Low-ranking individuals present high and unstable fecal cortisol levels in provisioned free-ranging adult male rhesus macaques (<Emphasis Type="Italic">Macaca mulatta</Emphasis>) during the birth season in a mountain area of northern China

Social hierarchy commonly exists in animal societies, affecting both the endocrine functioning and the behavior of animals. In nonhuman primates, the relationship between social rank and cortisol levels varies across species and even within species. Here, we assessed the relationships between social rank and fecal cortisol levels in adult male Taihangshan macaques (rhesus macaques, Macaca mulatta tcheliensis) from the provisioned, free-ranging Wulongkou-2 (WLK-2) group inhabiting Wulongkou Scenic Area, Jiyuan, China. From March to May 2014, we recorded 195 agonistic behaviors and collected 54 fresh fecal samples from eight adult male Taihangshan macaques. Males were assigned a social rank according to an agonistic behavior matrix, and enzyme-linked immunosorbent assay was then used to measure the cortisol concentration in the fecal samples. We found that social rank among the eight male Taihangshan macaques in WLK-2 group followed a strict linear hierarchy, and that fecal cortisol levels were significantly higher and more variable in low-ranking males than in more dominant individuals. Age was not significantly associated with social rank or fecal cortisol levels. Our results suggest that social rank and maintenance of the social hierarchy within the WLK-2 group is a chronic stressor, with low-ranking males maintaining heightened stress levels and enlarged reactive scope relative to dominant males. This provides new support for the theory that social environment can influence endocrine functioning.     

Comparison of Endocrine Response to Stress Between Captive-Raised and Wild-Caught Bighorn Sheep

ABSTRACT Stress hormones in Rocky Mountain bighorn sheep (Ovis canadensis canadensis), produced in response to environmental changes, road development, or high population density, may impact their immune systems to a threshold level that predisposes them to periodic, large-scale mortality. We compared the stress response to a novel environmental situation and repeated handling between bighorn sheep born and raised in captivity (CR) and bighorn sheep born in the wild (WC) and brought into captivity. We measured plasma epinephrine, norepinephrine, cortisol, and fecal glucocorticoid metabolites (FGM). Three weeks after each group's arrival we used a one-time drop-net event to elicit an acute stress response, and we collected blood samples from each sheep over 35 minutes, as well as one fecal sample. We collected blood and fecal samples from both groups on 7 other occasions over the subsequent 6 months. We also collected fecal samples from the pen at approximately 24-hour intervals for 3 days following every handling event to monitor the stress response to handling. We found that CR sheep had a stronger autonomic nervous system response than WC sheep, as measured by epinephrine and norepinephrine levels, but we found a very similar hypothalamic-pituitary–adrenal axis (HPA) response, measured by cortisol levels, to the acute stress event of a drop-net restraint. We also found that once the WC sheep had acclimated, as indicated by the return to the initial baseline FGM levels within 12 weeks, the CR and WC groups' HPA responses to sampling events were not significantly different from one another. Fecal samples can provide a noninvasive mechanism for managers to monitor baseline FGM for a given herd. Using long-term monitoring of FGM rather than values from a single point in time may allow managers to correlate these levels to outside influences on the herd and better understand the impacts of management changes, population density, or increased human developments on the health of the sheep population.     

Relocation and Hair Cortisol Concentrations in New Zealand White Rabbits

To investigate how long relocation modified hair cortisol concentrations in New Zealand white rabbits, 19 rabbits were subjected to a change in their breeding facility at the beginning of the trial and then were kept under stable environmental conditions. Hair samples were collected at the time of arrival to the nonhuman animal facility and at 40-day intervals from the same skin area for up to 440 days after the animals' arrival to the facility. A period effect on the hair cortisol concentration was found (p < .01). The transfer of the rabbits to the new facility might have induced an increase in the hypothalamic-pituitary-adrenal axis activity (p < .01). A second increase in hair cortisol concentration (p < .01) occurred at 320 days, after a change of personnel at the facility that occurred at 280 days, which was the only environmental change. The relocation of rabbits to the facility resulted in a stress response leading to elevated cortisol levels. The effect of relocation on mean cortisol concentrations was exhausted within 120 days when all environmental factors were kept stable.     

A Case Study: Fecal Corticosteroid and Behavior as Indicators of Welfare During Relocation of an Asian Elephant

This study was a preliminary investigation of an enzyme immunoassay for measuring fecal glucocorticoid metabolites in a male Asian elephant (Elephas maximus) by investigating changes in behavior and cortisol metabolite excretion associated with a putative stressful event. The study collected fecal samples for 10 days prior to, and 10 days after, 24-hr transport and relocation of the elephant to a new herd. The study measured cortisol metabolites using 2 enzyme immunoassays indicating a 389% and 340% increase in cortisol metabolite excretion following relocation. Maximal cortisol metabolite excretion occurred 2 days after relocation and remained elevated during establishment of the new herd. Stereotypic behavior increased approximately 400% after relocation. The relocation disturbed sleep patterns, the elephant spent less time sleeping during the night, and the elephant slept standing up. These results provide preliminary evidence that noninvasive monitoring of fecal cortisol metabolites can be used to investigate adrenal activity in Asian elephants and may be a safe, practical, and accurate welfare indicator.     

Familiarity may buffer the adverse effects of relocation on marmosets (Callithrix kuhlii): Preliminary evidence

The relocation of exotic species is increasingly crucial for the proper management of captive populations. We investigated the effects of relocation on urinary cortisol levels and proximity behavior in four Wied's marmosets (Callithrix kuhlii). Two of the relocated marmosets constituted a well‐established pair, whereas two (the newly established pair) were introduced when they arrived at the new facility. Four adult females served as control animals in which we assessed cortisol response to pair formation in the absence of relocation. Urinary cortisol in the relocated subjects was monitored for 4 weeks before and 12 weeks after the relocation event. Urinary cortisol in the control subjects was monitored for 4 weeks before and 8 weeks after the pairs were formed. Proximity was monitored in the relocated subjects for 12 weeks following the move. After the animals were relocated, the newly established pair had significantly elevated levels of urinary cortisol compared to the well‐established pair and control females, which lasted at least 8 weeks. For both pairs, cortisol levels decreased over time across the 12 weeks following the relocation event. The members of the well‐established pair were in contact with each other for 65% of the intervals sampled in the 2 weeks immediately following relocation, compared to only 12% in the newly established pair. These preliminary findings suggest that relocation is associated with significant adverse physiological and behavioral changes that may be ameliorated by the presence of a highly familiar social partner. The study highlights the importance of future research in this field. Zoo Biol 24:93–100, 2005. © 2005 Wiley‐Liss, Inc.     

A case study: fecal corticosteroid and behavior as indicators of welfare during relocation of an Asian elephant

This study was a preliminary investigation of an enzyme immunoassay for measuring fecal glucocorticoid metabolites in a male Asian elephant (Elephas maximus) by investigating changes in behavior and cortisol metabolite excretion associated with a putative stressful event. The study collected fecal samples for 10 days prior to, and 10 days after, 24-hr transport and relocation of the elephant to a new herd. The study measured cortisol metabolites using 2 enzyme immunoassays indicating a 389% and 340% increase in cortisol metabolite excretion following relocation. Maximal cortisol metabolite excretion occurred 2 days after relocation and remained elevated during establishment of the new herd. Stereotypic behavior increased approximately 400% after relocation. The relocation disturbed sleep patterns, the elephant spent less time sleeping during the night, and the elephant slept standing up. These results provide preliminary evidence that noninvasive monitoring of fecal cortisol metabolites can be used to investigate adrenal activity in Asian elephants and may be a safe, practical, and accurate welfare indicator.     

Response of fecal cortisol to stress in captive chimpanzees (Pan troglodytes)

This study examined whether fecal cortisol could be used as an index of stress responses. The stress responsiveness of fecal cortisol was tested with a stressor known to stimulate adrenal activity, the stress of anesthesia. Daily fecal and urine samples were collected from four captive chimpanzees (Pan troglodytes) before and after anesthetizations with Telazol^®/Ketasat^®. Tests of assay validity indicated that cortisol was measurable in chimpanzee fecal extracts. Fecal cortisol concentrations were significantly elevated 2 days after anesthetization, with elevations in seven of the eight treatments. The posttreatment peak was significantly greater than baseline values in three of the four subjects. Both fecal concentrations and proportionate increases in response to stress were significantly correlated with the corresponding values in urinary cortisol, confirming the stressfulness of these procedures and the stress responsiveness of fecal cortisol. These findings provide evidence for the application of fecal cortisol as a noninvasive index of physiologic stress in nonhuman primates. Am. J. Primatol. 44:57–69, 1998. © 1998 Wiley-Liss, Inc.     

Optimal group size and seasonal stress in ring-tailed lemurs (Lemur catta)

Adaptive explanations for social grouping assume that thereare fitness consequences associated with group size, and individualsmaintain membership in groups of favorable size to maximizefitness. Here I examine fecal cortisol concentrations as a hormonalmeasure of stress to assess the relative well-being of Lemurcatta in groups of different size and in seasons of normal andlow tamarind fruit availability. I test the hypotheses thatthere is an optimal group size at which cortisol is lowest andthat optimal group size changes in food-scarce conditions. Icollected 799 fecal samples from 87 individuals in seven free-rangingL. catta groups at Berenty Reserve, Madagascar, over a 1-yearperiod (August 1999–July 2000) and determined fecal cortisolconcentrations using a radioimmunoassay. Expressing these asresiduals from monthly population means to control for temporalfluctuations in cortisol concentration, I calculated mean fecalcortisol levels for each animal in seasons of normal and lowtamarind fruit abundance and over the entire year. Overall,females exhibited lowest mean cortisol levels in groups of intermediatesize, suggesting that there are benefits to maintaining membershipin these groups. Females in groups that were atypically largeor small for their habitat type had higher mean cortisol levelsthan typical groups. Cortisol levels increased in food-scarceconditions for larger groups, suggesting that intergroup competitiveadvantages do not outweigh intragroup feeding competition atthis time. Group size may be optimized for long-term averageconditions, and short-term stresses may intermittently alterthe costs associated with group size.     

Foraging Success,Agonism, and Predator Alarms: Behavioral Predictors of Cortisol in <Emphasis Type="Italic">Lemur catta</Emphasis>

Primate social grouping is understood as an adaptive strategy for mitigating environmental selection pressures, but the relative importance of various pressures may vary. Physiological measures of well-being can show their short-term impacts and suggest their relative importance and capacity to provide ultimate or proximate control of group size. I examined correlations between pressures commonly proposed as causes of social grouping (foraging success, intergroup and intragroup agonism, and predation risk) and individual levels of fecal cortisol, a hormonal stress measure, in a free-ranging population of Lemur catta. I collected behavioral data on 45 female Lemur catta at Berenty Reserve, Madagascar, over 3 seasons (August 1999-July 2000) and determined individual cortisol levels from 474 fecal samples. Neither predator alarm rates nor intragroup agonism rates correlated with cortisol levels in any season. However, females with low daily food intake and high rates of escalated intergroup defense exhibited higher cortisol levels. The data suggest that acquisition and defense of food resources are principal challenges in Lemur catta, and may be important factors determining social grouping and other behavioral or life history adaptations.     

Factors affecting fecal glucocorticoid levels in semi-free-ranging female mandrills (Mandrillus sphinx)

Subordinate female cercopithecine primates often experience decreased reproductive success in comparison with high-ranking females, with a later age at sexual maturity and first reproduction and/or longer interbirth intervals. One explanation that has traditionally been advanced to explain this is high levels of chronic social stress in subordinates, resulting from agonistic and aggressive interactions and leading to higher basal levels of glucocorticoids. We assessed the relationships among fecal cortisol levels and reproductive condition, dominance rank, degree of social support, and fertility in female mandrills (Mandrillus sphinx) living in a semi-free-ranging colony in Franceville, Gabon. Lower-ranking females in this colony have a reproductive disadvantage relative to higher-ranking females, and we were interested in determining whether this relationship between dominance rank and reproductive success is mediated through stress hormones. We analyzed 340 fecal samples from 19 females, collected over a 14-month period. We found that pregnant females experienced higher fecal cortisol levels than cycling or lactating females. This is similar to results for other primate species and is likely owing to increased metabolic demands and interactions between the hypothalamus-pituitary-adrenal axis, estrogen, and placental production of corticotrophin releasing hormones during pregnancy. There was no influence of dominance rank on fecal cortisol levels, suggesting that subordinate females do not suffer chronic stress. This may be because female mandrills have a stable social hierarchy, with low levels of aggression and high social support. However, we found no relationship between matriline size, as a measure of social support, and fecal cortisol levels. Subordinates may be able to avoid aggression from dominants in the large enclosure or may react only transiently to specific aggressive events, rather than continuously expecting them. Finally, we found no relationship between fecal cortisol levels and fertility. There was no difference in fecal cortisol levels between conceptive and nonconceptive cycles, and no significant relationship between fecal cortisol level and either the length of postpartum amenorrhea or the number of cycles before conception. This suggests that the influence of dominance rank on female reproductive success in this population is not mediated through chronic stress in subordinate females, and that alternative explanations of the relationship between social rank and reproduction should be sought.     

Sources of variation in fecal cortisol levels in howler monkeys in belize

High cortisol levels are known to cause low fecundity and increased mortality; thus, the prospect of using cortisol as a measure of population health is an exciting one. However, because so many factors can interact to influence cortisol release, it can be difficult to interpret what exactly is creating changes to cortisol levels. This study investigates variation in fecal cortisol levels in a population of black howlers (Alouatta pigra) from 350 fecal samples collected from 33 individuals in more than 4 years. A general linear mixed model revealed that cortisol varied significantly with fruit availability and contact with tourists. When fruit availability was low, cortisol increased, likely because when fruit availability is low monkeys eat less fruit, thus obtaining less sugar. This result may simply reflect cortisol's metabolic function of mobilizing glucose. It also indicates that these monkeys may be experiencing periods of food stress throughout the year, which was earlier thought to be minimal for a primarily folivorous species. Presence of tourists was the only other factor found to lead to high cortisol; with exposure to tourists increasing stress levels. These results highlight the importance of understanding how physiological factors can influence cortisol, making it easier to interpret results and determine the external social or ecological stressors that may increase cortisol. Am. J. Primatol. 72:600–606, 2010. © 2010 Wiley‐Liss, Inc.     

季节和等级顺位对雄性黄山短尾猴粪样皮质醇水平的影响

在长期的进化选择压力下,灵长类动物大多数群居生活,群体中的动物个体也面临来自群内成员强度不一的社会压力,压力大小的重要生理指标通常用其体内的皮质醇水平的变化来衡量.季节性繁殖的黄山短尾猴(Macaca thibetana),群体内的个体有严格的社会等级,是研究等级顺位与社会压力水平之间关系的理想研究对象.我们采集13只...     

Impact of zoo visitors on the fecal cortisol levels and behavior of an endangered species: Indian blackbuck (Antelope cervicapra L.)

This study investigated behavioral activities (resting, moving, aggressive, social, and reproductive behavior) and fecal cortisol levels in 8 individually identified adult male blackbucks during periods of varying levels of zoo visitors (zero, low, high, and extremely high zoo visitor density). This study also elucidated whether zoo visitor density could disturb nonhuman animal welfare. This study analyzed fecal cortisol from the samples of blackbuck by radioimmunoassay and found significant differences (p < .05) for time the animals devoted to moving, resting, aggressive, reproductive, and social behavior on days with high and extremely high levels of zoo visitors. The ANOVA with Duncan's Multiple Range Test test showed that the fecal cortisol concentration was higher (p < .05) during the extremely high (137.30 ± 5.88 ng/g dry feces) and high (113.51 ± 3.70 ng/g dry feces) levels of zoo visitor density. The results of the study suggest that zoo visitor density affected behavior and adrenocortical secretion in Indian Blackbuck, and this may indicate an animal welfare problem.     

Impact of Zoo Visitors on the Fecal Cortisol Levels and Behavior of an Endangered Species: Indian Blackbuck (Antelope cervicapra L.)

This study investigated behavioral activities (resting, moving, aggressive, social, and reproductive behavior) and fecal cortisol levels in 8 individually identified adult male blackbucks during periods of varying levels of zoo visitors (zero, low, high, and extremely high zoo visitor density). This study also elucidated whether zoo visitor density could disturb nonhuman animal welfare. This study analyzed fecal cortisol from the samples of blackbuck by radioimmunoassay and found significant differences (p < .05) for time the animals devoted to moving, resting, aggressive, reproductive, and social behavior on days with high and extremely high levels of zoo visitors. The ANOVA with Duncan's Multiple Range Test test showed that the fecal cortisol concentration was higher (p < .05) during the extremely high (137.30 ± 5.88 ng/g dry feces) and high (113.51 ± 3.70 ng/g dry feces) levels of zoo visitor density. The results of the study suggest that zoo visitor density affected behavior and adrenocortical secretion in Indian Blackbuck, and this may indicate an animal welfare problem.     

The influence of sex and relatedness on stress response in common marmosets (Callithrix jacchus)

Research in stress physiology has demonstrated the benefits of receiving social support during stressful conditions. However, recent data have shown that the efficacy of social support in buffering physiological and behavioral responses to stressor agents depends on species, sex, and relatedness among animals. This study investigated whether different kinds of social support (presence of same sex related or nonrelated conspecifics) have the same effect on hormonal (fecal cortisol levels) and behavioral responses (agonistic: scent-marking and individual piloerection; anxiety: locomotion; tension-reducing: autogrooming, allogrooming, and body contact). We used adult male and female isosexual dyads of Callithrix jacchus, a small Neotropical primate from the Callitrichidae family, widely used in the study of stress and related diseases. Following a 28-day baseline phase, dyads faced three challenging situations (phase 1: dyads were moved together from the baseline cage to a similar new cage; phase 2: each dyad member was moved alone to a new cage; and phase 3: dyad members were reunited in the same baseline cage). Type of social support was found to influence the response to stressors differently for each sex. Related male dyads did not change their hormonal or behavioral profile over the three experimental phases, when compared to the baseline phase. For nonrelated male dyads, social support buffered hormonal but not behavioral response. For females, the social support offered by a related and nonrelated animal, does not seem to buffer the stress response, as shown by correlations between agonistic behaviors versus cortisol and locomotion during all three experimental phases and a significant increase in fecal cortisol levels during phases 2 and 3, when compared with baseline levels. The results only partially support the buffering model theory and corroborate other studies reporting that the benefits of social support during a period of crisis arise only when it is adaptive for that species.     

Bigger groups have fewer parasites and similar cortisol levels: a multi-group analysis in red colobus monkeys

If stress and disease impose fitness costs, and if those costs vary as a function of group size, then stress and disease should exert selection pressures on group size. We assessed the relationships between group size, stress, and parasite infections across nine groups of red colobus monkeys (Procolobus rufomitratus) in Kibale National Park, Uganda. We used fecal cortisol as a measure of physiological stress and examined fecal samples to assess the prevalence and intensity of gastrointestinal helminth infections. We also examined the effect of behaviors that could potentially reduce parasite transmission (e.g., increasing group spread and reducing social interactions). We found that cortisol was not significantly related to group size, but parasite prevalence was negatively related to group size and group spread. The observed increase in group spread could have reduced the rate of parasite transmission in larger groups; however, it is not clear whether this was a density-dependent behavioral counter-strategy to infection or a response to food competition that also reduced parasite transmission. The results do not support the suggestion that gastrointestinal parasitism or stress directly imposed group-size-related fitness costs, and we cannot conclude that they are among the mechanisms limiting group size in red colobus monkeys.     

Fecal cortisol levels in free-ranging female chacma baboons: relationship to dominance, reproductive state and environmental factors

Savannah baboons are one of the few mammalian species that do not exhibit seasonal reproduction patterns and are therefore ideally suited to study the effect of female reproductive states (cycling, pregnant, lactating) on cortisol levels independent of seasonal factors. Fecal samples from 10 free-ranging female chacma baboons (Papio hamadryas ursinus), collected during a period of 17 months, were analyzed using a steroid-extraction method. Reproductive state had a significant effect on fecal cortisol, with lowest levels found in estrous females. Fertility was not related to fecal cortisol levels; we found no significant differences between samples collected on conceptive and nonconceptive cycles. Environmental factors explained most of the variance of fecal cortisol levels. Cortisol measures were strongly correlated with seasonal differences such as daylight duration, temperature and the amount of time that baboons spent resting. We measured higher cortisol levels during winter months and suggest that this could be related to shorter resting periods and to the cold minimum ambient temperatures at this study site. Finally, we found no relationship between social rank nor the rate of agonistic interactions with basal fecal cortisol levels.     

Urinary and fecal immunoglobulin A, cortisol and 11-17 dioxoandrostanes, and serum cortisol in metabolic cage housed female cynomolgus monkeys (Macaca fascicularis)

BACKGROUND AND METHODS: Quantitative enzyme-immunoassays of urinary and fecal immunoglobulin A (IgA), cortisol and 11-17-dioxoandrostanes (11,17-DOA), and serum cortisol in eight metabolic-cage-housed female cynomolgus monkeys were performed. The monkeys were divided into two groups, B and NB. Group B animals were blood sampled every 6 hours, whereas Group NB animals were not handled/blood sampled. RESULTS: No differences were recorded between the amounts of feces and urine excreted by the two groups. Group B animals excreted more urinary cortisol than did Group NB animals indicating that restraint-blood sampling resulted in a stress response. Excreted amounts of IgA and 11,17-DOA (urine and feces) did not differ between the groups. CONCLUSIONS: Urinary cortisol was a reliable marker of the stress associated with repeated blood sampling. Declining amounts of excreted urinary cortisol indicated that cynomolgus monkeys acclimated quickly to repeated blood sampling in metabolism cages. Within and between animal variation in amounts of feces voided demonstrated the importance of expressing fecal markers as 'amounts excreted per time unit per kg body weight' rather than just measuring the concentrations in fecal samples.     

Reproductive tactics influence cortisol levels in individual male gray-cheeked mangabeys (Lophocebus albigena)

Concentration of the hormone cortisol is often used as an indicator of stress, and chronically high cortisol levels are often associated with poor health. Among group living animals that compete for resources, agonistic social interactions can be expected to contribute to variation in cortisol levels within and among individuals over time. Reproductive tactics of males can change with individual quality, relatedness, and social structure, and affect cortisol levels. In gray-cheeked mangabey (Lophocebus albigena) groups, male rank is an important factor in social interactions, and males also move between groups while actively competing for females or sneaking copulations. During a 20-month study we observed the social behavior and collected 461 fecal samples from 24 adult male gray-cheeked mangabeys from five groups in Kibale National Park, Uganda. Aggressive interactions and the presence of females at the peak of sexual swelling were associated with elevated cortisol concentrations in all males. Independently, dominant (i.e., highest-ranking) males within groups had higher cortisol concentrations than subordinate males, and immigrant males had higher cortisol concentrations than dominant males.     

Is social dispersal stressful? A study in male crested macaques (Macaca nigra)

In gregarious species, dispersal events represent one of the most dramatic changes in social life and environment an animal will experience during life due to increased predation risk, aggression from unfamiliar conspecifics and the lack of social support. However, little is known about how individuals respond physiologically to dispersal and whether this process is stressful for the individuals involved. We therefore studied the physiological stress response during dispersal in the crested macaque, a primate species in which males often change groups. Over a period of 14 months and 14 dispersal events in 4 groups, we determined faecal glucocorticoid metabolite (FGCM) levels during the process of immigration into a new group and examined a variety of factors (e.g. male age, rank achieved, number of males in the group) potentially affecting FGCM levels during this process. We found that FGCM levels were significantly elevated in the first few days upon immigration, after which levels returned quickly to baseline. FGCM response levels upon immigration were significantly and positively influenced by the number of males in the group. The rank a male achieved upon immigration, aggression received, as well as the proximity to other males did not significantly influence FGCM levels. Our data confirm previous findings on other species demonstrating that in crested macaques immigration into a new social group is associated with an acute endocrine stress response. However, given that stress hormone levels remained elevated only for a short period of time, we do not expect males to experience high physiological costs during immigration. Given our limited knowledge on the physiological responses to dispersal in animals, this study contributes to our understanding of dispersal more generally, and particularly inter-individual differences in the stress response and the potential physiological costs associated with these.