A Novel Method for Assessing Enamel Thickness Distribution in the Anterior Dentition as a Signal for Gouging and Other Extractive Foraging Behaviors in Gummivorous Mammals.

Gummivory poses unique challenges to the dentition as gum acquisition may often require that the anterior teeth be adapted to retain a sharp edge and to resist loading because they sometimes must penetrate a highly obdurate substrate during gum extraction by means of gouging or scraping. It has been observed previously that the enamel on the labial surface of the teeth used for extraction is thicker relative to that on the lingual surface in taxa that extract gums, while enamel is more evenly distributed in the anterior teeth of taxa that do not regularly engage in extractive behaviors. This study presents a quantitative methodology for measuring the distribution of labial versus lingual enamel thickness among primate and marsupial taxa in the context of gummivory. Computed microtomography scans of 15 specimens representing 14 taxa were analyzed. Ten measurements were taken at 20% intervals starting from the base of the crown of the extractive tooth to the tip of the cutting edge across the lingual and labial enamel. A method for including worn or broken teeth is also presented. Mann-Whitney U tests, canonical variates analysis, and between-group principal components analysis were used to examine variation in enamel thickness across taxa. Our results suggest that the differential distribution of enamel thickness in the anterior dentition can serve as a signal for gouging behavior; this methodology distinguishes between gougers, scrapers, and nonextractive gummivores. Gouging taxa are characterized by significantly thicker labial enamel relative to the lingual enamel, particularly towards the crown tip. Examination of enamel thickness patterning in these taxa permits a better understanding of the adaptations for the extraction of gums in extant taxa and offers the potential to test hypotheses concerning the dietary adaptations of fossil taxa.

Extraordinary grip strength and specialized myology in the hyper-derived hand of Perodicticus potto?

Previous behavioral reports of the African lorisid, Perodicticus potto, have speculated that these animals have an extraordinary grip strength. This ability is hypothesized to be facilitated by a range of anatomical features within the forelimb, ranging from the presence of a retia mirabilia in its wrist to the hyper-abduction of its pollex. Despite numerous behavioral reports, however, this claim of extraordinary grip strength has not been empirically substantiated. This study quantifies the physiological cross-sectional area of the digital flexor muscles within P. potto. These data are compared with a broad primate sample, including several similarly sized strepsirrhines. Contrary to expectation, we found that P. potto actually has relatively below-average digital flexor PCSA. However, we identified other myological characteristics in the upper limb of P. potto that were unexpected, including the largest brachioradialis muscle (an elbow flexor) among our primate sample, and - despite P. potto having only a vestigial second digit - an independent digital extensor indicis that is absent in almost a quarter of our primate sample.

The Topographic Organization of Retinal Ganglion Cell Density and Spatial Resolving Power in an Unusual Arboreal and Slow-Moving Strepsirhine Primate, the Potto (Perodicticus potto).

The potto (Perodicticus potto) is an arboreal strepsirhine found in the rainforests of central Africa. In contrast to most primates, the potto shows slow-moving locomotion over the upper surface of branches, where it forages for exudates and crawling invertebrates with its head held very close to the substrate. Here, we asked whether the retina of the potto displays topographic specializations in neuronal density that correlate with its unusual lifestyle. Using stereology and retinal wholemounts, we measured the total number and topographic distribution of retinal ganglion cells (total and presumed parasol), as well as estimating the upper limits of the spatial resolution of the potto eye. We estimated ∼210,000 retinal ganglion cells, of which ∼7% (∼14,000) comprise presumed parasol ganglion cells. The topographic distribution of both total and parasol ganglion cells reveals a concentric centroperipheral organization with a nasoventral asymmetry. Combined with the upwardly shifted orbits of the potto, this nasoventral increase in parasol ganglion cell density enhances contrast sensitivity and motion detection skywards, which potentially assists with the detection of predators in the high canopy. The central area of the potto occurs ∼2.5 mm temporal to the optic disc and contains a maximum ganglion cell density of ∼4,300 cells/mm2. We found no anatomical evidence of a fovea within this region. Using maximum ganglion cell density and eye size (∼14 mm), we estimated upper limits of spatial resolving power between 4.1 and 4.4 cycles/degree. Despite their reported reliance on olfaction to detect exudates, this level of spatial resolution potentially assists pottos with foraging for small invertebrates and in the detection of predators.

Morphological convergence in the pubis of slow-moving primates and xenarthrans.

OBJECTIVES: Determining the functional significance of pubic rami is important for reconstructing locomotor behavior of fossil species. The slow loris pelvis, characterized by long pubic rami, is unusual among primates. Long pubic rami may be related to increasing the moment arm of the abdominal musculature during ventroflexion after the termination of hindlimb suspension, which is a common component of slow arboreal quadrupedalism (AQ). Some extant xenarthran species are also slow AQ taxa, and provide an ideal group to test hypotheses of morphologically convergent adaptations to slow AQ. MATERIALS AND METHODS: A model relating abdominal moment arms to pubic morphology is tested in three genera of slow-moving xenarthrans (Bradypus, Choloepus, and Cyclopes) and two species of slow loris (Nycticebus coucang and Perodicticus potto), using a comparative sample of 37 species of primates and xenarthrans. Phylogenetic analyses of variance and regression were performed on pubic dimensions (superior and inferior pubic ramus length, pubic symphysis length). RESULTS: As a locomotor group, slow-moving xenarthrans and lorises share superior pubic rami that are longer than all other locomotor groups; at the species level, there is some overlap among slow AQ and non-slow-AQ taxa. Inferior pubic ramus and pubic symphysis lengths also differ according to locomotor category, but multiple comparisons among locomotor groups are non-significant. DISCUSSION: These results support the hypothesis that superior pubic ramus length is functionally related to slow, suspensory locomotion by increasing the leverage of the ventral abdominal musculature, and demonstrates morphological convergence among two phylogenetically distant groups of mammals that have evolved adaptations for slow, suspensory locomotion.

Exudativory in the Asian loris, Nycticebus: Evolutionary divergence in the toothcomb and M3.

OBJECTIVES: Slow lorises (Nycticebus spp.) are obligate exudativores that gouge tree bark. Dental adaptations for gouging within marmosets, the only other known primate obligate exudativore, are well-known but dental adaptations in Nycticebus are largely unidentified. In an effort to more completely understand potential dental adaptions within Nycticebus and the evolution of this dietary niche within Primates as an order, the present study examined dental morphometrics in the Asian lorises (Nycticebus and Loris). MATERIALS AND METHODS: We compared dental morphometrics between Nycticebus and the insectivorous slender lorises (Loris). Measurements from the toothcomb and select other teeth were taken from 92 specimens. Each variable was scaled by the geometric mean and resulting mean ratios were statistically compared between groups. A biomechanical shape variable was also calculated to estimate the ability of the toothcomb to resist bending that may be experienced during gouging. RESULTS: Toothcombs in Nycticebus were significantly (P < 0.05) more narrow, shorter, and thicker than those in Loris and had a higher calculated ability to withstand bending forces. Nycticebus also had reduced size in the last lower molar relative to Loris. CONCLUSIONS: The more robust, "squared off" toothcomb in Nycticebus matches behavioral observations that these primates gouge to access exudates. Results of the present study indicate that the toothcomb is the likely candidate for the dental tool used in gouging. The size reduction of the lower last molar in Nycticebus, a trait also found in a previous study in exudativorous galagos, may indicate that there is reduced selective pressure in a diet where little mastication would be needed to mechanically process exudates. These results may indicate that reduction in molar size could be a potential dental signature for exudativory, but further studies on a wider phylogenetic range of exudativorous primates would be necessary.

Extreme primates: Ecology and evolution of Asian lorises.

Asia's slow and slender lorises (Nycticebus and Loris) are among nature's most extreme primates. Until recently, it was not understood why lorises have such huge forward-facing eyes, strange steady climbing locomotion, tiny dependent babies, and a bite that potentially can kill a human! Indeed, early studies described them as slow, solitary, and boring. Twenty years of field research now indicate that lorises are among the most intriguing mammal species.

The estimated mechanical advantage of the prosimian ankle joint musculature, and implications for locomotor adaptation.

In this study we compared the power arm lengths and mechanical advantages attributed to 12 lower leg muscles across three prosimian species. The origins and insertions of the lower leg muscles in Garnett's galago, the ring-tailed lemur, and the slow loris were quantified and correlated with positional behaviour. The ankle joint of the galago has a speed-oriented mechanical system, in contrast to that of the slow loris, which exhibits more power-oriented mechanics. The lemur ankle joint exhibited intermediate power arm lengths and an intermediate mechanical advantage relative to the other primates. This result suggests that the mechanical differences in the ankle between the galago and the lemur, taxa that exhibit similar locomotory repertoires, reflect a difference in the kinematics and kinetics of leaping (i.e. generalised vs. specialised leapers). In contrast to leaping primates, lorises have developed a more power-oriented mechanical system as a foot adaptation for positional behaviours such as bridging or cantilevering in their arboreal habitat.

Taxonomy of the Bornean slow loris, with new species Nycticebus kayan (Primates, Lorisidae).

More species of nocturnal primates are now recognized than in the past, because many are cryptic species. Subtle morphological disparities, such as pelage pattern and color variation, vocal cues, and genetics have aided in elucidating the number of diagnosable species in a genus. The slow lorises (genus Nycticebus) once included only two species, but recent taxonomic studies resulted in the description of three additional species; further incompletely explored variability characterizes each of the currently described species. The Bornean loris in particular is characterized by pelage and body size variation. In this study, we explored facemask variation in the Bornean loris (N. menagensis). Differing facemask patterns, particularly influenced by the amount of white on the face, significantly clustered together by geographic regions, separated by notable geographic boundaries. Our results support the recognition of four species of Bornean lorises: N. menagensis, N. bancanus, N. borneanus, and N. kayan. Genetic studies are required to support these findings and to refine further our understanding of the marked variability within the Bornean loris populations.

Molecular Phylogenetic Relationships and Unveiling Novel Genetic Diversity among Slow and Pygmy Lorises, including Resurrection of Xanthonycticebus intermedius.

Genetic analysis of historical museum collections presents an opportunity to clarify the evolutionary history of understudied primate groups, improve taxonomic inferences, and inform conservation efforts. Among the most understudied primate groups, slow and pygmy lorises (genera Nycticebus and Xanthonycticebus) are nocturnal strepsirrhines found in South and Southeast Asia. Previous molecular studies have supported five species, but studies using morphological data suggest the existence of at least nine species. We sequenced four mitochondrial loci, CO1, cytb, d-loop, and ND4, for a total of 3324 aligned characters per sample from 41 historical museum specimens for the most comprehensive geographic coverage to date for these genera. We then combined these sequences with a larger dataset composed of samples collected in Vietnam as well as previously published sequences (total sample size N = 62). We inferred phylogenetic relationships using Bayesian inference and maximum likelihood methods based on data from each locus and on concatenated sequences. We also inferred divergence dates for the most recent common ancestors of major lineages using a BEAST analysis. Consistent with previous studies, we found support for Xanthonycticebus pygmaeus as a basal taxon to the others in the group. We also confirmed the separation between lineages of X. pygmaeus from northern Vietnam/Laos/China and southern Vietnam/Cambodia and included a taxonomic revision recognizing a second taxon of pygmy loris, X. intermedius. Our results found support for multiple reciprocally monophyletic taxa within Borneo and possibly Java. The study will help inform conservation management of these trade-targeted animals as part of a genetic reference database for determining the taxonomic unit and provenance of slow and pygmy lorises confiscated from illegal wildlife trade activities.

Growth and the development of sexual size dimorphism in lorises and galagos.

Three fundamental ontogenetic pathways lead to the development of size differences between males and females. Males and females may grow at the same rate for different durations (bimaturism), grow for the same duration at different rates, or grow at a mix of rate and duration differences. While patterns of growth and the development of adult body size are well established for many haplorhines, the extent to which rate and duration differences affect strepsirrhine growth trajectories remains unclear. Here, we present iterative piecewise regression models that describe the ontogeny of adult body mass for males and females of five lorisoid species (i.e., lorises and galagos) from the Duke Lemur Center. We test the hypotheses that, like most haplorhines, sexual size dimorphism (SSD) is a result of bimaturism, and males and females of monomorphic species grow at the same rate for a similar duration. We confirm that the galagos in this sample (Galago moholi and Otolemur garnettii) show significant SSD that is achieved through bimaturism. Unlike monomorphic lemurids, the lorises in this sample show a diversity of ontogenetic patterns. Loris tardigradus does follow a lemur-like trajectory to monomorphism but Nycticebuscoucang and Nycticebus pygmaeus achieve larger adult female body sizes through a mixture of rate and duration differences. We show that contrary to previous assumptions, there are patterns of both similarity and difference in growth trajectories of comparably sized lorises and galagos. Furthermore, when ontogenetic profiles of lorisoid and lemurid growth are compared, it is evident that lorisoids grow faster for a shorter period of time.

Topsy-turvy locomotion: biomechanical specializations of the elbow in suspended quadrupeds reflect inverted gravitational constraints.

Some tetrapods hang upside down from tree branches when moving horizontally. The ability to walk in quadrupedal suspension has been acquired independently in at least 14 mammalian lineages. During the stance (supportive) phase of quadrupedal suspension, the elbow joint flexor muscles (not the extensors as in upright vertebrates moving overground) are expected to contract to maintain the flexed limb posture. Therefore muscular control in inverted, suspended quadrupeds may require changes of muscle control, and even morphologies, to conditions opposite to those in upright animals. However, the relationships between musculoskeletal morphologies and elbow joint postures during the stance phase in suspended quadrupeds have not been investigated. Our analysis comparing postures and skeletal morphologies in Choloepus (Pilosa), Pteropus (Chiroptera), Nycticebus (Primates) and Cynocephalus (Dermoptera) revealed that the elbow joints of these animals were kept at flexed angles of 70-100 ° during the stance phase of quadrupedal suspension. At these joint angles the moment arms of the elbow joint flexors were roughly maximized, optimizing that component of antigravity support. Our additional measurements from various mammalian species show that suspended quadrupeds have relatively small extensor/flexor ratios in both muscle masses and maximum moment arms. Thus, in contrast to the pattern in normal terrestrial quadrupeds, suspended quadrupeds emphasize flexor over extensor muscles for body support. This condition has evolved independently multiple times, attendant with a loss or reduction of the ability to move in normal upright postures.

Technical note: a landmark-based approach to the study of the ear ossicles using ultra-high-resolution X-ray computed tomography data.

Previous study of the ear ossicles in Primates has demonstrated that they vary on both functional and phylogenetic bases. Such studies have generally employed two-dimensional linear measurements rather than three-dimensional data. The availability of Ultra- high-resolution X-ray computed tomography (UhrCT) has made it possible to accurately image the ossicles so that broadly accepted methodologies for acquiring and studying morphometric data can be applied. Using UhrCT data also allows for the ossicular chain to be studied in anatomical position, so that it is possible to consider the spatial and size relationships of all three bones. One issue impeding the morphometric study of the ear ossicles is a lack of broadly recognized landmarks. Distinguishing landmarks on the ossicles is difficult in part because there are only two areas of articulation in the ossicular chain, one of which (the malleus/incus articulation) has a complex three-dimensional form. A measurement error study is presented demonstrating that a suite of 16 landmarks can be precisely located on reconstructions of the ossicles from UhrCT data. Estimates of measurement error showed that most landmarks were highly replicable, with an average CV for associated interlandmark distances of less than 3%. The positions of these landmarks are chosen to reflect not only the overall shape of the bones in the chain and their relative positions, but also functional parameters. This study should provide a basis for further examination of the smallest bones in the body in three dimensions.

Effects of seasonality on brain size evolution: evidence from strepsirrhine primates.

Seasonal changes in energy supply impose energetic constraints that affect many physiological and behavioral characteristics of organisms. As brains are costly, we predict brain size to be relatively small in species that experience a higher degree of seasonality (expensive brain framework). Alternatively, it has been argued that larger brains give animals the behavioral flexibility to buffer the effects of habitat seasonality (cognitive buffer hypothesis). Here, we test these two hypotheses in a comparative study on strepsirrhine primates (African lorises and Malagasy lemurs) that experience widely varying degrees of seasonality. We found that experienced seasonality is negatively correlated with relative brain size in both groups, controlling for the effect of phylogenetic relationships and possible confounding variables such as the extent of folivory. However, relatively larger-brained lemur species tend to experience less variation in their dietary intake than indicated by the seasonality of their habitat. In conclusion, we found clear support for the hypothesis that seasonality restricts brain size in strepsirrhines as predicted by the expensive brain framework and weak support for the cognitive buffer hypothesis in lemurs.

A digital collection of rare and endangered lemurs and other primates from the Duke Lemur Center.

Scientific study of lemurs, a group of primates found only on Madagascar, is crucial for understanding primate evolution. Unfortunately, lemurs are among the most endangered animals in the world, so there is a strong impetus to maximize as much scientific data as possible from available physical specimens. MicroCT scanning efforts at Duke University have resulted in scans of more than 100 strepsirrhine cadavers representing 18 species from the Duke Lemur Center. An error study of the microCT scanner recovered less than 0.3% error at multiple resolution levels. Scans include specimen overviews and focused, high-resolution selections of complex anatomical regions (e.g., cranium, hands, feet). Scans have been uploaded to MorphoSource, an online digital repository for 3D data. As captive (but free ranging) individuals, these specimens have a wealth of associated information that is largely unavailable for wild populations, including detailed life history data. This digital collection maximizes the information obtained from rare and endangered animals with minimal degradation of the original specimens.

[Hands and feet of prosimians primates. Attempts of morphologic characterization]. / Mains et pieds des primates prosimiens. Essai de caractérisation morphologique.

The length of the carpus and tarsus, the metacarpus and metatarsus, the fingers and toes of 142 prosimian apes was measured. The relationship expressed as a percentage was drawn up for each individual between the length of each osseous part and that of its third metacarpal in order to eliminate the differences related to the size of the rest of the body. This ratio was compared with that of man. The characteristic variations appeared at the level of the subfamilies. CONCERNING THE HAND: The carpus presented the same values as that of man except for that of the indris, which was shorter. The thumb had proportionally the same length as that of man, sometimes longer and sometimes smaller as in the Eulemurs, Hapalidea, Megalapidea, Indrises, Daubentonia and Perodictus. The different metacarpals, including the fourth, were a little shorter than the third. In these subfamilies, the second ray was also often shorter and even much shorter in the Megalapidea and the Perodictus. The other rays were a little longer, in particular the fourth which could exceed the third in rather many subfamilies. CONCERNING THE FOOT: The length of the tarsus was extremely variable. It was twice larger in the Galagoidae, definitely larger in the tarsius and discreetly in the Hapalidae, a little smaller in the other Lemurs and much smaller in the other Prosimian apes, joining in that the near totality of the simians. The hallux was proportionally as long as that of man and sometimes even longer. The metatarsals were sometimes a little longer, sometimes less long, but always appreciably of the same length between them. The other toes were short at the aye aye (daubentonia), of which the foot appeared even smaller than that of man. The toes of the other prosimious resembled much to the fingers and in the propithecus and the perodictus, the fourth took gigantic proportions. There has been establishment of an anatomical relation and functional calculus between the length of the last three rays of the hands and the feet of prosimian apes and the biomechanics of their trapezometacarpal and their first cuneometacarpal joints.

The Distribution of Ki-67 and Doublecortin-Immunopositive Cells in the Brains of Three Strepsirrhine Primates: Galago demidoff, Perodicticus potto, and Lemur catta.

This study investigated the pattern of adult neurogenesis throughout the brains of three prosimian primate species using immunohistochemical techniques for endogenous markers of this neural process. Two species, Galago demidoff and Perodicticus potto, were obtained from wild populations in the primary rainforest of central Africa, while one species, Lemur catta, was captive-bred. Two brains from each species, perfusion-fixed with 4% paraformaldehyde, were sectioned (50 µm section thickness) in sagittal and coronal planes. Using Ki-67 and doublecortin (DCX) antibodies, proliferating cells and immature neurons were identified in the two canonical neurogenic sites of mammals, the subventricular zone of the lateral ventricle (SVZ) giving rise to the rostral migratory stream (RMS), and the subgranular zone of the dentate gyrus of the hippocampus. In addition a temporal migratory stream (TMS), emerging from the temporal horn of the lateral ventricle to supply the piriform cortex and adjacent brain regions with new neurons, was also evident in the three prosimian species. While no Ki-67-immunoreactive cells were observed in the cerebellum, DCX-immunopositive cells were observed in the cerebellar cortex of all three species. These findings are discussed in a phylogenetic context.

Organization of cholinergic, catecholaminergic, serotonergic and orexinergic nuclei in three strepsirrhine primates: Galago demidoff, Perodicticus potto and Lemur catta.

The nuclear organization of the cholinergic, catecholaminergic, serotonergic and orexinergic systems in the brains of three species of strepsirrhine primates is presented. We aimed to investigate the nuclear complement of these neural systems in comparison to those of simian primates, megachiropterans and other mammalian species. The brains were coronally sectioned and immunohistochemically stained with antibodies against choline acetyltransferase, tyrosine hydroxylase, serotonin and orexin-A. The nuclei identified were identical among the strepsirrhine species investigated and identical to previous reports in simian primates. Moreover, a general similarity to other mammals was found, but specific differences in the nuclear complement highlighted potential phylogenetic interrelationships. The central feature of interest was the structure of the locus coeruleus complex in the primates, where a central compactly packed core (A6c) of tyrosine hydroxylase immunopositive neurons was surrounded by a shell of less densely packed (A6d) tyrosine hydroxylase immunopositive neurons. This combination of compact and diffuse divisions of the locus coeruleus complex is only found in primates and megachiropterans of all the mammalian species studied to date. This neural character, along with variances in a range of other neural characters, supports the phylogenetic grouping of primates with megachiropterans as a sister group.

Neuronal and synaptic arrangements of the lateral geniculate nucleus in night-active primates.

The lateral geniculate nucleus (LGN) of Aotus trivirgatus and Nycticebus coucang shows two types of neurons at the ultrastructural level: a large thalamo-cortical relay neuron (TCR) and a small neuron of Golgi type II, which is considered to be an interneuron. The interneuron contains small aggregations of synaptic vesicles in the perikaryon adjacent to the cell membrane in synaptic contact to a TCR neuron. Only in the perikaryon of the interneurons can cilia have their origin. After unilateral enucleation, neurofilamentous or dark degenerations of large boutons with round vesicles (type RL) occur in the outer layers of the contralateral LGN and in the inner layers of the ipsilateral LGN. The optic terminals establish synaptic contacts in the glomerulus with dendritic spines of the TCR neurons and with presynaptic dendrites (F2) of interneurons.

Histology of vermiform appendix-like organ in slow loris.

The vermiform appendix-like organ (VALO) of the slow loris was investigated for its histology and immunohistochemical characteristics. The VALO has a much thinner wall with flat folded mucosa and shallower crypts than the cecal mucosa, while cellular components and population of the mucosa were similar to those of the cecum. No coalescent lymph nodules were seen in the submucosa. Immunohistochemically 5-HT-positive cells in the crypts and CD3- and CD8-positive lymphocytes in the lymph nodules were shown in the VALO as well as in the cecum. These findings suggest that the VALO is a low-differentiated vermiform appendix of the slow loris.

Mapping the nasal airways: using histology to enhance CT-based three-dimensional reconstruction in Nycticebus.

Three-dimensional reconstructions of imaging data are an increasingly common approach for studying anatomical structure. However, certain aspects of anatomy, including microscopic structure and differentiating tissue types, continue to benefit from traditional histological analyses. We present here a detailed methodology for combining data from microCT and histological imaging to create 3D virtual reconstructions for visualization and further analyses. We used this approach to study the distribution of olfactory mucosa on ethmoturbinal I of an adult pygmy slow loris, Nycticebus pygmaeus. MicroCT imaging of the specimen was followed by processing, embedding, and sectioning for histological analysis. We identified corresponding features in the CT and histological data, and used these to reconstruct the plane of section in the CT volume. The CT volume was then digitally re-sliced, such that orthogonal sections of the CT image corresponded to histological sections. Histological images were annotated for the features of interest (in this case, the contour of soft tissue on ethmoturbinal I and the extent of olfactory mucosa), and annotations were transferred to binary masks in the CT volume. These masks were combined with density-based surface reconstructions of the skull to create an enhanced 3D virtual reconstruction, in which the bony surfaces are coded for mucosal function. We identified a series of issues that may be raised in this approach, for example, deformation related to histological processing, and we make recommendations for addressing these issues. This method provides an evidence-based approach to 3D visualization and analysis of microscopic features in an anatomic context.