Glucocorticoid Metabolite Concentrations Research Articles

Physiological stress response to urbanisation differs between native and invasive squirrel species

Novel pressures derived from urbanisation can alter native habitats and ultimately impact wildlife. Coping with such human-driven changes might induce shifts in species phenotypic traits, such as physiological responses to anthropogenic stressors. Preadaptation to face those challenges has been suggested to favour settlement and spread of invasive alien species in urbanised areas which, consequently, might respond differently than ecologically similar native species to stressors posed by urbanisation. The activation of the hypothalamic-pituitary-adrenal (HPA) axis and the subsequent release of glucocorticoids (GCs) has been suggested to mediate responses to anthropogenic disturbance in vertebrates. Furthermore, intraspecific competition, in conjunction with stressors related to urbanisation, might affect invasive and native species physiological stress responses differently. Using a parallel pseudo-experimental study system we measured faecal glucocorticoid metabolite (FGM) concentrations of the native Eurasian red squirrel and the invasive alien Eastern grey squirrel along a rural-urban gradient and in relation to conspecific density. The two species responded differently to challenges posed by the synergic effect of urbanisation and intraspecific competition. Association of FGMs and conspecific density in native red squirrels varied between rural and suburban sites, potentially depending on differential HPA axis responses. In urban sites, this relationship did not differ significantly from that in rural and suburban ones. Conversely, invasive grey squirrels' FGMs did not vary in relation to conspecific density, nor differed along the rural-urban gradient. Improving knowledge about native and competing invasive species' physiological responses to anthropogenic stressors can support conservation strategies in habitats altered by man. Our findings suggested that the invasive squirrels might be preadapted to cope with these challenges in urbanised areas, potentially increasing their success under the future global change scenario.

Mantled howler monkey males assess their rivals through formant spacing of long-distance calls.

Formant frequency spacing of long-distance vocalizations is allometrically related to body size and could represent an honest signal of fighting potential. There is, however, only limited evidence that primates use formant spacing to assess the competitive potential of rivals during interactions with extragroup males, a risky context. We hypothesized that if formant spacing of long-distance calls is inversely related to the fighting potential of male mantled howler monkeys (Alouatta palliata), then males should: (1) be more likely and (2) faster to display vocal responses to calling rivals; (3) be more likely and (4) faster to approach calling rivals; and have higher fecal (5) glucocorticoid and (6) testosterone metabolite concentrations in response to rivals calling at intermediate and high formant spacing than to those with low formant spacing. We studied the behavioral responses of 11 adult males to playback experiments of long-distance calls from unknown individuals with low (i.e., emulating large individuals), intermediate, and high (i.e., small individuals) formant spacing (n = 36 experiments). We assayed fecal glucocorticoid and testosterone metabolite concentrations (n = 174). Playbacks always elicited vocal responses, but males responded quicker to intermediate than to low formant spacing playbacks. Low formant spacing calls were less likely to elicit approaches whereas high formant spacing calls resulted in quicker approaches. Males showed stronger hormonal responses to low than to both intermediate and high formant spacing calls. It is possible that males do not escalate conflicts with rivals with low formant spacing calls if these are perceived as large, and against whom winning probabilities should decrease and confrontation costs increase; but are willing to escalate conflicts with rivals of high formant spacing. Formant spacing may therefore be an important signal for rival assessment in this species.

Kibble diet is associated with higher faecal glucocorticoid metabolite concentrations in zoo-managed red wolves (Canis rufus).

The red wolf (Canis rufus) is a critically endangered canid that exists solely because of the establishment of the ex situ population in the late 1980s. Yet, the population under human care suffers from gastrointestinal (GI) disease in captivity. While the cause of GI disease is unknown, it is speculated that environmental factors can influence GI health of zoo-managed red wolves. The goal of the present study was to investigate the relationship between faecal glucocorticoid metabolite (FGM) concentrations, a biomarker for stress, and environmental factors for zoo-managed red wolves. Faecal samples were collected from 14 adult wolves three times a week for 5 to 12months. Using a single-antibody cortisol enzyme immunoassay, FGM concentrations were quantified. Environmental factors were collected for each participating wolf on dietary type, sex, type of public access to enclosure, density (enclosure size [ft2]/number of wolves living in enclosure) and a monthly average status of GI health. Red wolves that ate a commercial kibble diet had both higher FGM concentrations over time and higher baseline FGM concentrations compared to individuals that received commercial kibble mixed with commercial meat. Density, public access or GI health were not related to FGM concentration; however, males had higher baseline FGM concentrations compared to female red wolves. Our findings suggest that management conditions, particularly diet, can strongly influence FGM concentration in the zoo-managed red wolf population. Findings from this study highlight the importance of management choices on individual welfare. Maintaining a healthy captive population of red wolves is imperative for the persistence of the species, including successful future reintroductions.

Feasibility of a Sprague-Dawley Rat Model for Investigating the Effects of Seated Whole-body Vibration.

Vehicular whole-body vibration (WBV) can have long-term adverse effects on human quality of life. Animal models can be used to study pathophysiologic effects of vibration. The goal of this study was to assess animal cooperation and well-being to determine the feasibility of a novel seated rat model for investigating the effects of WBV on biologic systems. Twenty-four male Sprague-Dawley rats were used. The experiment consisted of an acclimation phase, 2 training phases (TrP1 and TrP2), and a testing phase (TeP), including weekly radiographic imaging. During acclimation, rats were housed in pairs in standard cages without vibration. First, experimental (EG; n = 18) and control group 1 (C1; n = 3) rats were placed in a vibration apparatus without vibration, with increasing duration over 5 d during TrP1. EG rats were exposed to vertical random WBV that was increased in magnitude over 5 d during TrP2 until reaching the vibration signal used during TeP (15 min, 0.7 m·s-2 root mean square, unweighted). C1 rats were placed in the vibration apparatus but received no vibration during any phase. Control group 2 (C2; n = 3) rats remained in the home cages. Cooperation was evaluated with regard to rat-apparatus interactions and position compliance. Behavior, weight, and fecal glucocorticoid metabolite concentrations (fGCM) were used to evaluate animal well-being. We observed good cooperation and no behavioral patterns or weight loss between phases, indicating little or no animal stress. The differences in fGCM concentration between groups indicated that the EG rats had lower stress levels than the control rats in all phases except TrP1. Thus, this model elicited little or no stress in the conscious, unrestrained, seated rats.

Parity and housing effects on the behavioural and hypothalamic–pituitary–adrenal axis responses of pregnant ewes

It is common in many countries for sheep to be housed during winter from mid-gestation until lambing to protect ewes and lambs from adverse conditions and improve late gestation nutritional management. Keeping ewes indoors, however, has its own challenges as the animals may be mixed with unfamiliar conspecifics, have limited floor and feeding space, experience changes to their diet and increased handling by humans. Therefore, the objective of this study was to investigate the effect of variation in housing management (space allowance and social stability) on the behaviour and hypothalamic–pituitary–adrenal (HPA) axis responses of pregnant ewes from mid-to-late gestation (weeks 11–18 of pregnancy). Seventy-seven ewes (41 primiparous, 36 multiparous) were divided into two groups: ‘Control’ and ‘Restricted space and mixed’ (RS-Mix), where RS-Mix ewes were allocated half the amount of space (1.27 vs 2.5 m2 for RS-Mix and Control, respectively) and feedface (concentrate feeder space) allowance (36 vs 71 cm per ewe) given to the Control group and were also subjected to two social mixing events. Aggressive behaviour at the feedface and time spent standing, lying, walking, feeding and ruminating were recorded and faecal samples were collected for assessment of faecal glucocorticoid metabolite (FGM) concentrations. Higher aggression was observed in RS-Mix ewes during the first week of observation (P = 0.044), which gradually declined to the same level as Control ewes by the end of the study (P = 0.045). RS-Mix ewes were significantly less likely to be able to freely join the feedface compared to Controls (P = 0.022). No other significant treatment effects on aggressive behaviour or FGM during gestation were found. RS-Mix ewes displayed significantly higher ruminating behaviour at week 18 of gestation compared to Control ewes (P < 0.001), but no other effects were seen on general pen behaviour. However, the effect of indoor housing had a significant impact on primiparous ewes, who had lower weight gain (P = 0.015) and higher FGM concentrations (P = 0.014) compared to multiparous ewes regardless of treatment group. The data suggest that, although no sustained effects on behaviour or HPA axis responses were seen with the differences in space and feeder allowance or social stability at the levels used in this study, inexperienced (primiparous) ewes may find indoor housing more stressful; and are less able to adapt compared to multiparous ewes. These effects may influence the behaviour of the ewe at lambing time, and her offspring.

Responses to Reduced Feeding Frequency in Captive-Born Cheetahs (Acinonyx jubatus): Implications for Behavioural and Physiological Stress and Gastrointestinal Health.

Unnatural diet composition and frequent feeding regimes may play an aetiological role in the multiple diseases prevalent in captive cheetahs. This study investigated the responses of captive-born (hand-reared) cheetahs (n = 6) to a reduced feeding frequency schedule distinguished by offering larger quantities of food less frequently. The study cheetahs were fed four once-daily meals per week during the 3-week treatment period, followed by a 3-week control period in which they were fed two daily rations six days a week. Total weekly food intake was maintained throughout the study. Variations in behaviour, faecal consistency score (FCS), and faecal glucocorticoid metabolite concentration were measured. Less frequent feeding resulted in higher FCS (p < 0.01) and locomotory behaviour (p < 0.05) among the studied cheetahs. Faecal glucocorticoid metabolite concentration demonstrated an initial acute stress response to the change in feeding frequency (p < 0.05) and subsequent adaptation. The results of the FCS analysis suggest that the more natural feeding pattern could have benefited the studied cheetahs' gastrointestinal health without a significant behavioural or physiological stress response overall to the change in feeding frequency.

Linking glucocorticoid variations to monthly and daily behavior in a wild endangered neotropical primate.

Identifying the factors swaying physiological stress levels in wild animals can help depict how they cope with environmental and social stressors, shedding light on their feeding ecology, behavioral plasticity, and adaptability. Here, we used noninvasive methods to explore the link between glucocorticoid levels and behavior in an endangered neotropical primate facing habitat fragmentation pressure, the black lion tamarin (Leontopithecus chrysopygus). We investigated monthly and day-to-day glucocorticoid variations independently to attempt to disentangle the complex nature of the adrenocortical activity. Between May 2019 to March 2020, we followed two groups of black lion tamarins in two different areas, a continuous forest and a small fragment, and gathered behavioral data (over 95 days in total; 8.6 ± 3.9 days/month) and fecal samples (Nsamples = 468; 4.93 ± 3.5 samples/day) simultaneously. Preliminary analyses enabled us to identify circadian variations linked to the biological rhythm, which were taken into account in subsequent models. Monthly analyses revealed that black lion tamarin fecal glucocorticoid metabolite levels vary according to changes in activity budget associated with the fruit consumption, movement, and resting time of the groups. At a day-to-day level, while intergroup encounters led to increases in fecal glucocorticoid metabolite concentrations, we found that changes in food intake or activity level did not trigger physiological stress responses. These findings suggest that diet and ranging patterns, driven by food availability and distribution, influence physiological stress at a seasonal scale, while acute stressors such as interspecific competition trigger short-term stress responses. Exploring fecal glucocorticoid metabolite variations over different timescales can help uncover the predictive and reactive facets of physiological stress in wild species. Moreover, having a comprehensive understanding of the physiological state of species is a valuable conservation tool for evaluating how they cope in changing environments.

Dog barks influence the physiological stress and behavior of a wild primate

Non-lethal impacts of dogs on primates have seldom been assessed. We used an experimental approach to determine if mantled howler monkeys (Alouatta palliata) perceive dog barks as an aversive stimulus and thus display physiological and behavioral responses toward simulated barks. For one year (1754 h of observations) we studied 16 adult males belonging to five groups in Los Tuxtlas (Mexico), and recorded the occurrence of naturally occurring dog barks, their sound pressure level (SPL), and the behavioral responses of howler monkeys to barks. We then exposed males to bark playbacks at two SPL treatments, 40 and 80 dB in a total of 50 experiments. We assayed glucocorticoid metabolite concentrations in fecal samples (fGCM) as a marker of the physiological stress response of males. We also recorded the duration of vigilance, vocalizations, and flight in relation to playbacks. Naturally occurring barks were frequent and usually elicited behavioral responses by males. fGCM concentrations increased after bark playbacks and with stimuli intensity. Time spent vigilant increased following playbacks independently of stimuli intensity but both vocalizations and flight were linked to stimuli intensity: vocalizations were the longest after barks played-back at 80 dB, but males spent more time fleeing in response to 40 dB bark playbacks. These results provide evidence that dog barks are pervasive in the habitat of mantled howler monkeys living at Los Tuxtlas and disturb males, both physiologically and behaviorally. Although the potential costs of physiological and behavioral responses could not be determined, there is sufficient evidence to assume that they do have negative impacts on individuals. Therefore, our study provides avenues for future research on dog-wildlife interactions and valuable information for the design of conservation actions aimed at mitigating the impact of dogs on mantled howler monkeys.

Measures of Adrenal and Gonadal Hormones in Relation to Biological and Management Factors among Captive Red Pandas in Indian Zoos.

Animals in human care are affected by stressors that can ultimately reduce fitness. When reproduction is affected, endangered species conservation programs can be severely compromised. Thus, understanding factors related to stress and reproduction, and related hormones, is important to ensure captive breeding success. Red pandas (Ailurus fulgens) are endangered, and populations in the wild are threatened with extinction. A global captive breeding program has been launched to conserve the species with the goal of reintroduction. However, there is little information on how stressors impact physiological aspects of the species. This study measured fecal glucocorticoid (fGCM), progestagen (fPM), and androgen (fAM) metabolite concentrations in 12 female and 8 male red pandas at 3 zoos in India to determine predictors of adrenal and gonadal steroid activity, and the influence of fGCM on reproduction. Based on the generalized linear mixed model (GLMM), fGCM concentrations were positively correlated with the number of visitors, number of nests and enclosure areas, and negatively related to frequency of feedings, log density, and social time, while fPM concentrations were negatively associated with enclosure areas. A confounder for enclosure areas and number of nests was the fact that these spaces were relatively barren, with limited hiding spaces, compared to the smaller enclosures. By contrast, no significant relationships were found for fAM, perhaps due to the smaller sample size. A negative relationship between fGCM and fPM was observed, indicating increasing adrenal hormones may decrease reproductive function among female red pandas. Results suggest that zoo management should consider increasing feeding frequency, providing larger enclosures with more enrichment and more nests in larger spaces, and regulating visitor numbers to support good welfare and potentially improve reproductive fitness of red pandas in captivity.

Temperament traits and glucocorticoid metabolites in tropical screech owl (Megascops choliba)

Animals constantly respond to stimuli from their surroundings, showing individual behavioral and endocrine differences upon perceived challenging and/or threatening stimuli. The temperament of 13 captive tropical screech owls (Megascops choliba) was defined by performing three challenge tests (novel space, taxidermist predator and trapping) to identify inter-individual differences and their possible relationship with the production of glucocorticoid metabolites (GCM). Feces samples of each bird were taken pre-and-post predator challenge to quantify GCM concentration by means of enzymatic immunoassay (EIA). The intensity of 13 adjectives selected to describe the owls' temperament was determined by three judges during behavioral testing. Only 3 adjectives (passive, vulnerable, and attentive) that showed strong inter-test and intra-individual correlation coefficients were considered. These adjectives were later grouped in a single dimension named “shyness”, enabling establishment of individual scores. The temperament of the owls showed correlation to post-predator challenge GCM levels (ρ = 0.79; p < 0.001). However, no significant differences were found in GCM levels before and after predator challenge. Our results both confirm that nocturnal behavior raptorial birds show positive relation between shy temper and adrenal response, and offer the non-invasive herein implemented methods for establishment of new hypotheses that will assess welfare of species kept under human care.

A randomized controlled trial assessing the effect of intermittent and abrupt cessation of milking to end lactation on the well-being and intramammary infection risk of dairy cows

The objectives were to compare the effects of an intermittent milking schedule with a thrice daily milking schedule during the final week of lactation on the well-being, udder health, milk production, and risk of culling of dairy cows. We hypothesized that cows subjected to an intermittent milking schedule would experience less udder engorgement and pain, lower concentrations of fecal glucocorticoid metabolites (11,17-dioxoandrostanes; 11,17-DOA concentration) after dry-off, lower risk of an intramammary infection during the dry period, higher milk production and lower somatic cell count in the subsequent lactation, and lower culling risk compared with herd mates milked 3 times daily and dried off by abrupt cessation. In a randomized controlled field study, Holstein cows (n = 398) with a thrice daily milking schedule were assigned to treatment and control groups. The treatment consisted of an intermittent milking schedule for 7 d before dry-off (gradual cessation of milking, GRAD). Gradual-cessation cows were milked once daily until the day of dry-off, whereas cows in the control group (abrupt cessation of milking, APT) were milked 3 times daily until the day of dry-off. Udder firmness and pain responses of the udder 3 d after dry-off, as well as the percentage change in fecal 11,17-DOA concentration (3 d after dry-off compared with the dry-off day), were used to assess the well-being of the animals. Compared with cows in the GRAD group, the odds [95% confidence interval (CI)] of udder firmness were 1.55 (0.99-2.42) for cows in the APT group, and the odds of a pain response were 1.48 (0.89-2.44) for cows in the APT group. The least squares means (95% CI) of the percentage change in 11,17-DOA concentration were 129.3% (111.1-150.4) for the APT group and 113.6% (97.5-132.4) for the GRAD group. Quarter-level culture results from the periods before dry-off and after calving were compared, to assess the likelihoods of microbiological cure and new infection. Cows in the APT group had lower odds of a new intramammary infection in the dry period [odds ratio, 95% CI: 0.63 (0.37-1.05)], whereas we observed no meaningful differences in the microbiological cure likelihood among groups. The least squares means (95% CI) for somatic cell counts (log10-transformed) were 4.9 (4.8-5.0) in the APT group and 4.9 (4.8-5.0) in the GRAD group. The odds (95% CI) of clinical mastitis in the first 30 d postcalving were 1.32 (0.53-3.30) in the APT group compared with the GRAD group. We observed no meaningful differences in milk production at the first test date postcalving or the culling risk among groups. We conclude that the gradual-cessation protocol tested herein failed to significantly improve animal well-being, udder health, milk production, and survival in the tested study cohort. However, the observed differences in udder firmness, as well as the numerical differences in udder pain and the percentage change in fecal 11,17-DOA concentrations suggest that this line of research may be useful. Future research is needed to develop drying-off strategies that are appropriate for lowering milk production at the end of the lactation and improve animal well-being without compromising udder health.

Validation of complementary non-invasive tools for stress assessment in spotted paca (Cuniculus paca).

Monitoring the concentration of glucocorticoid metabolites (GCMs) in faecal samples is a non-invasive tool for physiological stress evaluation, particularly useful when studying wild species. However, both negative and positive stimuli (distress and eustress, respectively) can lead to a rise in glucocorticoids. Thus, besides validating whether GCM concentration in faeces reflects endogenous adrenal activity, we also need to identify behavioural indicators of distress to avoid misinterpretation. Therefore, we submitted four adult male spotted pacas (Cuniculus paca) to an exogenous adrenocorticotropic hormone (ACTH) challenge-test in a Latin square design (4 × 4) to monitor changes in the GCM concentration in faeces. We also aimed to describe behaviours potentially indicative of distress. We collected excreted faeces and video-recorded the animals' behaviours for five consecutive days, one day before and four days after application of the following four treatments: 1st control (no-handling); 2nd control (intra-muscular [IM] injection of saline solution); low-dose ACTH (IM injection of 0.18 ml ACTH); and high-dose ACTH (IM injection of 0.37 ml ACTH). There was a peak in the concentration of GCM in faeces collected 24 h after the injection of the high-dose ACTH treatment. Additionally, independent of the treatments, spotted pacas spent less time on exploration and feeding states, while spending more time in the inactive but awake (IBA) state following the treatment application (challenge day). The use of GCM concentration in faecal samples together with the behavioural changes (less exploration and feeding, and more IBA) showed to be efficient as a non-invasive tool for welfare assessment of farmed spotted paca.

Glucocorticoid concentration and parasitic load in a wild striped hyaena (Hyaena hyaena) population in Southern India

Glucocorticoid levels (GCs) are an important parameter that infers the pathological condition of an organism. GCs can vary with life history stages like gender, age, and reproductive stage and signify the degree of allostatic load experienced. Deleterious increases in GCs can severely affect the wildlife population by suppressing the reproductive viability of individuals and may cause immunosuppressive, making the population more susceptible to parasitic load. We have studied the stress physiology and parasitic load across the life history stages in the striped hyaena population confined to the Moyar Valley of Mudumalai and Sathyamangalam Tiger Reserves. The fecal glucocorticoid metabolite concentrations were measured using cortisol enzyme immunoassay from 30 scats. Collected scats were categorized into adults, sub-adults, and juvenile age groups based on the size and colour of droppings. Sex of the scat samples was identified post-DNA extraction from their epithelial lining cells. Parasitic load from scats was quantified as the number of different parasites recorded through sucrose floatation and sedimentation techniques. We found no significant difference in GCs concentration and parasitic load across the life history stages, but juveniles were recorded to have the least parasitic load. The highest GCs concentration and parasitic load were recorded from adults and female individuals, while both parameters were found least among juvenile individuals. Though we did not observe any significant differences across life-history stages, this study provides the first baseline information on stress physiology and parasitic load in striped hyaenas, which could help further studies assess the impact of external perturbations on stress physiology.

Assessing the drivers of gut microbiome composition in wild redfronted lemurs via longitudinal metacommunity analysis

The gut microbiome influences host’s immunity, development, and metabolism and participates in the gut–brain axis, thus impacting the health of the host. It is a dynamic community varying between individuals and within individuals at different time points. Hence, determining the factors causing this variability may elucidate their impact on host’s health. However, understanding the drivers of variation has proven difficult particularly as multiple interactions occur simultaneously in the gut microbiome. We investigated the factors shaping the gut microbiome by applying the metacommunity concept where the gut microbiome is considered as a microbial community shaped by the interactions within the community, with the host and microbial communities outside the host, this through a longitudinal study in a wild primate. Focal behavioral data were collected for 1 year in four groups of redfronted lemurs to determine individual social and feeding behaviors. In addition, regular fecal samples were collected to assess bacteria, protozoa, and helminths through marker gene analysis and to measure fecal glucocorticoid metabolite (fGCM) concentrations to investigate the impact of physiological stress on the gut microbiome. Higher consumption of leaves and elevated fGCM concentrations correlated with higher alpha diversity, which also differed among groups. The major drivers of variation in beta diversity were group membership, precipitation and fGCM concentrations. We found positive and negative associations between bacterial genera and almost all studied factors. Correlations between bacterial indicator networks and social networks indicate transmission of bacteria between interacting individuals. We detected that processes occurring inside the gut environment are shaping the gut microbiome. Host associated factors such as, HPA axis, dietary changes, and fluctuations in water availability had a greater impact than interactions within the microbial community. The interplay with microbial communities outside the host also shape the gut microbiome through the exchange of bacteria through social relationships between individuals and the acquisition of microorganisms from environmental water sources.

Longitudinal Analysis of Variability in Fecal Glucocorticoid Metabolite Concentrations in Three Orangutans (Pongo pygmaeus pygmaeus and Pongo pygmaeus abelii) before, during, and after Transition from a Regular Habitat Environment to Temporary Housing in Indoor Holding Facilities.

Considerable research has been conducted on the effects of inter-institutional transfers, but far less consideration has been given to intra-institutional transfers and extended housing in off-habitat holding. On 15 May 2018, The Oregon Zoo's orangutans (n = 3) were moved from the Red Ape Reserve (RAR) to the Veterinary Medical Center (VMC) indoor holding areas and remained there until 22 December 2020, resulting in over two years of housing in a facility not specifically designed for orangutans. This study aimed to quantify potential changes in fecal glucocorticoid metabolites (fGM) typically associated with increased adrenal activity as a result of transfers, as well as potential differences in fGM concentrations associated with housing in the two different types of locations. We collected fecal samples from all orangutans during three time periods: the initial housing at RAR (RAR1), the time spent at VMC holding (VMC), and the return to RAR (RAR2). Samples were analyzed using enzyme-immunoassay (EIA) analyses and compared using two-way ANOVA tests with Games-Howell post-hoc evaluations. The results of our analyses showed the following: (1) significant differences in fGM concentrations based on location in two orangutans, with the highest fGM concentration occurring in fecal samples collected at the VMC; and (2) a lack of significant fGM peaks following multiple intra-institutional transfers for all three orangutans. Though requiring further corroboration through future studies, we speculated that pre-transfer behavior training and intensive, continued care by familiar animal care staff may have helped to mitigate the stress responses commonly associated with transfers and major changes in housing. Furthermore, this study highlights the individualistic nature of the stress response, as illustrated by the substantial variation in fGM concentrations across different housing regimens in the three orangutans.

Effects of vessel traffic and ocean noise on gray whale stress hormones

Human use of marinescapes is rapidly increasing, especially in populated nearshore regions where recreational vessel traffic can be dense. Marine animals can have a physiological response to such elevated human activity that can impact individual health and population dynamics. To understand the physiological impacts of vessel traffic on baleen whales, we investigated the adrenal stress response of gray whales (Eschrichtius robustus) to variable vessel traffic levels through an assessment of fecal glucocorticoid metabolite (fGC) concentrations. This analysis was conducted at the individual level, at multiple temporal scales (1–7 days), and accounted for factors that may confound fGC: sex, age, nutritional status, and reproductive state. Data were collected in Oregon, USA, from June to October of 2016–2018. Results indicate significant correlations between fGC, month, and vessel counts from the day prior to fecal sample collection. Furthermore, we show a significant positive correlation between vessel traffic and underwater ambient noise levels, which indicates that noise produced by vessel traffic may be a causal factor for the increased fGC. This study increases knowledge of gray whale physiological response to vessel traffic and may inform management decisions regarding regulations of vessel traffic activities and thresholds near critical whale habitats.

Diurnal pastoralism does not reduce juvenile recruitment nor elevate allostatic load in spotted hyenas

Abstract Anthropogenic activity can have substantial effects on wildlife. These effects may vary according to the characteristics of the activity and the species involved. Although effects on behaviour are well studied, studies of effects on fitness and physiology are scarce, particularly for group‐living species. We exploited a natural experimental setup to investigate the effect of diurnal pastoralism on juvenile recruitment and allostatic load in a population of free‐ranging spotted hyenas in the Ngorongoro Crater, Tanzania, over a 24‐year period. Pastoralism was restricted to the territories of two of the eight study clans, allowing us to compare juvenile recruitment in exposed and unexposed clans. We also compared faecal glucocorticoid metabolite concentrations (fGMC)—a biomarker of an organism's allostatic load—between exposed and unexposed clans using 975 faecal samples from 475 hyenas. We found no detectable difference in juvenile recruitment nor fGMC between the exposed and unexposed clans, indicating that the pastoralism had no substantial deleterious effect on the spotted hyenas. The lack of a deleterious effect likely stems from the combined effect of the predictable and undisruptive nature of the pastoralism, the socio‐ecology of spotted hyenas and the Ngorongoro Crater's consistently abundant prey. Our findings demonstrate that exposure to anthropogenic activity may be compatible with the persistence of certain group‐living species, especially if the overlap between the species' critical behaviours and the activity is limited. Our study thereby provides new perspectives for ecologists, conservation biologists and stakeholders who seek to assess human–wildlife conflicts and balance the needs of local human communities and wildlife.

Native species exhibit physiological habituation to invaders: a reason for hope.

Animals cope with environmental perturbations through the stress response, a set of behavioural and physiological responses aimed to maintain and/or return to homeostasis and enhance fitness. Vertebrate neuroendocrine axis activation in response to environmental stressors can result in the secretion of glucocorticoids (GCs), whose acute increases may be adaptive, while chronic elevation may be detrimental. Invasive grey squirrels (Sciurus carolinensis) act as a stressor eliciting elevation of GCs in native red squirrels (Sciurus vulgaris). Here we used 6-year data of variation in faecal glucocorticoid metabolite (FGM) concentrations following invasion by grey squirrels in three red squirrel populations, to identify if red squirrels showed physiological habituation to this stressor. The decrease in FGMs over time was more pronounced shortly after invasion and at high densities of grey squirrels while it decreased less strongly and was no longer influenced by the invader density as time since invasion elapsed. At the individual level, FGMs also decreased more markedly as each red squirrel experienced prolonged contact with the invader. Our study provides compelling new data suggesting that native species in the wild can habituate to prolonged contact with invasive species, showing that they may avoid the potentially harmful effects of chronic elevations in GCs.

Can stress and anxiety be assessed in African elephants (Loxodonta africana) using self-directed behaviour?

Captive African elephants used in the tourism industry face numerous welfare issues which are often stress related and linked to high numbers of tourists or human-elephant interactions. Elephant welfare is commonly assessed by quantifying faecal glucocorticoid metabolite (fGCM) concentrations, and monitoring stereotypic behaviour, which are either costly or problematic in identifying underlying causes. Self-directed behaviours (SDBs), a form of displacement activity, have been substantially linked with stress and anxiety in primates, and could be a potential but not yet utilised behavioural marker in elephants. We thus explored the usage of several trunk, tail, and body related SDBs in a group of 7 semi-captive African elephants maintained at the Knysna Elephant Park, a tourist destination offering a variety of close contact experiences. Using continuous behavioural sampling (focal following), SDBs were recorded as they occurred, along with the numbers of nearby tourists (<5 m to focal; with 3 levels: None, Low 1–5, and High >5), and the nature of the tourist interaction (5 levels: None, Feed, Touch, Walk, and Ride). We then compared SDB rates with fGCM concentrations (n = 115) determined following observations (1 day and 2 days after behavioural sampling). Data were analysed using Generalized Linear Mixed effects Models. SDB rates (p/min) significantly increased during elephant Walks (p < 0.001) and Rides (p < 0.001), indicating a correlation with their usage during potentially stressful scenarios. The Touch interaction significantly decreased SDBs (p < 0.05), whilst the Feed interaction had a non-significant effect (p > 0.05), which could indicate the element of control in terms of perceived stress is an important component in welfare, as elephants were free to move away during these interactions. Interestingly, SDBs significantly decreased when tourist numbers were High (p = < 0.05), potentially due to greater quantities of high value food. Additionally, no correlation was found between SDBs and fGCM concentrations determined on day one (p > 0.05), and day two (p > 0.05). Hence, SDBs may be correlated more with anxiety or lower-level stress, which is not significant enough to activate GC production, and may therefore act as a coping strategy utilised to maintain physiological homeostasis during anxiety-inducing situations. Additional research would benefit from coupling SDB observations with different forms of physiological assessments to better understand the internal motivations, and formally establish SDBs as a reliable, cost-effective, and non-invasive welfare index to identify stressed individuals in real time and ensure optimum husbandry.

Exploring the effect of auditory stimuli on activity levels, milk yield and faecal glucocorticoid metabolite concentrations in Holstein cows

Health and welfare are inextricably linked within efficient and sustainable dairy production, and several potential risk factors may affect the well-being of dairy cows, including chronic stress. Although auditory stimuli could be used as a tool to decrease the potential stress that cows might experience, it is seldom applied to livestock production systems due to the perception that enrichment is an unnecessary expense. This study aimed to explore the effect of auditory stimuli as a form of enrichment in a Holstein herd by monitoring fecal glucocorticoid metabolite (fGCM) concentrations (a non-invasive, stress-associated biomarker). Cow activity level and milk yield were also measured. Nine cows in their second and third lactation were divided into 3 groups, using a Latin Square experimental design, exposing each cow group to each of the 3 treatments, namely constant exposure (CE), limited exposure (LE), and no exposure (NE) to classical music. FGCMs were quantified using a group-specific enzyme immunoassay detecting 11,17-dioxoandrostanes. Compared to LE and NE animals, cows exposed to constant music had significantly lower fGCM concentrations (P = 0.012), as well as higher milk yields (P < 0.0001) and lowered activity levels during the morning (P = 0.005) and the evening activity period (P = 0.048). These findings indicate that auditory stimuli in the form of classical music may have a positive effect on the welfare of cows as well as milk yield, which hold economic benefits for the producer and potentially reduces the number of cows needed for profitable production.