDH/UFRN; Dra. Maria Bernardete Cordeiro de Souza for opening her laboratory to us; CETAS employees: Dr. Paulo, Michele, Mércia, Ronny, Adair, Hércules, Aluilson, Zé, and Dinho for help throughout the experiment; Dra. Renata Ferreira for sample transportation; Edith Klobetz-Rassam for help with steroid analyses and Gisele Zago for logistic help throughout this research. We also thank Dr. Alex Hubbe, Dr. Brandon Wheeler, Dr. James Higham, Dr. Joanna Setchell, and an anonymous reviewer for helpful comments on previous drafts of this manuscript. This research was funded by FAPESP (O. Mendonça-Furtado: 08/52293-3 and P. Izar: 08/55684-3) and Capes (OMF: 13/1537). Permits to collect data were granted from ICMBio to P. Izar (21406).

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Authors and Affiliations

  1. Institute of Psychology, University of São Paulo, São Paulo, Brazil

    Olivia Mendonça-Furtado & Patricia Izar

  2. Department of Biomedical Sciences, University of Veterinary Medicine, Vienna, Austria

    Rupert Palme

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  1. Olivia Mendonça-Furtado
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  2. Patricia Izar
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Correspondence to Olivia Mendonça-Furtado.

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Handling Editor: Joanna M. Setchell

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Mendonça-Furtado, O., Izar, P. & Palme, R. Validation of an Enzyme Immunoassay for Measuring Fecal Cortisol Metabolites of Bearded (Sapajus libidinosus) and Black (Sapajus nigritus) Capuchins. Int J Primatol 38, 1002–1016 (2017). https://doi.org/10.1007/s10764-017-9993-6

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  • DOI: https://doi.org/10.1007/s10764-017-9993-6

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