Corticosterone Metabolite Concentration Is Not Related to Problem Solving in the Fawn-Footed Mosaic-Tailed Rat Melomys cervinipes

Animals can respond physiologically, such as by adjusting glucocorticoid hormone concentrations, to sudden environmental challenges. These physiological changes can then affect behavioural and cognitive responses. While the relationships between adrenocortical activity and behaviour and cognition are well documented, results are equivocal, suggesting species-specific responses. We investigated whether adrenocortical activity, measured using corticosterone metabolite concentration, was related to problem solving in an Australian rodent, the fawn-footed mosaic-tailed rat (Melomys cervinipes). Mosaic-tailed rats live in complex environments that are prone to disturbance, suggesting a potential need to solve novel problems, and have been found to show relationships between physiology and other behaviours. We measured problem solving using five food-baited puzzles (matchbox and cylinder in the home cage, and activity board with pillars to push, tiles to slide and levers to lift in an open field), and an escape-motivated obstruction task in a light/dark box. Faecal samples were collected from individuals during routine cage cleaning. Adrenocortical activity was evaluated non-invasively by measuring faecal corticosterone metabolites using an enzyme immunoassay, which was biochemically and biologically validated. Despite varying over time, adrenocortical activity was not significantly related to problem solving success or time spent interacting for any task. However, as adrenocortical activity is reflective of multiple physiological processes, including stress and metabolism, future studies should consider how other measures of physiology are also linked to problem solving.

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

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

Fecal glucocorticoid metabolites as an indicator of adrenocortical activity in Arctic foxes (Vulpes lagopus) and recommendations for future studies

Measuring 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.

‘Kindergarten’ keeping-system in farrowing house: effect the socialization of piglets on weight performances, fecal cortisol metabolite level and post-weaning behavior

This study aimed to examine whether litters’ let-together system before weaning (‘kindergarten-system’) has any stress effect and effect on post-weaning behavior, respectively. One week before weaning piglets were socialized by removing four adjacent farrowing crate walls. After weaning, piglets from the same experimental groups were housed in the same rearing crates. The piglet’s body weight was measured at birth than weekly till the 7th life-week. Fecal cortisol metabolites were measured for evaluation of the adrenocortical activity. Personal observation and evaluation were carried out for behavioral analyses after weaning. There were no significant differences in weight development and cortisol metabolite levels between the control and experimental groups. Behavioral analysis showed that pigs grow up in a socialized system, rapidly evolve ranking in the rearing crates, and the self-maintenance and social behavior were more pronounced. In control groups, the activity involving movement (exploring, walking, and running) was much more decisive. Recent work suggested the beneficial effect of the ‘kindergarten’ system in the farrowing house in animal welfare aspects.

Association between stress and bilateral symmetrical alopecia in free-ranging Formosan macaques in Mt. Longevity, Taiwan

Since 2013, a high incidence of bilateral symmetrical alopecia has been observed in free-ranging Formosan macaques (Macaca cyclopis) in Mt. Longevity, Taiwan. We hypothesized that stress induces alopecia in this population. To verify our hypothesis, we evaluated the histopathological characteristics of skin biopsy and used a validated enzyme immunoassay (EIA) for fecal glucocorticoid metabolite (FGM) analysis, which act as an indicator of stress experienced by the individual. Follicular densities were lower (2.1–3.0 mm2) in individuals with symmetrical alopecia than in those with normal hair conditions (4.7 mm2). Furthermore, anagen to catagen/telogen ratios were lower in individuals with alopecia (0–1.4) than in those with normal hair (4.0). The histopathological characteristics of alopecia were similar to those of telogen effluvium, which indicates that stress is one of the possible etiologies. On the basis of the analytical and biological validation of EIAs for FGM analysis, 11β-hydroxyetiocholanolone was considered suitable for monitoring adrenocortical activity in both sexes of Formosan macaques. The mean concentrations (standard error; sample size) of 11β-hydroxyetiocholanolone were 2.02 (0.17; n = 10) and 1.41 (0.10; n = 31) μg/g for individuals with and without alopecia, respectively. Furthermore, the results of logistic regression analysis show that 11β-hydroxyetiocholanolone (p = 0.012) concentration was positively associated with alopecia. Thus, stress was the most likely to trigger symmetrical alopecia in Formosan macaques in Mt. Longevity. Although stress can decrease the fitness of an individual, considering the population status of Formosan macaques in Taiwan is stable and alopecia was only observed in our study area, which is isolated from other populations, the impact on the total population of Formosan macaque in Taiwan is limited. Nonetheless, stress-induced immunosuppression and alopecia might affect the local abundance and increase zoonosis risk due to frequent human–macaque contact in Mt. Longevity. Future studies are suggested to focus on the causative factor of stress and the effects of stress and alopecia on the health and welfare in the Formosan macaques.

Adrenocortical Activity and Aggressive Behavior in Children: A Longitudinal Study on Risk and Protective Effects

Most research on aggression and delinquency concentrates on risk factors. There has been less attention for protective factors and mechanisms, in particular with regard to biosocial influences. Based on theories of autonomous arousal and stress reactance the present study addresses the influence of adrenocortical activity as a risk and/or protective factor in the development of antisocial behavior in children. We also investigated relations to anxiousness and family stressors. In a prospective longitudinal study of 150 German boys, the first measurement took place at preschool age and contained an assessment of cortisol after waking up and 30 min later. Aggressiveness and anxiousness of the children were assessed by the kindergarten teachers with the Social Behavior Questionnaire. After 6 years, the children's behavior was rated by the teachers in middle school. Variable-oriented data analyses revealed a significant correlation between the total amount of cortisol after waking up and 30 min later (AUCG) and anxiousness both cross-sectionally and longitudinally, but not with aggressiveness. A family stress index correlated positively with aggressiveness but neither with cortisol nor with anxiousness. There were significant correlations between aggressiveness and anxiousness at kindergarten age and the respective behavior problems 6 years later. In a linear regression analysis on aggression only family stress had a significant effect but anxiousness not. Moderator analyses on aggressiveness with anxiousness and AUCG or on AUCG with anxiousness and aggressiveness did not show any significant interactions. Longitudinally, only aggression significantly predicted aggression 6 years later in a linear regression. In addition to variable-oriented analyses, we also applied a person-oriented approach to investigate specific patterns of behavior. Children who were high in both aggressiveness and anxiousness had the highest cortisol level and those with low anxiousness and high aggressiveness the lowest. The groups with different patterns of externalizing and internalizing problems at preschool age showed significant differences in aggression 6 years later. Our results underline the need for complex pattern analyses on cortisol, aggression, and anxiousness in children and for a differentiated consideration of emotional reactive aggression and unemotional instrumental aggression.

Association Between Stress and Bilateral Symmetrical Alopecia in Free-Ranging Formosan Macaques (Macaca Cyclopis) in A High-Population-Density Area

Since 2013, a high incidence of bilateral symmetrical alopecia has been observed in free-ranging Formosan macaques (Macaca cyclopis) in Mt. Longevity, Taiwan. We hypothesized that stress induces alopecia in this population. To verify our hypothesis, we evaluated the histopathological characteristics of skin biopsy and used a validated enzyme immunoassay (EIA) for fecal glucocorticoid metabolite (FGM) analysis, which act as an indicator of stress experienced by the individual. Follicular densities were lower (2.1–3.0 mm2) in individuals with symmetrical alopecia than in those with normal hair conditions (4.7 mm2). Furthermore, anagen to catagen/telogen ratios were lower in individuals with alopecia (0–1.4) than in those with normal hair (4.0). The histopathological characteristics of alopecia were similar to those of telogen effluvium, which indicates that stress is one of the possible etiologies. On the basis of the analytical and biological validation of EIAs for FGM analysis, 11β-hydroxyetiocholanolone was considered suitable for monitoring adrenocortical activity in both sexes of Formosan macaques. The mean concentrations (standard error; sample size) of 11β-hydroxyetiocholanolone were 2.02 (0.17; n = 10) and 1.41 (0.10; n = 31) μg/g for individuals with and without alopecia, respectively. Furthermore, the results of logistic regression analysis show that 11β-hydroxyetiocholanolone (p = 0.012) concentration was positively associated with alopecia. Thus, stress was the most likely to trigger symmetrical alopecia in Formosan macaques in Mt. Longevity. Although stress can decrease the fitness of an individual, it should not impact the total population of Formosan macaque in Taiwan. Nonetheless, stress-induced immunosuppression might increase zoonosis risk due to frequent human–macaque contact in Mt. Longevity.

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

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