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Cortisol in hair: a comparison between wild and feral cats in the north-eastern Alps

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Marcello Franchini at University of Udine
Marcello Franchini
Alberto Prandi
Alberto Prandi
Alberto Prandi
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Stefano Filacorda at University of Udine
Stefano Filacorda
Eva Nilanthi Pezzin
Eva Nilanthi Pezzin
Eva Nilanthi Pezzin
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Abstract and Figures

The quantification of glucocorticoid metabolites in hair is a non-invasive tool that provides important information regarding the endocrine status and represents a valuable method for studying potential stressors that may affect carnivores under both natural and non-natural conditions. Cortisol is the main glucocorticoid hormone of the hypothalamic-pituitary-adrenal gland axis and is considered a standard stress indicator for animal welfare. The current study aimed to compare cortisol levels extracted from hair of both dead, frozen European wildcats (Felis silvestris silvestris) and living feral individuals (Felis silvestris catus) living in different environmental conditions. The results obtained revealed that wild individuals exhibited a significantly (p < 0.001) higher cortisol concentration (n = 15, mean ± SD = 8.91 ± 4.48 pg/mg) than feral ones (n = 10, mean ± SD = 3.57 ± 1.25 pg/mg), probably as a result of both the physiological and/or environmental factors to which each subspecies was subject. This is the first study in which cortisol concentrations have been compared within the Felis silvestris subspecies, thus enriching the scarce information available for the Felidae. Nevertheless, further research is needed to better understand the various physiological and ecological factors affecting the adrenocortical activity of species or populations living in different environmental contexts.
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9 /Published online: 9 November 2019
European Journal of Wildlife Research (2019) 65: 90
ORIGINAL ARTICLE
Cortisol in hair: a comparison between wild and feral cats
in the north-eastern Alps
Marcello Franchini
1
&Alberto Prandi
1
&Stefano Filacorda
1
&Eva Nilanthi Pezzin
2
&Yannick Fanin
1
&Antonella Comin
1
#Springer-Verlag GmbH Germany, part of Springer Nature 2019
Abstract
The quantification of glucocorticoid metabolites in hair is a non-invasive tool that provides important information regarding the
endocrine status and represents a valuable method for studying potential stressors that may affect carnivores under both natural
and non-natural conditions. Cortisol is the main glucocorticoid hormone of the hypothalamic-pituitary-adrenal gland axis and is
considered a standard stress indicator for animal welfare. The current study aimed to compare cortisol levels extracted from hair
of both dead, frozen European wildcats (Felis silvestris silvestris) and living feral individuals (Felis silvestris catus)livingin
different environmental conditions. The results obtained revealed that wild individuals exhibited a significantly (p<0.001)
higher cortisol concentration (n= 15, mean ± SD = 8.91 ± 4.48 pg/mg) than feral ones (n= 10, mean ± SD = 3.57 ± 1.25 pg/mg),
probably as a result of both the physiological and/or environmental factors to which each subspecies was subject. This is the first
study in which cortisol concentrations have been compared within the Felis silvestris subspecies, thus enriching the scarce
information available for the Felidae. Nevertheless, further research is needed to better understand the various physiological
and ecological factors affecting the adrenocortical activity of species or populations living in different environmental contexts.
Keywords Wildcat .Feral cat .Felis silvestris .Hair cortisol .Adrenocortical activity
Introduction
The ability of an organism to adapt to changes in environmental
conditions has been receiving increased attention in recent years
(Koolhaas et al. 1999; Janczak et al. 2003;Natolietal.2005;
David et al. 2011;Ruiz-Gomezetal.2011;Montiglioetal.
2012; Dingemanse and Reale 2005). Abiotic and biotic changes
are common within the environment, and animals may respond
through temporal variation in their vital rate and/or an alteration
in their physiological response (Darlingtont et al. 1990). The
activity of the hypothalamic-pituitary-adrenal (HPA) gland axis
leads to the release of glucocorticoids (GCs), which are com-
monly used as indicators of physiological stress (Romero and
Wingfield 2001;WikelskiandCooke2006). When animals are
subjected to a stressor, the hypothalamus releases corticotropin-
releasing hormones (CRHs) which signal to the adrenal cortex
to release steroid hormones (such as GCs) to overcome stressful
situations (Sapolsky et al. 2000) and restore homeostasis (Möstl
and Palme 2002). This said, prolonged exposure to stressors
mayleadtochronicGCaccumulation leading to chronic stress
(Romero 2004) which is detrimental. Chronic stress negatively
affects individuals’fitness in terms of the depression of immune
responses, reduced reproductive success, and growth suppres-
sion (Romero 2004). The quantification of glucocorticoid me-
tabolites is a non-invasive tool that provides important informa-
tion regarding endocrine status, and is a valuable method to
study the potential stressors that may affect carnivores under
natural conditions (Barja et al. 2007;Sheriffetal.2011;Piñeiro
et al. 2012;Schelletal.2017). Cortisol is the main glucocorti-
coid hormone of the HPA gland axis and is considered the
standard stress indicator for animal welfare (Mormède et al.
2007). Indeed, despite the various autonomic and endocrine
responses that occur when an organism faces a stressful situa-
tion, cortisol has become commonly known as the stress hor-
mone. As part of the stress response, cortisol acts on various
metabolic pathways providing energy during a stressful situa-
tion. Cortisol’s role in the endocrine system is metabolic how-
ever, and it is also released in response to arousal situations such
as during sexual activity (Hamilton et al. 2008). Cortisol levels
can be measured using both invasive (e.g. blood) and non-
*Stefano Filacorda
stefano.filacorda@uniud.it
1
Dipartimento di Scienze Agroalimentari, Ambientali e Animali, Via
Sondrio 2/A, 33100 Udine, Italy
2
Dipartimento di Biologia, Via Ugo Bassi 58/B, 35131 Padua, Italy
https://doi.org/10.1007/s10344-019-1330-2
Received: 30 June 2018 /Revised: 7 August 2019 /Accepted: 15 October 2019
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
... In this context, when long-term steroid hormone changes have to be investigated, hair/coat and nails/claws represent the first choice of tissue materials. Many studies have in fact proved the reliability of hair/coat and nails/claws for the retrospective evaluation of long-term hormonal accumulation in both humans and animals [5][6][7][8][9], including felids [10][11][12][13][14][15], especially for cortisol and dehydroepiandrosterone (DHEA). Other than their reliability, the non-invasive collection of these matrices has the advantage of respecting animal welfare [9,16]. ...
... The same authors also reported a growth rate in the front nails of cats of 2.4 mm in 21 days, even if the portion of trimmed nails may contain the hormones accumulated in a prior time period (at least 1 month) [13]. Hair was also used for cortisol measurement in cats in other studies [5,15], and in feral and wild felids [11]. Very recently, [14] investigated the usefulness of seasonal coat collection in wild felids and domestic cats for the measurement of not only cortisol, but also testosterone concentrations. ...
... To the authors' knowledge, this study reports, besides the confirmed usefulness of coat and claws, the first use of dewclaws as a possible matrix for the measurement of E2 and T in postpubertal domestic cats during the breeding season. The results, in fact, demonstrated once more the usefulness of coat for studies monitoring function for an extended period, as previously suggested in humans and animals, and specifically in felids [5,11,14,15], even if more studies are needed to discover the exact timing and percentage of hormone accumulation from peripheral blood. Besides coat, the present study also confirmed that claws could be suitable for measuring sexual steroids in postpubertal cats, supporting the first reports about the usefulness of claws for non-invasive and objective hormonal assessment in cats [12,13]. ...
Coat, Claw and Dewclaw 17-β-Estradiol and Testosterone Concentrations in Male and Female Postpubertal Cats: Preliminary Results
Article
Full-text available
  • Feb 2023
In the recent past, tissue materials such as hair/coat and nails/claws have proved to be useful for the study of long-term hormonal changes in humans and animals and shown to be advantageous in terms of being collectable without invasiveness, with a benefit in terms of animal welfare. However, studies using these tissue materials in cats are scarce, especially on sexual hormone measurement. In this study, the concentrations of 17-β-estradiol (E2) and testosterone (T) were assessed in 20 male and 18 female domestic postpubertal cats at the time of neutering/spaying during the breeding season. Hormones were measured in coat shaved from the forearm (ACOAT) and from the surgical area (SCOAT); claws were collected from the forearms (CLAWS) and the dewclaws (DCLAWS). Although all these tissue materials were shown to be useful for E2 and T long-term measurement, only T concentrations were higher (p < 0.001) in males from both ACOAT and SCOAT samples when compared to females and therefore useful for distinguishing between the two sexes. Within each sex, E2 and T concentrations can be assessed on coat, but also on the dewclaws, providing an alternative, practical, matrix for sexual steroid measurement in postpubertal cats during the breeding season.
View
... The measurement of hair hormone levels has also been used for carnivores [10], to estimate interspecific [21] and intraspecific [22] differences in felines, and sometimes with large datasets such as those in bears [7,23,24], indicating that the determined level of the hormone (cortisol) characterizes the physiological state of animals quite well. The changes in hair glucocorticoids (both cortisol and corticosterone [25]) may be attributed to the effect of age and pregnancy, body region, sex and season of year, but these results are not very consistent [11,13]. ...
... In temperate climates and in outdoor enclosures, domestic cats showed a fairly clear breeding season (spring-summer), as was previously revealed in our colony [33] and in other published studies from the northern hemisphere [34,35]. Sampling in mid-January reflected hormone levels at the beginning of the year (assuming a two-week time lag Animals 2022, 12, 2792 9 of 14 between increases in blood and hair hormones [21]), when, apparently, the activity of males' reproductive system was at the low level based on hormonal data and sperm analysis [33]. ...
Application of Felid Hair for Non-Invasive Tracking of Animal Reproductive Status and Adrenal Activity
Article
Full-text available
  • Oct 2022
Hair can be a useful matrix to examine the hormonal status of an animal, although it is difficult to correlate the results to a specific time point. The aim of this study was to evaluate seasonal changes in cortisol and testosterone levels in the hair of four feline species (lynx, Lynx lynx, n = 8; Amur wildcat, Prionailurus bengalensis euptilurus, n = 8; caracal, Caracal caracal, n = 6 and domestic cat, Felis catus, n = 17) with different breeding strategies. Animals of both sexes were sampled over the year, once per season (every three months), and the concentrations of hair testosterone and cortisol were measured by EIA. Both hormones showed annual dynamic changes, which coincided with the reproductive seasonality of the studied species. Sexual differences in testosterone level were found only during the mating season (spring for lynx and Amur wildcats, spring–summer for domestic cats), when testosterone levels were higher in males than in females. Cortisol levels were higher in males than in females in domestic cats and Amur wildcat, but also only during the mating season. Seasonal increases in testosterone were observed in three seasonal breeders (lynx, Amur wildcat and domestic cat) but not in caracal, which had high testosterone levels over the whole year. In lynx and Amur wildcat, it decreased sharply in the summer. Cortisol levels increased during the mating period in domestic cat males and lynx of both sexes; in caracal, an increase in cortisol was related to the transfer of animals to smaller winter cages. Measurements of steroids in hair can provide a reliable method for evaluating the reproductive status and the activity of the hypothalamus–pituitary–adrenal axis in several felid species.
View
... Hair cortisol concentration is a minimally invasive, retrospective measure of cumulative HPA activity over the preceding 3 months and is increasingly viewed as an emergent biomarker of HPA activation and response to stressors [70][71][72]. Hair cortisol measurements have been reported across wild felid species [73,74] and are considered valid and reliable measures in domestic cats [75,76]. ...
Understanding the Influence of Early-Life Stressors on Social Interaction, Telomere Length, and Hair Cortisol Concentration in Homeless Kittens
Article
Full-text available
  • Feb 2025
The early postnatal period is a critical neurodevelopmental stage characterized by rapid neural maturation and is adversely affected by early-life stressors. This study explored the behavioural, physiological, and epigenetic consequences of early-life stress in a population of homeless rescue kittens. This longitudinal study included 50 kittens rescued and placed into foster care by the Prince Edward Island Humane Society. They underwent behavioural testing at 8, 10, and 12 weeks of age. Hair cortisol concentration was measured at 8 weeks and served as a physiological marker of the previous 3 months’ cumulative stress response, which, for these kittens, included the late gestation period. A blood sample for relative telomere length measurement was taken at 10–12 weeks to estimate epigenetic changes as young kittens. Data were analyzed with respect to age and performance in all repeated measures tests, status as a stray or a surrender, and the presence of the dam in their foster homes. As expected, the performance of kittens in all tests changed over the 5 weeks of testing. Kittens separated from their mothers exhibited significantly higher hair cortisol concentrations (p = 0.02) and elongated relative telomere lengths (p = 0.04). No correlation was found between hair cortisol concentration and relative telomere lengths (p = 0.99). These results support the need for further study on the effects of epigenetics and early-life stress, both in kittens and across species.
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... The majority of intact females (15/16) without owner-reported estrus or pregnancy were at least one year old at the time of hair sampling, suggesting potential estrus experiences not reported by the owner 64 . In a study of wildcats and feral cats, higher HCC was observed in male cats compared to females, although this difference was not statistically significant, possibly due to a small sample size 65 . In our study, we found an association between male sex and HCC. ...
Hair cortisol concentrations in clipped and combed hair and associations with characteristics, health status and stress in domestic cats
Article
Full-text available
  • Sep 2024
Hair cortisol concentrations (HCC) are measured to assess long-term HPA-axis activity and may represent a valuable non-invasive tool to evaluate chronic stress in cats. This study investigated combing as a novel, low-stress method for HCC assessment, as well as possible associations between HCC and cat characteristics in 167 owned cats. Hair was sampled at veterinary clinics through clipping and/or combing the cat, or at home by the owner combing the cat. A questionnaire was sent to cat owners, including inquiries about the cat’s sex, health status, and exposure to stress. HCC was quantified using a commercial cortisol assay kit. Despite variations within and between sampling methods, Spearman’s correlation and Bland–Altman plots revealed a moderate correlation between clipped and combed samples (rs = 0.61, LOA -5.51 ± 22.54). In multiple linear regression, variations in HCC were observed based on sex, health status and cat group size. No associations were found between HCC and stress as assessed by owners. Despite study limitations and remaining uncertainty regarding factors influencing HCC, combing presents a convenient approach for evaluating long-term HPA-axis activity in clinical settings. The association between health and HCC suggests alterations in cortisol levels that are related to disease processes and stress-inducing events associated with the disease.
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... Cortisol concentrations are mainly measured in blood (Gardiner & Hall, 1997) and faeces (Fattorini et al., 2018). However, compared to other matrices, measuring cortisol cumulated in hair (Filacorda et al., 2021;Franchini et al., 2019;Prandi et al., 2018) represents a valuable method to assess chronic stress in wild species, since it is incorporated into the hair shaft during the anagen phase (i.e. active growth; Montillo et al., 2014) and over an extended time period (Meyer & Novak, 2012). ...
You're stressing me out! Effect of interspecific competition from red deer on roe deer physiological stress response
Article
Full-text available
Cortisol is the main glucocorticoid hormone commonly measured to assess adrenocortical activity in vertebrates in response to potential stressors. Competition and habitat quality may act as chronic stressors leading to an increase in cortisol accumulation. Moreover, chronic cortisol concentrations may seriously hamper the reproductive potential, health status and body condition of a species. Using hair collected from the rump of 88 adult roadkill roe deer Capreolus capreolus in the north‐eastern Italy, we aimed to assess the effect of both ungulate competitor population densities and habitat suitability on the long‐term physiological stress response of roe deer, using hair cortisol as a reference parameter. Our findings revealed that roe deer showed higher cortisol concentrations in response to higher red deer Cervus elaphus population density, most likely as a consequence of direct interspecific competition. No significant cortisol changes were however recorded in response to both wild boar Sus scrofa and roe deer population densities, probably as a result of reduced competition because of abundant feeding resources, different species population densities and spatial/temporal avoidance among species. Moreover, the absence of a significant effect as far as habitat suitability is concerned, suggest that roe deer are well adapted to live even in poor quality habitats most likely as long as they are close to better quality habitats which provide access to food resources and refuge sites. Our results highlight the importance of hair cortisol as a suitable parameter to assess physiological chronic stress response towards different environmental/ecological conditions.
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... Studies on the HCL of dogs were conducted, among others, by Accorsi et al. [24] and Direksin et al. [25]. The level of HCL of cats has been studied by Accorsi et al. [24] and Franchini et al. [26]. As for other animal species, cortisol has been studied in, among others, sheep wool [27], pig hair [28], dairy cows [29], horses [30], and captive wild animals [31,32]. ...
Are Hair Cortisol Levels of Humans, Cats, and Dogs from the Same Household Correlated?
Article
Full-text available
  • Jun 2022
Human–animal interactions and the emotional relationship of the owner with the pet are the subjects of many scientific studies and the constant interest of not only scientists but also pet owners. The aim of this study was to determine and compare the hair cortisol levels of dogs, cats, and their owners living in the same household. The owners were asked to complete a questionnaire concerning the frequency of their interactions with pets and emotional relationship with each of their cats and each of their dogs. The study involved 25 women who owned at least one dog and at least one cat. In total, 45 dogs and 55 cats from 25 households participated in the study. The average level of hair cortisol of the owners was 4.62 ng/mL, of the dogs 0.26 ng/mL, and in the hair of cats 0.45 ng/mL. There was no significant correlation between the hair cortisol level of the owner and dog or the owner and the cat and between dogs and cats living together. A significant positive correlation was observed between the hair cortisol level in the owner and the pet, for dogs in which the owner performs grooming treatments once a week and for cats which are never kissed. Although our study did not find many significant correlations, studies using other stress markers might have yielded different results.
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... Extensive descriptions of the study areas located in Sicily (Etna Regional Park), Apulia (Gargano National Park), and Friuli (points) and of the road-killed wildcats (triangles) analysed in the current study. European wildcat distribution range across the Italian peninsula is shown in light grey Venezia Giulia are provided elsewhere (Anile et al. 2019;Franchini et al. 2019;Filacorda et al. 2021;Gaudiano et al. 2021). In Basilicata region, a variety of habitats (e.g. ...
Show me your tail, if you have one! Is inbreeding depression occurring in wildcats (Felis silvestris silvestris) from Italy?
Article
  • Mar 2022
Knowledge of genetic diversity is important to wildlife conservation because genetically depleted populations experience an increased risk of extinction. Mammalian carnivores are characterized by small and fragmented populations and low dispersal, so that genetic erosion can lead to the fixation of deleterious genes relatively quickly, leading to morphological abnormalities. Kinked tails and cowlicks are indicative of inbreeding depression and have been described in two wild cat species so far, the puma (Puma concolor) and the cheetah (Acinonyx jubatus). Here we report the first records of morphological abnormalities in five populations of the European wildcat (Felis silvestris silvestris) in Italy by using (1) camera-trapping and (2) necropsy of road-killed individuals assessed through genetic analysis. We collected 24,055 trap-nights from 251 cameras and recorded 566 wildcat detections, from which we identified 148 wildcats. Among these, 11 individuals had a kinked tail and four displayed brachyuria, whereas three wildcats from Sicily had cowlicks on the thorax. We recovered 28 road-killed wildcats and two of them (from Sicily and Friuli Venezia Giulia) had a kinked tail. Among these, one female with a kinked tail had a male foetus with a kinked tail, which proved that this characteristic was genetically inherited. We are unsure why brachyuria or cowlicks were not detected across all monitored wildcat populations, given we found kinked tails throughout Italy. The frequencies at which we have detected these abnormalities in wildcats are far lower than reports from Florida panthers (Puma concolor). Future research is needed to verify whether these abnormalities are also associated with low genetic diversity or other morphological defects which might lower fitness. We recommend a nationwide effort, using these techniques within a standardized sampling design, to further understand the status of the wildcat in Italy.
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Potential of Nail Cortisol for Welfare Assessment in Shelter and Owned Cats
Article
  • Oct 2024
  • APPL ANIM BEHAV SCI
Shelter cats often experience considerable stressors, such as confinement and unfamiliar surroundings, leading to welfare challenges. Evaluating and enhancing their well-being is crucial, and measuring cortisol levels in cats’ nails offers a minimally invasive method for assessing stress and welfare. This study compared nail cortisol concentrations (NCCs) between owned cats (OC), and shelter cats in favorable conditions (FS) and unfavorable conditions (UFS) in Thailand and Vietnam. Nails from 31 OC, 29 FS, and 27 UFS cats were collected twice, approximately 28 days apart, with details of living conditions recorded at the second collection. Nail clippings were processed for NCC measurement using the enzyme-linked immunosorbent assay (ELISA) technique. NCC levels of UFS cats were significantly higher than those of FS and OC cats in both collections, with no gender-based differences. NCCs positively correlated with dog exposure, and negatively correlated with space allowance, number of litterboxes, free-roaming time, air conditioning, and the presence and variety of enrichment (hiding places, elevated areas, outdoor views, scratching structures, and toys). These findings indicate that improved living conditions positively impact welfare, as indicated by NCCs. Nail cortisol can serve as a minimally invasive method for assessing cat welfare, particularly considering long-term factors, and offers valuable insights into the well-being of cats in various conditions.
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Stress in wildlife: comparison of the stress response among domestic, captive, and free-ranging animals
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  • Apr 2023
The stress response, which involves joint activity of the nervous and endocrine systems, is one of the basic adaptive mechanisms that ensures the survival of the individual. The activation of the sympathetic nervous system, the sympathetic-adrenal-medullary axis, and the hypothalamic–pituitary–adrenal axis enables organisms to respond to endogenous and exogenous challenges. Repeated short-term stress leads to long-term stress, which disrupts physiological homeostasis. Unlike domestic animals, wild animals are not protected from environmental and weather influences or treated for diseases. In addition, climate change, habitat fragmentation and loss, and urban stressors (such as light, noise and chemical pollution; xenobiotics; traffic; and buildings) affect individual wildlife and populations. In this review, we have attempted to depict the magnitude of the stress response in wildlife and related domestic animals as well as in captive and free-ranging animals. The intensity of the stress response can be estimated by determining the concentration of glucocorticoids in body fluids, tissues, and excreta. A comparison of results from different studies suggests that domestic animals have lower fecal and hair glucocorticoid concentrations than related wild animals. Additionally, fecal and hair glucocorticoid concentrations in captive animals are higher than in free-ranging animals of the same species. As there are limited data on this topic, we cannot draw definitive conclusions about glucocorticoid concentration and stress response. Further studies are needed to clarify these issues.
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Correlations between behavior and hormone concentrations or gut microbiome imply that domestic cats (Felis silvestris catus) living in a group are not like ‘groupmates’
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Domestic cats (Felis silvestris catus) can live in high densities, although most feline species are solitary and exclusively territorial animals; it is possible that certain behavioral strategies enable this phenomenon. These behaviors are regulated by hormones and the gut microbiome, which, in turn, is influenced by domestication. Therefore, we investigated the relationships between the sociality, hormone concentrations, and gut microbiome of domestic cats by conducting three sets of experiments for each group of five cats and analyzing their behavior, hormone concentrations (cortisol, oxytocin, and testosterone), and their gut microbiomes. We observed that individuals with high cortisol and testosterone concentrations established less contact with others, and individuals with high oxytocin concentrations did not exhibit affiliative behaviors as much as expected. Additionally, the higher the frequency of contact among the individuals, the greater the similarity in gut microbiome; gut microbial composition was also related to behavioral patterns and cortisol secretion. Notably, individuals with low cortisol and testosterone concentrations were highly tolerant, making high-density living easy. Oxytocin usually functions in an affiliative manner within groups, but our results suggest that even if typically solitary and territorial animals live in high densities, their oxytocin functions are opposite to those of typically group-living animals.
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Comparison of tigers' fecal glucocorticoids level in two extreme habitats
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Application of different antibodies and extraction methods results in a wide range of steroid metabolite concentrations obtained during noninvasive hormones monitoring. It makes regional comparisons of steroid concentration very difficult. We compared three methods for extraction of glucocorticoids metabolites in tiger feces to examine correct stress level in Bengal and Amur tigers in India and Russia respectively. The results obtained with three different extraction methods correlate with each other positively and significantly. The highest concentration of fecal glucocorticoids metabolites (FGCM) was found after the extraction of wet feces samples with 90% methanol. The level of FGCM was significantly higher in Bengal tigers in India than in Amur tigers in Russian Far East. The reasons might be related to tigers’ density or anthropogenic pressure.
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A first survey on hair cortisol of an alpine ibex (Capra ibex ibex) population
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  • Jan 2018
  • ANIM SCI PAP REP
Biometric measurements of horn, body and physiological variables of 35 Alpine ibex, Capra ibex ibex, (14 females and 21 males, age range: 2-15 years) were analysed in relation to gender and area of origin. Moreover, for the first time hair cortisol concentration was evaluated in this species. The ibexes were live-captured in spring in the Maritime Alps Natural Park. All the horn and body morphometric measurements were significantly different between genders and a statistical difference was found between horn and body data considering the area of origin of ibexes, with the exception of horn circumference and height as well of body and hock length. Among the physiological variables, female ibex showed higher heart rate and glycemia, but tended to have lower respiratory rate than male. The mean hair cortisol concentration was 22.40±1.44 pg/mg (±SE). A different distribution was described for the male hair cortisol concentrations compared to that in females. Obtained data indicate that the geographical area where Alpine ibex lives could affect the endurance of an ibex population but further investigations are needed to understand if intra- and inter-specific competition and different environment could be the basis of the HPA axis activation. Hair cortisol evaluation together with biometric values could be an interesting way to obtain information on the adaptation that the animal attempts to reach in relation to environmental factors and pursuing organism homeostasis. This feature could be important in attempting to preserve wildlife biodiversity that also depends from the state of animal welfare.
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Hair cortisol and testosterone concentrations and semen production of Bos taurus bulls
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  • Mar 2017
The experiment was aimed to evaluate the usefulness of hair cortisol concentrations (HCC) in revealing the activation of the hypothalamic-pituitary-adrenal (HPA) axis due to the passage of young bulls from genetic rearing stations to artificial insemination (AI) centre and the influence of this passage on hair testosterone concentrations (HTC). Hair samples on 33 yearling bulls were collected at the time of arrival (ST1) to the AI centre and 3 more hair samples (ST2, ST3, ST4) were collected at monthly intervals. After the end of quarantine each animal was submitted to the standard procedures for collection and production of semen. Overall HCC were significantly affected by period of collection (p<.01). In particular, HCC was higher in samples obtained after finishing the quarantine period (ST2) compared with those obtained on ST1. After ST2, the HCC decreased reaching at ST4 a value similar to that recorded at ST1. An effect of sampling time on HTC was not found (p>.05). The effect of breed was observed only for HTC that was higher in PRI than BS (p<.01). The HCC group have not influenced the semen variables (p>.05). On the contrary, the effect of breed was evident. The results of this study replicated the effects of a major environmental stressor on mean cortisol concentrations assessed in hair, and add to the growing body of evidence that HCC is an effective and simply collected marker for long-term activity of the HPA system in response to persistent environmental challenge.
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Investigation of techniques to measure cortisol and testosterone concentrations in coyote hair
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  • Mar 2017
  • ZOO BIOL
Long-term noninvasive sampling for endangered or elusive species is particularly difficult due to the challenge of collecting fecal samples before hormone metabolite desiccation, as well as the difficulty in collecting a large enough sample size from all individuals. Hair samples may provide an environmentally stable alternative that provides a long-term assessment of stress and reproductive hormone profiles for captive, zoo, and wild mammals. Here, we extracted and analyzed both cortisol and testosterone in coyote (Canis latrans) hair for the first time. We collected samples from 5-week old coyote pups (six female, six male) housed at the USDA-NWRC Predator Research Facility in Millville, UT. Each individual pup was shaved in six different locations to assess variation in concentrations by body region. We found that pup hair cortisol (F5,57.1 = 0.47, p = 0.80) and testosterone concentrations (F5,60 = 1.03, p = 0.41) did not differ as a function of body region. Male pups generally had higher cortisol concentrations than females (males = 17.71 ± 0.85 ng/g, females = 15.48 ± 0.24 ng/g; F1,57.0 = 5.06, p = 0.028). Comparatively, we did not find any differences between male and female testosterone concentrations (males = 2.86 ± 0.17 ng/g, females = 3.12 ± 0.21 ng/g; F1,60 = 1.42, p = 0.24). These techniques represent an attractive method in describing long-term stress and reproductive profiles of captive, zoo-housed, and wild mammal populations.
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Home-range size of the European wildcat (Felis silvestris silvestris): A report from two areas in Central Italy
Article
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  • Jan 2017
Although the behavioural ecology of the European wildcat (Felis silvestris silvestris) has been studied in several European countries, its home-range size is still poorly known due to elusive behaviour of the species living in typically low population densities. In our study, 11 wildcats from two distinct areas, the Maremma Regional Park (Tuscany; 3 males and 1 female) and the Paradiso di Pianciano Estate (Umbria; 6 males and 1 female), both located in Central Italy, were studied by means of classical VHF radio-tracking. Home ranges were calculated by means of the Brownian bridge movement model (BBMM). Variables significantly affecting home-range size were: study area, gender and the interaction between gender*age. The potential effect of the reproductive season and the nocturnal period was not supported. The main findings indicate that: (1) home-range sizes in Tuscany were larger than those in Umbria; (2) home-range size was positively related to the age of individuals; (3) males exploited larger home ranges than females in all age-classes except for the sub-adult age-class; this latter pattern has never emerged from any previous wildcat radio-telemetry study. Population density, different management regimes in the areas considered and the local abundance of prey may explain differences in home-range sizes between the study areas. The estimated home ranges of wildcats in Umbria were slightly larger than those reported across Europe, while those calculated in the Tuscan study area were significantly greater.
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Use of hair cortisol analysis for comparing population status in wild red deer (Cervus elaphus) living in areas with different characteristics
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  • Dec 2016
  • EUR J WILDLIFE RES
We tested a method to measure Hair Cortisol Concentration (HCC) in 174 red deer (Cervus elaphus) culled in the hunting season 2011/12 in four areas of Central Italian Alps, with different population densities (SPN, 11.8 deer/km2; HD-AV, 3.6 deer/km2; HD-SO, 2.1 deer/km2; HD-MO, 2.0 deer/km2) and environmental conditions. Our hypothesis was that higher population densities, associated with more difficult environmental conditions, may result in higher allostatic load for these wild ungulates. No significant differences in HCC were detected between sexes (males, 4.77 ± 0.69 pg/mg; females, 5.75 ± 0.63 pg/mg) nor among age classes (calves, 6.17 ± 0.66 pg/mg; yearlings, 4.47 ± 0.83 pg/mg; adults, 5.15 ± 0.74 pg/mg; least square mean ± SE), but HCC difference between calves and yearlings was close to statistical significance (P = 0.059). HCC showed high individual variation, but on average it was higher in areas with higher deer density (SNP, 7.45 ± 1.01 pg/mg; HD-AV, 6.07 ± 0.89 pg/mg; HD-SO, 4.67 ± 1.14 pg/mg; HD-MO, 2.87 ± 1.56 pg/mg), with significant differences between HD-AV and HD-MO (P = 0.01). Carcass weight was significantly lower in SNP (46.74 ± 1.49 kg) than in HD-MO (62.71 ± 4.01 kg), HD-SO (61.73 ± 2.9 kg) and HD-AV (62.07 ± 2.04 kg) (P < 0.001). These results seem to confirm our hypothesis that allostatic load is higher in areas with higher density and harder environmental conditions. We suggest that the methodology used in this study to measure HCC provides good information on long-term HPA axis activity and allostatic load and constitutes a highly promising, reliable and non-invasive method in wildlife management for assessing HPA axis activity over extended time periods.
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Hair cortisol concentrations in New Zealand white rabbits subjected to surgery
Article
  • Feb 2018
The aim of this study was to assess hair cortisol concentrations in New Zealand white rabbits (Oryctolagus cuniculus) that were subjected to relocation and surgery to evaluate HPA-axis activity; in addition, we used this marker of cortisol secretion to evaluate the allostatic load of animals undergoing surgery. After a period of acclimatisation, which lasted 40 days from their arrival at the enclosure, 19 rabbits were subjected to T1-T12 dorsal arthrodesis (RS), 19 were sham-operated (SS), and 19 were non-operated (CON). Hair samples were collected at the time of arrival (ST1) at the animal facility, and seven other sets of hair samples were collected at 40- day intervals from the same area of skin for a period of 240 days as re-shaved hair (anagen phase): immediately before surgery (ST2) and after the surgery (ST3, ST4, ST5, ST6, ST7, and ST8). The transition from the rabbitry to the animal breeding facility led to a significant increase in cortisol concentration (ST2) in all of the groups. At ST3, the RS group presented higher cortisol concentrations than those of the SS group and the CON group. At ST4, the experimental groups showed similar values that remained constant until ST8. The results show that the management of rabbits undergoing surgery should be evaluated very carefully, and hair cortisol concentrations may provide a means of avoiding the dangerous cumulative effects of additional stressors close to surgery.
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Cortisol and DHEA concentrations in the hair of dairy cows managed indoor or on pasture
Article
  • May 2017
  • LIVEST SCI
This study was conducted in order to assess the changes of cortisol and dehydroepiandrosterone (DHEA) concentrations in hair of Italian Simmental dairy cows managed indoor in tie-stall barns or grazing on high mountain pasture. In experiment I, animals were kept in tie-stalls throughout the year (n = 27). In experiment II, animals remained on mountain pastures during summer (from 20th June to 20th September) and then kept indoors in tie-stalls for the rest of the year (n = 18). At the end of July, following vertical transhumance, dairy cows were moved to a pasture located at a higher altitude. In the two studies, hair samples, obtained using clippers from the animal's forehead, were collected on the 20th June (P1), 20th July (P2), 20th August (P3), 20th September (P4) and 20th October (P5) and analysed for cortisol and DHEA. In experiment I, DHEA concentration was unaffected by sampling time (P > 0.05), whereas, cortisol/DHEA ratio was similar between P1 and P2 (P > 0.05) and then increased during the experimental period reaching the highest value at P5 (P < 0.05). In experiment II, dairy cows had higher concentration of DHEA during grazing compared to cows kept indoors (P < 0.05), conversely cortisol/DHEA ratio decreased from P1 to P2 (P < 0.05), remained constant from P2 to P3 (P > 0.05) and then increased from P3 to P4 (P < 0.05), the highest value was recorded at P5. In conclusion, cortisol and DHEA concentrations and their ratio measured in the hair of dairy cows can be useful to evaluate the allostatic load in animals managed indoor in tie-stall barn or grazing on mountain pasture.
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Relocation and Hair Cortisol Concentrations in New Zealand White Rabbits
Article
  • Jan 2017
To investigate how long relocation modified hair cortisol concentrations in New Zealand white rabbits, 19 rabbits were subjected to a change in their breeding facility at the beginning of the trial and then were kept under stable environmental conditions. Hair samples were collected at the time of arrival to the nonhuman animal facility and at 40-day intervals from the same skin area for up to 440 days after the animals' arrival to the facility. A period effect on the hair cortisol concentration was found (p < .01). The transfer of the rabbits to the new facility might have induced an increase in the hypothalamic-pituitary-adrenal axis activity (p < .01). A second increase in hair cortisol concentration (p < .01) occurred at 320 days, after a change of personnel at the facility that occurred at 280 days, which was the only environmental change. The relocation of rabbits to the facility resulted in a stress response leading to elevated cortisol levels. The effect of relocation on mean cortisol concentrations was exhausted within 120 days when all environmental factors were kept stable.
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