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Comparing plasma and faecal measures of steroid hormones in Adelie penguins Pygoscelis adeliae

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Abstract

Physiological measurements of both stress and sex hormones are often used to estimate the consequences of natural or human-induced change in ecological studies of various animals. Different methods of hormone measurement exist, potentially explaining variation in results across studies; methods should be cross-validated to ensure that they correlate. We directly compared faecal and plasma hormone measurements for the first time in a wild free-living species, the Adelie penguin (Pygoscelis adeliae). Blood and faecal samples were simultaneously collected from individual penguins for comparison and assayed for testosterone and corticosterone (or their metabolites). Sex differences and variability within each measure, and correlation of values across measures were compared. For both hormones, plasma samples showed greater variation than faecal samples. Males had higher mean corticosterone concentrations than females, but the difference was only statistically significant in faecal samples. Plasma testosterone, but not faecal testosterone, was significantly higher in males than females. Correlation between sample types was poor overall, and weaker in females than in males, perhaps because measures from plasma represent hormones that are both free and bound to globulins, whereas measures from faeces represent only the free portion. Faecal samples also represent a cumulative measure of hormones over time, as opposed to a plasma ‘snapshot’ concentration. Our data indicate that faecal sampling appears more suitable for assessing baseline hormone concentrations, whilst plasma sampling may best define immediate responses to environmental events. Consequently, future studies should ensure that they select the most appropriate matrix and method of hormone measurement to answer their research questions.

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References

  • Adkins-Regan E (2005) Hormones and animal social behavior. Princeton University Press, Princeton

    Google Scholar 

  • Ainley DG (2002) The Adelie penguin: bellwether of climate change. Columbia University Press, New York

    Google Scholar 

  • Ainley DG, Emison W (1972) Sexual size dimorphism in Adelie penguins. Ibis 114:267–271

    Article  Google Scholar 

  • Angelier F, Shaffer SA, Weimerskirch H, Chastel O (2006) Effect of age, breeding experience and senescence on corticosterone and prolactin levels in a long-lived seabird: the wandering albatross. Gen Comp Endocrinol 149:1–9

    Article  CAS  PubMed  Google Scholar 

  • Angelier F, Bost CA, Giraudeau M, Bouteloup G, Dano S, Chastel O (2008) Corticosterone and foraging behavior in a diving seabird: the Adelie penguin, Pygoscelis adeliae. Gen Comp Endocrinol 156:134–144

    Article  CAS  PubMed  Google Scholar 

  • Arlettaz R, Patthey P, Baltic M, Leu T, Schaub M, Palme R, Jenni-Eiermann S (2007) Spreading free-riding snow sports represent a novel serious threat for wildlife. Proc R Soc Biol Sci B 274:1219–1224

    Article  Google Scholar 

  • Astheimer LB, Buttemer WA, Wingfield JC (1994) Gender and seasonal differences in the adrenocortical-response to ACTH challenge in an arctic passerine, Zonotrichia leucophrys gambelii. Gen Comp Endocrinol 94:33–43

    Article  CAS  PubMed  Google Scholar 

  • Bears H, Smith JNM, Wingfield JC (2003) Adrenocortical sensitivity to stress in dark-eyed juncos (Junco hyemalis oregonus) breeding in low and high elevation habitat. Ecoscience 10:127–133

    Google Scholar 

  • Breuner CW, Orchinik M (2002) Plasma-binding proteins as mediators of corticosteroid action in vertebrates. J Endocrinol 175:99–112

    Article  CAS  PubMed  Google Scholar 

  • Broom DM, Johnson KG (1993) Stress and animal welfare. Chapman & Hall, London

    Google Scholar 

  • Cekan SZ (1979) On the assessment of validity of steroid radioimmunoassays. J Steroid Biochem 11:1629–1634

    Article  CAS  PubMed  Google Scholar 

  • Chomczynski P, Rymaszewski M (2006) Alkaline polyethylene glycol-based method for direct PCR from bacteria, eukaryotic tissue samples, and whole blood. Biotechniques 40:454–458

    Article  CAS  PubMed  Google Scholar 

  • Cockrem JF, Potter MA, Candy EJ (2006) Corticosterone in relation to body mass in Adelie penguins (Pygoscelis adeliae) affected by unusual sea ice conditions at Ross Island, Antarctica. Gen Comp Endocrinol 149:244–252

    Article  CAS  PubMed  Google Scholar 

  • Cyr NE, Romero LM (2008) Fecal glucocorticoid metabolites of experimentally stressed captive and free-living starlings: implications for conservation research. Gen Comp Endocrinol 158:20–28

    Article  CAS  PubMed  Google Scholar 

  • Dallman MF, Bhatnager S (2001) Chronic stress and energy balance: role of the hypothalamic–pituitary–adrenal axis. In: McEwen BS, Goodman HM (eds) Handbook of physiology, sect 7: the endocrine system, vol IV, coping with the environment: neural and endocrine mechanisms. Oxford University Press, New York, pp 197–210

  • Dehnhard M, Schreer A, Krone O, Jewgenow K, Krause M, Grossmann R (2003) Measurement of plasma corticosterone and faecal glucocorticoid metabolites in the chicken (Gallus domesticus), the great cormorant (Phalacrocorax carbo), and the goshawk (Accipiter gentilis). Gen Comp Endocrinol 131:345–352

    Article  CAS  PubMed  Google Scholar 

  • Ellenberg U, Setiawan AN, Cree A, Houston DM, Seddon PJ (2007) Elevated hormonal stress response and reduced reproductive output in yellow-eyed penguins exposed to unregulated tourism. Gen Comp Endocrinol 152:54–63

    Article  CAS  PubMed  Google Scholar 

  • Fowler GS (1999) Behavioral and hormonal responses of Magellanic penguins (Spheniscus magellanicus) to tourism and nest site visitation. Biol Conserv 90:143–149

    Article  Google Scholar 

  • Goymann W (2005) Noninvasive monitoring of hormones in bird droppings: physiological validation, sampling, extraction, sex differences, and the influence of diet on hormone metabolite levels. Ann N Y Acad Sci 1046:35–53

    Article  CAS  PubMed  Google Scholar 

  • Griffiths R, Double MC, Orr K, Dawson RJG (1998) A DNA test to sex most birds. Mol Ecol 7:1071–1075

    Article  CAS  PubMed  Google Scholar 

  • Groscolas R, Jallageas M, Goldsmith A, Assenmacher I (1986) The endocrine control of reproduction and molt in male and female emperor (Aptenodytes forsteri) and Adelie (Pygoscelis adeliae) penguins. Gen Comp Endocrinol 62:43–53

    Article  CAS  PubMed  Google Scholar 

  • Harper JM, Austad SN (2000) Fecal glucocorticoids: a noninvasive method of measuring adrenal activity in wild and captive rodents. Physiol Biochem Zool 73:12–22

    Article  CAS  PubMed  Google Scholar 

  • Harvey S, Phillips JG, Rees A, Hall TR (1984) Stress and adrenal function. J Exp Zool 232:633–645

    Article  CAS  PubMed  Google Scholar 

  • Hayward LS, Richardson JB, Grogan MN, Wingfield JC (2006) Sex differences in the organizational effects of corticosterone in the egg yolk of quail. Gen Comp Endocrinol 146:144–148

    Article  CAS  PubMed  Google Scholar 

  • Holberton RL, Helmuth B, Wingfield JC (1996) The corticosterone stress response in gentoo and king penguins during the non-fasting period. Condor 98:850–854

    Article  Google Scholar 

  • Hood LC, Boersma PD, Wingfield JC (1998) The adrenocortical response to stress in incubating Magellanic penguins (Spheniscus magellanicus). Auk 115:76–84

    Google Scholar 

  • Jackson S (1992) Do seabird gut sizes and mean retention times reflect adaptation to diet and foraging method? Physiol Zoo 65:674–697

    Google Scholar 

  • Klasing KC (2005) Potential impact of nutritional strategy on noninvasive measurements of hormones in birds. Ann N Y Acad Sci 1046:5–16

    Article  CAS  PubMed  Google Scholar 

  • Klukowski LA, Cawthorn JM, Ketterson ED, Nolan V (1997) Effects of experimentally elevated testosterone on plasma corticosterone and corticosteroid-binding globulin in dark-eyed juncos (Junco hyemalis). Gen Comp Endocrinol 108:141–151

    Article  CAS  PubMed  Google Scholar 

  • Knapp R, Moore MC (1997) Male morphs in tree lizards have different testosterone responses to elevated levels of corticosterone. Gen Comp Endocrinol 107:273–279

    Article  CAS  PubMed  Google Scholar 

  • Kralj-Fišer S, Scheiber IBR, Blejec A, Moestl E, Kotrschal K (2007) Individualities in a flock of free-roaming greylag geese: behavioral and physiological consistency over time and across situations. Horm Behav 51:239–248

    Article  PubMed  Google Scholar 

  • Marchant S, Higgins PJ (1990) Handbook of Australian, New Zealand and Antarctic birds. Oxford University Press, Melbourne

    Google Scholar 

  • McQueen SM, Davis LS, Young G (1998) The reproductive endocrinology of Fiordland crested penguins Eudyptes pachyrhynchus. Emu 98:127–131

    Article  Google Scholar 

  • McQueen SM, Davis LS, Young G (1999) Sex steroid and corticosterone levels of Adelie penguins (Pygoscelis adeliae) during courtship and incubation. Gen Comp Endocrinol 114:11–18

    Article  CAS  PubMed  Google Scholar 

  • Millspaugh JJ, Washburn BE (2004) Use of fecal glucocorticoid metabolite measures in conservation biology research: considerations for application and interpretation. Gen Comp Endocrinol 138:189–199

    Article  CAS  PubMed  Google Scholar 

  • Moberg GP (2000) Biological response to stress: implications for animal welfare. In: Moberg GP, Mench JA (eds) The biology of animal stress: basic principles and implications for animal welfare. CABI Publishing, Wallingford, pp 1–21

    Chapter  Google Scholar 

  • Möhle U, Heistermann M, Palme R, Hodges JK (2002) Characterization of urinary and fecal metabolites of testosterone and their measurement for assessing gonadal endocrine function in male nonhuman primates. Gen Comp Endocrinol 129:135–145

    Article  PubMed  Google Scholar 

  • Möstl E, Palme R (2002) Hormones as indicators of stress. Domest Anim Endocrinol 23:67–74

    Article  PubMed  Google Scholar 

  • Möstl E, Messmann S, Bagu E, Robia C, Palme R (1999) Measurement of glucocorticoid metabolite concentrations in faeces of domestic livestock. J Vet Med A 46:621–631

    Article  Google Scholar 

  • Möstl E, Rettenbacher S, Palme R (2005) Measurement of corticosterone metabolites in birds’ droppings: an analytical approach. Ann N Y Acad Sci 1046:17–34

    Article  PubMed  Google Scholar 

  • Nakagawa S, Cuthill IC (2007) Effect size, confidence interval and statistical significance: a practical guide for biologists. Biol Rev 82:1–15

    Article  Google Scholar 

  • Nakagawa S, Möstl E, Waas JR (2003) Validation of an enzyme immunoassay to measure faecal glucocorticoid metabolites from Adelie penguins (Pygoscelis adeliae): a non-invasive tool for estimating stress? Polar Biol 26:491–493

    Google Scholar 

  • Palme R, Möstl E (1993) Biotin-streptavidin enzyme immunoassay for the determination of oestrogens and androgens in boar faeces. In: Proceedings of the 5th symposium on the analysis of steroids, pp 111–117

  • Palme R, Möstl E (1997) Measurement of cortisol metabolites in faeces of sheep as a parameter of cortisol concentration in blood. Zeitschrift Saugetierkunde Int J Mammal Biol 62(Suppl 2):192–197

    Google Scholar 

  • Palme R, Fischer P, Schildorfer H, Ismail MN (1996) Excretion of infused C-14-steroid hormones via faeces and urine in domestic livestock. Anim Reprod Sci 43:43–63

    Article  CAS  Google Scholar 

  • Palme R, Robia Ch, Messmann S, Hofer J, Möstl E (1999) Measurement of faecal cortisol metabolites in ruminants: a non-invasive parameter of adrenocortical function. Wien Tierärztl Mschr 86:237–241

    CAS  Google Scholar 

  • Palme R, Rettenbacher S, Touma C, El-Bahr SM, Möstl E (2005) Stress hormones in mammals and birds: comparative aspects regarding metabolism, excretion, and noninvasive measurement in fecal samples. Ann N Y Acad Sci 1040:162–171

    Article  CAS  PubMed  Google Scholar 

  • Quillfeldt P, Möstl E (2003) Resource allocation in Wilson’s storm-petrels Oceanites oceanicus determined by measurement of glucocorticoid excretion. Acta Ethol 5:115–122

    Article  Google Scholar 

  • Raouf SA, Smith LC, Brown MB, Wingfield JC, Brown CR (2006) Glucocorticoid hormone levels increase with group size and parasite load in cliff swallows. Anim Behav 71:39–48

    Article  Google Scholar 

  • Rettenbacher S, Möstl E, Hackl R, Ghareeb K, Palme R (2004) Measurement of corticosterone metabolites in chicken droppings. Br Poult Sci 45:704–711

    Article  CAS  PubMed  Google Scholar 

  • Ringer RK (1976) Adrenals. In: Sturkie PD (ed) Avian physiology. Springer, New York, pp 372–382

    Google Scholar 

  • Roberts ML, Buchanan KL, Hasselquist D, Evans MR (2007) Effects of testosterone and corticosterone on immunocompetence in the zebra finch. Horm Behav 51:126–134

    Article  CAS  PubMed  Google Scholar 

  • Romero LM (2002) Seasonal changes in plasma glucocorticoid concentrations in free-living vertebrates. Gen Comp Endocrinol 128:1–24

    Article  CAS  PubMed  Google Scholar 

  • Romero LM (2004) Physiological stress in ecology: lessons from biomedical research. Trends Ecol Evol 19(5):249–255

    Article  PubMed  Google Scholar 

  • Sapolsky RM (1992) Neuroendocrinology of the stress-response. In: Becker JB, Breedlove SM, Crews D (eds) Behavioral endocrinology. MIT Press, Cambridge, pp 287–324

    Google Scholar 

  • Scheiber IBR, Kralj S, Kotrschal K (2005) Sampling effort/frequency necessary to infer individual acute stress responses from fecal analysis in greylag geese (Anser anser). Ann N Y Acad Sci 1046:154–167

    Article  CAS  PubMed  Google Scholar 

  • Siegel HS (1980) Physiological stress in birds. Bioscience 30:529–534

    Article  CAS  Google Scholar 

  • Sorato E, Kotrschal K (2006) Hormonal and behavioural symmetries between the sexes in the northern bald ibis. Gen Comp Endocrinol 146:265–274

    Article  CAS  PubMed  Google Scholar 

  • Stevens CE, Hume ID (1995) Comparative physiology of the vertebrate digestive system. Cambridge University Press, Cambridge

  • Swett MB, Breuner CW (2008) Interaction of testosterone, corticosterone and corticosterone binding globulin in the white-throated sparrow (Zonotrichia albicollis). Comp Biochem Physiol A Mol Integr Physiol 151:226–231

    Article  CAS  PubMed  Google Scholar 

  • Touma C, Palme R (2005) Measuring fecal glucocorticoid metabolites in mammals and birds: the importance of validation. Ann N Y Acad Sci 1046:54–74

    Article  CAS  PubMed  Google Scholar 

  • Vleck CM, Van Hook JA (2002) Absence of daily rhythms of prolactin and corticosterone in Adelie penguins under continuous daylight. Condor 104:667–671

    Article  Google Scholar 

  • Vleck CM, Vertalino N, Vleck D, Bucher TL (2000) Stress, corticosterone, and heterophil to lymphocyte ratios in free-living Adelie penguins. Condor 102:392–400

    Article  Google Scholar 

  • von Holst D (1998) The concept of stress and its relevance for animal behavior. Adv Stud Behav 27:1–131

    Article  Google Scholar 

  • Wada M, Shimizu T, Kobayashi S, Yatani A, Sandaiji Y, Ishikawa T, Takemure E (1999) Behavioral and hormonal basis of polygynous breeding in male bush warblers (Cettia diphone). Gen Comp Endocrinol 116:422–432

    Article  CAS  PubMed  Google Scholar 

  • Wada H, Moore IT, Breuner CW, Wingfield JC (2006) Stress responses in tropical sparrows: comparing tropical and temperate Zonotrichia. Physiol Biochem Zool 79:784–792

    Article  PubMed  Google Scholar 

  • Walker BG, Boersma PD, Wingfield JC (2005a) Physiological and behavioral differences in Magellanic penguin chicks in undisturbed and tourist-visited locations of a colony. Conserv Biol 19:1571–1577

    Article  Google Scholar 

  • Walker BG, Wingfield JC, Boersma PD (2005b) Age and food deprivation affects expression of the glucocorticosteroid stress response in Magellanic penguin (Spheniscus magellanicus) chicks. Physiol Biochem Zool 78:78–89

    Article  CAS  PubMed  Google Scholar 

  • Walker BG, Boersma PD, Wingfield JC (2006) Habituation of adult Magellanic penguins to human visitation as expressed through behavior and corticosterone secretion. Conserv Biol 20:146–154

    Article  PubMed  Google Scholar 

  • Washburn BE, Millspaugh JJ, Schulz JH, Jones SB, Mong T (2003) Using fecal glucocorticoids for stress assessment in mourning doves. Condor 105:696–706

    Article  Google Scholar 

  • Wikelski M, Cooke SJ (2006) Conservation physiology. Trends Ecol Evol 21:38–46

    Article  PubMed  Google Scholar 

  • Wingfield JC (1994) Hormone–behavior interactions and mating systems in male and female birds. In: Short RV, Balaban E (eds) The differences between the sexes. Cambridge University Press, New York, pp 303–330

    Google Scholar 

  • Wingfield JC, Smith JP, Farner DS (1982) Endocrine responses of white-crowned sparrows to environmental stress. Condor 84:399–409

    Article  Google Scholar 

  • Wingfield JC, Vleck CM, Moore MC (1992) Seasonal changes of the adrenocortical response to stress in birds of the Sonoran Desert. J Exp Zool 264:419–428

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We thank Antarctica New Zealand, especially the staff at Scott Base and the field support staff. Our thanks are also due to the staff at the Institute for Biochemistry, University of Veterinary Medicine, Vienna, who helped conduct the EIAs. We thank Dick Wilkins and Raewyn Towers for help with the molecular sexing of samples; Karen Nutt for providing the necessary primers; and Bruce Patty for the technical support. We thank Education New Zealand, the Perry Foundation and the University of Waikato for the financial assistance. We also thank Rupert Palme, Lance McLeay, Lloyd Davis, and anonymous reviewers for their helpful comments on this manuscript.

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Authors and Affiliations

  1. Department of Biological Sciences, University of Waikato, Private Bag 3105, Hamilton, New Zealand

    Calum Edward Ninnes, J. R. Waas, N. Ling, S. Nakagawa, J. C. Banks, D. G. Bell, A. Bright, J. Chandler, Q. J. Hudson, J. R. Ingram, K. Lyall, D. K. J. Morgan & J. Wallace

  2. Department of Zoology, University of Otago, PO Box 56, Dunedin, New Zealand

    S. Nakagawa

  3. SubAntarctic Foundation for Ecosystems Research, 8 Estuary Road, Christchurch, 8061, New Zealand

    P. W. Carey

  4. Allan Wilson Centre for Molecular Ecology and Evolution, Massey University, PB 11-222, Palmerston North, New Zealand

    M. I. Stevens

  5. School of Biological Sciences, Flinders University of South Australia, GPO Box 2100, Adelaide, SA, 5001, Australia

    M. I. Stevens

  6. Department of Natural Sciences, Institute of Biochemistry, University of Veterinary Medicine, 1210, Vienna, Austria

    E. Möstl

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  1. Calum Edward Ninnes
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Correspondence to Calum Edward Ninnes.

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Communicated by G. Heldmaier.

Authors 5-15 are listed alphabetically.

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Ninnes, C.E., Waas, J.R., Ling, N. et al. Comparing plasma and faecal measures of steroid hormones in Adelie penguins Pygoscelis adeliae . J Comp Physiol B 180, 83–94 (2010). https://doi.org/10.1007/s00360-009-0390-0

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