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Use of fecal glucocorticoid metabolite measures in conservation biology research: considerations for application and interpretation
Abstract
Fecal glucocorticoid metabolite analyses are increasingly being used by a variety of scientists (e.g., conservation biologists, animal scientists) to examine glucocorticoid (i.e., stress hormone) secretion in domestic and wild vertebrates. Adrenocortical activity (i.e., stress response) is of interest to conservation biologists because stress can alter animal behavior, reduce resistance to disease, and affect population performance. The noninvasiveness of fecal-based assessments is attractive, particularly when studying endangered species, because samples can often be obtained without disturbing the animal. Despite such advantages, many confounding factors inhibit the utility of this technique in addressing conservation problems. In particular, interpretation of fecal glucocorticoid metabolite (FGM) measures may be confounded by the length of time animals are held in captivity, normal seasonal and daily rhythms, body condition, sample storage and treatment techniques, diet of the animal, assay selection, animal status (i.e., social ranking, reproductive status), sample age and condition, and sample mass. Further complicating interpretation and utility of these measures is the apparent species-specific response to these factors. The purpose of this paper is to discuss the factors that confound interpretation of FGM measures, summarize research that addresses these issues, and offer an agenda for future research and interpretation. We urge conservation biologists to carefully c…
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“Sample collection took place immediately after defecation to avoid bacterial and microbial degradation (Millspaugh and Washburn, 2004).”
From Effect of weather conditions and presence of visitors on adrenocortical activity in captive African penguins (Spheniscus demersus) (2017) · cites this paper as Millspaugh and Washburn, 2004
In the citing paper
“This non-invasive method permits the monitoring of the animal's endocrine status without the necessity to capture or handle the animals, which may compromise the accurate assessment of stress (Millspaugh and Washburn, 2004;Goymann, 2012).”
From Effect of weather conditions and presence of visitors on adrenocortical activity in captive African penguins (Spheniscus demersus) (2017) · cites this paper as Millspaugh and Washburn, 2004
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“Samples were gathered in cryovials and frozen immediately after collection at À20 °C (Millspaugh and Washburn, 2004;Palme et al., 2005;Sheriff et al., 2011).”
From Effect of weather conditions and presence of visitors on adrenocortical activity in captive African penguins (Spheniscus demersus) (2017) · cites this paper as Millspaugh and Washburn, 2004
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Establishing Stress Behaviors in Response to Manual Restraint in Cockatiels (Nymphicus hollandicus)
Turpen, K., Welle, K., Trail, J., Patel, S., & Allender, M. (2019) · Journal of avian medicine and surgery · doi:10.1647/2017-315 · Cited by 9
Glucs, Z. E., Smith, D. R., Tubbs, C. W., Scherbinski, J. J., Welch, A., Burnett, J., Clark, M., Eng, C., & Finkelstein, M. E. (2018) · PLoS ONE · doi:10.1371/journal.pone.0205565 · Cited by 11
Ahuja-Aguirre, C., López-deBuen, L., Rojas-Maya, S., & Hernández-Cruz, B. (2017) · Animal Reproduction Science · doi:10.1016/j.anireprosci.2017.03.008 · Cited by 9
Khonmee, J., Vorawattanatham, N., Pinyopummin, A., Thitaram, C., Somgird, C., Punyapornwithaya, V., & Brown, J. L. (2016) · Conservation Physiology · doi:10.1093/conphys/cow021 · Cited by 11
“…Morgan and Tromborg, 2007; Moreira et al., 2007; Scarlata et al., 2013; Khonmee et al., 2014a), many of which are associated with increased glucocorticoid production (Liptrap, 1993; Möstl and Palme, 2002; Millspaugh and Washburn, 2004; Touma and Palme, 2005; Brown, 2006; Khonmee et al., 2014b).”
BackgroundMethodInfluentialNarayan, E. J., Evans, N., & Hero, J. (2014) · European Journal of Wildlife Research · doi:10.1007/s10344-014-0842-z · Cited by 9
“…analysed bilby fecal extracts for concentrations of FCM using enzyme immunoassay (EIA) previously described for other mammals (Wielebnowski et al. 2002; Millspaugh and Washburn 2004), and recently used for a captive subpopulation of the greater bilby (Narayan et al. 2012; Evans et al. 2013).”
MethodTemporal stability of fecal cortisol metabolites in mountain-dwelling ungulates
Donini, V., Iacona, E., Pedrotti, L., Macho‐Maschler, S., Palme, R., & Corlatti, L. (2022) · Die Naturwissenschaften · doi:10.1007/s00114-022-01792-y · Cited by 12
“As high levels of stress may affect individual fitness (Millspaugh and Washburn 2004), e.”
BackgroundMethodInfluentialLong, C., Tordiffe, A. S. W., Sauther, M. L., Cuozzo, F. P., Millette, J. B., Ganswindt, A., & Scheun, J. (2021) · Conservation Physiology · doi:10.1093/conphys/coab081 · Cited by 11
“…metabolic processes and reproductive activities .......................................................................................................................................................... could be factors contributing to these changes (Millspaugh and Washburn, 2004; Goymann, 2012).”
BackgroundKachamakova, M., Koshev, Y., & Millesi, E. (2020) · Mammalian Biology · doi:10.1007/s42991-020-00066-2 · Cited by 8
Lind, M.-A., Hõrak, P., Sepp, T., & Meitern, R. (2019) · Hormones and Behavior · doi:10.1016/j.yhbeh.2019.104642 · Cited by 8
Escobar-Ibarra, I., Mayagoitia-Novales, L., Alcántara-Barrera, A., Cerda‐Molina, A. L., Mondragón‐Ceballos, R., Ramı́rez-Necoechea, R., & Alonso-Spilsbury, M. (2017) · The European Zoological Journal · doi:10.1080/24750263.2017.1332111 · Cited by 10
“Samples were kept in hermetically sealed plastic bags, correctly identified and frozen unpreserved within a few hours after collection, and stored at −20°C until analysis, as recommended by Millspaugh and Washburn (2004).”
MethodHUMAN DISTURBANCE AND THE PHYSIOLOGICAL RESPONSE OF ELK IN EASTERN WASHINGTON
Jachowski, D. S., McCorquodale, S. M., Washburn, B. E., & Millspaugh, J. J. (2015) · Wildlife Biology in Practice · doi:10.2461/wbp.2015.11.3 · Cited by 9
Self-citationAlbano, N., Santiago‐Quesada, F., Masero, J. A., Sánchez‐Guzmán, J. M., & Möstl, E. (2015) · General and Comparative Endocrinology · doi:10.1016/j.ygcen.2015.02.019 · Cited by 10
“The adrenocortical response of individuals prepare them to deal with a stressor; however, taking into account the potential adverse effects of chronic stress, these studies are particularly relevant in endangered species (Millspaugh and Washburn, 2004).”
BackgroundInfluentialGrotta‐Neto, F., Christofoletti, M. D., Piovezan, U., Herédias-Ribas, C. M., & Duarte, J. M. (2024) · Journal of Zoology · doi:10.1111/jzo.13165 · Cited by 7
Physiological and behavioural adjustment of a wild rodent to laboratory conditions
Carrilho, M., Monarca, R. I., Aparício, G., Mathias, M. D. L., Tapisso, J. T., & von Merten, S. (2023) · Physiology & Behavior · doi:10.1016/j.physbeh.2023.114385 · Cited by 10
“Two typical physiological reactions are changes in body mass and stress response [9], the latter usually being assessed by measuring the levels of glucocorticoid hormones, such as corticosterone (commonly in rodents, birds and reptiles) and cortisol (commonly in fish and most mammals) [10, 11].”
BackgroundBritton, J., Andreou, D., López-Béjar, M., & Carbajal, A. (2022) · bioRxiv · doi:10.1101/2022.11.15.516354 · Cited by 7
“Although the scale mass required to provide reliable cortisol content estimates from this method has not been determined specifically, in other taxa, material from feathers and faeces has required a minimum of =/> 20 mg of material (e.g. Millspaugh and Washburn, 2004; Lattin et al., 2011).”
MethodSocial roles influence cortisol levels in captive Livingstone's fruit bats (Pteropus livingstonii)
Edwards, M. J., Stanley, C. R., Hosie, C. A., Richdon, S., Price, E. C., Wormell, D., & Smith, T. E. (2022) · Hormones and Behavior · doi:10.1016/j.yhbeh.2022.105228 · Cited by 7
“Faecal Glucocorticoid Metabolites (FGMs) also provide a more complete picture of chronic stress than other substrates, as they are less impacted by instantaneous, acute events experienced by individuals (Millspaugh and Washburn, 2004; Sheriff et al., 2011).”
Chancellor, S., Abbo, L. A., Grasse, B., Sakmar, T., Brown, J. S., Scheel, D., & Santymire, R. M. (2021) · General and Comparative Endocrinology · doi:10.1016/j.ygcen.2021.113903 · Cited by 11
Zane, L., Ensminger, D. C., & Vázquez-Medina, J. (2021) · bioRxiv · doi:10.1371/journal.pone.0257370 · Cited by 7
Laver, P. N., Ganswindt, A., Ganswindt, S. B., & Alexander, K. A. (2020) · BMC Ecology · doi:10.1186/s12898-020-00280-z · Cited by 10
Validation of a noninvasive technique to quantify stress in northern bobwhite (Colinus virginianus)
Mohlman, J. L., Navara, K. J., Sheriff, M. J., Terhune, T. M., & Martin, J. A. (2020) · Conservation Physiology · doi:10.1093/conphys/coaa026 · Cited by 6
“While the use of fecal sampling is less invasive than plasma, relationships to blood glucocorticoid concentrations can require species-specific validation (Millspaugh and Washburn, 2004; Möstl et al., 2005: Palme, 2018).”
BackgroundStress in captive Blue-fronted parrots (Amazona aestiva): the animalists’ tale
Vidal, A. C., Roldán, M., Christofoletti, M. D., Tanaka, Y., Galindo, D. J., & Duarte, J. M. B. (2019) · Conservation Physiology · doi:10.1093/conphys/coz097 · Cited by 9
“Although in some occasions it is possible to separate these two fractions, using them together as a single sample has been recommended to achieve a more integrated estimate of the total hormones (Millspaugh and Washburn, 2004).”
BackgroundInfluentialLima, M., Scalercio, S., Lopes, C. T. A., Martins, N. D., Oliveira, K. G., Caldas-Bussiere, M. C., Santos, R., & Domingues, S. F. S. (2019) · Heliyon · doi:10.1016/j.heliyon.2019.e02166 · Cited by 7
“This technique is applied especially in pregnant females or new-born animals [1, 2], wild animals, or those of small size [3].”
MethodMolina‐García, L., Pérez, J. M., Sarasa, M., Ureña‐Gutiérrez, B., Espinosa, J., & Azorit, C. (2018) · Ecology and Evolution · doi:10.1002/ece3.4285 · Cited by 6
Bienboire‐Frosini, C., Alnot-Perronin, M., Chabaud, C., Asproni, P., Lafont‐Lecuelle, C., Cozzi, A., & Pageat, P. (2018) · Animals · doi:10.3390/ani8070105 · Cited by 10
“But because clear differences regarding the metabolism and excretion of GCM exist between species and sexes, a careful evaluation of the method used to assay GCM in feces or droppings is required for each species and sex investigated, as highlighted by some authors [24,26,36,37].”
BackgroundMethodResultInfluentialLiu, G., Hu, X., Kessler, M., Gong, M., Wang, Y., Li, H., Dong, Y., Yang, Y., & Li, L. (2018) · General and Comparative Endocrinology · doi:10.1016/j.ygcen.2018.02.014 · Cited by 9
“Improvements in noninvasive sampling methods to quantify stress offer opportunities to more easily carry out repeated sampling as well as reduce stress to study organisms (Millspaugh and Washburn, 2004).”
BackgroundCornale, P., Macchi, E., Renna, M., Prola, L., Perona, G. E., & Mimosi, A. (2015) · Journal of Applied Animal Welfare Science · doi:10.1080/10888705.2015.1072468 · Cited by 8
Determining hormone metabolite concentrations when enzyme immunoassay accuracy varies over time
Davidian, E., Benhaiem, S., Courtiol, A., Hofer, H., Höner, O. P., & Dehnhard, M. (2015) · Methods in Ecology and Evolution · doi:10.1111/2041-210x.12338 · Cited by 8
Azorit, C., Castro, J., Carrasco, R., Tellado, S., Órpez, R., & Moro, J. (2012) · doi:10.1071/an12022 · Cited by 7
Meij, B. P., & Mol, J. A. (2008) · Clinical Biochemistry of Domestic Animals · doi:10.1016/b978-0-12-370491-7.00019-2 · Cited by 7
Bertrand, D., Berman, C. M., Heistermann, M., Agil, M., Sutiah, U., & Engelhardt, A. (2023) · Animals · doi:10.3390/ani13182842 · Cited by 6
“…addition to real or perceived risks of tourism, FGCM levels in our macaque study groups could increase with environmental pressures, e.g., food resources [39], extreme weather [40,41], reproductive hormones (estradiol: [33,42,43]; testosterone: [44,45]), vigorous physical activity [46] and others.”
BackgroundAssessing stress in wild black-and-white colobus monkeys non-invasively
King, A. G., Edwards, P. D., Cote, S. M., Palme, R., Boonstra, R., & Sicotte, P. (2023) · General and Comparative Endocrinology · doi:10.1016/j.ygcen.2023.114212 · Cited by 8
Hariohay, K. M., Hunninck, L., Ranke, P., Fyumagwa, R., Palme, R., & Røskaft, E. (2023) · Conservation Physiology · doi:10.1093/conphys/coad002 · Cited by 6
“Following Millspaugh and Washburn (2004), we collected fecal samples rapidly (here, within 25 minutes) after defe-cation to avoid environmental degradation of FGMs.”
MethodImmune and stress physiology of two captively‐housed tortoise species
Hartzheim, A. M., Terry, J., Field, E. K., Haydt, N. T., Poo, S., & Neuman‐Lee, L. A. (2022) · Journal of Experimental Zoology Part A Ecological and Integrative Physiology · doi:10.1002/jez.2674 · Cited by 10
Fink, L. B., Mukobi, A., Gruber, L., Reed, C., DeLibero, J., Jackson, S., Neill, S., Walz, J., Sines, C., VanBeek, B., et al. (2022) · Animals · doi:10.3390/ani12233303 · Cited by 7
“Care staff were instructed to avoid samples contaminated by urine as glucocorticoid metabolites are secreted in both urine and feces but to maintain consistency, only one medium should be analyzed at once [13,14].”
BackgroundRosenbaum, S., Eckardt, W., Stoinski, T. S., Umuhoza, R., Kuzawa, C. W., & Santymire, R. M. (2021) · American Journal of Primatology · doi:10.1002/ajp.23295 · Cited by 7
“…samples contain hormone “pools” that represent hours or even days, there is no relationship between collection time and fecal steroid metabolite concentration in mountain gorillas or other species with long gut passage times (Eckardt et al., 2016; Millspaugh & Washburn, 2004; Nizeyi et al., 2011).”
BackgroundChen, C., Chang, A., Tsai, M.-S., Huang, Y., Pei, K. J., & Li, Y.-C. (2021) · Scientific Reports · doi:10.1038/s41598-021-90725-2 · Cited by 7
Christianson, D., Coleman, T. H., Doan, Q., & Haroldson, M. A. (2021) · Conservation Physiology · doi:10.1093/conphys/coab029 · Cited by 6
“Weathering is known to influence faecal cortisol concentrations, possibly due to microbial metabolism of cortisol in the feces after defecation (Millspaugh and Washburn, 2004; Stetz et al., 2013; Dalerum et al., 2020).”
BackgroundCondition of mule deer during winter: stress and spatial overlap with North American elk
Atwood, M., Kie, J., Millspaugh, J., Matocq, M., & Bowyer, R. (2019) · Mammal Research · doi:10.1007/s13364-019-00474-x · Cited by 6
Self-citationKrone, O., Bailey, L. D., Jähnig, S., Lauth, T., & Dehnhard, M. (2019) · General and Comparative Endocrinology · doi:10.1016/j.ygcen.2019.113223 · Cited by 6
Juliana, J. R. S., Bryant, J. L., Wielebnowski, N. C., & Kotler, B. P. (2019) · Israel Journal of Ecology and Evolution · doi:10.1163/22244662-20191043 · Cited by 7
Kvasha, I. G., Laktionova, T. K., & Voznessenskaya, V. V. (2018) · Biology Bulletin · doi:10.1134/s1062359018030081 · Cited by 5
“Samples under 0.02 g were eliminated from the analysis due to a nonlinear upward skew of the hormone concentration (Millspaugh and Washburn, 2004).”
MethodInfluentialBryant, J. L. (2018) · International Journal of Avian & Wildlife Biology · doi:10.15406/ijawb.2018.03.00064 · Cited by 8
Within-sample variation in snowshoe hare faecal glucocorticoid metabolite measurements
Lafferty, D. J. R., Kumar, A. V., Whitcher, S., Hackländer, K., & Mills, L. S. (2017) · Conservation Physiology · doi:10.1093/conphys/cox068 · Cited by 6
“For example, faeces collection is efficient and cost-effective relative to other non-invasive stress hormone sampling techniques (i.e. hair cortisol extraction) (Möstl and Palme, 2002; Millspaugh and Washburn, 2004; Palme et al., 2005; Sheriff et al., 2011a; Goymann, 2012; Dantzer et al., 2014).”
BackgroundInfluentialWelfare of a pair of captive tigers - a hand-reared female and a parent - reared male
Bertocchi, M., Caterina, S., Francesco, D., Elisabetta, M., Enrico, P., Camillo, S., Patrizia, P., & Fausto, Q. (2015) · doi:10.24297/jaa.v5i1.4508 · Cited by 6
“All the above leads to a greater difficulty in interpreting results (34).”
BackgroundCalifornia gull chicks raised near colony edges have elevated stress levels
Herring, G., & Ackerman, J. T. (2011) · General and Comparative Endocrinology · doi:10.1016/j.ygcen.2011.04.029 · Cited by 7
Blas, J., & Fairhurst, G. D. (2021) · Elsevier eBooks · doi:10.1016/b978-0-12-819770-7.00003-7 · Cited by 13
Narayan, E. J., Baskaran, N., & Vaz, J. (2017) · InTech eBooks · doi:10.5772/intechopen.69780 · Cited by 7
“Millspaugh and Washburn [24] reviewed the factors that can influence GC levels, such as season, food and habitat availability, reproductive variability, age and gender.”
BackgroundWatanabe, T. T., Lolli, S., Ferrari, L., & Ferrante, V. (2013) · doi:10.2298/bah1304727w · Cited by 6
Barukčić, V., Berlinguer, F., Pasciu, V., Sotgiu, F. D., & Šprem, N. (2025) · BMC Veterinary Research · doi:10.1186/s12917-025-04678-z · Cited by 6
“One reason for the frequent use of this matrix could be that the collection of feces is not associated with stress for the animals and that it provides information on long-term hormone levels [24, 28, 50].”
Commentary: Environmental RNA and the assessment of organismal function in the field
Glover, C. N., Veilleux, H. D., & Misutka, M. D. (2024) · Comparative Biochemistry and Physiology Part B Biochemistry and Molecular Biology · doi:10.1016/j.cbpb.2024.111036 · Cited by 8
Physiological response after translocation differs between source populations in a threatened mammal
Williams-Kelly, K. S., Berry, L. E., Branch, K., Cowen, S., Garretson, S., Holland, G. J., Ladd, R., Olds, L. G. M., Rayner, K., Sims, C., et al. (2023) · Royal Society Open Science · doi:10.1098/rsos.230836 · Cited by 5
“Furthermore, blood sampling gives a snapshot of glucocorticoid levels at the specific time of sampling, while faecal samples contain average levels of hormone metabolites and provide information about the animal ’ s experience over a period of time, dependent on defecation rates [22].”
BackgroundFokidis, H. B., Brock, T., Newman, C., Macdonald, D. W., & Buesching, C. D. (2023) · Conservation Physiology · doi:10.1093/conphys/coad024 · Cited by 9
“However, all of these represent concentrations at short timescales (minutes to hours) that can be impacted by many concomitant factors like recent capture or evasion of a predator (Millspaugh and Washburn, 2004; Fokidis, 2016).”
BackgroundD'Agostino, V. C., Ajó, A. F., Degrati, M., Krock, B., Hunt, K., Uhart, M., & Buck, C. (2021) · Oecologia · doi:10.1007/s00442-021-05078-4 · Cited by 5
“Based on data from other species, the lag time for right whales is usually estimated at 24 h, i.e., fGCm concentrations in a given fecal sample likely reflects circulating levels in plasma of the day prior to fecal sample collection (Millspaugh and Washburn 2004; Rolland et al.”
BackgroundMétodos não-invasivos para análises hormonais aplicadas aos estudos de ecologia e etologia
Pereira, R. (2007) · doi:10.1590/s1516-35982007001000008 · Cited by 5
“…estudos recentes demonstraram que o simples congelamento de amostras fecais em freezer convencional (-20°C) poucas horas após a sua defecação é uma das melhores opções para preservação dos níveis hormonais por períodos prolongados (Millspaugh & Washburn, 2004; Palme, 2005; Wasser et al., 2002).”
BackgroundClegg, S. R., & Millson, S. H. (2021) · Comparative Cognition & Behavior Reviews · doi:10.3819/ccbr.2021.160002 · Cited by 6
“As such, it is important that faecal samples are collected rapidly postdefecation and frozen to prevent further bacterial degradation of hormones (Hodges & Heistermann, 2011; Millspaugh & Washburn, 2004), as this will affect the results (Palme, 2005, 2019).”
BackgroundMethodTrends in Wildlife Research: A Bibliometric Approach
Arroyo, B., Mateo, R., & García, J. T. (2016) · Wildlife research monographs · doi:10.1007/978-3-319-27912-1_1 · Cited by 5
Keech, A. L. (2008) · cIRcle (University of British Columbia) · doi:10.14288/1.0066818 · Cited by 2
Why not both? Measuring cortisol and corticosterone in poison frogsPreprint
Westrick, S. E., Paitz, R. T., & Fischer, E. K. (2023) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2023.06.19.545597 · Cited by 7
“…t e r one r e l ea s e r a t e , pg / h r) species D. tinctorius E. anthonyi P. terribilis inform research and conservation efforts and improve our understanding of how animals are impacted by and respond to environmental changes and threats (Busch and Hayward, 2009; Millspaugh and Washburn, 2004).”
Osburn, K., Crossey, B., Majelantle, T. L., & Ganswindt, A. (2025) · Journal of Zoology · doi:10.1111/jzo.13257 · Cited by 6
Agustí, C., Manteca, X., García‐Párraga, D., & Talló-Parra, O. (2024) · Animals · doi:10.3390/ani14091377 · Cited by 4
“In particular, the interpretation of cetacean epidermal cortisol concentrations may be confounded by sample storage and treatment techniques, the sex and age of the individual, variations in matrix growth rates, the location of the body and the epidermal layers sampled, among others [44,46–49].”
BackgroundGort‐Esteve, A., Carbajal, A., López, M., Manteca, X., Ruiz‐Olmo, J., & Riera, J. L. (2024) · Journal of Zoology · doi:10.1111/jzo.13149 · Cited by 6
Majelantle, T. L., Ganswindt, A., Ganswindt, S. B., Hagenah, N., Hart, D. W., & Bennett, N. C. (2023) · Animals · doi:10.3390/ani13193039 · Cited by 4
“Consequently, samples less than 0.0150 g were considered too small [31] and were not used in any further analyses ( n = 30).”
InfluentialMajelantle, T. L., Bennett, N. C., Ganswindt, S. B., Hart, D. W., & Ganswindt, A. (2023) · Animals · doi:10.3390/ani13081424 · Cited by 4
“0150 g were considered too small [31] and not used in any further analyses (n = 30).”
BackgroundHartell‐DeNardo, J., Kozlowski, C. P., Baskir, E. A., Macek, M., Dorsey, C. L., & Powell, D. (2022) · Zoo Biology · doi:10.1002/zoo.21735 · Cited by 6
Monitoring of Cortisol Levels in Hog Deer with Varying Environment Exposure
Shah, M. H. A., Rafi, U., Yasmeen, R., & Ahmad, M. (2022) · Vol 4 Issue 3 · doi:10.33411/ijist/2022040321 · Cited by 4
Racine, H., Guthrie, K., Hill, T., & Loughman, Z. (2022) · Animals · doi:10.3390/ani12111410 · Cited by 4
“However, there are a considerable number of benefits to this methodology, including a noninvasive method of sample collection, as well as the means to assess net effects of stress accumulated over time [8,9,24,25].”
BackgroundValerio, A., Borrego, C. S., Boitani, L., Casadei, L., Giuliani, A., Wielgus, R. B., Simek, S. L., & Valerio, M. (2021) · Scientific Reports · doi:10.1038/s41598-021-85600-z · Cited by 4
Can antibody-based assays consistently detect differences in feather corticosterone?
Fischer, C. P., Vitousek, M., & Romero, L. (2021) · Journal of Ornithology · doi:10.1007/s10336-021-01866-0 · Cited by 4
Spatial and temporal trends in mercury levels in the down of black stork chicks in central Europe.
Kucharska, K., Binkowski, Ł., & Dudzik, K. (2021) · Environmental Pollution · doi:10.1016/j.envpol.2021.116571 · Cited by 4
Kroska, A. C., Wolf, N., Planas, J. V., Baker, M. R., Smeltz, T. S., & Harris, B. P. (2021) · Conservation Physiology · doi:10.1093/conphys/coab001 · Cited by 6
“For example, for organisms in natural settings, the capture and sampling processes may themselves be reflected in the plasma cortisol levels, thereby resulting in potential overestimation of poststressor cortisol levels (Millspaugh and Washburn, 2004; Romero and Reed, 2005; Touma and Palme, 2005; Bertotto et al., 2010…”
BackgroundGholib, G., Pampang, F. H. R. A., Lubis, T. M., Adam, M., Jalaluddin, M., Razali, R., Azhar, A., & Karmil, T. (2020) · E3S Web of Conferences · doi:10.1051/e3sconf/202015101061 · Cited by 4
“The last few decades have developed a non-invasive method for measuring cortisol hormone from feces or urine samples [5,6].”
MethodGlucocorticoid Profiles in Frugivorous Bats on Wind Farms in the Mexican Tropics
Medina-Cruz, G. E., Méndez, A. S., & Briones-Salas, M. A. (2020) · Acta Chiropterologica · doi:10.3161/15081109acc2020.22.1.013 · Cited by 7
Grotto, C. E., Wolf, T. E., Berkeley, E. V., Lee, S. J., & Ganswindt, A. (2020) · African Zoology · doi:10.1080/15627020.2020.1776635 · Cited by 3
Lavin, S. R., Woodruff, M. C., Atencia, R., Cox, D., Woodruff, G. T., Setchell, J. M., & Wheaton, C. J. (2019) · Conservation Physiology · doi:10.1093/conphys/coz032 · Cited by 4
“, 2018), but no comprehensive set of validations (Millspaugh and Washburn, 2004; Shutt et al., 2012; Behringer and Deschner, 2017) for quantifying FGMs in mandrills has been published.”
BackgroundMethodGöttert, T., Grothmann, P., & Zeller, U. (2015) · The Journal of zoo and aquarium research · doi:10.19227/jzar.v3i1.92 · Cited by 4
“02g) resulted in proportionally higher FCM levels (Millspaugh and Washburn 2004; Goymann 2005).”
BackgroundResultParnell, T., Narayan, E. J., Magrath, M. J. L., Roe, S., Clark, G., Nicolson, V. N., Martin-Vegue, P., Mucci, A., & Hero, J. (2014) · Conservation Physiology · doi:10.1093/conphys/cou038 · Cited by 13
“…as diet, seasonal rhythms, sample storage and preservation techniques, reproductive status, social arrangements and exposure to visitors are widely discussed in published literature but are yet to be explored rigorously (Sapolsky et al. , 2000; Goymann et al. , 2001; Millspaugh and Washburn, 2004).”
The pattern of ovulation in the southern African spiny mouse (Acomys spinosissimus)
de Bruin, P., Ganswindt, A., Bennett, N., & Medger, K. (2014) · doi:10.1016/j.mambio.2014.05.003 · Cited by 4
“Hormone analysis The minimal appropriate mass for hormone measurement (Millspaugh and Washburn, 2004) was determined in a preceding study (de Bruin et al.”
MethodZimmerman, G. S., Millspaugh, J., Link, W., Woods, R., & Gutiérrez, R. J. (2013) · General and Comparative Endocrinology · doi:10.1016/j.ygcen.2013.08.010 · Cited by 4
Self-citationAgricultural Expansion and the Future of the Maned Wolf
Vynne, C. (2013) · doi:10.1201/b15607-16 · Cited by 4
Non-invasive endocrine monitoring using fecal steroid analysis: opportunities and challenges
Schwarzenberger, F. (2007) · Revista Brasileira de Zootecnia · doi:10.1590/s1516-35982007001000010 · Cited by 3
“Because of species specific differences in hormone metabolite excretion, validation should be done separately for even closely related species (Graham, 2004; Keay et al., 2006; Millspaugh & Washburn, 2004; Möstl & Palme, 2002; Palme et al., 2005; Schwarzenberger et al., 1997; Touma & Palme, 2005).”
BackgroundResultGenetic Analyses of Non-invasively Collected Samples Aids in the Conservation of Elephants
Ahlering, M., Budd, K., Schuttler, S., & Eggert, L. S. (2020) · Springer eBooks · doi:10.1007/978-3-030-33334-8_11 · Cited by 5
Piñero, J. A., Jansen, H. T., Robbins, C. T., Vincent, E. P., & Lafferty, D. J. R. (2025) · Conservation Physiology · doi:10.1093/conphys/coae093 · Cited by 3
Pritchard, A. J., Vogel, E. R., Blersch, R., & Palombit, R. A. (2024) · Primates · doi:10.1007/s10329-024-01172-2 · Cited by 3
Granweiler, J., Cristóbal‐Azkarate, J., Morton, N., Palme, R., & Shultz, S. (2024) · Hormones and Behavior · doi:10.1016/j.yhbeh.2024.105526 · Cited by 3
“…metabolites in faeces, cross-species and populations comparisons, assay reactivity) and they require careful interpretation of their biological and ecological significance (Millspaugh and Washburn, 2004; M ¨ ostl and Palme, 2002; Palme, 2019, 2005; Romero and Beattie, 2022; Sheriff et al., 2011).”
BackgroundDriscoll, M. V., Tuttle, A. D., & Romano, T. A. (2022) · General and Comparative Endocrinology · doi:10.1016/j.ygcen.2022.114147 · Cited by 3
Santymire, R. M., Young, M., Lenihan, E., & Murray, M. J. (2022) · Animals · doi:10.3390/ani12202868 · Cited by 3
“Acclimation to a new environment is a proven stressor in cephalopods [36,54–56] and mammalian species [16,57].”
BackgroundRamos, A., Robin, J., Manizan, L., Audroin, C., Rodríguez, E., Kemp, Y. J. M., & Sueur, C. (2022) · Animals · doi:10.3390/ani12070849 · Cited by 5
Kummrow, M., Pimm, R. H., Mackie, P. M., Tabh, J. K. R., & Mastromonaco, G. F. (2021) · General and Comparative Endocrinology · doi:10.1016/j.ygcen.2021.113822 · Cited by 4
“Glucocorticoids also increase in response to external stressors as animals respond behaviorally and physiologically to conserve energy and increase their chances of survival (Moore and Jessop, 2003; Millspaugh and Washburn, 2004; Palme, 2018).”
BackgroundVolfová, M., Machovcová, Z., Voslářová, E., Bedáňová, I., & Večerek, V. (2020) · Animals · doi:10.3390/ani10061013 · Cited by 3
“Given their small body size and fast metabolism, changes in FGMs were expected to be measurable within one to two days after exposure to a stressor [1,10] as confirmed by several studies assessing changes in FGM concentrations resulting from stress, for example, in zoo-living orangutans (Pongo spp.”
BackgroundKhonmee, J., Wathirunwong, T., Yano, T., Somgird, C., Brown, J. L., & Yamsakul, P. (2018) · Veterinary Medicine International · doi:10.1155/2018/2762153 · Cited by 3
Botelho, G. G., Falbo, M. K., Ost, P. R., Czekoski, Z. M., Raviolo, A., Giotto, F., Goldoni, E., & Morais, R. N. (2015) · Journal of animal physiology and animal nutrition · doi:10.1111/jpn.12257 · Cited by 3
Fecal glucocorticoid metabolites associated with nutrition in American plains bison
Metrione, L. C., Kruse, C. G., Wesley, B., Mowle, A. K., Hunter, D. L., & Penfold, L. M. (2026) · Frontiers in Ecology and Evolution · doi:10.3389/fevo.2026.1683923 · Cited by 2
Huang, R., Chen, H., Guo, Q., Lu, M., Wu, S., Nong, D., Li, C., Ma, Y., & Zhou, Q. (2025) · Journal of Hazardous Materials · doi:10.1016/j.jhazmat.2025.140162 · Cited by 2
Green, J., Schmidt-Burbach, J., & Hartley-Backhouse, L. (2025) · Frontiers in Ethology · doi:10.3389/fetho.2025.1532995 · Cited by 2
Bisheh, A. Y., Amininasab, S. M., Pirsaraei, Z. A., & Jafarian, Z. (2025) · Global Ecology and Conservation · doi:10.1016/j.gecco.2025.e03526 · Cited by 2
Quirici, V., Kilgour, D. A. V., Cuevas, É., Fletcher, S., Sarmiento, C., González‐Lagos, C., & Romero, L. M. (2025) · Frontiers in Physiology · doi:10.3389/fphys.2025.1523983 · Cited by 2
Serres‐Corral, P., Almagro, V., Ensenyat, C., Carbonell, L., Borragan, S., Martínez‐Nevado, E., Quevedo, M. Á., Fernández-Bellón, H., Carbajal, A., & López‐Béjar, M. (2025) · Zoo Biology · doi:10.1002/zoo.21891 · Cited by 4
“Hormone measurements in feces are particularly advantageous as they allow for long ‐ term monitoring without disturbing the animals, reflecting hormonal activity over a species ‐ specific intestinal transit time (Möstl and Palme 2002; Millspaugh and Washburn 2004; Palme 2019).”
Bartolomé, A., Palme, R., Macho‐Maschler, S., Carazo, P., & Font, E. (2024) · Physiology & Behavior · doi:10.1016/j.physbeh.2024.114751 · Cited by 5
“Non-invasive techniques to measure stress-related hormonal levels, such as faecal cortisol/corticosterone metabolites (FCMs), have become popular tools to assess the welfare state for mammals (particularly farm animals) and birds, but their use in reptiles is still very limited [18 – 20].”
Barbosa-Moyano, H., Frediani, M., Prosperi, M., Titon, S. C. M., de Figueiredo, A. D., Gomes, F. R., & de Oliveira, C. D. (2024) · Global Ecology and Conservation · doi:10.1016/j.gecco.2024.e02845 · Cited by 2
Metadata
- Authors
- Millspaugh, J. J., & Washburn, B. E.
- Affiliations
- University of Missouri (US)
- Animal and Plant Health Inspection Service (US)
- Corresponding author
- Millspaugh, Joshua J.
- Venue
- General and Comparative Endocrinology
- Publisher
- Elsevier BV
- Published
- 2004-09
- Volume
- 138
- Issue
- 3
- Pages
- 189-199
- ISSN
- 0016-6480 (ISSN-L)
- OpenAlex
- W2168394255
- Cited by
- 553
- References
- 0
- Citations by year
- 2026: 19 · 2025: 20 · 2024: 24 · 2023: 25 · 2022: 18 · 2021: 43 · 2020: 25 · 2019: 25 · 2018: 32 · 2017: 25 · 2016: 30 · 2015: 25 · 2014: 43 · 2013: 37 · 2012: 31
- FWCI
- 3.53
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- General and Comparative Endocrinology (publishedVersion)
- PubMed (publishedVersion)
- Lincoln (University of Nebraska) (submittedVersion)
- Funders
- University of Missouri
- Topics
- Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Animal Behavior and Welfare Studies
- Physical Sciences › Environmental Science › Ecology › Wildlife Ecology and Conservation
- Life Sciences › Neuroscience › Behavioral Neuroscience › Stress Responses and Cortisol
- Keywords
- fecal glucocorticoid metabolites, stress hormones, glucocorticoid secretion, adrenocortical function, conservation biology, seasonal rhythms, body condition, assay selection, social ranking, reproductive status, threatened species, wildlife disturbance, surrogate species
- MeSH terms
- Age Factors, Animals, Conservation of Natural Resources, Ecology, Feces, Female, Glucocorticoids, Male, Reproduction, Seasons, Sex Factors, Specimen Handling, Zoology, Reproducibility of Results, Clinical Laboratory Techniques
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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