Faecal Cortisol Metabolites as an Indicator of Adrenocortical Activity in Farmed Blue Foxes

Welfare studies of blue foxes would benefit from a measurement of faecal cortisol metabolites (FCMs) as a non-invasive, physiological stress parameter reflecting hypothalamus–pituitary–adrenal (HPA) axis activity. Before implementation, a species-specific validation of such a method is required. Therefore, we conducted a physiological validation of an enzyme immunoassay (EIA) to measure FCMs in blue foxes. Twenty individuals (nine males and eleven females) were injected with synthetic adrenocorticotrophic hormone (ACTH) and faecal samples were collected every third h for two days. The FCM baseline levels were assessed based on the first sampling day (control period, 144 samples), followed by the ACTH injection and the second day of sampling (treatment period, 122 samples). FCMs were analysed with a 5α-pregnane-3ß,11ß,21-triol-20-one EIA. We compared the estimated mean FCM concentrations of the treatment samples to the baseline average. All samples for the two periods were collected at the same time of the day, which enabled to test the data also with an hourly pairwise comparison. With the two statistical approaches, we tested whether a possible diurnal fluctuation in the FCM concentrations affected the interpretation of the results. Compared to the baseline levels, both approaches showed 2.4–3.2 times higher concentrations on time points sampled 8–14 h after the ACTH injection (p < 0.05). The estimated FCM concentrations also fluctuated slightly within the control period (p < 0.01). Inter-individual variations in FCM levels were marked, which highlights the importance of having a sufficient number of animals in experiments utilising FCMs. The sampling intervals of 3 h enabled forming of informative FCM curves. Taken together, this study proves that FCM analysis with a 5α-pregnane-3ß,11ß,21-triol-20-one EIA is a valid measurement of adrenocortical activity in the farmed blue foxes. Therefore, it can be utilised as a non-invasive stress indicator in future animal welfare studies of the species.

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Measuring Faecal Glucocorticoid Metabolites to Assess Adrenocortical Activity in Reindeer

Animals ◽

10.3390/ani9110987 ◽

2019 ◽

Vol 9 (11) ◽

pp. 987

Author(s):

Şeyda Özkan Gülzari ◽

Grete Helen Meisfjord Jørgensen ◽

Svein Morten Eilertsen ◽

Inger Hansen ◽

Snorre Bekkevold Hagen ◽

...

Keyword(s):

Stress Responses ◽

Dna Analysis ◽

Stress Hormones ◽

Invasive Technique ◽

Animal Conservation ◽

Adrenocortical Activity ◽

Non Invasive ◽

Faecal Glucocorticoid Metabolites ◽

Faecal Samples ◽

Species Specific

Several non-invasive methods for assessing stress responses have been developed and validated for many animal species. Due to species-specific differences in metabolism and excretion of stress hormones, methods should be validated for each species. The aim of this study was to conduct a physiological validation of an 11-oxoaetiocholanolone enzyme immunoassay (EIA) for measuring faecal cortisol metabolites (FCMs) in male reindeer by administration of adrenocorticotrophic hormone (ACTH; intramuscular, 0.25 mg per animal). A total of 317 samples were collected from eight male reindeer over a 44 h period at Tverrvatnet in Norway in mid-winter. In addition, 114 samples were collected from a group of reindeer during normal handling and calf marking at Stjernevatn in Norway. Following ACTH injection, FCM levels (median and range) were 568 (268–2415) ng/g after two hours, 2718 (414–8550) ng/g after seven hours and 918 (500–6931) ng/g after 24 h. Levels were significantly higher from seven hours onwards compared to earlier hours (p < 0.001). The FCM levels at Stjernevatn were significantly (p < 0.001) different before (samples collected zero to two hours; median: 479 ng/g) and after calf marking (eight to ten hours; median: 1469 ng/g). Identification of the faecal samples belonging to individual animals was conducted using DNA analysis across time. This study reports a successful validation of a non-invasive technique for measuring stress in reindeer, which can be applied in future studies in the fields of biology, ethology, ecology, animal conservation and welfare.

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Examination the stresseffect of litter’s let-together system (“kindergarten”) by measuring faecal cortisol metabolites of sows and piglets

Acta Agraria Debreceniensis ◽

10.34101/actaagrar/69/1794 ◽

2016 ◽

pp. 87-90

Author(s):

Zsolt Győri ◽

Margit Kulcsár ◽

Péter Balogh ◽

László Huzsvai ◽

Gabriella Novotniné Dankó

Keyword(s):

Stress Effect ◽

Sample Collection ◽

Blood Samples ◽

Adrenocortical Activity ◽

Non Invasive ◽

Invasive Method ◽

Corticosterone Metabolites ◽

Pig Husbandry ◽

Cortisol Metabolites

Piglets in commercial intensive pig husbandry are often abruptly weaned between 3 and 4 weeks for economic reasons. The process of weaning is a multifactorial stressor, in which nutritional, social, physical and psychologic stressors are combined. Piglets are often exposed to unfamiliar piglets around weaning which results in a period of vigorous fighting. Stress plays an important part in welfare research. Traditionally glucocorticoids are measured in blood samples but their use is often limited as the act of sample collection may stress an animal. Measurement of faecal cortisol/corticosterone metabolites is a non-invasive method for evaluation adrenocortical activity. The aim of this study was to examine the effect of litter’s let-together system (“kindergarten”) in the farrowing house by measuring faecal cortisol metabolites. According to our results the “kindergarten” system has no stress effect on sows and piglets, respectively.

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Non-invasive monitoring of adrenocortical activity in European badgers (Meles meles) and effects of sample collection and storage on faecal cortisol metabolite concentrations

Animal Biology ◽

10.1163/157075612x642914 ◽

2012 ◽

Vol 62 (4) ◽

pp. 419-432 ◽

Cited By ~ 15

Author(s):

Isabel Barja ◽

Gema Escribano-Ávila ◽

Carlos Lara-Romero ◽

Emilio Virgós ◽

Javier Benito ◽

...

Keyword(s):

Time Lapse ◽

Meles Meles ◽

Sample Collection ◽

Acth Stimulation ◽

Adrenocortical Activity ◽

Non Invasive ◽

Faecal Samples ◽

European Badgers ◽

And Storage ◽

Cortisol Metabolites

Due to the increasing demand for methods to quantify adrenal activity in response to stressors in wild animals, we evaluated whether stimulation with adrenocorticopic hormone (ACTH) increases faecal cortisol metabolite levels in European badgers. We also conducted several experiments to evaluate the effect of storage time and collection methods on cortisol metabolite levels of faecal samples. Faecal samples were collected from five captive European badgers (Meles meles) and cortisol metabolites were quantified by an enzyme immunoassay. Faecal cortisol metabolite levels increased 1-2 days after ACTH injection and dropped to pre-treatment levels 3 days after ACTH stimulation. We found that cortisol metabolite levels were not affected by the 8-hour time lapse from collection in the field until freezing of faecal samples, environmental exposure (temperature 25.66 ± 2.55°C; range 21.20-32.20) or mean relative humidity 62.77 ± 9.90% (range 42.00-79.00). Cortisol metabolites were not homogenously distributed in the scats. Cortisol metabolite levels were significantly lower when faecal extracts and samples were frozen for long periods of time (14 weeks or more). Thus, the measurement of faecal cortisol metabolites is a suitable method for the non-invasive evaluation of adrenocortical activity in European badgers. However, some aspects related to the collection and storage of faecal samples should be considered in future studies conducted with this species.

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Assessment of adrenocortical activity by non-invasive measurement of faecal cortisol metabolites in dromedary camels (Camelus dromedarius)

Tropical Animal Health and Production ◽

10.1007/s11250-013-0374-7 ◽

2013 ◽

Vol 45 (6) ◽

pp. 1453-1458 ◽

Cited By ~ 6

Author(s):

Omer-Elfaroug Sid-Ahmed ◽

Ahmed Sanhouri ◽

Badr-Eldin Elwaseela ◽

Imad Fadllalah ◽

Galal-Eldin Elazhari Mohammed ◽

...

Keyword(s):

Camelus Dromedarius ◽

Dromedary Camels ◽

Adrenocortical Activity ◽

Non Invasive ◽

Cortisol Metabolites ◽

Invasive Measurement

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Non-Invasive Monitoring of Adrenocortical Activity in Three Sympatric Desert Gerbil Species

Animals ◽

10.3390/ani11010075 ◽

2021 ◽

Vol 11 (1) ◽

pp. 75

Author(s):

Álvaro Navarro-Castilla ◽

Mario Garrido ◽

Hadas Hawlena ◽

Isabel Barja

Keyword(s):

Small Sample ◽

Post Injection ◽

Control Groups ◽

Treatment Groups ◽

Adrenocortical Activity ◽

Non Invasive ◽

Endocrine Status ◽

Corticosterone Metabolites ◽

Small Sample Sizes ◽

Fecal Corticosterone Metabolites

The study of the endocrine status can be useful to understand wildlife responses to the changing environment. Here, we validated an enzyme immunoassay (EIA) to non-invasively monitor adrenocortical activity by measuring fecal corticosterone metabolites (FCM) in three sympatric gerbil species (Gerbillus andersoni, G. gerbillus and G. pyramidum) from the Northwestern Negev Desert’s sands (Israel). Animals included into treatment groups were injected with adrenocorticotropic hormone (ACTH) to stimulate adrenocortical activity, while control groups received a saline solution. Feces were collected at different intervals and FCM were quantified by an EIA. Basal FCM levels were similar in the three species. The ACTH effect was evidenced, but the time of FCM peak concentrations appearance differed between the species (6–24 h post-injection). Furthermore, FCM peak values were observed sooner in G. andersoni females than in males (6 h and 18 h post-injection, respectively). G. andersoni and G. gerbillus males in control groups also increased FCM levels (18 h and 48 h post-injection, respectively). Despite the small sample sizes, our results confirmed the EIA suitability for analyzing FCM in these species as a reliable indicator of the adrenocortical activity. This study also revealed that close species, and individuals within a species, can respond differently to the same stressor.

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Stress and the costs of extra-territorial movement in a social carnivore

Biology Letters ◽

10.1098/rsbl.2009.0032 ◽

2009 ◽

Vol 5 (4) ◽

pp. 439-441 ◽

Cited By ~ 55

Author(s):

Andrew J Young ◽

Steven L Monfort

Keyword(s):

Chronic Stress ◽

Physiological Stress ◽

Negative Effects ◽

Delayed Dispersal ◽

Non Invasive ◽

Suricata Suricatta ◽

Related Distribution ◽

Selection For ◽

Animal Societies ◽

Pituitary Adrenal Axis

Costs associated with extra-territorial movement are believed to have favoured the evolution of delayed dispersal and sociality across a range of social vertebrates, but remain surprisingly poorly understood. Here we reveal a novel mechanism that may contribute substantially to the costs of extra-territorial movement: physiological stress. We show that subordinate male meerkats, Suricata suricatta , exhibit markedly elevated faecal glucocorticoid metabolite levels (a non-invasive measure of hypothalamic–pituitary–adrenal axis activity) while conducting extra-territorial prospecting forays. While brief increases in glucocorticoid levels are unlikely to be costly, chronic elevations, arising from prolonged and/or frequent forays, are expected to compromise fitness through their diverse negative effects on health. Our findings strongly suggest that prolonged extra-territorial movements do result in chronic stress, as the high glucocorticoid levels of prospectors do not diminish on longer forays and are no lower among males with greater prospecting experience. A generalized ‘stress’ of extra-territorial movement may therefore have strengthened selection for delayed dispersal and sociality in this and other species, and favoured the conduct of brief forays over extended periods of floating. Our findings have implications too for understanding the rank-related distribution of physiological stress in animal societies, as extra-territorial movements are often conducted solely by subordinates.

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A Review of Murine Cytomegalovirus as a Model for Human Cytomegalovirus Disease—Do Mice Lie?

International Journal of Molecular Sciences ◽

10.3390/ijms22010214 ◽

2020 ◽

Vol 22 (1) ◽

pp. 214

Author(s):

Michelle A. Fisher ◽

Megan L. Lloyd

Keyword(s):

Human Cytomegalovirus ◽

Murine Cytomegalovirus ◽

Clinical Samples ◽

Mouse Strains ◽

Cytomegalovirus Disease ◽

Wild Mice ◽

Congenital Cytomegalovirus ◽

Non Invasive ◽

Species Specific ◽

Disseminated Disease

Since murine cytomegalovirus (MCMV) was first described in 1954, it has been used to model human cytomegalovirus (HCMV) diseases. MCMV is a natural pathogen of mice that is present in wild mice populations and has been associated with diseases such as myocarditis. The species-specific nature of HCMV restricts most research to cell culture-based studies or to the investigation of non-invasive clinical samples, which may not be ideal for the study of disseminated disease. Initial MCMV research used a salivary gland-propagated virus administered via different routes of inoculation into a variety of mouse strains. This revealed that the genetic background of the laboratory mice affected the severity of disease and altered the extent of subsequent pathology. The advent of genetically modified mice and viruses has allowed new aspects of disease to be modeled and the opportunistic nature of HCMV infection to be confirmed. This review describes the different ways that MCMV has been used to model HCMV diseases and explores the continuing difficulty faced by researchers attempting to model HCMV congenital cytomegalovirus disease using the mouse model.

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Fecal cortisol metabolites as indicators of stress in crab-eating-fox (Cerdocyoun thous) in captivity

Pesquisa Veterinária Brasileira ◽

10.1590/s0100-736x2015001000007 ◽

2015 ◽

Vol 35 (10) ◽

pp. 859-862 ◽

Cited By ~ 3

Author(s):

Regina C.R. Paz ◽

Nely P. Souza ◽

Janine L. Brown

Keyword(s):

Fold Increase ◽

Reproductive Activity ◽

Biological Research ◽

Non Invasive ◽

Fecal Cortisol ◽

Fecal Glucocorticoid Metabolites ◽

Physiological Relevance ◽

In Captivity ◽

Invasive Method ◽

Cortisol Metabolites

Abstract: Blood samples collection is a common method in biological research using domestic animals. However, most blood sampling techniques are complicated and highly invasive and may therefore not be appropriate for wildlife animals in research concerning stress. Thus, a non-invasive method to measure steroid hormones is critically needed. The first goal of this study was to determine how glucocorticoids concentrations are impacted by translocation and reproductive activity in crab-eating-fox (Cerdocyoun thous) in captivity. The physiological relevance of fecal glucocorticoid metabolites was further validated by demonstrating: (1) The translocation of a male to a females enclosure resulted in a 3.5-fold increase compared to baseline concentrations, (2) changes in adrenocortical activity, as reflected in concentrations of fecal cortisol metabolites during reproduction, gestation and lactation in females foxes, indicating that social interactions resulted in large increases of fecal glucocorticoids metabolites during the reproductive season. From these findings we conclude that fecal samples can be used for the non-invasive assessment of adrenocortical status in crab-eating-fox.

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Temporal profiles of cortisol accumulation and clearance support scale cortisol content as an indicator of chronic stress in fish

Conservation Physiology ◽

10.1093/conphys/coz052 ◽

2019 ◽

Vol 7 (1) ◽

Cited By ~ 2

Author(s):

Frédéric Laberge ◽

Irene Yin-Liao ◽

Nicholas J Bernier

Keyword(s):

Chronic Stress ◽

The Body ◽

High Dose ◽

Stress Hormone ◽

Stress Indicators ◽

Stress Indicator ◽

Terrestrial Vertebrates ◽

Non Invasive ◽

Goldfish Carassius Auratus ◽

Temporal Profiles

Abstract The development of chronic stress indicators for fish is of great interest, but appropriate non-invasive methods are lagging those used in terrestrial vertebrates. Here, we explore the possibility that levels of the stress hormone cortisol in scales could be used as a chronic stress indicator. Three experiments were conducted to assess the temporal profiles of cortisol rise and fall in plasma and scales of goldfish (Carassius auratus) in response to stressors of varying intensity and duration. Results show that a single acute air emersion stressor does not influence scale cortisol content. In contrast, relative to plasma levels, the fall in scale cortisol content following a high-dose cortisol implant is delayed by at least 8 days, and the rise and fall in scale cortisol content in response to unpredictable chronic stress are delayed by at least 7 days. Also, scale cortisol content is spatially heterogeneous across the body surface of goldfish. Overall, since high and sustained circulating cortisol levels are needed to influence scale cortisol content and the rates of cortisol accumulation and clearance are much slower in scales than in plasma, our results show that scales can provide an integrated measure of cortisol production and serve as a chronic stress indicator.

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