scholarly journals Measuring Faecal Glucocorticoid Metabolites to Assess Adrenocortical Activity in Reindeer

Animals ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 987
Author(s):  
Şeyda Özkan Gülzari ◽  
Grete Helen Meisfjord Jørgensen ◽  
Svein Morten Eilertsen ◽  
Inger Hansen ◽  
Snorre Bekkevold Hagen ◽  
...  

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.

2019 ◽  
Vol 65 (1-2) ◽  
pp. 21-27 ◽  
Author(s):  
Justin R. St. Juliana ◽  
Jocelyn L. Bryant ◽  
Nadja Wielebnowski ◽  
Burt P. Kotler

We evaluated the suitability of a corticosterone enzyme immunoassay (EIA) to monitor excretion of fecal glucocorticoid metabolites (FGM) in response to Adrenocorticotropic hormone (ACTH) and saline injections in three desert rodent species (Gerbillus andersoni allenbyi (GA), Gerbillus nanus (GN), and Gerbilis piridium (GP). We exposed 24 gerbils (N = 9 for GA, N = 7 for GN, N = 8 for GP) to an ACTH and a saline injection at different times. Fecal samples were collected hourly for 24 hours after injection. The average starting concentration (baseline) FGM concentration was 797 ng/g for GA, 183 ng/g for GN, and 749 ng/g for GP. The average peak concentration was 2377 ng/g for GA, 589 ng/g for GN, and 1987 ng/g for GP. We were able to provide a physiological validation for the chosen assay in GAs and GPs, however, our results for GNs were less clear. We found an increase in FGM concentrations on average after 5.5 hours in GA, 3.1 hours in GN, and 3.8 hours in GP. We found a peak in FGM concentration on average after 8.8 hours in GA, 5.6 hours in GN, and 10.3 hours in GP. We determined that FGM concentration returned to starting value on average after 14.4 hours in GA, 9.1 hours in GN, and 15.1 hours in GP. The outcomes of this study can help establish trapping protocols and time frames for FGM monitoring of these wild small mammal populations. The time course for excretion of FGM is similar between the species in this study, and comparable to some non-desert rodents. We found high variation in the time course of excretion within species. This variation needs to be taken into account when monitoring stress responses in the field. By assessing adrenocortical activity using FGM monitoring, stress responses to varying ecological and environmental factors can be reliably examined in the field.


Author(s):  
D. Chharang ◽  
S. Choudhary ◽  
V. Kumar ◽  
S. Sharma ◽  
P. C. Sharma ◽  
...  

Background: Wildlife tourism has been shown to increase stress in a variety of species and can negatively affect the survival, reproduction, welfare, and behaviour of individuals. The use of faecal glucocorticoid metabolites has favoured the development of non-invasive methods to study physiological conditions and could aid in successful and effective strategies for the management and conservation of endangered species. Methods: An experiment was conducted to determine the effect of dietary inclusion of live microbial cultures to find patterns of physiological stress hormone in faeces of 18 safari Asian elephants of 30-62 years age and body weight (3495 ± 133.34 Kg) for two months. They were randomly divided into three groups, with six elephants in each, i.e. control with no probiotic (T1) and the other two with probiotics, which contained 1 × 109 CFU/g concentrate of Lactobacillus acidophilus (T2) and Saccharomyces cerevisiae (T3) @ every 50 kg body weight per day. Faecal samples were taken on 0, 30th, 40th and 50th day of the experiment. Result: It was observed that the mean faecal glucocorticoid metabolites, demonstrated by the elephants of T2, and T3 groups were not significantly (P > 0.05) different from the control group (T1) and remained unaffected on the supplementation of live microbial cultures.


Polar Biology ◽  
2021 ◽  
Author(s):  
Malin Larm ◽  
Anne Lene Hovland ◽  
Rupert Palme ◽  
Anne-Mathilde Thierry ◽  
Andrea L. Miller ◽  
...  

AbstractMeasuring fecal glucocorticoid metabolites (fGCMs) is a widely used, non-invasive method for studies of stress in vertebrates. To study physiological responses in wild Arctic foxes (Vulpes lagopus) to perceived stressors such as fluctuating food availability, occurrence of competitors and predators and disturbance from human activities, a species-specific physiological validation of a method to evaluate adrenocortical activity is needed. Here we used 15 captive Arctic foxes (both males and females and juveniles and adults) to investigate fGCM concentrations following ACTH injection (physiological validation), or handling alone and compared them with their respective baseline concentrations prior to the treatments. A 5α-pregnane-3ß,11ß,21-triol-20-one enzyme immunoassay measured significant fGCM increases following both treatments. The time lags to reach peak fGCM values were 9.3 ± 1.3 h and 12.8 ± 1.7 h for ACTH and handling treatment, respectively. Concentrations of fGCMs varied a lot between individuals, but not attributed to sex nor age of the foxes. However, we found a negative relationship between boldness and fGCM concentrations. Faecal glucocorticoid metabolites concentrations did not change significantly over a period of 48 h in samples kept at temperatures reflecting winter and summer means. This would allow the collection of samples up to two days old in the wild regardless of the season. We conclude that our successfully validated method for measuring fGCMs can be used as a non-invasive tool for studies exploring various stressors both in wild and captive Arctic foxes.


Animals ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 75
Author(s):  
Gabina V. Eguizábal ◽  
Mariella Superina ◽  
Rupert Palme ◽  
Camila J. Asencio ◽  
Daniel P. Villarreal ◽  
...  

Management procedures affect behavioural and physiological stress responses of wild mammals under human care. According to the Reactive Scope Model, normal values are presumed to exist within predictive and reactive ranges. First, stress parameters of zoo-housed adult Tamandua tetradactyla were evaluated in winter and summer (29 days each), determining the level of behaviour and/or physiological parameters needed to respond to predictable environmental changes. Secondly, the effects of veterinary procedures and transportation were studied in both seasons. Non-invasive methods were applied, assessing behaviour through videos and adrenocortical activity by faecal glucocorticoid metabolites (FGMs). Lesser anteaters exhibited seasonality (summer > winter) in some behavioural parameters, such as nocturnal activities, as well as in the activity cycle (e.g., acrophase) and FGMs. A veterinary check elicited an increase in total activity (TA), natural behaviours and repetitive locomotion and affected the activity cycle, particularly in summer. Transport produced changes in TA, nocturnal and natural activity and some variables of the activity cycle, mostly during summer. Although the effects of routine management procedures were different from each other and presumably stressful, they elicited changes only at the behavioural level, which was greater during summer. The differences observed according to non-invasive methodologies highlight the importance of a multidisciplinary approach in this context and suggest that it is unlikely that individual welfare was affected.


Animals ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 2631
Author(s):  
Eeva A. Ojala ◽  
Mika Kurkilahti ◽  
Anne Lene Hovland ◽  
Rupert Palme ◽  
Jaakko Mononen

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.


2012 ◽  
Vol 62 (4) ◽  
pp. 419-432 ◽  
Author(s):  
Isabel Barja ◽  
Gema Escribano-Ávila ◽  
Carlos Lara-Romero ◽  
Emilio Virgós ◽  
Javier Benito ◽  
...  

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.


Author(s):  
S.K. Stead ◽  
D.G.A. Meltzer ◽  
R. Palme

Conventionally, the assessment of adrenal responses to stress relies on blood sample collection. However, blood collection from animals is impossible without restraint or immobilisation that influences results. This study was undertaken to validate recently established enzyme immunoassays that measure faecal glucocorticoid metabolites in elephants, and to perform a preliminary investigation into the biological relevance of this non-invasive method for use in assessing the degree of stress in this species. Four juvenile African elephants were injected i.m. with 2.15 mg synthetic adrenocorticotrophic hormone (Synacthén, Novartis, Switzerland). Blood and faecal samples were collected over 4 h and 7 d respectively. Concentrations of serum cortisol and faecal cortisol metabolites were determined using immunoassay. Variability of basal and peak values in blood and faeces was observed among the elephants. After ACTH injection, serum cortisol concentrations increased by 400-700 %. An 11-oxoaetiocholanolone enzyme immunoassay (EIA) proved best suited to measure cortisol metabolites (11,17-dioxoandrostanes) when compared to a cortisol and corticosterone EIA in faecal samples. Concentrations of faecal 11,17-dioxoandrostanes increased by 570-1070 %, reaching peak levels after 20.0-25.5 h. Greater levels of glucocorticoid metabolites were measured in faecal samples from elephants kept in small enclosures compared to levels in the faeces of animals ranging over a larger area. The results of this preliminary study suggest that non-invasive faecal monitoring of glucocorticoid metabolites is useful in investigating adrenal activity in African elephants.


2019 ◽  
Vol 59 ◽  
pp. e20195911
Author(s):  
Márcia Fontes Figueiredo Lima ◽  
Cristiano Schetini de Azevedo ◽  
Robert John Young ◽  
Priscila Viau

Distress can be defined as a biological response of an individual to long-term threats to its homeostasis and it should be avoided from an animal welfare perspective. High levels of stress hormones and the expression of abnormal behaviours are responses normally observed in distressed animals. Captive environments can provoke distress, especially when inappropriate stimuli are provided to the animals. The concomitant use of behavioural and non-invasive hormonal measures is a means to evaluate captive animal welfare. Environmental enrichment is a tool that can reduce distress and minimize the expression of abnormal behaviour in captive animals. The aim of this study was to evaluate greater rheas’ responses (behavioral and hormonal) to food-based enrichment. Three birds from the Belo Horizonte Zoo, Brazil were studied. The study was divided into three phases (baseline, enrichment and post-enrichment): fruits scattered around the birds’ enclosure were used as enrichment. Behaviour and faecal sampling were undertaken in all phases of the study. Abnormal behaviours and faecal glucocorticoid metabolites (GCM) levels showed significant reduction during the environmental enrichment phase, and a significative positive correlation between GCM production and abnormal pacing was observed. From the results of this study, we conclude that the use of food as environmental enrichment should be encouraged because of its positive effects on animal welfare. Besides, studies with larger groups of greater rheas, with individuals of both sexes, should also be encouraged to evaluate if the results found in this pilot study are consistent and can be generalized to the species.


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