Hematology and serum biochemistry of free-ranging mantled howler monkeys (Alouatta palliata) at La Pacifica, Costa Rica.

BACKGROUND: Hematologic and blood biochemical values are key tools for assessing primate health. A long-term behavioral study of howler monkeys at a single site (La Pacífica, Guanacaste, Costa Rica), afforded the opportunity to develop baseline values for a large group of animals, evaluating differences between adult males and females and comparing to a report in the same population two decades later. METHODS: In 1998, 64 free-ranging mantled howler monkeys were anesthetized and sampled for hematologic and biochemical analysis. RESULTS: Blood analysis is reported for 29 adult females, 9 juvenile females, 19 adult males and 3 juvenile males. Four adults were excluded due to external injury or disease. There were few significant differences between adult females, juvenile females, and adult males. CONCLUSIONS: Baseline blood parameters are useful for determining normal values for howler monkey populations. The values for total protein, blood urea nitrogen, glucose, liver enzymes and potassium differed from a later study in 2019 may indicate changes that are influencing howler monkey health.

Sexual dimorphism and growth in Alouatta palliata based on 20+ years of field data.

OBJECTIVES: Alouatta palliata patterns of growth and sexual dimorphism are evaluated using 20 plus years of field data. Comparisons are made to other species of Alouatta and other New World primates. MATERIALS AND METHODS: Records of 92 A. palliata from Guanacaste Province, Costa Rica, were used to generate growth curves for body mass and linear measurements. Timing of growth for the properties was compared, and males and females were contrasted. Slopes and elevations for periods of rapid growth were evaluated. Growth allometry and proportion ratios were also explored. RESULTS: Body mass growth is rapid during the first 2 years. Males and females begin to diverge around a year of age as male growth increases and female growth slows. Adult mass for both is reached about 4 years of age. Linear measurements show rapid growth the first 18 months for both sexes. Differences develop as males continue the same rate of linear growth while female growth slows. Adult size is reached for head and body length around 3 years, and for hind-foot and tail lengths around 2 years. DISCUSSION: A. palliata males grow in mass more rapidly than females, while both grow similarly in linear dimensions, so that dimorphism is more pronounced in mass. This pattern is seen in other dimorphic New World primates. Male A. palliata may grow more rapidly than A. seniculus, reflecting earlier emigration for A. palliata males. Linear dimensions reach adult proportions earlier than body mass. For hind-foot and tail, this is probably an adaptation for gripping.

Captivity humanizes the primate microbiome.

The primate gastrointestinal tract is home to trillions of bacteria, whose composition is associated with numerous metabolic, autoimmune, and infectious human diseases. Although there is increasing evidence that modern and Westernized societies are associated with dramatic loss of natural human gut microbiome diversity, the causes and consequences of such loss are challenging to study. Here we use nonhuman primates (NHPs) as a model system for studying the effects of emigration and lifestyle disruption on the human gut microbiome. Using 16S rRNA gene sequencing in two model NHP species, we show that although different primate species have distinctive signature microbiota in the wild, in captivity they lose their native microbes and become colonized with Prevotella and Bacteroides, the dominant genera in the modern human gut microbiome. We confirm that captive individuals from eight other NHP species in a different zoo show the same pattern of convergence, and that semicaptive primates housed in a sanctuary represent an intermediate microbiome state between wild and captive. Using deep shotgun sequencing, chemical dietary analysis, and chloroplast relative abundance, we show that decreasing dietary fiber and plant content are associated with the captive primate microbiome. Finally, in a meta-analysis including published human data, we show that captivity has a parallel effect on the NHP gut microbiome to that of Westernization in humans. These results demonstrate that captivity and lifestyle disruption cause primates to lose native microbiota and converge along an axis toward the modern human microbiome.

The gut microbiome of nonhuman primates: Lessons in ecology and evolution.

The mammalian gastrointestinal (GI) tract is home to trillions of bacteria that play a substantial role in host metabolism and immunity. While progress has been made in understanding the role that microbial communities play in human health and disease, much less attention has been given to host-associated microbiomes in nonhuman primates (NHPs). Here we review past and current research exploring the gut microbiome of NHPs. First, we summarize methods for characterization of the NHP gut microbiome. Then we discuss variation in gut microbiome composition and function across different NHP taxa. Finally, we highlight how studying the gut microbiome offers new insights into primate nutrition, physiology, and immune system function, as well as enhances our understanding of primate ecology and evolution. Microbiome approaches are useful tools for studying relevant issues in primate ecology. Further study of the gut microbiome of NHPs will offer new insight into primate ecology and evolution as well as human health.

Thyroid hormone fluctuations indicate a thermoregulatory function in both a tropical (Alouatta palliata) and seasonally cold-habitat (Macaca fuscata) primate.

Thyroid hormones boost animals' basal metabolic rate and represent an important thermoregulatory pathway for mammals that face cold temperatures. Whereas the cold thermal pressures experienced by primates in seasonal habitats at high latitudes and elevations are often apparent, tropical habitats also display distinct wet and dry seasons with modest changes in thermal environment. We assessed seasonal and temperature-related changes in thyroid hormone levels for two primate species in disparate thermal environments, tropical mantled howlers (Alouatta palliata), and seasonally cold-habitat Japanese macaques (Macaca fuscata). We collected urine and feces from animals and used ELISA to quantify levels of the thyroid hormone triiodothyronine (fT3 ). For both species, fT3 levels were significantly higher during the cooler season (wet/winter), consistent with a thermoregulatory role. Likewise, both species displayed greater temperature deficits (i.e., the degree to which animals warm their body temperature relative to ambient) during the cooler season, indicating greater thermoregulatory pressures during this time. Independently of season, Japanese macaques displayed increasing fT3 levels with decreasing recently experienced maximum temperatures, but no relationship between fT3 and recently experienced minimum temperatures. Howlers increased fT3 levels as recently experienced minimum temperatures decreased, although demonstrated the opposite relationship with maximum temperatures. This may reflect natural thermal variation in howlers' habitat: wet seasons had cooler minimum and mean temperatures than the dry season, but similar maximum temperatures. Overall, our findings support the hypothesis that both tropical howlers and seasonally cold-habitat Japanese macaques utilize thyroid hormones as a mechanism to boost metabolism in response to thermoregulatory pressures. This implies that cool thermal pressures faced by tropical primates are sufficient to invoke an energetically costly and relatively longer-term thermoregulatory pathway. The well-established relationship between thyroid hormones and energetics suggests that the seasonal hormonal changes we observed could influence many commonly studied behaviors including food choice, range use, and activity patterns.

An assessment of skin temperature gradients in a tropical primate using infrared thermography and subcutaneous implants.

Infrared thermography has become a useful tool to assess surface temperatures of animals for thermoregulatory research. However, surface temperatures are an endpoint along the body's core-shell temperature gradient. Skin and fur are the peripheral tissues most exposed to ambient thermal conditions and are known to serve as thermosensors that initiate thermoregulatory responses. Yet relatively little is known about how surface temperatures of wild mammals measured by infrared thermography relate to subcutaneous temperatures. Moreover, this relationship may differ with the degree that fur covers the body. To assess the relationship between temperatures and temperature gradients in peripheral tissues between furred and bare areas, we collected data from wild mantled howling monkeys (Alouatta palliata) in Costa Rica. We used infrared thermography to measure surface temperatures of the furred dorsum and bare facial areas of the body, recorded concurrent subcutaneous temperatures in the dorsum, and measured ambient thermal conditions via a weather station. Temperature gradients through cutaneous tissues (subcutaneous-surface temperature) and surface temperature gradients (surface-ambient temperature) were calculated. Our results indicate that there are differences in temperatures and temperature gradients in furred versus bare areas of mantled howlers. Under natural thermal conditions experienced by wild animals, the bare facial areas were warmer than temperatures in the furred dorsum, and cutaneous temperature gradients in the face were more variable than the dorsum, consistent with these bare areas acting as thermal windows. Cutaneous temperature gradients in the dorsum were more closely linked to subcutaneous temperatures, while facial temperature gradients were more heavily influenced by ambient conditions. These findings indicate that despite the insulative properties of fur, for mantled howling monkeys surface temperatures of furred areas still demonstrate a relationship with subcutaneous temperatures. Given that most mammals possess dense fur, this provides insight for using infrared imaging in thermoregulatory studies of wild animals lacking bare skin.

Dust in the wind: How climate variables and volcanic dust affect rates of tooth wear in Central American howling monkeys.

OBJECTIVES: Two factors have been considered important contributors to tooth wear: dietary abrasives in plant foods themselves and mineral particles adhering to ingested food. Each factor limits the functional life of teeth. Cross-population studies of wear rates in a single species living in different habitats may point to the relative contributions of each factor. MATERIALS AND METHODS: We examine macroscopic dental wear in populations of Alouatta palliata (Gray, 1849) from Costa Rica (115 specimens), Panama (19), and Nicaragua (56). The sites differ in mean annual precipitation, with the Panamanian sites receiving more than twice the precipitation of those in Costa Rica or Nicaragua (∼3,500 mm vs. ∼1,500 mm). Additionally, many of the Nicaraguan specimens were collected downwind of active plinian volcanoes. Molar wear is expressed as the ratio of exposed dentin area to tooth area; premolar wear was scored using a ranking system. RESULTS: Despite substantial variation in environmental variables and the added presence of ash in some environments, molar wear rates do not differ significantly among the populations. Premolar wear, however, is greater in individuals collected downwind from active volcanoes compared with those living in environments that did not experience ash-fall. DISCUSSION: Volcanic ash seems to be an important contributor to anterior tooth wear but less so in molar wear. That wear is not found uniformly across the tooth row may be related to malformation in the premolars due to fluorosis. A surge of fluoride accompanying the volcanic ash may differentially affect the premolars as the molars fully mineralize early in the life of Alouatta.

A comparison of activity patterns for captive Propithecus tattersalli and Propithecus coquereli.

The activity patterns and social interactions of two species of captive sifaka were observed during a 2-year period. Allogrooming was not observed in golden-crowned sifaka and they spent significantly more time resting than the Coquerel's sifaka. Females of both species were found to be dominant to males. The golden-crowned sifaka (Propithecus tattersalli) spent significantly less time feeding than the Coquerel's sifaka. Temperature, time of day, species, and interpair comparisons for the golden-crowned sifaka were found to affect activity and social interactions, while gender did not. Like the Coquerel's sifaka, the golden-crowned sifaka was found to be diurnal; however, they differed in that the golden-crowned sifaka did not descend to the ground.

Testing parallel laser image scaling for remotely measuring body dimensions on mantled howling monkeys (Alouatta palliata).

Body size is a fundamental variable for many studies in primate biology. However, obtaining body dimensions of wild primates through live capture is difficult and costly, so developing an alternative inexpensive and non-invasive method is crucial. Parallel laser image scaling for remotely measuring body size has been used with some success in marine and terrestrial animals, but only one arboreal primate. We further tested the efficacy of this method on the arboreal mantled howling monkey (Alouatta palliata) in La Pacifica, Costa Rica. We calculated interobserver error, as well as the method's repeatability when measuring the same animal on different occasions. We also compared measurements obtained physically through live capture with measurements obtained remotely using parallel laser image scaling. Our results show that the different types of error for the remote technique are minimal and comparable with the error rates observed in physical methods, with the exception of some dimensions that vary depending on the animals' posture. We conclude that parallel laser image scaling can be used to remotely obtain body dimensions if careful consideration is given to factors such as species-specific morphology and postural habits.

Daily activity and light exposure levels for five species of lemurs at the Duke Lemur Center.

Light is the primary synchronizer of all biological rhythms, yet little is known about the role of the 24-hour luminous environment on nonhuman primate circadian patterns, making it difficult to understand the photic niche of the ancestral primate. Here we present the first data on proximate light-dark exposure and activity-rest patterns in free-ranging nonhuman primates. Four individuals each of five species of lemurs at the Duke Lemur Center (Eulemur mongoz, Lemur catta, Propithecus coquereli, Varecia rubra, and Varecia variegata variegata) were fitted with a Daysimeter-D pendant that contained light and accelerometer sensors. Our results reveal common as well as species-specific light exposure and behavior patterns. As expected, all five species were more active between sunrise and sunset. All five species demonstrated an anticipatory increase in their pre-sunrise activity that peaked at sunrise with all but V. rubra showing a reduction within an hour. All five species reduced activity during mid-day. Four of the five stayed active after sunset, but P. coquereli began reducing their activity about 2 hours before sunset. Other subtle differences in the recorded light exposure and activity patterns suggest species-specific photic niches and behaviors. The eventual application of the Daysimeter-D in the wild may help to better understand the adaptive evolution of ancestral primates.

Body temperature and thermal environment in a generalized arboreal anthropoid, wild mantled howling monkeys (Alouatta palliata).

Free-ranging primates are confronted with the challenge of maintaining an optimal range of body temperatures within a thermally dynamic environment that changes daily, seasonally, and annually. While many laboratory studies have been conducted on primate thermoregulation, we know comparatively little about the thermal pressures primates face in their natural, evolutionarily relevant environment. Such knowledge is critical to understanding the evolution of thermal adaptations in primates and for comparative evaluation of humans' unique thermal adaptations. We examined temperature and thermal environment in free-ranging, mantled howling monkeys (Alouatta palliata) in a tropical dry forest in Guanacaste, Costa Rica. We recorded subcutaneous (Tsc ) and near-animal ambient temperatures (Ta ) from 11 animals over 1586.5 sample hours during wet and dry seasons. Howlers displayed considerable variation in Tsc , which was largely attributable to circadian effects. Despite significant seasonal changes in the ambient thermal environment, howlers showed relatively little evidence for seasonal changes in Tsc . Howlers experienced warm thermal conditions which led to body cooling relative to the environment, and plateaus in Tsc at increasingly warm Ta . They also frequently faced cool thermal conditions (Ta < Tsc ) in which Tsc was markedly elevated compared with Ta . These data add to a growing body of evidence that non-human primates have more labile body temperatures than humans. Our data additionally support a hypothesis that, despite inhabiting a dry tropical environment, howling monkeys experience both warm and cool thermal pressures. This suggests that thermal challenges may be more prevalent for primates than previously thought, even for species living in nonextreme thermal environments.

Calibrating single-ended fiber-optic Raman spectra distributed temperature sensing data.

Hydrologic research is a very demanding application of fiber-optic distributed temperature sensing (DTS) in terms of precision, accuracy and calibration. The physics behind the most frequently used DTS instruments are considered as they apply to four calibration methods for single-ended DTS installations. The new methods presented are more accurate than the instrument-calibrated data, achieving accuracies on the order of tenths of a degree root mean square error (RMSE) and mean bias. Effects of localized non-uniformities that violate the assumptions of single-ended calibration data are explored and quantified. Experimental design considerations such as selection of integration times or selection of the length of the reference sections are discussed, and the impacts of these considerations on calibrated temperatures are explored in two case studies.

Dietary choices by four captive slender lorises (Loris tardigradus) when presented with various insect life stages.

The slender loris (Loris tardigradus) is a rare, nocturnal prosimian found only in the tropical rainforest of southern India and Sri Lanka. Little is known about their diet, though it is assumed that insects comprise a majority of their wild diet. Based on this assumption, captive lorises are offered a variety of insects or insect life stages; the species of insect or the life stage is often determined by what is easiest to buy or rear. Captive lorises at the Duke Lemur Center (DLC) were offered the opportunity to choose which life stage of mealworms (Tenebrio molito), superworms (Zophobus morio), or waxworms (Galleria mellonella) they preferred. The DLC captive lorises did not select the largest life stages of any insect offered. They preferred the larvae stage to the adult stage in all three insect species, and males and females had different insect species and life stage preferences.

Chemical characterization of oligosaccharides in the milk of six species of New and Old World monkeys.

Human and great ape milks contain a diverse array of milk oligosaccharides, but little is known about the milk oligosaccharides of other primates, and how they differ among taxa. Neutral and acidic oligosaccharides were isolated from the milk of three species of Old World or catarrhine monkeys (Cercopithecidae: rhesus macaque (Macaca mulatta), toque macaque (Macaca sinica) and Hamadryas baboon (Papio hamadryas)) and three of New World or platyrrhine monkeys (Cebidae: tufted capuchin (Cebus apella) and Bolivian squirrel monkey (Saimiri boliviensis); Atelidae: mantled howler (Alouatta palliata)). The milks of these species contained 6-8% total sugar, most of which was lactose: the estimated ratio of oligosaccharides to lactose in Old World monkeys (1:4 to 1:6) was greater than in New World monkeys (1:12 to 1:23). The chemical structures of the oligosaccharides were determined mainly by (1)H-NMR spectroscopy. Oligosaccharides containing the type II unit (Gal(ß1-4)GlcNAc) were found in the milk of the rhesus macaque, toque macaque, Hamadryas baboon and tufted capuchin, but oligosaccharides containing the type I unit (Gal(ß1-3)GlcNAc), which have been found in human and many great ape milks, were absent from the milk of all species studied. Oligosaccharides containing Lewis x (Gal(ß1-4)[Fuc(α1-3)]GlcNAc) and 3-fucosyl lactose (3-FL, Gal(ß1-4)[Fuc(α1-3)]Glc) were found in the milk of the three cercopithecid monkey species, while 2-fucosyl lactose (5'-FL, Fuc(α1-2)Gal(ß1-4)Glc) was absent from all species studied. All of these milks contained acidic oligosaccharides that had N-acetylneuraminic acid as part of their structures, but did not contain oligosaccharides that had N-glycolylneuraminic acid, in contrast to the milk or colostrum of great apes which contain both types of acidic oligosaccharides. Two GalNAc-containing oligosaccharides, lactose 3'-O-sulfate and lacto-N-novopentaose I (Gal(ß1-3)[Gal(ß1-4)GlcNAc(ß1-6)]Gal(ß1-4)Glc) were found only in the milk of rhesus macaque, hamadryas baboon and tufted capuchin, respectively. Further research is needed to determine the extent to which the milk oligosaccharide patterns observed among these taxa represent wider phylogenetic trends among primates and how much variation occurs among individuals or species.

Getting Humans Off Monkeys' Backs: Using Primate Acclimation as a Guide for Habitat Management Efforts.

Wild primates face grave conservation challenges, with habitat loss and climate change projected to cause mass extinctions in the coming decades. As large-bodied Neotropical primates, mantled howling monkeys (Alouatta palliata) are predicted to fare poorly under climate change, yet are also known for their resilience in a variety of environments, including highly disturbed habitats. We utilized ecophysiology research on this species to determine the morphological, physiological, and behavioral mechanisms howlers employ to overcome ecological challenges. Our data show that howlers at La Pacifica, Costa Rica are capable of modifying body size. Howlers displayed reduced mass in warmer, drier habitats, seasonal weight changes, frequent within-lifetime weight fluctuations, and gradual increases in body mass over the past four decades. These within-lifetime changes indicate a capacity to modify morphology in a way that can impact animals' energetics and thermodynamics. Howlers are also able to consume foods with a wide variety of food material properties by altering oral processing during feeding. While this capability suggests some capacity to cope with the phenological shifts expected from climate change and increased habitat fragmentation, data on rates of dental microwear warn that these acclimations may also cost dental longevity. Lastly, we found that howlers are able to acclimate to changing thermal pressures. On shorter-term daily scales, howlers use behavioral mechanisms to thermoregulate, including timing activities to avoid heat stress and utilizing cool microhabitats. At the seasonal scale, animals employ hormonal pathways to influence heat production. These lines of evidence cumulatively indicate that howlers possess morphological, physiological, and behavioral mechanisms to acclimate to environmental challenges. As such, howlers' plasticity may facilitate their resilience to climate change and habitat loss. While habitat loss in the tropics is unlikely to abate, our results point to a potential benefit of active management and selective cultivation to yield large, interconnected forest fragments with targeted phenology that provides both a complex physical structure and a diversity of food sources. These steps could assist howlers in using their natural acclimation potential to survive future conservation threats.

Morphometric signals of population decline in diademed sifakas occupying degraded rainforest habitat in Madagascar.

Anthropogenic habitat change can have varied impacts on primates, including both negative and positive outcomes. Even when behavioural shifts are seen, they may reflect decreased health, or simply behavioural flexibility; understanding this distinction is important for conservation efforts. This study examines habitat-related variation in adult and immature morphometrics among diademed sifakas (Propithecus diadema). We collected morphometric data from sifakas at Tsinjoarivo, Madagascar (19 years, 188 captures, 113 individuals). Captures spanned 12 groups, five within continuous forest ("CONT"), and seven in degraded fragments ("FRAG") where sifakas have lower nutritional intakes. Few consistent differences were found between CONT and FRAG groups. However, using home range quality as a covariate rather than a CONT/FRAG dichotomy revealed a threshold: the two FRAG groups in the lowest-quality habitat showed low adult mass and condition (wasting), and low immature mass and length (stunting). Though less-disturbed fragments apparently provide viable habitat, we suggest the sifakas in the most challenging habitats cannot evolve fast enough to keep up with such rapid habitat change. We suggest other long-lived organisms will show similar morphometric "warning signs" (wasting in adults, stunting in immatures); selected morphometric variables can thus be useful at gauging vulnerability of populations in the face of anthropogenic change.

Fatty acid composition of wild anthropoid primate milks.

Fatty acids in milk reflect the interplay between species-specific physiological mechanisms and maternal diet. Anthropoid primates (apes, Old and New World monkeys) vary in patterns of growth and development and dietary strategies. Milk fatty acid profiles also are predicted to vary widely. This study investigates milk fatty acid composition of five wild anthropoids (Alouatta palliata, Callithrix jacchus, Gorilla beringei beringei, Leontopithecus rosalia, Macaca sinica) to test the null hypothesis of a generalized anthropoid milk fatty acid composition. Milk from New and Old World monkeys had significantly more 8:0 and 10:0 than milk from apes. The leaf eating species G. b. beringei and A. paliatta had a significantly higher proportion of milk 18:3n-3, a fatty acid found primarily in plant lipids. Mean percent composition of 22:6n-3 was significantly different among monkeys and apes, but was similar to the lowest reported values for human milk. Mountain gorillas were unique among anthropoids in the high proportion of milk 20:4n-6. This seems to be unrelated to requirements of a larger brain and may instead reflect species-specific metabolic processes or an unknown source of this fatty acid in the mountain gorilla diet.

Associations Between Nutrition, Gut Microbiome, and Health in A Novel Nonhuman Primate Model.

Red-shanked doucs (Pygathrix nemaeus) are endangered, foregut-fermenting colobine primates which are difficult to maintain in captivity. There are critical gaps in our understanding of their natural lifestyle, including dietary habits such as consumption of leaves, unripe fruit, flowers, seeds, and other plant parts. There is also a lack of understanding of enteric adaptations, including their unique microflora. To address these knowledge gaps, we used the douc as a model to study relationships between gastrointestinal microbial community structure and lifestyle. We analyzed published fecal samples as well as detailed dietary history from doucs with four distinct lifestyles (wild, semi-wild, semi-captive, and captive) and determined gastrointestinal bacterial microbiome composition using 16S rRNA sequencing. A clear gradient of microbiome composition was revealed along an axis of natural lifestyle disruption, including significant associations with diet, biodiversity, and microbial function. We also identified potential microbial biomarkers of douc dysbiosis, including Bacteroides and Prevotella, which may be related to health. Our results suggest a gradient-like shift in captivity causes an attendant shift to severe gut dysbiosis, thereby resulting in gastrointestinal issues.

Food choice from endemic North Carolina tree species by captive prosimians (Lemur fulvus).

Seven captive-born lemurs (Lemur fulvus) at the Duke University Primate Center were presented with leaves of different maturity from five species of North Carolina trees. The animals demonstrated three distinct behaviors toward the novel plant material. They sniffed, tasted, and/or ingested it. New leaves were sniffed, but little tasting and ingestion was observed. Intermediate pine needles were sniffed and ingested but little tasted. Mature leaves were sniffed equally, but the mature leaves of tulip trees and honeysuckle were tasted significantly more than pine, sweetgum, and red maple. Pine, sweetgum, and red maple were ingested significantly more than tulip trees and honeysuckle. Male lemurs ate significantly more mature pine needles and new sweetgum leaves than did the females. Chemical analysis of these plant materials indicated that the new and mature leaves of tulip tree and honeysuckle contained alkaloids. Captive-born lemurs apparently use their sense of smell and taste in choosing what to eat and seem just as capable as free-ranging animals in finding food when faced with the chemical defenses that protect trees from insect predation.

Adoption of infant howling monkeys (Alouatta palliata).

During 3 years of continuous field observations on mantled howlers (Alouatta palliata Gray) in Costa Rica we observed five infants without mothers in the main study group. Four of these infants solicited care and two were adopted (one permanently, one temporarily) by lactating females. The other two were carried but not adopted. The fifth neither solicited nor received care. An infant must solicit care to receive care, and female howlers apparently can suckle more than one infant at a time.