Zoo studies in primate physiology, health, and welfare.

Researchers have been studying primates in zoos for more than half a century. There are numerous benefits to conducting research with zoo collections, such as access to a variety of species, ease of sample collection, and the potential to manipulate some research variables. While much of the primate research conducted in zoos is behavioral, there also is a tradition of research focused on reproduction and endocrinology, especially in North America. The contributions to this special issue exemplify how this tradition continues today through a collection of articles on basic and applied research on reproduction and using physiological measures of health and welfare that could be beneficial across primate taxa. As is the case for primatological research in zoos more broadly, the articles in this special issue reflect a taxonomic bias for great apes despite the high species diversity found across zoo collections. Given this bias as well as the threat of extinction faced by many species, there remains a pressing need to increase primatology in zoos through research dedicated to both conservation in the wild and wellbeing in human care.

Evaluating individual biomarkers for predicting health risks in zoo-housed chimpanzees (Pan troglodytes) and bonobos (Pan paniscus).

Although biomarkers are often used for predicting morbidity and mortality in humans, similar data are lacking in our closest relatives. This study analyzed 16 biomarkers in zoo-housed chimpanzees and bonobos from serum samples collected during both routine and nonroutine veterinary immobilizations. Generalized linear and generalized linear mixed models were used to determine the efficacy of each biomarker to predict all-cause morbidity, defined as the presence of at least one chronic condition, or cardiac disease as a subset of all-cause morbidity. Cox proportional hazards models were used to examine associations between biomarkers and mortality risk from any cause. Analyses were conducted using two data sets for each species, one with all values retained (chimpanzees: n = 148; bonobos: n = 33) and the other from samples collected during routine immobilizations only (chimpanzees: n = 95; bonobos: n = 23). Consistent results across both data sets in chimpanzees included associations of higher cortisol with all-cause morbidity risk, lower creatinine with cardiac disease risk, and higher creatinine with mortality risk, and in bonobos were increased cardiac disease risk with higher cortisol and lower dehydroepiandrosterone-sulfate, fructosamine, and triglycerides. However, there were some inconsistencies between data sets, such as tumor necrosis factor-α predicting mortality risk positively in chimpanzees when all values were retained, but negatively for routine values only. Despite the close evolutionary relationships between chimpanzees and bonobos, the only result observed in both species was a negative association between albumin and mortality risk in the all values retained data sets. Thus, data suggest some biomarkers may be useful predictors of future health outcomes, although a better understanding of both individual and species variation in biomarkers and their contribution to health risks is needed.

Value Ranges and Clinical Comparisons of Serum DHEA-S, IL-6, and TNF-α in Western Lowland Gorillas.

Physiological data can provide valuable information about the health and welfare of animals. Unfortunately, few validated assays and a lack of information on species-typical levels of circulating biomarkers for wildlife make the measurement, interpretation, and practical application of such data difficult. We validated commercially available kits and calculated reference intervals (herein called "value ranges") for dehydroepiandrosterone-sulfate (DHEA-S), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) in a sample of zoo-housed western lowland gorillas due to the roles these biomarkers play in stress and immune responses. For each biomarker, we present species-specific value ranges for a sample of gorillas in human care (n = 57). DHEA-S did not vary significantly by sex or age, while IL-6 was higher in males and older gorillas and TNF-α was higher in females but not associated with age. We also compared non-clinical with clinical samples (n = 21) to explore whether these biomarkers reflect changes in health status. There was no significant difference between clinical and non-clinical samples for DHEA-S, but both IL-6 and TNF-α were significantly higher in gorillas showing clinical symptoms or prior to death. Additional work is needed to improve our understanding of normal versus clinical variation in these biomarkers, and we encourage continued efforts to identify and validate additional biomarkers that can be used to inform assessments of health and welfare in wildlife.

The application of allostasis and allostatic load in animal species: A scoping review.

Principles of allostasis and allostatic load have been widely applied in human research to assess the impacts of chronic stress on physiological dysregulation. Over the last few decades, researchers have also applied these concepts to non-human animals. However, there is a lack of uniformity in how the concept of allostasis is described and assessed in animals. The objectives of this review were to: 1) describe the extent to which the concepts of allostasis and allostatic load are applied theoretically to animals, with a focus on which taxa and species are represented; 2) identify when direct assessments of allostasis or allostatic load are made, which species and contexts are represented, what biomarkers are used, and if an allostatic load index was constructed; and 3) detect gaps in the literature and identify areas for future research. A search was conducted using CABI, PubMed, Agricola, and BIOSIS databases, in addition to a complementary hand-search of 14 peer-reviewed journals. Search results were screened, and articles that included non-human animals, as well as the terms "allostasis" or "allostatic" in the full text, were included. A total of 572 articles met the inclusion criteria (108 reviews and 464 peer-reviewed original research). Species were represented across all taxa. A subset of 63 publications made direct assessments of allostatic load. Glucocorticoids were the most commonly used biomarker, and were the only biomarker measured in 25 publications. Only six of 63 publications (9.5%) constructed an allostatic load index, which is the preferred methodology in human research. Although concepts of allostasis and allostatic load are being applied broadly across animal species, most publications use single biomarkers that are more likely indicative of short-term rather than chronic stress. Researchers are encouraged to adopt methodologies used in human research, including the construction of species-specific allostatic load indexes.

Individual measures of social support may have limited impact on physiological parameters among elderly Kuwaitis.

OBJECTIVES: Social support can buffer physiological stress responses, reducing morbidity and mortality risk, but research has occurred primarily in western populations. We examined whether social support was associated with physiological biomarkers in a non-western sample. METHODS: We predicted evidence of increased physiological dysregulation in those with less social support among elderly Kuwaitis (≥60 years, n = 253). Measures of social support included marital status (married/unmarried), religiosity (low/high), whether adult children lived at home (yes/no), and perceived social support (low/medium/high). Using linear regression, we tested relationships between each social support measure and 17 biomarkers: cortisol, dehydroepiandrosterone-sulfate (DHEA-S), epinephrine, norepinephrine, systolic blood pressure (SBP), diastolic blood pressure (DBP), waist-hip ratio (WHR), total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), very-low-density lipoprotein (VLDL), TC/HDL, LDL/HDL, glycated hemoglobin (HbA1c), C-reactive protein (CRP), and resting heart rate (RHR). We analyzed two models for each, one with the independent variable only and a second including age, smoking, and education as covariates. RESULTS: Married participants had lower norepinephrine, but higher TC/HDL and LDL/HDL. Higher religiosity was negatively associated with TC (men only) but higher LDL, TC/HDL, and LDL/HDL. Participants with low self-reported social support had higher DBP and HbA1c than those reporting medium or high levels. CONCLUSIONS: Relatively few biomarkers associated significantly with individual social support measures in a way that suggests improved health for those with more support. As such, some measures of social support may not be universally beneficial across cultures. Additionally, the high degree of respect for and integration of elders in Kuwait society may collectively buffer against negative effects. Cross-cultural comparisons are critical to better understand how social support influences morbidity and mortality across populations.

Age, sex, and inflammatory markers predict chronic conditions, cardiac disease, and mortality among captive western lowland gorillas (Gorilla gorilla gorilla).

In humans, inflammatory markers predict health risks. As great apes experience many similar conditions, measuring inflammation may provide valuable health information. We examined four serum inflammatory markers in zoo-housed gorillas (n = 48): albumin, CRP, IL-6, and TNF-α. We first analyzed age- and sex-associated patterns, then used multimodel inference to evaluate models with age, sex, and inflammatory markers as predictors of all-cause morbidity, cardiac disease, and mortality. Older gorillas had lower albumin and higher IL-6, and males had higher albumin, lower CRP, and lower TNF-α. All-cause morbidity was best predicted by age, sex, and TNF-α, but the second model containing only age and sex was equivalent. Cardiac disease was best predicted by TNF-α alongside age and sex, with lower levels associated with increased risk. When outliers were removed, the model with TNF-α was second to the model containing only age and sex. Finally, mortality risk was best predicted by the model with only age and sex. Other models containing individual inflammatory markers were within top model sets for each health outcome. Our results indicate that age and sex are robust for predicting all-cause morbidity and mortality risk in gorillas; while models which include individual inflammatory markers also predict risk, they may not improve predictions over age and sex alone. However, given the prevalence of cardiac disease in great apes, these results suggest that TNF-α warrants further investigation. With their potential to provide valuable health information, data on inflammatory markers may contribute to the care and management of gorillas in human care.

Testing a method to improve predictions of disease and mortality risk in western lowland gorillas (gorilla gorilla gorilla) using allostatic load.

Allostatic load is the wear-and-tear organisms accumulate due to senescence and stress; it is measured by combining biomarkers from multiple somatic systems into allostatic load indices (ALIs). Frequently used in human research, ALIs have shown consistent results across samples despite different biomarkers and methods. However, determining optimal models likely is necessary if ALIs are to be feasible research tools in other species. Herein, we build on prior research in western lowland gorillas to explore one potential method for determining which biomarkers may be best for estimating allostatic load. After narrowing down which biomarkers to include using a combination of forward stepwise regression and independent biomarker associations with project variables, we estimated allostatic load using both the traditional one-tailed quartile method as well as a multi-method approach. There was a significant positive association between allostatic load and triglycerides, but not cholesterol, both of which are commonly used as diagnostic markers of poor health. Using binomial generalized linear models, a one-unit increase in allostatic load was associated with increased risk of all-cause morbidity and mortality, but reduced risk of cardiac disease. Although conclusions were similar, compared to our original ALIs, these new ALIs had weaker effect sizes and poorer relative goodness of fit, suggesting this method for identifying the best possible list of biomarkers to include in an index was not effective. This report continues the development of ALIs as a clinical tool in wildlife while systematically testing one possible method for determining an optimal ALI for a particular species.

Allostatic Load Indices With Cholesterol and Triglycerides Predict Disease and Mortality Risk in Zoo-Housed Western Lowland Gorillas (Gorilla gorilla gorilla).

Allostatic load, or the physiological dysregulation accumulated due to senescence and stress, is an established predictor of human morbidity and mortality and has been proposed as a tool for monitoring health and welfare in captive wildlife. It is estimated by combining biomarkers from multiple somatic systems into allostatic load indices (ALIs), providing a score representing overall physiological dysregulation. Such ALIs have been shown to predict disease and mortality risk in western lowland gorillas. In these prior analyses, we were unable to include lipid markers, a potential limitation as they are key biomarkers in human models. Recently, we were able to assay serum cholesterol and triglycerides and add them to our previous ALI. We then re-examined associations with health outcomes using binomial generalized linear models. We constructed ALIs using 2 pooling strategies and 2 methods. By itself, a 1-unit increase in allostatic load was associated with higher odds of all-cause morbidity and mortality, but results were mixed for cardiac disease. However, the best fit models for all-cause morbidity and cardiac disease included only age and sex. Allostatic load was retained alongside age in the best fit models for mortality, with a 1-unit increase associated with 23% to 45% higher odds of death. Compared with previous results, ALIs containing cholesterol and triglycerides better predict disease risk in zoo-housed western lowland gorillas, as evidenced by larger effect sizes for some models and better goodness of fit for all ALIs. Based on these results, we address methodology for future allostatic load research on wildlife.

Testing lipid markers as predictors of all-cause morbidity, cardiac disease, and mortality risk in captive western lowland gorillas (Gorilla gorilla gorilla).

Great apes and humans develop many of the same health conditions, including cardiac disease as a leading cause of death. In humans, lipid markers are strong predictors of morbidity and mortality risk. To determine if they similarly predict risk in gorillas, we measured five serum lipid markers and calculated three lipoprotein ratios from zoo-housed western lowland gorillas (aged 6-52 years, n = 61 , subset with routine immobilizations only: n = 47 ): total cholesterol (TC), triglycerides (TGs), high-density lipoprotein (HDL), low-density lipoprotein (LDL), apolipoprotein A1 (apoA1), TC / HDL , LDL / HDL , and TG / HDL . We examined each in relation to age and sex, then analyzed whether they predicted all-cause morbidity, cardiac disease, and mortality using generalized linear models (GLMs). Older age was significantly associated with higher TG, TC / HDL , LDL / HDL , and TG / HDL , and lower HDL and apoA1. With all ages combined, compared to females, males had significantly lower TG, TC / HDL , LDL / HDL , and TG / HDL , and higher HDL. Using GLMs, age, sex, and lower LDL / HDL were significant predictors of all-cause morbidity; this is consistent with research demonstrating lower LDL in humans with arthritis, which was the second most prevalent condition in this sample. In contrast to humans, lipid markers were not better predictors of cardiac disease and mortality risk in gorillas, with cardiac disease best predicted by age and sex alone, and mortality risk only by age. Similar results were observed when multimodel inference was used as an alternative analysis strategy, suggesting it can be used in place of or in addition to traditional methods for predicting risk.

Stress, Well-Being and Reproductive Success.

To reverse the trend of declining wildlife populations globally, individuals must be provided with conditions that allow them to not just survive, but to thrive. It is no longer only the remit of captive breeding programs to ensure animal well-being; in situ conservation efforts also must consider how environmental and anthropogenic pressures impact wild populations, and how to mitigate them-especially with regards to reproduction and survival. Stress and welfare are complex concepts that necessitate an understanding of how stressors affect animals on both individual and population levels. There are species differences in how factors impact well-being, related in part to natural history, which also are shaped by individual perceptions and coping abilities. A multitude of stress-related responses then have the potential to disrupt fertility on many levels, and ultimately fitness. A major limitation to advancing welfare science is the lack of definitive tests to verify welfare status; i.e., is the animal happy or not? While analyses of circulating or excreted glucocorticoids have for decades been the primary method of assessing stress, today we recognize the need for more objective indicators that incorporate multiple physiological systems, including behavior, to assess both negative and positive welfare states. In this chapter, we discuss the potential for stress to disrupt, and sometimes facilitate reproduction, including the key role that glucocorticoids play. We then discuss a number of physiological biomarkers, which in addition to glucocorticoids, have the potential to assess well-being and the role of stress on reproduction. Finally, we discuss allostatic load, a method by which multiple physiological markers are used to inform on morbidity and mortality risk in humans, which if applied to wildlife, could be a powerful tool for conservation.

EVALUATING ALLOSTATIC LOAD: A NEW APPROACH TO MEASURING LONG-TERM STRESS IN WILDLIFE.

Animal welfare, conservation, and stress assessment are all critical components of species survival. As organisms experience stressors, they accumulate physiologic dysregulation, leading to multiple negative health outcomes. This brief review suggests measuring the degree of stress-induced damage, known as allostatic load, and then using allostatic load to evaluate changes implemented to improve animal welfare and conservation efforts. Over the past two decades, human clinical research has developed multiple allostatic load indices constructed from composites of neuroendocrine, cardiovascular, metabolic, and immune biomarkers. These indices are designed to estimate allostatic load in hopes of ameliorating or even negating damaging effects of stress. Among humans, allostatic load is associated with a variety of factors such as age, sex, stressful experiences, personality, social position, and early life history. Despite conservation of stress responses throughout mammalian species, reported allostatic load indices for animals are rare. Because many zoo researchers and field scientists already collect data on multiple biomarkers, constructing allostatic load indices may be a relatively affordable, easily implemented, and powerful tool for assessing relative risks of morbidity and mortality within wildlife. As an example, in a study among zoo-housed gorillas, an allostatic load index constructed using seven biomarkers was associated significantly with age, sex, stressful experiences, rearing history, markers of poor health, and mortality risk. Such results evidence that allostatic load is as applicable to animal populations as it is to humans. By using allostatic load as a predictive tool, human caretakers will be better informed of individuals at greatest risk for health declines. Most importantly, allostatic load may provide earlier opportunity for preemptive care while contributing a transformational tool to animal welfare research. Additionally, allostatic load may be compared between individuals and groups within the same population and allow comparisons of health between and across populations, consequently informing habitat and population protection efforts.

The first multi-zoo application of an allostatic load index to western lowland gorillas (Gorilla gorilla gorilla).

Vertebrate stress responses are highly adaptive biological functions, maximizing survival probability in life-threatening situations. However, experiencing repeated and/or chronic stressors can generate physiological dysregulation and lead to disease. Because stress responses are multi-systemic and involve a wide range of physiological functions, identifying responses to stressors is best accomplished using integrated biomarker models. Allostatic load, defined as the physiological dysregulation that accumulates over the lifespan due to stressful experiences, is one such model. Allostatic load is measured using allostatic load indices, which are composites of biomarkers from multiple somatic systems. Previously, we reported the use of a 7-biomarker allostatic load index (albumin, CRH, cortisol, DHEA-S, glucose, IL-6, TNF-α) in western lowland gorillas housed at a single zoo. Herein, this index is expanded to examine allostatic load responses to lifetime stressors in gorillas from two additional zoos (n = 63) as well as two pooled samples. The index was created using quartile cut-points for each biomarker. Significant associations were observed between multiple predictor variables and allostatic load, including sex, age, number of stressful events (anesthetic events, zoo transfers, agonistic interactions with wounding, pregnancies), and rearing history (mother-reared, nursery-reared, wild-caught). Additionally, allostatic load was associated with indicators of morbidity (creatinine, cholesterol, triglycerides), age at death, and mortality risk. These results are consistent with those reported in human allostatic load research, suggesting allostatic load indices have potential as an investigative and clinical tool for gorillas and other great apes.

Dehydroepiandrosterone-sulfate (DHEA-S), sex, and age in zoo-housed western lowland gorillas (Gorilla gorilla gorilla).

Among humans, dehydroepiandrosterone-sulfate (DHEA-S) declines with age and is hypothesized to be involved in somatic maintenance and healthy aging. Men have significantly higher DHEA-S than women, contradicting longer lifespans in the latter. Declines of DHEA-S with age also are observed in chimpanzees. In both chimpanzees and bonobos, males and females show no differences in DHEA-S production. Based on human and chimpanzee data, gorillas were predicted to show declining DHEA-S with age. Similar to chimpanzees and bonobos, it also was predicted DHEA-S would not be significantly different between males and females. DHEA-S was assayed from serum banked during physical examinations of gorillas housed at three North American zoos (n = 63). Gorillas ranged from 6 to 52 years of age. Differences between males and females were examined using t tests. Linear regression was used to determine the relationship of DHEA-S with age. There was no significant difference in DHEA-S between males and females. Additionally, there was no significant relationship between DHEA-S and age. As predicted, there were no sex-based differences in DHEA-S in gorillas, which is similar to chimpanzees and bonobos but different from modern humans. Unlike chimpanzees and humans, there was no significant relationship between DHEA-S and age in gorillas. The absence of a relationship between age and DHEA-S may be due to the lack of gorillas under age 6 years in this sample as declines in chimpanzees occur prior to age 5 years, more rapid growth and development among gorillas compared with other African hominoids, or a unique pattern of DHEA-S production.

Allostatic load and biological anthropology.

Multiple stressors affect developing and adult organisms, thereby partly structuring their phenotypes. Determining how stressors influence health, well-being, and longevity in human and nonhuman primate populations are major foci within biological anthropology. Although much effort has been devoted to examining responses to multiple environmental and sociocultural stressors, no holistic metric to measure stress-related physiological dysfunction has been widely applied within biological anthropology. Researchers from disciplines outside anthropology are using allostatic load indices (ALIs) to estimate such dysregulation and examine life-long outcomes of stressor exposures, including morbidity and mortality. Following allostasis theory, allostatic load represents accumulated physiological and somatic damage secondary to stressors and senescent processes experienced over the lifespan. ALIs estimate this wear-and-tear using a composite of biomarkers representing neuroendocrine, cardiovascular, metabolic, and immune systems. Across samples, ALIs are associated significantly with multiple individual characteristics (e.g., age, sex, education, DNA variation) of interest within biological anthropology. They also predict future outcomes, including aspects of life history variation (e.g., survival, lifespan), mental and physical health, morbidity and mortality, and likely health disparities between groups, by stressor exposures, ethnicity, occupations, and degree of departure from local indigenous life ways and integration into external and commodified ones. ALIs also may be applied to similar stress-related research areas among nonhuman primates. Given the reports from multiple research endeavors, here we propose ALIs may be useful for assessing stressors, stress responses, and stress-related dysfunction, current and long-term cognitive function, health and well-being, and risk of early mortality across many research programs within biological anthropology.

Rearing history and allostatic load in adult western lowland gorillas (Gorilla gorilla gorilla) in human care.

Disrupted rearing history is a psychological and physical stressor for nonhuman primates, potentially resulting in multiple behavioral and physiological changes. As a chronic, soma-wide stressor, altered rearing may be best assessed using a holistic tool such as allostatic load (AL). In humans, AL estimates outcomes of lifetime stress-induced damage. We predicted mother-reared gorillas would have lower AL than nursery-reared and wild-caught conspecifics. We estimated AL for 27 gorillas housed at the Columbus Zoo and Aquarium between 1956 and 2014. AL estimates were calculated using biomarkers obtained during previous anesthetic events. Biomarkers in the high-risk quartile were counted toward a gorilla's AL. Rearing history was categorized as mother-reared, nursery-reared, and wild-caught. Using ANCOVA, rearing history and AL are significantly associated when age and sex are entered as covariates. Wild-caught gorillas have significantly higher AL than mother-reared gorillas. Neither wild-caught nor mother-reared gorillas are significantly different from nursery-reared gorillas. When examined by sex, males of all rearing histories have significantly lower AL than females. We suggest males face few stressors in human care and ill effects of rearing history do not follow. Wild-caught females have significantly higher AL than mother-reared females, but neither is significantly different from nursery-reared females. Combined with our previous work on AL in this group, wherein females had twofold higher AL than males, we suggest females in human care face more stressors than males. Disrupted rearing history may exacerbate effects of these stressors. Providing opportunities for females to choose their distance from males may help reduce their AL.