Fecal Glucocorticoid Metabolites Research Articles

The dissection of a despotic society: exploration, dominance and hormonal traits.

Naked mole-rats (Heterocephalus glaber) live in large colonies with one breeding female (queen), one to three breeding males (BMs) and the remainder are non-reproductive subordinates. The animals have a linear dominance rank with the breeders at the top of the hierarchy. We investigated how dominance rank in naked mole-rats differs with exploration (the propensity to explore a novel environment) and related endocrine markers. Exploration behaviour, faecal progestagen metabolite (fPM), faecal glucocorticoid metabolite (fGCM), faecal androgen metabolite (fAM) and plasma prolactin concentrations were quantified in breeding, high-, middle- and low-ranked females and males from five naked mole-rat colonies. There were no significant differences between the dominance rank and exploration behaviour. Interestingly, the queens and high-ranking females had higher fGCM and fAM concentrations compared with middle- and low-ranked females. The queens had significantly higher fPM concentrations than all other ranked females, since they are responsible for procreation. In the males, the BMs had higher fGCM concentrations compared with high- and low-ranked males. In addition, BMs and middle-ranking males had overall higher prolactin levels than all other ranked males, which could be linked to cooperative care. Overall, the results suggest that physiological reproductive suppression is linked to high dominance rank.

Validation of Fecal Glucocorticoid Metabolites as Non-Invasive Markers for Monitoring Stress in Common Buzzards (Buteo buteo).

For wild animals, being in captivity in wildlife centers can cause considerable stress. Therefore, it is necessary to establish and validate non-invasive tools to measure chronic stress during rehabilitation. Eight Common Buzzards which lived in permanent husbandry were placed individually into prepared aviaries and their feces were collected before, during and after a stress event for biological validation over a period of seven days. The extracted fecal glucocorticoid metabolites (fGCMs) were analyzed with three different enzyme immune assays (EIA) to find the most suitable one. Additionally, we aimed to investigate the stability of fGCM levels after defecation because further metabolization by bacterial enzymes can lead to changed results. The Cortisone-EIA performed best in males and females and showed that the stress event led to an fGCM increase of 629% (557% in females and 702% in males) in relation to basal values. We found no significant differences between the sexes, but observed significant differences between different times of day. FGCM concentration significantly changed after eight hours at room temperature. Our study successfully validated the non-invasive measurement of fGCM as a stress indicator in Common Buzzards and could therefore lay the foundation for future studies providing new insights for animal welfare research in Buzzards.

Effects of reproductive seasonality on the excretion of fecal glucocorticoid metabolites in free‐ranging Pampas Deer

AbstractUnderstanding how fecal glucocorticoid (GC) concentration is associated with reproduction in wild animals allows us to associate physiological stress with the costs of reproduction. Glucocorticoids are among the main stress‐related hormones, and their secretion is strongly associated with reproductive seasonality. Using non‐invasive methods (thereby avoiding causing stress), we used fecal GC metabolites (FGC) to test the hypothesis that the reproductive phase (mating, non‐mating, gestation, and lactation) influences stress levels of the seasonally reproductive Pampas Deer (Ozotoceros bezoarticus). Furthermore, we compared FGC between sexes and between males of different antler statuses (velvet, hard, cast). During 1 year, in the Pantanal of Brazil, we collected 621 fresh fecal samples (327 from females, 294 from males) from which we estimated FGC using enzyme immunoassay (EIA). FGC concentrations varied by reproductive phase and antler status. Agonistic and courtship interactions associated with mating (i.e., fights between males, attempts to mount females), suggest that they influence FGC concentrations most strongly in both sexes. Females also had greater FGC concentrations during lactation, suggesting that this phase and parental care are also associated with increased physiological stress. In males, the association of FGC concentrations with antler status may be, in part, associated with photoperiod and testosterone secretion, both of which can trigger reproductive and agonistic behaviors. Finally, reproductive synchrony of the sexes causes similar FGC over time and suggests that environmental factors contribute as well. We show that non‐invasive monitoring of glucocorticoid metabolites levels is an effective tool for detecting changes in the physiological stress response in Pampas Deer, suggesting that this tool will be useful for detecting changes in physiological stress caused by human disturbance, such as cattle ranching in Pantanal and similar disturbances elsewhere.

Asian elephant calf physiology and mahout perspectives during taming in Myanmar.

A quarter of Asian elephants are captive, with greater than 90% of these tamed and cared for by handlers (mahouts) in Asia. Although taming is a much-discussed welfare issue, no studies to our knowledge have empirically assessed its impact on calves, and dialogue surrounding taming often lacks perspectives of those involved. Here, we interviewed mahouts involved in taming and monitored five physiological measures (faecal glucocorticoid metabolites (FGMs), serum cortisol, glucose, creatine kinase (CK) and heterophil:lymphocyte (H:L)) over the first 10 days of taming and following six months in 41 calves undergoing taming and 16 control individuals. These measures assess the duration and intensity of stress during taming. Interviews suggested mahouts had major concerns for their safety when discussing changing taming practices, an important consideration for future management. Calf physiological measures were elevated by 50-70% (FGMs/cortisol/glucose), 135% (H:L) and greater than 500% (CK) over the first few days of taming, indicative of elevated stress, not seen to the same extent in control adults. Some measures stabilized sooner (glucose/cortisol/CK/FGM: 7-10 days) than others (H:L: one-two months), indicating mostly acute stress. Our findings inform the welfare of approximately 15 000 captive elephants around the world. Future studies should compare taming in different populations and consider calf and mahout welfare.

Faecal glucocorticoid analysis as indicator of stress and its effect on the reproduction hormones of the female Malayan sun bear (Helarctus malayanus) in captivity.

Faecal glucocorticoid metabolite (FGM) is frequently used as a non-invasive tool to assess adrenocortical activity in animal conservation and welfare studies. However, studies assessing the stress level and its effect on the reproductive hormones of female Malayan sun bear is lacking. This study aims to assess the level of stress of the female Malayan sun bear in captivity and its effect on the female reproductive hormones of the female Malayan sun bears in captivity in Malaysian Borneo. Through FGM analysis via the enzyme-linked immunoassay (ELISA) method, it was found that the bear living in BSBCC has the lowest level of glucocorticoid. as for the relationship between the level of stress and the female reproductive hormones, the level of estrogen which was indicated by estradiol, decreases as the level of FGM increases. In contrast, the level of progesterone increase as the level of FGM increases. It can be concluded that the level of stress in captivity could affect the level of estrogen and progesterone and a high level of progesterone does not always indicate a good reproductive function in female mammals.

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.

The Comparative Effect of Occupational and Musical Enrichment on Fecal Glucocorticoid Metabolite Levels in a Captive Colony of Stumptail Macaques (Macaca arctoides).

Environmental enrichment improves captive animal welfare by reducing stress-related behaviors. Previous studies in a captive colony of stumptail macaques (Macaca arctoides) reported a reduction of aggression, coprophilia, and stereotypic behaviors after an occupational enrichment program; however, the effect on stress hormones such as glucocorticoids has not been investigated yet. The goal of this study was to compare the effect of sex, age, and social rank on changes in fecal glucocorticoid metabolites (fGCM) after applying two kinds of enrichments (occupational vs. musical) in a captive colony of stumptail macaques. We collected 234 fecal samples from 25 stumptail macaques under the following conditions: (1) basal (no enrichment), (2) three weeks of occupational enrichment, and (3) three weeks of relaxing/classical music. The Generalized Estimated Equation Model showed an increase in fGCM levels after the occupational enrichment only in adult subjects (p = 0.003 compared to basal). The fGCM levels reached by the adults after the occupational enrichment was higher than that of juveniles (p = 0.002) and subadults (p = 0.02). Occupational and musical enrichment decreased fGCM levels only in middle-ranking individuals (p < 0.001 and p = 0.013, respectively). No sex differences were found. In conclusion, there were age and rank differences in individuals' physiological reactivity to the effects of environmental enrichment which need to be considered when planning enrichment programs.

Physiological changes in captive elephants in northern Thailand as a result of the COVID-19 tourism ban-stress biomarkers.

The international travel ban instituted by the Thai government in March 2020 in response to the COVID-19 pandemic greatly affected how tourist camp elephants were managed, with reductions in exercise opportunities, longer chaining hours, and diminished food provisioning. This study was conducted to determine how those changes affected health and welfare biomarkers in individual elephants over the 2 years of the countrywide lockdown (April 2020-April 2022). Blood and fecal samples were collected from 58 elephants at six camps (monthly in Year 1, quarterly in Year 2) and analyzed for stress biomarkers - fecal glucocorticoid metabolites (fGCM), serum oxidative stress [malondialdehyde (MDA) and 8-hydroxy-2'-deoxyguanosine (8-OHdG)], and stress leukograms. Overall, fGCM concentrations increased within the first few months and remained higher than pre-COVID levels, as did the H/L ratio, a measure affected by cortisol. Serum 8-OHdG, an indicator of DNA oxidative damage, also increased over time, while monocytosis and lymphopenia further suggested alterations in immune function as a result of stress. By contrast, another marker of oxidative stress, serum MDA, declined, possibly in response to reduced roughage and supplement intake. A notable finding was a seasonal pattern of fGCM that was significantly different from previous studies. Whereas higher fGCM during the rainy season were observed in this study, previously, concentrations were highest during the winter, high tourist season. Thus, ironically, both the presence and absence of tourists have been associated with increased fGCM concentrations, albeit for different reasons. Camp management factors negatively affecting stress outcomes included shorter chain lengths, longer chain hours, lack of exercise, and reduced roughage and supplements. Overall, it was clear that camps struggled to maintain adequate care for elephants during the COVID-19 pandemic, highlighting the importance of tourist income and need for contingency plans to cope with potential future disruptions to tourism.

Little agreement among methodologies to determine fecal glucocorticoid metabolites in a mountain ungulate

Faecal glucocorticoid metabolites (FGMs) have gained relevance in ecological studies and population monitoring, allowing non-invasive remote sampling without the need to capture and handle animals. Enzyme immunoassays (EIAs) and radioimmunoassays (RIAs) are commonly used for FGM determination. Although these methods should be validated for each species, non-validated tests are still widely used to determine FGMs in wildlife. Near-infrared reflectance spectroscopy (NIRS) is a predictive method requiring calibration against reference methods used to assess FGMs in wildlife. EIAs and RIAs have been utilized to determine FGMs in chamois (Rupicapra spp.), a medium-sized mountain ungulate.This study aims to assess the potential of NIRS to determine FGMs and to evaluate the correlation among analytical methods used to determine FGMs in chamois. Faecal samples from 125 Alpine chamois (Rupicapra rupicapra rupicapra) and 125 Pyrenean chamois (R. pyrenaica pyrenaica) were collected from the field, frozen at − 20 °C, lyophilized, grounded, and scanned using a NIRSystems 5000 monochromator over a 1108–2492 nm wavelength. After this non-destructive NIRS analysis, FGMs were extracted and analysed using four immunoassays previously used in chamois studies: a 125-I-corticosterone RIA, a cortisol EIA, and two 11-oxoetiocholanolone EIAs (72a and 72T). Only the 11-oxoetiocholanolone 72T EIA has been validated for Alpine chamois. NIRS predictions were calibrated and cross-validated for each of the four immunoassays. The correlation among the four immunoassays was assessed using Spearman's rank.The coefficient of determination for NIRS calibration (R²) values ranged from 0.37 to 0.75, and the ratio of performance to deviation values from 1.2 to 1.6. Therefore, NIRS could not predict FGM concentration in chamois faeces. This could be due to the complexity and variability of the FGM detected by the immunoassays as reference methods, and by potential interference of other compounds in the faecal matrix. The correlation among the immunoassays was low overall.As a conclusion, NIRS cannot be recommended for measuring FGMs in chamois. The low correlation among the immunoassays used for FGM determination raises concern about the reliability of previous studies using non-validated methods in chamois. Only biologically validated tests should be used to assess FGMs to avoid incorrect inferences about biological responses in physiological, conservational, and ecological studies or population monitoring. These conclusions are applicable beyond the species studied here.

Analytical and biological validation of a noninvasive measurement of glucocorticoid metabolites in feces of Geoffroy's spider monkeys (Ateles geoffroyi).

We report on an analytical and biological validation of a commercial cortisol enzyme immunoassay (EIA)to measure glucocorticoids (GC)in feces of Geoffroy's spider monkeys (Ateles geoffroyi). Validation of endocrinological methods for each sample matrix and study species is crucial to establish that the methods produce reliable results. For the analytical validation of the EIA, we assessed parallelism, accuracy, and precision. We carried out a biological validation based on three well-studied GC patterns with the following predictions: (1) increased fecal GC metabolite (fGCM)concentrations after veterinary intervention; (2) increased fGCM concentrations during early morning hours; and (3) higher fGCM concentrations during gestation than in other female reproductive states. For the first prediction, we sampled feces of two zoo-housed females 2 days before, the day of, and 2 days after a veterinary intervention. For the second prediction, we analyzed 284 fecal samples collected from 12 wild males using a linear mixed model (LMM).For the third prediction, we analyzed 269 fecal samples of eight wild females using an LMM. Analytical validation revealed that the EIA showed parallelism, was accurate, and precise within each assay. However, there was elevated variation in between-assay precision. The biological validation supported all predictions: (1) the two zoo-housed females showed a substantial increase in fGCM concentrations 2.5and 11 h after veterinary intervention; (2) there was a negative effect of sample collection time on fGCM concentrations (i.e., higher concentrations during early morning); (3) gestating females had significantly higher fGCM concentrations than lactating females. Thus, we analytically validated the commercial EIA and, despite between-assay variation, we were able to find three biologically relevant GC signals in captive and wild settings, and in males and females. We are therefore confident that the method can be used to noninvasively address behavioral endocrinology questions in Geoffroy's spider monkeys.

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.

Physiological stress in eastern black rhinoceros (Diceros bicornis michaeli) as influenced by their density, climatological variables and sexes

AbstractIt is important to understand the physiological stressors in animals especially for threatened species or intensively managed to improve their conservation and optimise their reproduction. We sought to understand changes in stress hormones (faecal glucocorticoid metabolites) in black rhinoceros (Diceros bicornis michaeli) in relation to population density and sex (intrinsic factors) and plant minerals, rainfall and land surface temperature (extrinsic factors). We used non‐invasive faecal sampling techniques on animals of known sex, age and dominance in seven populations of contrasting population densities over 1 year. We measured variability in faecal corticosterone metabolites through radioimmunoassay and related them to population density, sex and faecal calcium, phosphorus, copper, zinc and potassium as characteristic of plant minerals, rainfall and temperature. We used linear mixed models (LMM) to analyse the data. We did not detect a significant relationship between physiological stress parameters and population density. However, we have indications that stress levels increased as rainfall and temperature increased and were correlated negatively and positively with concentration of faecal phosphorus and copper respectively; we found higher stress levels in females than in males. These results suggest that both intrinsic and extrinsic factors explain the variation in physiological stress observed in black rhinoceros.

Physiological response after translocation differs between source populations in a threatened mammal

Conservation translocations are an important tool in the prevention of species loss, but the translocation process is associated with numerous stressors. Non-invasively monitoring stress physiology via faecal glucocorticoid metabolites (FGMs) can provide valuable insights into factors impacting translocation success and how to mitigate negative impacts. After validating an assay to measure FGMs in greater stick-nest rats ( Leporillus conditor ), we examined whether translocation caused a predictable change in physiology. We compared longer-term (one to five months post-translocation) physiological responses across three source populations (remnant-wild, reintroduced-wild, captive-bred), and investigated effects of body condition and sex on FGMs. Notably, FGMs of the remnant-wild population did not significantly change post-translocation, while the reintroduced-wild population exhibited a significant decrease and the captive-bred population a significant increase. Individuals in lower body condition had the highest FGMs in both wild-type populations, whereas the captive-bred population showed the opposite relationship. There was no difference in FGMs between the sexes. Our work highlights that physiological responses after translocation may not be uniform and source population history is an important factor to be considered, emphasizing the need for novel ideas that facilitate successful adaptation. By better understanding how species and individuals respond to translocation, we can improve translocation outcomes.

Changes in Fecal Glucocorticoid Metabolites in Captive Coyotes (Canis latrans): Influence of Gender, Time, and Reproductive Status.

Reproduction is considered an energetically and physiologically demanding time in the life of an animal. Changes in physiological stress are partly reflected in changes in glucocorticoid metabolites and can be measured from fecal samples. We examined levels of fecal glucocorticoid metabolites (fGCMs) in 24 captive coyotes (Canis latrans) to investigate responses to the demands of reproduction. Using 12 pairs of coyotes (five pairs produced pups, seven pairs did not), we analyzed 633 fecal samples covering 11 biological periods (e.g., breeding, gestation, and lactation). Levels of fGCMs showed high individual variability, with females having higher fGCM levels than males. The production of pups showed no statistical effect on fGCM levels among females or males. Among females, fGCM levels were highest during 4-6 weeks of gestation compared to other periods but were not significantly different between pregnant and nonpregnant females. Among males, the highest fGCM levels were during 1-3 weeks of gestation compared to other periods, but were not significantly different between males with a pregnant mate versus nonpregnant mate. Of females producing pups, litter size did not influence fGCM levels. Given that they were fed ample food throughout the year, we found that the demands of producing pups did not appear to statistically influence measures of fGCM concentrations in captive coyotes.

Reaction to Novel Objects and Fecal Glucocorticoid Metabolite Levels in Two Species of Nocturnal Geckos.

Many reptiles are maintained in captivity and heavily traded, although welfare measures for many species are not well established and are under-researched compared to other animals. In this study, we focused on two of these species: crested geckos (Correlophus ciliatus) and leopard geckos (Eublepharis macularius). To better interpret their behavior in captivity, the individual reaction to novel objects and the fecal glucocorticoid metabolite levels were measured in an attempt to identify the potential correlation between them. Also, we explored if some characteristic of the objects (e.g., color, shape, or smell) resulted in being more attractive to some species and/or individuals. Equivalent responses to different objects were not obtained for all the geckos, the behavioral response being highly individual and context-dependent, although modulated by the species. Individuals which manipulated earlier and interacted longer with novel objects showed lower basal fecal corticosterone metabolite (FCM) levels. Differences according to the species suggested that crested geckos have significantly greater and more variable FCM levels than leopard geckos. Our results can help to understand the reaction of geckos to novelty and have the potential to serve in their welfare assessment, although more studies are needed to proper establish welfare protocols.

Noise intensity modulates the responses of mantled howler monkeys to anthropophony.

Anthropogenic noise is a major global pollutant but its effects on primates are poorly understood, limiting our ability to develop mitigation actions that favor their welfare and conservation. In this study, we used an experimental approach to determine the impact of variation in noise intensity on mantled howler monkeys (Alouatta palliata). We conducted the study at Los Tuxtlas (México), where we studied the physiological stress (proxied via fecal glucocorticoid metabolites, fGCM) and behavioral responses of 16 males. We played back chainsaw noise at two intensities (40 and 80 dB) and used days in which groups were not exposed to noise as matched controls. With increased noise intensity fGCM increased, vigilance and vocalizations were longer, and vigilance, vocalizations, and flight occurred quicker. Physiological and behavioral responses occurred even after low-intensity noise playbacks (i.e., 40 dB). Therefore, noise intensity is a significant factor explaining the responses of mantled howler monkeys to anthropogenic noise. These results imply that management actions aimed at eradicating anthropogenic noise are required for the conservation and welfare of mantled howler monkeys at Los Tuxtlas.

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.

Identifying potential measures of stress and disturbance during a captive to wild African elephant reintegration.

There is increasing evidence of compromised welfare for elephants managed in captivity. Should such facilities eventually close, more elephants will need to be rehabilitated and reintegrated into the wild. The goal of such reintegration would be to restore any physical or psychological aspects of the elephant that may have been compromised in captivity, followed by introduction into a free-roaming system where they can interact with other elephants. However, to achieve this goal, the reintegration methods implemented need to be assessed to ensure that welfare remains the priority. The objective of this study was to test whether parameters generally associated with stress and disturbance in African elephants, respond to changes in potentially stressful environmental conditions, assessed at multiple temporal scales ranging from minutes to months. The main changes in environmental conditions that were investigated included the different phases of reintegration of a group of elephants from captivity into the wild. Stress and disturbance related parameters used for comparisons included physiological responses, namely the extent of temporal gland secretions (eTGS) and faecal glucocorticoid metabolite (fGCM) levels as well as behavioural responses, namely the display of stereotype and stress-related behaviours. Results showed that eTGS significantly increased during the initial release of the elephants compared to when in captivity. Stereotypic behaviours were only recorded during the captive phase and immediately ceased after release. Faecal GCM levels spiked in the first year after release before decreasing back to pre-reintegration levels during the third year. These findings indicate that fGCM levels, the eTGS and disturbance related behaviours all proved effective in explaining the changes in stress and disturbance experienced by elephants during the initial years after being reintegrated from captivity into the wild.