Faecal Cortisol Metabolites Levels Research Articles

The impact of net cleaning on fecal cortisol metabolites in farmed Atlantic Salmon

Stress assessment is crucial in aquaculture to ensure and improve the health and well-being of production animals including fish. The primary objective of this study was to utilize the assessment of fecal cortisol metabolites (FCMs) as a low-invasive stress determination method for farmed Atlantic salmon (Salmo salar L.) and to suggest an appropriate sampling timeframe for this measurement.Blood, mid-intestinal contents, and feces samples were collected before, during, and at 2 and 20 h after a stress-inducing event, namely, net cleaning. Plasma cortisol (PC) and its metabolite levels in the mid-intestinal contents (MICMs) and FCMs were assessed by an enzyme-linked immunosorbent assay (ELISA). The research reveals a distinctive temporal profile, with a sequential elevation in levels of PC, MICMs and FCMs in response to the effects of net cleaning. Peak FCMs levels were observed approximately 20 h after net cleaning and were significantly higher than peak PC levels (P < 0.05). The findings suggest that FCMs are a promising indicator for acute stress assessment. To enhance the understanding and improve the applicability of FCMs as stress indicators in fish farming practices the critical challenges regarding feces sampling and FCMs analyses are further discussed. Future research aimed at developing practical protocols and industry standards will emphasize method validation and include a greater number of sampling points as well as an expanded experimental period.This study signifies a crucial step toward understanding the role of FCMs as a reliable stress indicator in fish. It underscores the importance of accounting for the delayed and extended sampling patterns when assessing stress in aquatic settings. Moreover, it offers a less invasive and more labor-efficient approach for monitoring the overall health and welfare of salmonids than PC analysis, thereby promoting sustainability in this vital industry.

Season-dependent impact of forage quality on stress in alpine chamois.

Chronically heightened stress levels in wildlife species may have detrimental effects on individual life history traits, for example, through the increased likelihood of disease, parasitic infections, and overall reduced fitness. Understanding the drivers of stress may thus have great potential for informing wildlife conservation. Although the role of climate and individual status is well studied in stress ecology, the impact of related stressors such as dietary quality is of increasing interest to wildlife research and conservation. In this study, fecal cortisol metabolites (FCMs) in Alpine chamois Rupicapra r. rupicapra used as bioindicators of stress, and their relationship with forage quality-measured as the percentage of fecal crude protein (CP)-were investigated. Data collection took place in 2011 and 2012 in the Gran Paradiso National Park (Western Italian Alps), on 22 individually marked adult males. The relationship between FCMs and CPs was analyzed through linear models and separated between winter and summer months, accounting for the effect of potentially confounding exogenous and endogenous variables. After AICc-based model selection, we found that forage quality was negatively related to FCM levels in Alpine chamois during the summer months, meaning that higher quality forage was associated with the decreased expression of stress hormones. However, during the winter months, we did not find a significant relationship, potentially as a result of forage quality being ubiquitously poor. Although the mechanisms through which dietary variations impact FCM concentrations in wildlife populations are largely unknown, the occurrence of significant relationships between forage quality and stress levels supports potentially important implications for the long-term effect of climatic changes on the fitness of wildlife populations.

Discussion paper on ‘Comparing the agreement of a commercial cortisol kit with a biologically validated assay in evaluating faecal cortisol metabolite levels in koala joeys’

Discussion paper on ‘Comparing the agreement of a commercial cortisol kit with a biologically validated assay in evaluating faecal cortisol metabolite levels in koala joeys’

Discussion paper on “Comparing the agreement of a commercial cortisol kit with a biologically validated assay in evaluating faecal cortisol metabolite levels in koala joeys”

Discussion paper on “Comparing the agreement of a commercial cortisol kit with a biologically validated assay in evaluating faecal cortisol metabolite levels in koala joeys”

Behavioral and hormonal assessment of stress in foals (Equus caballus) throughout the weaning process.

This study had the aim to demonstrate the midterm effects (three weeks) of weaning on foals' welfare. For this purpose, foals' behavioral changes and fecal levels metabolites of cortisol were evaluated. The observations took place at the state stud farm of Baden-Wuerttemberg in Germany. Ten foals (six colts and four fillies) were observed from one day before weaning up until three weeks after weaning. Weaning was divided into three blocks, the first in September, the second in October, and the last in November. The behavioral observation was done during an eight-hour period between 7:00 a.m. and 5:00 p.m. The observer documented the exact behavior shown by the foal every five minutes during the eight hours. To scale the stress experienced by the foal, the glucocorticoid metabolite 11,17-dioxoandrostane was measured with the 11-oxoetiocholanolone enzyme immunoassay, which allows assessing the foal's plasma cortisol level changes throughout the trail through fecal samples. All foals displayed a distinct hormonal stress response to the weaning process through increased fecal cortisol metabolite levels. Their body posture distribution took a shift from mainly moving before weaning to mainly standing during the three weeks after weaning. Compared with the day before weaning, the foals showed less active behavior and significantly increased their resting behavior. Regarding the overall resting behavior, the weaned foals initially increased their time spent resting in a lying position during daytime and then started to decrease the time lying. After weaning, the foals showed a significant increase in resting while standing. In conclusion, the foals showed an expected behavioral development and an expected curve of cortisol metabolite values throughout the study. However, it seemed that the changes had not returned "back to normal" at three weeks after weaning. Therefore, we suggest that weaned foals need a minimum of three weeks to acclimate to the new situation.

Seasonal variation in molecular and physiological stress markers in Asian elephants.

Free-living species exhibit seasonal variation in various life history traits, including vital rates such as birth and death patterns. Different physiological mechanisms are thought to underlie the expression of life history traits that contribute to lifetime fitness. However, although the broad impacts of seasonality on life history traits and trade-offs is well established in many systems, the exact physiological mechanisms responsible for driving differences within and between individuals are poorly understood. Among them, molecular and physiological stress pathways, such as stress hypothalamic-pituitary-adrenal axis and oxidative stress, have potential to mediate relationships between individual survival, reproduction and environmental seasonality. Here, we determine how different physiological markers of stress including faecal cortisol metabolites (FCMs), heterophils/lymphocytes (H/L) ratio, two markers indicating oxidative balance including a marker of oxidative damage (reactive oxygen metabolites, ROM) and a marker of antioxidant defences (superoxide dismutase, SOD) and body weight vary in a large semi-captive population of Asian elephants (Elephas maximus) exposed to extreme seasonality (e.g. elevated temperatures). Individuals showed higher FCM levels and H/L ratios during cold season, indicating increased stress, and the lowest FCM levels during monsoon season and H/L ratios during hot and dry season, but we found no pattern in oxidative stress (ROM and SOD) levels. Hot season also associated with a decline in body weight. The present study shows how different physiological parameters (FCM levels and H/L ratio), molecular (oxidative stress) and body condition vary with seasonal changes, and how these parameters might allow individuals to adapt to such variations. Our results on an endangered long-lived species are crucial in indicating the most productive timing for conservation efforts, predicting how individuals cope with environmental changes, and allow for a more accurate representation of how animal physiology operates in nature.

Comparing the agreement of a commercial cortisol kit with a biologically validated assay in evaluating faecal cortisol metabolite levels in koala joeys

Orphaned koala joeys constitute a substantial number of wildlife rescues. Mortality is highly prevalent in rehabilitating joeys, with little knowledge about the causes of mortality. The hypothalamic–pituitary–adrenal axis plays a vital role in mediating stress by producing glucocorticoids (e.g. cortisol), however, no studies have quantified glucocorticoids in koala joeys. Traditional cortisol enzyme immuno-assay (e.g. R4866) are limited in supply and are process intensive, whereas, modern enzyme immuno-assay (EIA) kits (e.g. Arbor Assay cortisol kit) are available world-wide and provide rapid results. Biological validation is unsuitable to be performed in recuperating joeys due to ethical considerations, hence, we compared the results from biologically validated R4866 assay with the commercially available Arbor Assay cortisol kit. Thirty-four faecal samples were collected, processed and analysed for faecal cortisol metabolites (FCM) using both, R4866 assay and Arbor Assay kit. The joeys presented a suite of clinical conditions which provided the natural variation in stress response for comparing the assay sensitivity and range. The results indicated that there were no significant differences between the FCM values measured by both the assays. Furthermore, the Bland-Altman plot indicated a very strong agreement between the FCM concentrations measured by the two assays. This study is only a step towards recommending the routine use of commercial kit in clinical settings with basic resources, for rapid quantification of stress in koala patients. It is crucial for future studies to perform laboratory validation procedures to confirm the efficacy of the commercial kit before practical use for FCM monitoring in koalas.

The use of fecal cortisol for enrolment of free-roaming dogs in Singapore to a national rehabilitation-rehoming program: A possible indicator of allostasis

Singapore's trap-neuter-release-management program strives to rehome as many free-roaming dogs as possible. However, before rehoming, most free-roaming dogs require rehabilitation and this requires sheltering them for some time. Therefore, an individual's suitability for rehoming hinges on his or her adaptability to shelter conditions. Currently, this is being assessed qualitatively across behavioral contexts. Augmenting the current assessment with a known bioindicator of stress, such as cortisol, would increase its robustness. However, the serum measurement of cortisol can be invasive. Canine fecal samples have presented an attractive alternative for measuring cortisol indirectly in various contexts. Nevertheless, it is unclear whether the resolution of fecal cortisol metabolites (FCM) is sufficient for evaluating Singapore free-roaming dogs’ adaptability to shelter conditions. To elucidate this, we performed linear mixed-effects model on 27 candidate models to compare FCM between pet and recently sheltered dogs, with sex, state of the feces, and time as covariates. Subsequently, we ran generalized mixed-effected model on 12 candidate models to examine the influence of between- and within-subject variations in FCM on the prognosis of sheltered dogs—based on our current assessment. Our analyses showed that FCM levels of recently sheltered dogs were higher than pet individuals. However, unlike several studies, FCM concentrations of recently sheltered dogs continued to rise, indicating that individuals have yet to adapt. Furthermore, the difference in FCM levels between newly sheltered individuals and pet dogs was moderated by sex. Unexpectedly, we found the average FCM levels of dogs with a good prognosis to be higher than those that received a poor prognosis, with sex and state of the fecal samples as important covariates. This may be due to the difference in allostasis between the 2 groups of dogs. Thus, our results showed FCM not only has sufficient resolution but is useful in evaluating dogs’ adaptability to shelter conditions.

Cortisol metabolites vary with environmental conditions, predation risk, and human shields in a wild primate, Cercopithecus albogularis

Environmental challenges are often associated with physiological changes in wildlife that allow animals to maintain homeostasis. Among these, scarcity in resources, and risks from predators, competitors, and humans can all result in psychological and physiological stress. Yet, for habituated species, it is not clear whether this relationship with humans still holds to a lesser degree or is outweighed by the benefits of human presence – such as serving as a buffer from competitors or predators. We investigated how human presence and environmental challenges such as resource availability, weather, predation, and competition may be associated with variation in fecal cortisol metabolite levels (FCMs) in a group of samango monkeys (Cercopithecus albogularis) in the Soutpansberg Mountains, South Africa. FCMs can often broadly track environmental challenges and perturbations. Initially, we employed an exploratory analysis comparing candidate models representing biological hypotheses and found that those incorporating information on human presence had less weight than models for food availability, thermoregulation, and water scarcity. When we examined a subset of the data that included information on intergroup competition and predator alarm calls, we found that FCMs were higher on the day following potential predator encounters but not competitive interactions. As observer numbers increased, responses to predators flattened, indicating that the presence of several humans might deter predators and/or affect samangos' perception of danger – yet we could not distinguish between these possibilities. Together, these results suggest that ecological perturbations track with FCMs in this study population and challenge long-held assumptions that human presence has negligible effects on habituated study animals.

How environmental conditions affect sleep? An investigation in domestic dogs (Canis lupus familiaris)

Studies with humans and some other animal species have shown that sleep is compromised when the presence of external factors such as light, sound, and temperature surpass normal levels. This study investigated the effects of these environmental conditions on 13 kennelled laboratory dogs, assessing whether each variable interfered with their sleep behaviour and/or increased stress responses, which could further compromise sleep quality. The behaviour of dogs was video recorded for eight months. Diurnal and nocturnal behaviour were recorded, along with naturally occurring levels of temperature, light and sound in the dogs’ kennel environment. Faecal cortisol metabolites (FCM), from samples collected every morning, were used to monitor the dogs’ adrenocortical activity. GLMM models and non-parametric tests were conducted to evaluate the relationship between sleeping patterns, environmental variables, and stress on the studied dogs. Nocturnal sleep decreased in response to increases in temperature and in day light duration. No effects of sound and FCM levels on dogs’ sleep were observed. However, diurnal sleep was affected by sound and FCM levels, decreasing when both factors increased. Additionally, noisier days increased stress responses, especially in male dogs. Increased FCM levels were associated with changes in the diurnal behaviour of dogs; for example, decreased activity. The decrease in daily activities and increased physiological stress responses could be associated with maladaptation to the environment, which could indicate poor welfare. Our study suggests that mitigating the impact of environmental conditions in the kennels could improve sleep quality and the overall quality of life of the dogs.

Covariation between glucocorticoids, behaviour and immunity supports the pace-of-life syndrome hypothesis: an experimental approach.

The biomedical literature has consistently highlighted that long-term elevation of glucocorticoids might impair immune functions. However, patterns are less clear in wild animals. Here, we re-explored the stress-immunity relationship considering the potential effects of behavioural profiles. Thirteen captive roe deer (Capreolus capreolus) were monitored over an eight-week period encompassing two capture events. We assessed how changes in baseline faecal cortisol metabolite (FCM) concentrations following a standardized capture protocol and an immune challenge using anti-rabies vaccination affected changes in 13 immune parameters of innate and adaptive immunity, and whether these changes in baseline FCM levels and immune parameters related to behavioural profiles. We found that individuals with increased baseline FCM levels also exhibited increased immunity and were characterized by more reactive behavioural profiles (low activity levels, docility to manipulation and neophilia). Our results suggest that the immunity of large mammals may be influenced by glucocorticoids, but also behavioural profiles, as it is predicted by the pace-of-life syndrome hypothesis. Our results highlight the need to consider covariations between behaviour, immunity and glucocorticoids in order to improve our understanding of the among-individual variability in the stress-immunity relationships observed in wildlife, as they may be underpinned by different life-history strategies.

Effect of intensity and duration of anthropic noises on European mink locomotor activity and fecal cortisol metabolite levels.

Human activities involving noise emission can affect wild animals. European mink was exposed to road noise and human voice playbacks to analyze how sound intensity level and duration of both noises altered the time that individuals were active and if their fecal cortisol metabolite (FCM) levels varied. A Hierarchical Analysis Cluster was performed to establish 2 mink groups with respect to both noise source type: short duration/low intensity (SL) and long duration/high intensity (LH). We performed general linear mixed models to evaluate the variation in locomotor activity duration (s) and FCM (nanogram per gram) levels, respectively. The results showed both road noise and human voices decreased locomotor activity duration in SL more sharply compared with LH, and human voices were the triggers that induced the most pronounced response to both exposure conditions. FCM (ng/g) levels increased in SL compared with LH during road noise while the opposite happened during human voices. Differences based on sex and age of individuals were observed. In conclusion, noise characteristics given by the sound type determined the variations in locomotor activity duration while noise exposure level determined the variations in FCM (ng/g) levels. Attention should be paid to noisy activities (e.g., recreational activities for visitors in protected natural areas) and loud groups of people to conserve wildlife, especially noise sensitive species.

Over Time Decay of Cortisol Metabolites in Faecal Pellets of Koalas in Central Queensland.

Simple SummaryFaecal cortisol metabolites (FCMs) are a useful, non-invasive tool for the assessment of stress in koalas. However, FCM stability after defecation is a critical issue. Therefore, we exposed faecal pellets of koalas to three different environmental conditions and measured FCMs with three recently evaluated enzyme immunoassays (EIAs). Because water loss over time exerted the greatest influence on FCMs, we strongly recommend collecting freshly defecated pellets in koalas.Faecal material can be a valuable source of information for a range of animal health aspects and can be used to measure faecal cortisol metabolites (FCMs). FCM values can relate to physiological stress responses. However, freshly defecated pellets are not always available and environmental conditions, such as temperature and humidity, might affect faecal pellet consistency and FCM levels. Therefore, the impact of environmental conditions on FCMs needs to be evaluated. We collected 107 pellets from two female and two male koalas, exposed them to three types of treatments, and analysed FCMs in these samples with three enzyme immunoassays (EIAs). After analysis, the original FCM values were mathematically corrected for water loss. Results show that the FCMs were more stable when measured using tetrahydrocorticosterone (50c) and 5α-pregnane-3β,11β,21-triol-20-one (37e) EIAs, and were less stable when measured with the cortisol EIA. With 50c, the FCM values did not vary significantly over time either before or after the adjustment with water in the environment treatment group. For samples kept under constant low (25 °C) and high (35 °C) temperatures, the 50c FCM values did not vary significantly over time, after adjustments were made for water loss. Thus, this study highlights the importance of considering the suitability of faecal field samples for FCM analysis. Because water loss was the main driver of FCM changes, we strongly recommend collecting koala pellets that are freshly defecated, despite the effort and time it might take to collect such pellets.

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

Simple SummaryThe measurement of faecal cortisol metabolites (FCMs) is increasingly used to monitor physiological stress responses in different animal species. Before this method is applied in coming stress- and welfare-related studies of farmed blue foxes (Vulpes lagopus), a species-specific validation is first required. In the current study, a 5α-pregnane-3ß,11ß,21-triol-20-one enzyme immunoassay was found suited to measure FCMs and thus hypothalamus–pituitary–adrenal (HPA) axis activity in farmed blue foxes. FCMs can therefore serve as a valid indicator of stress in future welfare studies of 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.

Give Them a Toy or Increase Time out of Kennel at Lawn Areas: What Is the Influence of These Interventions on Police Dogs' Welfare?

Simple SummaryPoor environments such as kennels can lead to compromised welfare, as they usually lack many kinds of stimuli. Working dogs are not only kept in kennels, but they are also often housed without any kind of environmental enrichment, allegedly due to motivational reasons, which is conducive to all kinds of behavioural issues. Thus, this study aimed to evaluate the impact of two interventions, a toy and going out to a lawn area, on a group of police dogs. Behavioural and physiological data were used to evaluate the animals’ responses to treatments. We identified a large variety of behavioural profiles amongst the animals that presented different levels of stereotypies, which is an abnormal behaviour pattern that can be associated with stress. A tendency to a reduction in these behaviours in high-stereotyping individuals was observed after dogs were able to go to the lawn area, indicating beneficial effects of this intervention. The physiological data indicated that the cumulative effect of interventions can also have a beneficial impact and that environmental enrichment plays an important role in kennelled police dogs’ welfare.This work was aimed at identifying the effectiveness of two interventions applied to a group of eight kennelled police dogs. Interventions consisted of access to a lawn area (350 m2) and “toy” (a 30 cm jute rag roll, hanging from the kennel ceiling), both available for 15 min a day, for four days in a row. We collected behavioural data and faecal samples for cortisol metabolites evaluation before, during and after interventions. Faecal cortisol metabolites levels were significantly reduced (Friedman, X2(3) = 12.450; p = 0.006) during the second round of intervention, regardless of the type of intervention, indicating that the interventions can have a beneficial cumulative effect. Regarding the frequency of stereotyped behaviour, cluster analysis identified two groups of individuals: (1) high-stereotyping individuals (n = 3) that had a tendency to reduce stereotyping behaviours in the lawn intervention when compared to toy intervention (Friedman, X2(3) = 2.530; p = 0.068), and (2) low-stereotyping animals (n = 5) that did not present significant behavioural changes during the experiment. The variety of behavioural and endocrine parameters evaluated highlights the need to account for the individual in behaviour and welfare research. Overall, our results suggest that even simple environmental enrichment can be an effective method to mitigate behaviour and physiological signs of stress.

Evaluation of Fecal Glucocorticoid Metabolite Levels in Response to a Change in Social and Handling Conditions in African Lions (Panthera leo bleyenberghi).

Simple SummaryNon-invasive determination of cortisol metabolite concentrations in feces is widely used to evaluate the influence of housing and handling conditions on the stress physiology of wildlife in captivity. The present study aimed to assess the physiological response of a lion pride to a change in management and social conditions after the death of the dominant male of the pride. Before the dominant male died, weekly management routines between the indoor and outdoor enclosures were conducted to avoid cohabitation problems between the two males of the pride. After the death of the dominant male, these weekly management dynamics ceased, leading to a decrease in the daily management routine of the lion pride. An individualized sampling of the animals through the utilization of indigestive markers was conducted, and fecal samples were collected before and after the death of the dominant male. Significant lower cortisol metabolite concentrations in feces were detected after the death of the dominant male, suggesting a positive impact of a decrease in daily management routines, together with a more stable social environment. In addition, assessment of individualized hormone concentrations throughout the study revealed variable physiological responses among lions, providing evidence of the importance of monitoring hormonal profiles individually.Monitoring the hypothalamic–pituitary–adrenal (HPA) axis through determination of fecal cortisol metabolite (FCM) levels is a non-invasive method useful for understanding how handling and social conditions may affect the physiological status of zoo animals. The present study used FCM analysis to evaluate whether the HPA axis activity of a lion pride was modified by a change in social and handling conditions after the death of the dominant male. Five African lions (Panthera leo bleyenberghi), two males and three females, were included in the study. Fecal samples were collected before and after the death of the dominant male. To avoid cohabitation conflicts between males before the dominant male died, subgroups were established and subjected to weekly changes between indoor and outdoor facilities. After the death of the dominant male, these management dynamics ceased, and the remaining four lions were kept together outdoors. Significant lower group FCM concentrations (p < 0.001) were detected after the decease of the dominant male, probably associated with a decrease in daily handling, together with a more stable social environment. Overall, the present study indicates the effect of different management scenarios on the HPA axis activity and differentiated physiological responses to the same situation between individuals.

Intestinal Parasites and Fecal Cortisol Metabolites in Multi-Unowned-Cat Environments: The Impact of Housing Conditions.

Simple SummaryMulti-unowned-cat environments can be highly stressful for cats, and infectious and parasite diseases spread quickly and are difficult to prevent. This study aimed to determine the occurrence of intestinal parasites and fecal cortisol metabolites (FCM) in cat feces collected from different multi-cat environments and assess the effect of housing conditions on intestinal parasites and FCM levels in order to develop more efficient control strategies. Cat fecal samples from rescue shelters, catteries and feline colonies were analyzed with coprological methods to detect intestinal parasite patency and determine FCM. Helminth infection was mainly detected in free-roaming cats. In confined cats, protozoa infections were more likely detected in shelter cats than in cattery cats. Exposure to dogs was associated with parasite infection and cats highly exposed to dogs with visual contact and audible barking showed higher intestinal protozoa prevalence than cats that were not exposed to dogs. FCM levels were correlated with enclosure size and protozoa infection. Reducing stress by improving housing in terms of enclosure size and avoiding exposure to dogs may have an impact on the occurrence of intestinal parasites, especially protozoa.Housing conditions were assessed in different unowned multi-cat management models in order to evaluate their impact on the occurrence of intestinal parasites and fecal cortisol metabolite (FCM) levels. Fresh stool fecal samples were collected from rescue shelters, catteries and feline colonies for coprological analyses in order to detect intestinal parasite patency and fecal cortisol metabolites. A questionnaire provided information about the facilities, management and housing conditions of cats, including information about dog exposure, enclosure size, environment enrichment and changes to group composition. Overall, intestinal parasite infection was detected in 58.2% of fecal samples collected. The occurrence of intestinal parasites detected in free-roaming cats was 82.2%, mainly due to helminth infection. The parasite infection rate was 57.3% in rescue shelters and 34.6% in catteries. In confined cats, protozoa infection was more likely detected in rescue shelters than in catteries (RR = 2.02 (1.30–3.14), p = 0.0012). Although the FCM values were very variable between cats, the enclosure size and parasite infection were correlated with the average FCM. A small enclosure size was correlated with high fecal cortisol metabolites (p = 0.016). Protozoa-positive samples showed higher FCM levels than negative samples (p = 0.0150). High dog exposure was statistically associated with protozoa infection (p = 0.0006). The results indicated that improving housing, especially in terms of floor space and avoiding dog exposure, reduces stress and can thus be applied to make control strategies in multi-unowned-cat environments more efficient, especially when cats are confined.

Social environment with high intrasexual competition enhances the positive relationship between faecal testosterone and cortisol metabolite levels in red deer

The relationship between reproductive and physiological stress hormones in vertebrates is poorly understood. In many species of mammals and especially in humans, the most widespread idea is that there is a negative relationship between them, i.e. higher stress levels are associated with lower testosterone levels. Likewise, the subordination stress paradigm supports that subordinates suffer greater stress in a competitive situation, while the dominant ones have higher levels of testosterone. However, this predominant idea of a negative relationship between testosterone and cortisol concentrations may be influenced by unnatural circumstances, such as chronic stress in humans or forced interactions between subordinates and dominants in laboratory or captivity. Some studies have reported that dominant males under natural conditions may show higher physiological stress and also higher testosterone levels than subordinates. But for this positive relationship, the question is whether there is a causal link or whether both hormones only coincide due to other factors. We hypothesized that testosterone should be related to physiological stress only as a result of individual males being subjected to stressful situations, such as intrasexual competition. We studied this topic in Iberian red deer (Cervus elaphus hispanicus) males in two types of populations with high and low levels of intrasexual competition. We found a positive relationship between faecal testosterone and cortisol metabolite levels, but also a significant interaction showing that this relationship occurs more intensely in populations with high competition level for mating. These results reinforce the positive relationship between both hormones under natural conditions and support the hypothesis that it is mediated by male-male competition for mates.

The cost of associating with males for Bornean and Sumatran female orangutans: a hidden form of sexual conflict?

Sexual coercion, in the form of forced copulations, is relatively frequently observed in orangutans and generally attributed to their semi-solitary lifestyle. High ecological costs of association for females may be responsible for this lifestyle and may have prevented the evolution of morphological fertility indicators (e.g., sexual swellings), which would attract (male) associates. Therefore, sexual conflict may arise not only about mating per se but also about associations, because males may benefit from associations with females to monitor their reproductive state and attempt to monopolize their sexual activities. Here, we evaluate association patterns and costs for females when associating with both males and females of two different orangutan species at two study sites: Suaq, Sumatra (Pongo abelii), and Tuanan, Borneo (Pongo pygmaeus wurmbii). Female association frequency with both males and females was higher in the Sumatran population, living in more productive habitat. Accordingly, we found that the cost of association, in terms of reduced feeding to moving ratio and increased time being active, is higher in the less sociable Bornean population. Males generally initiated and maintained such costly associations with females, and prolonged associations with males led to increased female fecal cortisol metabolite (FCM) levels at Tuanan, the Bornean population. We conclude that male-maintained associations are an expression of sexual conflict in orangutans, at least at Tuanan. For females, this cost of association may be responsible for the lack of sexual signaling, while needing to confuse paternity.Significance statementSocioecological theory predicts a trade-off between the benefits of sociality and the ecological costs of increased feeding competition. Orangutans’ semi-solitary lifestyle has been attributed to the combination of high association costs and low predation risk. Previous work revealed a positive correlation between association frequencies and habitat productivity, but did not measure the costs of association. In this comparative study, we show that females likely incur costs from involuntary, male-maintained associations, especially when they last for several days and particularly in the population characterized by lower association frequencies. Association maintenance therefore qualifies as another expression of sexual conflict in orangutans, and especially prolonged, male-maintained associations may qualify as an indirect form of sexual coercion.

Losing seasonal patterns in a hibernating omnivore? Diet quality proxies and faecal cortisol metabolites in brown bears in areas with and without artificial feeding.

Bears are omnivores particularly well-adapted to variations in the nutritional composition, quality and availability of food resources. Artificial feeding practices have been shown to strongly influence diet composition and seasonality, as well as to cause alterations in wintering and movement in brown bears (Ursus arctos). In this study, we investigated seasonal differences (hypophagia vs hyperphagia) in food quality of two brown bear subpopulations in the Polish Carpathians using faecal nitrogen (FN) and carbon (FC) estimates. The subpopulations inhabit areas that differ in artificial feeding practices: no artificial feeding occurs in the western subpopulation (Tatra Mountains), while artificial food targeted to ungulates is provided and used year-round in the eastern subpopulation (Bieszczady Mountains). We also compared these results with faecal cortisol metabolites (FCM) to explore how FN and FC correlate with the hypothalamic-pituitary-adrenal axis activity and if the seasonal patterns are apparent. We found that in Tatra Mts bears fed on significantly higher quality diet, as shown by FN and FC values, and had significantly higher FC levels in hyperphagia, when they accumulate fat reserves for wintering. The pattern in FCM levels for Tatra subpopulation followed the changes in energy intake during the seasons of hypo- and hyperphagia, while in Bieszczady Mts, the area with intensive feeding, no seasonal patterns could be observed. Artificial feeding practices may disrupt nutrient phenology and seasonality, relative to subpopulations with natural diets. We showed that the availability of human-provided foods may alter not only the overall dietary quality, but also hormonal patterns linked to seasonal nutritional requirements. Combining FN, FC and FCM proved to be a useful tool for reconstructing diet quality and related physiological patterns.