FIRST REPORT OF CHEMICAL IMMOBILIZATION AND PHYSIOLOGICAL PARAMETERS FOR FREE-RANGING MANED SLOTH (BRADYPUS TORQUATUS), USING A COMBINATION OF KETAMINE AND MEDETOMIDINE.

The maned sloth (Bradypus torquatus) is an endemic and endangered species of two Brazilian states, with much unknown biological information needed to direct conservation actions. Other sloth species have been studied regarding anesthesia; however, there is a lack of anesthesia research for the maned sloth. Anesthetic data were collected from 12 free-range maned sloths that were immobilized for a field examination. Individuals were anesthetized using a combination of ketamine (4.0 mg/kg) and medetomidine (0.03 mg/kg), and antagonized with atipamezole (0.1 mg/kg). Time to induction and recovery were recorded and compared with sex and age classes. After the induction and until antagonist administration, physiological parameters (rectal temperature, heart rate, respiratory rate, and oxygen saturation) were recorded every 10 min during anesthesia and were statistically evaluated over time. Induction was fast (3.21 ± 0.76), but recovery was longer (113.3 ± 18) when compared to other studies. Induction and recovery times were not different across sex or age classes. Rectal temperature, heart rate, and oxygen saturation remained stable throughout the procedure. Respiratory rate significantly decreased over time, from 18.25 ± 7.03 to 13.17 ± 3.66 movements per minute. Our results indicate that the described combination of ketamine and medetomidine is a safe and effective choice for anesthesia of maned sloths.

Aortic branches and rete mirabile of the limbs of two-toed sloth (Choloepus didactylus).

Choloepus didactylus has reduced metabolism and difficulty in thermoregulation owing to its low body mass, and there are few studies related to the vascularization of abdominal and thoracic organs in this species. Therefore, we macroscopically described the arteries that comprise the aortic arch, thoracic aorta, and abdominal aorta. Six specimens were used, and their arterial systems filled with red latex before fixation in 10% formaldehyde, and fragments of the rete mirabile were processed for histological analysis using light and scanning electron microscopy. In these species, the aortic arch had two branches: the brachiocephalic trunk and left subclavian artery. The initial portion of the abdominal aorta presented four different ramifications, besides to the peculiarities of the adrenal, renal, and iliac arteries. Microscopy of the rete mirabile revealed a muscular artery surrounded by smaller muscular arteries, veins, nerves, and lymphatic tissue joined by connective tissue. Thus, the data obtained have clinical and surgical importance, with applicability in procedures involving vascularization of the thoracic and abdominal organs. We suggest that the rete mirabile is an efficient thermoregulatory structure because it allows the accumulation of blood and the countercurrent heat exchange, as there is no blood mixing.

Muscle architectural properties indicate a primary role in support for the pelvic limb of three-toed sloths (Bradypus variegatus).

Tree sloths evolved below-branch locomotion making them one of few mammalian taxa beyond primates for which suspension is nearly obligatory. Suspension requires strong limb flexor muscles that provide both propulsion and braking/support, and available locomotor kinetics data indicate that these roles differ between fore- and hindlimb pairs. Muscle structure in the pelvic limb is hypothesized to be a key anatomical correlate of function in braking/support during suspensory walking and propulsion and/or support during vertical climbing. This expectation was tested by quantifying architecture properties in the hindlimb limb musculature of brown-throated three-toed sloths (Bradypus variegatus: N = 7) to distinguish the roles of the flexor/extensor functional muscle groups at each joint. Measurements of muscle moment arm (rm ), mass, belly length, fascicle length, pennation angle, and physiological cross-sectional area (PCSA) were taken from n = 45 muscles. Overall, most muscles studied show properties for contractile excursion and fast joint rotational velocity. However, the flexor musculature is more massive (p = 0.048) and has larger PCSA (p = 0.003) than the extensors, especially at the knee joint and digits where well-developed and strong flexors are capable of applying large joint torque. Moreover, selected hip flexors/extensors and knee flexors have modified long rm that can amplify applied joint torque in muscles with otherwise long, parallel fascicles, and one muscle (m. iliopsoas) was capable of moderately high power in B. variegatus. The architectural properties observed in the hip flexors and extensors match well with roles in suspensory braking and vertical propulsion, respectively, whereas strong knee flexors and digital flexors appear to be the main muscles providing suspensory support in the pelvic limb. With aid in support by the forelimbs and the use of adaptive slow locomotion and slow muscle fiber recruitment patterns, structure-function in the tensile limb systems of sloths appears to collectively represent an additional mechanism for energy conservation.

FATAL GASTRIC DILATATION AND VOLVULUS IN THREE CAPTIVE JUVENILE LINNAEUS'S TWO-TOED SLOTHS (CHOLOEPUS DIDACTYLUS).

Linnaeus's two-toed sloth (Choloepus didactylus) is one of two extant neotropical species of the family Megalonychidae. Despite their being commonly kept under managed care, the digestive physiology of sloths remains poorly understood. Gastrointestinal disease has been reported as a primary or contributing cause of morbidity and mortality in captive two-toed and three-toed (Bradypus spp.) sloths. Gastric dilatation due to gas accumulation ("bloat") has been reported in sloths; however, a literature search failed to yield any published mention of gastric volvulus in any sloth species. Following an inquiry sent to the electronic mailing lists of the American Association of Zoo Veterinarians, the European Association of Zoo and Wildlife Veterinarians, and the LatinVets community, three cases of fatal gastric dilatation and volvulus (GDV) were identified in one male and two female Linnaeus's two-toed sloths from institutions in the United States, Canada, and Germany. All cases occurred in juvenile sloths <1 yr of age. Two animals were primarily hand reared, whereas one was reared primarily by its dam. Two animals were found dead with no overt premonitory signs, whereas one animal died following a 3-wk course of waxing and waning clinical signs consistent with gastric gas accumulation. In all cases, GDV was diagnosed on postmortem examination. Similar to other species, the condition is likely subsequent to a combination of host- and husbandry-related contributing factors. Further research into sloth husbandry is required in order to take an evidence-based approach to their management.

Pump the brakes! The hindlimbs of three-toed sloths decelerate and support suspensory locomotion.

Modern tree sloths are one of few mammalian taxa for which quadrupedal suspension is obligatory. Sloth limb musculature is specialized for slow velocity, large force contractions that stabilize their body below branches and conserve energy during locomotion. However, it is unknown whether two- and three-toed sloths converge in their use of limb kinetics and if these patterns are comparable to how primates perform arboreal suspensory locomotion. This study addressed this need by collecting limb loading data in three-toed sloths (Bradypus variegatus; N=5) during suspensory walking. Sloths performed locomotor trials at their preferred speed on an instrumented beam apparatus with a force platform as the central supporting segment. Peak forces and impulses of the forelimb and hindlimb were recorded and analyzed in three dimensions. The hindlimbs of B. variegatus apply large braking forces greater in magnitude than peak forces generated by the forelimbs in propulsion, a pattern consistent with that observed in two-toed sloths. However, B. variegatus exhibits hindlimb-biased body weight support in vertical peak forces and impulse, with appreciable laterally directed forces in each limb pair, both of which vary from limb loading distributions in two-toed sloths. Moreover, body weight distribution between limb pairs is opposite to that employed by primates during quadrupedal suspension. Thus, there appear to be multiple strategies for achieving suspensory locomotion in arboreal mammals. These differences may be attributable to anatomical variation or phylogenetic position, but as of yet an explanation remains unknown. Future EMG analyses are expected to provide insight into how specific hindlimb muscle groups contribute to braking forces and stabilizing the center of mass of sloths during suspension.

Three toes and three modes: Dynamics of terrestrial, suspensory, and vertical locomotion in brown-throated three-toed sloths (Bradypodidae, Xenarthra).

Living sloths exhibit numerous anatomical specializations towards inverted quadrupedalism, however, previous studies have noted a more varied locomotor repertoire than previously anticipated. In this study, we present spatiotemporal gait characteristics and triaxial kinetic data from the brown-throated three-toed sloth (Bradypus variegatus) across three locomotor modes: terrestrial quadrupedal "crawling", suspensory walking, and vertical climbing. Compared to quadrupedal crawling and suspensory walking, B. variegatus adopted longer contact times and stride durations, larger duty factors, and greater speed during vertical climbing. Net fore-aft impulses were significantly greater during vertical climbing in both limb pairs than in quadrupedal crawling and suspensory walking. Functionally, during quadrupedal crawling and vertical climbing, both limb pairs served propulsive roles, while differentiation between a propulsive forelimb and braking hindlimb was observed during suspension. Net tangential forces differentiated vertical climbing kinetics from the other modes of locomotion, with the introduction of bidirectional pulling and pushing forces in the forelimb and hindlimb, respectively. The net mediolateral impulses were similar in vertical climbing and quadrupedal crawling as both limb pairs directed forces in one direction, whereas during suspensory walking, the laterally dominant forelimb was opposed by the medially dominant hindlimb. In total, this study provides novel data on the diverse locomotor dynamics in a slow-moving arboreal tetrapod and posits new testable hypotheses about the neuroplasticity and ease of transitioning between locomotor behaviors. The strikingly similar kinetic profiles of quadrupedal crawling and suspensory walking compared to vertical climbing suggest shared neuromuscular and mechanical demands between these mirrored locomotor modes.

Testing mechanisms for weight support distribution during inverted quadrupedalism in primates.

A key characteristic of primate above-branch arboreal locomotion is hindlimb-biased weight support, subverting the typical mammalian condition in which the majority of the body weight is supported by the forelimb. This shift is thought to reflect an adaptation toward the arboreal niches exploited by early primates. However, above-branch quadrupedalism represents only one locomotor mode employed by primates in arboreal contexts. Inverted quadrupedal gaits, in which primates are suspended beneath branches by their hands and feet, have been documented in more than 50 primate taxa. This gait is characterized by a return to forelimb-biased weight distributions and a transition from peak vertical forces being greatest in the hindlimb to being greatest in the forelimb, which may occur to protect the hindlimb from high magnitudes of tensile loading when inverted. In this study, we compare kinetic and kinematic data during upright and inverted quadrupedalism in Lemur catta, Varecia variegata, Cebus capucinus, and Saimiri sciureus. These data are referenced against a classical inverted quadrupedal model: the two-toed sloth (Choloepus didactylus). Our findings show that inverted quadrupedalism in primates is differentiated from above-branch quadrupedalism by increases in forelimb weight support, forelimb contact times, and both forelimb and hindlimb joint excursions. Previously postulated biomechanical models outlining mechanisms relating to the control of weight support during upright walking do not translate well to inverted quadrupedal walking. We suggest that inverted primates may simply be adopting basal neuromuscular gait characteristics and applying them facultatively to this infrequent locomotor behavior.

Myology of the pelvic limb of the brown-throated three-toed sloth (Bradypus variegatus).

Tree sloths rely on their limb flexors for bodyweight support and joint stability during suspensory locomotion and posture. This study aims to describe the myology of three-toed sloths and identify limb muscle traits that indicate modification for suspensorial habit. The pelvic limbs of the brown-throated three-toed sloth (Bradypus variegatus) were dissected, muscle belly mass was recorded, and the structural arrangements of the muscles were documented and compared with the available myological accounts for sloths. Overall, the limb musculature is simplified by containing muscles with generally long and parallel fascicles. A number of specific and informative muscle traits are additionally observed in the pelvic limb of B. variegatus: well-developed hip flexors and hip extensors each displaying several fused bellies; massive knee flexors; two heads of the m. adductor longus and m. gracilis; robust digital flexors and flexor tendons; m. tibialis cranialis muscle complex originating from the tibia and fibula and containing a modified m. extensor digitorum I longus; appreciable muscle mass devoted to ankle flexion and hindfoot supination; only m. extensor digitorum brevis acts to extend the digits. Collectively, the findings for tree sloths emphasize muscle mass and organization for suspensory support namely by the hip flexors, knee flexors, and limb adductors, for which the latter two groups may stabilize suspensory postures by exerting appreciable medially-directed force on the substrate. Specializations in the distal limb are also apparent for sustained purchase of the substrate by forceful digital flexion coupled with strong ankle flexion and supination of the hind feet, which is permitted by the reorganization of several digital extensors. Moreover, the reduction or loss of other digital flexor and ab-adductor muscles marks a dramatic simplification of the intrinsic foot musculature in B. variegatus, the extent to which varies across extant species of two- and three-toed tree sloths and likely is related to substrate preference/use.

A novel method for quantifying femoral neck anteversion: A case study in extinct and extant sloths.

Extinct sloths represent a wide range of morphological, locomotor, and body size variation. Researchers have examined femoral neck angle in two dimensions to hypothesize locomotor behaviors in this group; however, this measure does not account for femoral neck anteversion. Here, we present a new method for quantifying femoral neck anteversion angle, in addition to femoral neck angle, to capture the 3D position of the femoral head/neck. Femora of extant (n = 21; Bradypus and Choloepus) and extinct (n = 49; Acratocnus, Megalocnus, Neocnus, and Parocnus) sloths were surface scanned and their surface models used to calculate three angles of femoral neck anteversion and femoral neck angle. Femoral neck anteversion was calculated as the angle between the femoral neck axis and the geometric axis of the femoral condyles (GA), the 35% cross section axis, and a trochanter axis. Femoral neck angle was calculated as the angle between the femoral neck and shaft axes. Genera were compared using ANOVAs with post hoc multiple comparisons for each angle. Femoral neck angle and femoral neck anteversion relative to the cross section were also analyzed. Significant differences among genera exist for all angles, (p < .001) but not all angles separate all genera. Femoral neck and anteversion angles typically yield different results, demonstrating the utility of analyzing both angles. The GA and cross section angles are highly correlated in sloths, with the exception of comparisons among Megalocnus, Parocnus, and Neocnus, suggesting morphological variation in the distal femur. While this method was applied to sloths, it has broad applicability to mammalian groups.

Anatomia macroscópica interna e avaliação elétrica do coração de Bradypus variegatus / Internal macroscopic anatomy and electrical evaluation of the Bradypus variegatus heart

The Bradypus variegatus species presents peculiar anatomophysiological properties and many aspects of its organic systems still need to be clarified, especially regarding the cardiovascular system, given its participation in vital activities. Disorderly anthropic action has had drastic consequences in sloth populations and the need to treat sick and injured animals is increasingly common. To this end, the importance of knowing its characteristics is emphasized. Therefore, this study proposed to describe the internal macroscopic structures of the sloth's heart, as well as to measure the ventricular walls and indicate the electrical activity of the organ. For the dissections, 15 Bradypus variegatus cadavers were used (1 young female, 9 adults females and 5 adult males) belonging to the Área de Anatomia of the Departamento de Morfologia e Fisiologia Animal (DMFA), Universidade Federal Rural de Pernambuco (UFRPE), Recide, PE, Brazil. After they were fixed and preserved, the specimens received a midsagittal incision in the chest, followed by soft tissue folding and removal of ribs to access the heart. The organ was derived from the cavity and sectioned sagittal medially to identify its internal anatomy. Ventricular walls and interventricular septum were measured with a steel caliper (150 mm / 0.02 mm). An electrocardiogram was performed to determine the electrical profile on 5 healthy B. variegatus sloths, living under semi-livestock conditions at the Recife Zoo, PE, Brazil. The electrodes were taken from the regions, scapular and glutes of the animals that were called hugging a keeper during the procedure, carried out in the Zoo itself, using a portable device. Based on the data obtained, sloths have cardiac chambers separated by septa, however between atria and ventricles, in both antimeres, there are atrioventricular ostia, where valves are found, consisting of 3 valves on the right and 2 on the left. The atria are practically smooth inside and have their cavity enlarged by the atria, the right being larger than the left, these having a greater amount of pectineal muscles in relation to the atria. The ventricles have trabeculae and papillary muscles, 3 on the right and 2 on the left. These muscles hold the tendinous chords that connect the valves. The existence of trabeculae marginal septum was not evidenced. The thickness of the wall of the left ventricle, as well as that of the interventricular septum, proved to be greater than the thickness of the wall of the right ventricle, regardless of the age or sex of the animals. Based on the electrocardiographic recordings, the sloths presented sinus rhythm, with a heart rate between 67 and 100 bpm. The electrical axis ranged from -60º to -90º. The P wave is smoother than the QRS complex. While the S-T segment was classified as isoelectric. The T wave was shown to be + and predominantly > or = at 25% of the S wave, which characterized an rS type QRS deflection in both females and males. The general characteristics of the cardiac chambers in sloths are similar to those observe in other domestic and wild mammals. However, the presence of pectineal muscles associated with the atria and auricles differs from that observed in mammals such as the paca and raccoon and in birds such as the ostrich, which have trabecular structures in these cavities. The number of valves in sloths is equal to the anteater. However, it has a marginal trabeculae septum, not seen in Bradypus variegatus. According to the electrocardiographic findings, the rhythm was sinus, but much lower than that observed in the capuchin monkey, which also maintains arboreal habits.(AU)

RENAL EVALUATION IN CHOLOEPUS SPECIES.

Two species of sloths in the family Megalonychidae, Hoffmann's (Choloepus hoffmanni) and Linnaeus's (Choloepus didactylus) two-toed sloths, are commonly held in zoological institutions. Despite frequent published reports of urinary tract disease in these species, reports of diagnostics are mostly limited to descriptions of hematology and serum chemistry. In this study, repeated urinalysis, urinary chemistry, serum chemistry, and radiographs were collected opportunistically from six Hoffmann's and five Linnaeus's sloths. Proteinuria, bacteriuria, low urine specific gravity, and crystalluria were common in the absence of other signs of urinary tract pathology.

Ancient Mitogenomes Reveal the Evolutionary History and Biogeography of Sloths.

Living sloths represent two distinct lineages of small-sized mammals that independently evolved arboreality from terrestrial ancestors. The six extant species are the survivors of an evolutionary radiation marked by the extinction of large terrestrial forms at the end of the Quaternary. Until now, sloth evolutionary history has mainly been reconstructed from phylogenetic analyses of morphological characters. Here, we used ancient DNA methods to successfully sequence 10 extinct sloth mitogenomes encompassing all major lineages. This includes the iconic continental ground sloths Megatherium, Megalonyx, Mylodon, and Nothrotheriops and the smaller endemic Caribbean sloths Parocnus and Acratocnus. Phylogenetic analyses identify eight distinct lineages grouped in three well-supported clades, whose interrelationships are markedly incongruent with the currently accepted morphological topology. We show that recently extinct Caribbean sloths have a single origin but comprise two highly divergent lineages that are not directly related to living two-fingered sloths, which instead group with Mylodon. Moreover, living three-fingered sloths do not represent the sister group to all other sloths but are nested within a clade of extinct ground sloths including Megatherium, Megalonyx, and Nothrotheriops. Molecular dating also reveals that the eight newly recognized sloth families all originated between 36 and 28 million years ago (mya). The early divergence of recently extinct Caribbean sloths around 35 mya is consistent with the debated GAARlandia hypothesis postulating the existence at that time of a biogeographic connection between northern South America and the Greater Antilles. This new molecular phylogeny has major implications for reinterpreting sloth morphological evolution, biogeography, and diversification history.

The demography of a resource specialist in the tropics: Cecropia trees and the fitness of three-toed sloths.

Resource specialists persist in a narrow range of resources. Consequently, the abundance of key resources should drive vital rates, individual fitness, and population viability. While Neotropical forests feature both high levels of biodiversity and numbers of specialist species, no studies have directly evaluated how the variation of key resources affects the fitness of a tropical specialist. Here, we quantified the effect of key tree species density and forest cover on the fitness of three-toed sloths ( Bradypus variegatus), an arboreal folivore strongly associated with Cecropia trees in Costa Rica, using a multi-year demographic, genetic, and space-use dataset. We found that the density of Cecropia trees was strongly and positively related to both adult survival and reproductive output. A matrix model parametrized with Cecropia-demography relationships suggested positive growth of sloth populations, even at low densities of Cecropia (0.7 trees ha-1). Our study shows the first direct link between the density of a key resource to demographic consequences of a tropical specialist, underscoring the sensitivity of tropical specialists to the loss of a single key resource, but also point to targeted conservation measures to increase that resource. Finally, our study reveals that previously disturbed and regenerating environments can support viable populations of tropical specialists.

Interacting circadian and homeostatic processes with opportunity cost: A mathematical model of sleep with application to two mammalian species.

This paper introduces a new model of sleep for mammals. It extends the classic 'two-process' model of sleep to account for differences in external circumstances. We apply this model to previously-collected data on elephants and sloths, comparing sleep patterns in the wild with sleep patterns in captivity. We find that the model does very well in explaining sleeping patterns for both types of animals, in both the captive state and in the wild state.

Bone cortical compactness in 'tree sloths' reflects convergent evolution.

Bone remodeling, one of the main processes that regulate bone microstructure, consists of bone resorption followed by the deposition of secondary bone at the same location. Remodeling intensity varies among taxa, but a characteristically compact cortex is ubiquitous in the long bones of mature terrestrial mammals. A previous analysis found that cortical bone in a few 'tree sloth' (Bradypus and Choloepus) specimens is heavily remodeled and characterized by numerous immature secondary osteons, suggesting that these animals were remodeling their bones at high rate until late in their ontogeny. This study aims at testing if this remodeling is generally present in 'tree sloths', using a quantitative analysis of the humeral cortical compactness (CC) among xenarthrans. The results of the investigation of humeral diaphyseal cross-sections of 26 specimens belonging to 10 xenarthran species including specimens from both extinct and extant species indicate that in 'tree sloths' the CC is significantly lower than in the other sampled xenarthrans. No significant difference was found between the CC of the two genera of 'tree sloths'. Our results are consistent with the hypothesis that the cortical bone of 'tree sloths' in general undergoes intense and balanced remodeling that is maintained until late (possibly throughout) in their ontogeny. In the light of xenarthran phylogeny, low CC represents another convergence between the long-separated 'tree sloth' lineages. Although the exact structural and/or functional demands that are associated with this trait are hitherto unknown, several hypotheses are suggested here, including a relationship to their relatively low metabolism and to the mechanical demands imposed upon the bones by the suspensory posture and locomotion, which was independently acquired by the two genera of 'tree sloths'.

A suspensory way of life: Integrating locomotion, postures, limb movements, and forces in two-toed sloths Choloepus didactylus (Megalonychidae, Folivora, Pilosa).

Over the last decade, we have learned much about the anatomy, evolutionary history, and biomechanics of the extant sloths. However, most of this study has involved studying sloths in controlled conditions, and few studies have explored how these animals are behaving in a naturalistic setting. In this study, we integrate positional activities in naturalistic conditions with kinematic and kinetic observations collected on a simulated runway to best capture the biomechanical behavior of Linnaeus's two-toed sloths. We confirm that the dominant positional behaviors consist of hanging below the support using a combination of forelimbs and hindlimbs, and walking quadrupedally below the branches. The majority of these behaviors occur on horizontal substrates that are approximately 5-10 cm in diameter. The kinematics of suspensory walking observed both in the naturalistic settings and on simulated arboreal runways are dominated by movement of the proximal limb elements, while distal limb elements tend to show little excursion. Joint kinematics are similar between the naturalistic setting and the simulated runway, but movements of the shoulder and hip tend to be exaggerated while moving in simulated conditions. Kinetic patterns of the two-toed sloth can be explained almost entirely by considering them as an inverted linked strut. However, medially directed forces toward the substrate were more frequent than expected in the forelimb, which may help sloths maintain a better "grip" on the substrate. This study serves as a model of how to gain a comprehensive understanding of the functional-adaptive profile of a particular species.

Evolutionary adaptation to aquatic lifestyle in extinct sloths can lead to systemic alteration of bone structure.

Through phenotypic plasticity, bones can change in structure and morphology, in response to physiological and biomechanical influences over the course of individual life. Changes in bones also occur in evolution as functional adaptations to the environment. In this study, we report on the evolution of bone mass increase (BMI) that occurred in the postcranium and skull of extinct aquatic sloths. Although non-pathological BMI in postcranial skeleton has been known in aquatic mammals, we here document general BMI in the skull for the first time. We present evidence of thickening of the nasal turbinates, nasal septum and cribriform plate, further thickening of the frontals, and infilling of sinus spaces by compact bone in the late and more aquatic species of the extinct sloth Thalassocnus Systemic bone mass increase occurred among the successively more aquatic species of Thalassocnus, as an evolutionary adaptation to the lineage's changing environment. The newly documented pachyostotic turbinates appear to have conferred little or no functional advantage and are here hypothesized as a correlation with or consequence of the systemic BMI among Thalassocnus species. This could, in turn, be consistent with a genetic accommodation of a physiological adjustment to a change of environment.

Cheap labor: myosin fiber type expression and enzyme activity in the forelimb musculature of sloths (Pilosa: Xenarthra).

Sloths are canopy-dwelling inhabitants of American neotropical rainforests that exhibit suspensory behaviors. These abilities require both strength and muscular endurance to hang for extended periods of time; however, the skeletal muscle mass of sloths is reduced, thus requiring modifications to muscle architecture and leverage for large joint torque. We hypothesize that intrinsic muscle properties are also modified for fatigue resistance and predict a heterogeneous expression of slow/fast myosin heavy chain (MHC) fibers that utilize oxidative metabolic pathways for economic force production. MHC fiber type distribution and energy metabolism in the forelimb muscles of three-toed ( Bradypus variegatus, n = 5) and two-toed ( Choloepus hoffmanni, n = 4) sloths were evaluated using SDS-PAGE, immunohistochemistry, and enzyme activity assays. The results partially support our hypothesis by a primary expression of the slow MHC-1 isoform as well as moderate expression of fast MHC-2A fibers, whereas few hybrid MHC-1/2A fibers were found in both species. MHC-1 fibers were larger in cross-sectional area (CSA) than MHC-2A fibers and comprised the greatest percentage of CSA in each muscle sampled. Enzyme assays showed elevated activity for the anaerobic enzymes creatine kinase and lactate dehydrogenase compared with low activity for aerobic markers citrate synthase and 3-hydroxyacetyl CoA dehydrogenase. These findings suggest that sloth forelimb muscles may rely heavily on rapid ATP resynthesis pathways, and lactate accumulation may be beneficial. The intrinsic properties observed match well with suspensory requirements, and these modifications may have further evolved in unison with low metabolism and slow movement patterns as means to systemically conserve energy. NEW & NOTEWORTHY Myosin heavy chain (MHC) fiber type and fiber metabolic properties were evaluated to understand the ability of sloths to remain suspended for extended periods without muscle fatigue. Broad distributions of large, slow MHC-1 fibers as well as small, fast MHC-2A fibers are expressed in sloth forelimbs, but muscle metabolism is generally not correlated with myosin fiber type or body size. Sloth muscles rely on rapid, anaerobic pathways to resist fatigue and sustain force production.

Digestive physiology, metabolism and methane production of captive Linné's two-toed sloths (Choloepus didactylus).

Sloths are renowned for their low metabolic rate, low food intake and low defecation frequency. We investigated factors of digestive physiology and energy metabolism in four captive individuals (mean body mass 10.0 ± SD 3.7 kg) of a hitherto mostly unstudied sloth species, Linné's two-toed sloth (Choloepus didactylus), in a 2-week digestion recording and 23-h respiration experiment on animals fed a standard zoo diet of vegetables and starchy components. Dry matter intake, defecation frequency and particle mean retention time (MRT) in the gastrointestinal tract (GIT) were 12 ± 3 g/(kg(0.75)  day), once every 5 days and >140 h in three individuals, but 53 g/(kg(0.75)  day), daily and 82 h in one individual that was apparently compensating for a period of weight loss prior to the experiment. In all animals, solute marker was eliminated at a faster rate than the particle marker, indicating 'digesta washing' in the sloths' GIT. The overall metabolic rate calculated from oxygen consumption matched the metabolisable energy intake in three individuals [173 ± 22 vs. 168 ± 44 kJ/(kg(0.75)  day)] but not in the fourth one [225 vs. 698 kJ/(kg(0.75)  day)], supporting the interpretation that this animal was replenishing body stores. In spite of the low food intake and the low-fibre diet (209 ± 26 g neutral detergent fibre/kg dry matter), methane production was rather high accounting for 9.4 ± 0.8% of gross energy intake (2.7% in the fourth individual), which exceeded literature data for ruminants on forage-only diets. These results corroborate literature reports on low intake, low defecation frequency, low metabolic rate and long MRT in other sloth species. The long MRT is probably responsible for the comparatively high methane production, providing more opportunity for methanogenic archaea than in other non-ruminant mammals to produce significant amounts of methane.