Sloth metabolism may make survival untenable under climate change scenarios.

Although climate change is predicted to have a substantial effect on the energetic requirements of organisms, the longer-term implications are often unclear. Sloths are limited by the rate at which they can acquire energy and are unable to regulate core body temperature (Tb) to the extent seen in most mammals. Therefore, the metabolic impacts of climate change on sloths are expected to be profound. Here we use indirect calorimetry to measure the oxygen consumption (VO2) and Tb of highland and lowland two-fingered sloths (Choloepus hoffmanni) when exposed to a range of different ambient temperatures (Ta) (18 °C -34 °C), and additionally record changes in Tb and posture over several days in response to natural fluctuations in Ta. We use the resultant data to predict the impact of future climate change on the metabolic rate and Tb of the different sloth populations. The metabolic responses of sloths originating from the two sites differed at high Ta's, with lowland sloths invoking metabolic depression as temperatures rose above their apparent 'thermally-active zone' (TAZ), whereas highland sloths showed increased RMR. Based on climate change estimates for the year 2100, we predict that high-altitude sloths are likely to experience a substantial increase in metabolic rate which, due to their intrinsic energy processing limitations and restricted geographical plasticity, may make their survival untenable in a warming climate.

Variations in dietary patterns of living sloths revealed by finite element analysis of jaws.

Although extinct sloths exhibited a wide range of dietary habits, modes of locomotion, and occupied various niches across the Americas, modern sloths are considered quite similar in their habits. The dietary habits of living sloths can be directly observed in the wild, and understanding the mechanical behavior of their jaws during chewing through finite element analysis (FEA) provides a valuable validation tool for comparative analysis with their extinct counterparts. In this study, we used FEA to simulate the mechanical behavior of sloth mandibles under lateral mastication loads, using it as a proxy for oral processing. Our research focused on the six extant sloth species to better understand their diets and validate the use of FEA for studying their extinct relatives. We found that all living sloths have the predominancy of low-stress areas in their mandibles but with significant differences. Choloepus didactylus had larger high-stress areas, which could be linked to a reduced need for processing tougher foods as an opportunistic generalist. Bradypus variegatus and Choloepus hoffmanni are shown to be similar, displaying large low-stress areas, indicating greater oral processing capacity in a seasonal and more competitive environment. Bradypus torquatus, Bradypus pygmaeus, and Bradypus tridactylus exhibited intermediary processing patterns, which can be linked to a stable food supply in more stable environments and a reduced requirement for extensive oral processing capacity. This study sheds light on extant sloths' dietary adaptations and has implications for understanding the ecological roles and evolutionary history of their extinct counterparts.

Evolutionary Specializations in the Venous Anatomy of the Two-Toed Sloth (Choloepus didactylus): Insights from CT-scan 3D Reconstructions.

The venous anatomy of the two-toed sloth (Choloepus didactylus) remains poorly understood, particularly in living specimens due to the limitations of traditional cadaveric studies. This study aims to describe the unique venous structures of Choloepus didactylus using computed tomography, enhancing our understanding of this species in a live setting. Three living Choloepus didactylus underwent CT scans as part of routine clinical assessments. The images were reconstructed using 3D Slicer software (version 5.6.2), focusing on the caudal vena cava anatomy. The reconstructions confirmed the presence of a significant intravertebral vein, showing complex venous connections through the ventral sacral foramen and vertebral foramina. These findings highlight notable anatomical variations and challenge existing literature on the species' venous architecture. By employing modern imaging technologies, this research provides new insights into the venous anatomy of Choloepus didactylus, demonstrating the value of non-invasive techniques in studying the anatomical features of live animals, thereby offering a foundation for further comparative and evolutionary studies.

Taphonomy of harpy eagle predation on primates and other mammals.

The goal of this study is to provide a taphonomic analysis of bone fragments found in harpy eagle nests in the Brazilian Amazonia, utilizing the largest sample of prey remains collected to date. Harpy eagle kill samples were collected from nine nests, between June 2016 and December 2020 in Mato Grosso, Brazil. We identified the specimens, calculated the number of identified specimens (NISP) and minimum number of individuals (MNI). These metrics were used to estimate bone survivability and fragmentation. A total of 1661 specimens (NISP) were collected, representing a minimum number of 234 individuals (MNI). We identified at least nine species of primates, which represent 63.8% of the individuals in the kill sample. Harpy eagles preyed mostly on the medium-sized capuchin and bearded saki monkeys (28.2% of the MNI), and two-toed sloths (17.7% of the MNI). The large woolly monkeys also represented a significant portion of the sample (11.5% of the MNI). Three distinct patterns of bone survivability were found, one characterizing two-toed sloths, another characterizing medium-sized monkeys, and a third typical of woolly monkeys. We conclude that harpy eagle predation leaves an identifiable signature on the prey with a bone survivability pattern specific to each taxon. The intertaxon variations observed in the taphonomic signatures of harpy eagle kills should be taken into account when evaluating the potential influence of these raptors as accumulators of bone material in both paleontological and neontological assemblages.

The Speaker Method: A Novel Release Method for Offspring Mammals and 5-Year Study on Three Costa Rican Mammals.

Nowadays, wild animals are threatened by humans, with the number of species and individuals decreasing during recent years. Wildlife rescue centers play a vital role in the conservation of wildlife populations. This study aims to describe a new release technique, the Speaker Method, to rescue and facilitate the reunion of different baby mammals that arrived at a wildlife rescue center with their mothers within their natural habitat, avoiding the need for captivity. This method is based on a recorded baby's cry played on a speaker to make a "call effect" in the mother. The efficacy of the Speaker Method for babies' reunion with their mothers was 45.8% in Hoffmann's two-toed sloths (Choloepus hoffmanni) and 91.9% in brown-throated sloths (Bradypus variegatus). Among the mantled howler monkeys (Alouatta palliata), 50% of the babies could be released using this new technique. The findings suggest that the method could be helpful in the early release of young individuals, highlighting higher release outcomes in these three species compared to traditional nursery care provided by human caretakers, who face inherent difficulties in raising young animals without their mothers.

The behaviour and activity budgets of two sympatric sloths; Bradypus variegatus and Choloepus hoffmanni.

It is usually beneficial for species to restrict activity to a particular phase of the 24-hour cycle as this enables the development of morphological and behavioural adaptations to enhance survival under specific biotic and abiotic conditions. Sloth activity patterns are thought to be strongly related to the environmental conditions due to the metabolic consequences of having a low and highly variable core body temperature. Understanding the drivers of sloth activity and their ability to withstand environmental fluctuations is of growing importance for the development of effective conservation measures, particularly when we consider the vulnerability of tropical ecosystems to climate change and the escalating impacts of anthropogenic activities in South and Central America. Unfortunately, the cryptic nature of sloths makes long term observational research difficult and so there is very little existing literature examining the behavioural ecology of wild sloths. Here, we used micro data loggers to continuously record, for the first time, the behaviour of both Bradypus and Choloepus sloths over periods of days to weeks. We investigate how fluctuations in the environmental conditions affect the activity of sloths inhabiting a lowland rainforest on the Caribbean coast of Costa Rica and examined how this might relate to their low power lifestyle. Both Bradypus and Choloepus sloths were found to be cathemeral in their activity, with high levels of between-individual and within-individual variation in the amounts of time spent active, and in the temporal distribution of activity over the 24-hour cycle. Daily temperature did not affect activity, although Bradypus sloths were found to show increased nocturnal activity on colder nights, and on nights following colder days. Our results demonstrate a distinct lack of synchronicity within the same population, and we suggest that this pattern provides sloths with the flexibility to exploit favourable environmental conditions whilst reducing the threat of predation.

Antimicrobial resistance and virulence profiles of Enterobacterales isolated from two-finger and three-finger sloths (Choloepus hoffmanni and Bradypus variegatus) of Costa Rica.

Background: Wildlife has been recently recognized as an environmental reservoir for antimicrobial resistance (AMR). However, less information on this topic is available in animals released back into the wild after rehabilitation in wildlife facilities, compared with studies performed exclusively in captive or free-ranging wildlife. This study aimed to evaluate the potential influence of captivity and/or treatment while in captivity of wild sloths on the AMR and virulence profiles of sloths' Enterobacterales. Methods: Oral and rectal swab samples were collected from 39 two-finger (Choloepus hoffmanni) and three-finger sloths (Bradypus variegatus) of Costa Rica (n = 78) and analyzed using conventional bacteriological techniques. A generalized linear mixed model was applied to estimate the isolates' multiple antimicrobial resistance and virulence indices as a function of animal status. Results: A considerable level of resistance was detected, especially for Citrobacter youngae and Escherichia coli, with 17.5% of isolates classified as multidrug-resistant. Virulence indices of isolates from rehabilitated sloths were significantly higher than the ones from sloths being hand-reared for shorter periods. Conclusions: To our knowledge, this is the first description of sloths' antimicrobial resistant Enterobacterales, suggesting that sloths' rehabilitation and consequent exposure to humans, may promote the selection of bacteria with higher virulence. Ultimately, these bacteria may represent a threat to human and animal health due to their zoonotic potential and AMR and virulence profiles.

Does a suspensory lifestyle result in increased tensile strength? Organ-level material properties of sloth limb bones.

The material composition of vertebrate connective tissue is highly conserved across taxa. Existing data suggest that the compressive and tensile strength of limb bones are very similar despite marked variation in limb posture and locomotor patterns. However, the material properties of limb bone tissue from suspensory taxa have not been formally evaluated. Sloths are nearly obligatory in their use of below-branch suspensory locomotion and posture, thus placing their limb bones and associated soft tissue structures under routine tensile loading. It is possible that sloth limb bones are modified for enhanced tensile strength, perhaps at the expense of compressive strength. Forelimb and hindlimb bones of two-toed (Choloepus hoffmanni) and three-toed (Bradypus variegatus) sloths were tested in compression and bending to evaluate this hypothesis. Strength and elastic (Young's) modulus were similarly lower in sloth limb bones during both compression and bending, as compared with pronograde taxa. Ratios of peak bending strength to compressive strength additionally were elevated (sloths: 1.4-1.7; upright taxa: 0.6-1.2) for sloth limb bones. Overall, the material properties measured from the limb bones of tree sloths support our hypothesis of predicted function in a tensile limb system. Future studies should aim to directly test bones in tension to confirm indications of elevated axial tensile strength. Nevertheless, the results herein expand understanding of functional adaptation in mammalian tissue for a range of locomotor/postural behaviors that were previously unexplored.

Bifidobacteria in two-toed sloths (Choloepus didactylus): phylogenetic characterization of the novel taxon Bifidobacterium choloepi sp. nov.

Seven bifidobacterial strains were isolated from the faeces of two adult males of the two-toed sloth (Choloepus didactylus) housed in Parco Natura Viva, in Italy. Comparative sequence analysis of 16S rRNA and of five housekeeping (hsp60, rpoB, clpC, dnaJ, dnaG) genes revealed that these strains were classified into two clusters. On the basis of 16S rRNA gene sequence similarity, the type strain of Bifidobacterium catenulatum subsp. kashiwanohense DSM 21854T (95.4 %) was the closest neighbour to strain in Cluster I (BRDM 6T), whereas the type strain of Bifidobacterium dentium DSM 20436T (values were in the range of 98‒99.8 %) was the closest neighbour to the other six strains in Cluster II. The average nucleotide identity (ANI) values of BRDM 6T and of strains in Cluster II with the closely related type strains were 76.0 and 98.9 % (mean value) respectively. Therefore, genotyping based on the genome sequence of the strain BRDM 6T combined with phenotypic analyses clearly revealed that the strain BRDM 6T represents a novel species for which the names Bifidobacterium choloepi sp. nov. (BRDM 6T=NBRC 114053T=BCRC 81222T) is proposed.

High moon brightness and low ambient temperatures affect sloth predation by harpy eagles.

BACKGROUND: Climate plays a key role in the life histories of tropical vertebrates. However, tropical forests are only weakly seasonal compared with temperate and boreal regions. For species with limited ability to control core body temperature, even mild climatic variation can determine major behavioural outcomes, such as foraging and predator avoidance. In tropical forests, sloths are the arboreal vertebrate attaining the greatest biomass density, but their capacity to regulate body temperature is limited, relying on behavioural adaptations to thermoregulate. Sloths are largely or strictly nocturnal, and depend on crypsis to avoid predation. The harpy eagle (Harpia harpyja) is a sloth-specialist and exerts strong top-down control over its prey species. Yet the role of environmental variables on the regulation of predator-prey interactions between sloths and harpy eagles are unknown. The harpy eagle is considered Near Threatened. This motivated a comprehensive effort to reintroduce this species into parts of Mesoamerica. This effort incidentally enabled us to understand the prey profile of harpy eagles over multiple seasons. METHODS: Our study was conducted between 2003 and 2009 at Soberanía National Park, Panamá. Telemetered harpy eagles were seen hunting and feeding on individual prey species. For each predation event, field assistants systematically recorded the species killed. We analysed the effects of climatic conditions and vegetation phenology on the prey species profile of harpy eagles using generalised linear mixed models. RESULTS: Here we show that sloth predation by harpy eagles was negatively affected by nocturnal ambient light (i.e. bright moonshine) and positively affected by seasonally cool temperatures. We suggest that the first ensured low detectability conditions for sloths foraging at night and the second posed a thermally unsuitable climate that forced sloths to forage under riskier daylight. We showed that even moderate seasonal variation in temperature can influence the relationship between a keystone tropical forest predator and a dominant prey item. Therefore, predator-prey ecology in the tropics can be modulated by subtle changes in environmental conditions. The seasonal effects shown here suggest important demographic consequences for sloths, which are under top-down regulation from harpy eagle predation, perhaps limiting their geographic distribution at higher latitudes.

EVALUATION OF 25-HYDROXYVITAMIN D IN HOFFMANN'S TWO-TOED SLOTHS (CHOLOEPUS HOFFMANNI) USING DRIED BLOOD SPOTS ANALYZED BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY.

Although biochemical analytes have typically been measured using serum or whole blood samples, an increasing number of assays are validated for measurement of analytes from dried blood spots (DBS) on filter paper. DBS techniques are minimally invasive, require only a small sample volume, and simplify processing, storage, and shipment of samples. These qualities make DBS-based assays ideal for sampling of wildlife species in both captive and field settings. In this study, a liquid chromatography-tandem mass spectrometry assay was evaluated for measurement of 25-hydroxyvitamin D in sloths. Paired serum and DBS samples were collected from nine healthy captive Hoffmann's two-toed sloths (Choloepus hoffmanni). Statistical analysis using Passing-Bablok regression analysis, Bland-Altman plots, and the Wilcoxon signed-rank tests found good agreement between 25-hydroxyvitamin D3 measurements in serum and DBS samples. Constant and proportional bias were absent. Results from this study support the use of DBS samples for the evaluation of vitamin D status in Hoffmann's two-toed sloths and provide a foundation for further studies to validate this technique.

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.

URINARY BLADDER TRANSITIONAL CELL CARCINOMA AND CARCINOMATOSIS IN A LINNAEUS'S TWO-TOED SLOTH, CHOLOEPUS DIDACTYLUS.

A 19-yr-old female Linnaeus's two-toed sloth (Choloepus didactylus) with a history of urinary incontinence, ascites, and behavioral changes was euthanized after diagnostic imaging revealed a large bladder mass. On gross necropsy, the sloth had a severely thickened bladder mucosa, partial urinary obstruction, and nonseptic exudate in the peritoneal cavity. Histopathology showed a malignant and highly invasive transitional cell (urothelial) carcinoma with transmural and intra-abdominal invasion and diffuse carcinomatosis. Immunohistochemistry for expression of pancytokeratin (AE1/AE3), cytokeratin 7 (CK7), cytokeratin 20 (CK20), and uroplakin III was performed to confirm the diagnosis of transitional cell carcinoma. Neoplastic cells had a strong cytoplasmic immunoreactivity with the antipancytokeratin antibody clone AE1/AE3, which was consistent with a neoplasm of epithelial origin. Neoplastic cells were negative for expression of CK20. This is the first detailed report describing the antemortem diagnosis of urinary bladder transitional cell carcinoma with carcinomatosis in a two-toed sloth.

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'.

Mitogenomics phylogenetic relationships of the current sloth's genera and species (Bradypodidae and Megalonychidae).

We sequenced the complete mitogenome of 39 sloths (19 Bradypus variegatus, 4 B. tridactylus, 1 B. pygmaeus, 1 B. torquatus, 4 Choloepus didactylus, and 10 C. hoffmanni). A Bayesian tree (BI) indicated a temporal split between Bradypus and Choloepus around 31 million years ago (MYA, Oligocene) and the other major splits within each genera during the Miocene and Pliocene. A haplotype network (MJN) estimated a lower temporal split between the sloth genera (around 23.5 MYA). Both methods detected the ancestor of B. torquatus as the first to diverge within Bradypus (21 for BI and 19 MJN), followed by that of the ancestor of B. tridactylus. The split of B. pygmaeus from the common ancestor with B. variegatus was around 12 MYA (BI) or 4.3 MYA (MJN). The splits among the previous populations of B. variegatus began around 8 MYA (BI) or 3.6 MYA (MJN). The trans-Andean population was the first to diverge from the remaining cis-Andean populations of B. variegatus. The genetic differentiation of the trans-Andean B. variegatus population relative to the cis-Andean B. variegatus is similar to that found for different species of sloths. The mitogenomic analysis resolved the differentiation of C. hoffmanni from the C. didactylus individuals of the Guiana Shield. However, one C. didactylus from the Colombian Amazon specimen was inside the C. hoffmanni clade. This could be the first example of possible natural hybridization in the Amazon of both Choloepus taxa or the existence of un-differentiable phenotypes of these two species in some Amazonian areas.

Cophylogenetics and biogeography reveal a coevolved relationship between sloths and their symbiont algae.

Specialized species, like arboreal folivores, often develop beneficial relationships with symbionts to exploit ecologically constrained lifestyles. Although coevolution can drive speciation by specialization of a symbiont to a host, a symbiotic relationship is not indicative of coevolution between host and symbiont. We tested for coevolved relationships between highly specialized two- and three-toed sloths (Choloepus spp. and Bradypus spp., respectively) and their symbiotic algae using cophylogenies and phylogeography. Our phylogeographic analysis showed a biogeographic pattern for the sloth distribution that was not found in the algal phylogeny. We found support for congruence between the sloth and algae phylogenies, implying cospeciation, only in the Bradypus lineage. Algae host-switching occurred from Bradypus spp. to Choloepus spp. Our results support a previously hypothesized symbiotic relationship between sloths and the algae in their fur and indicate that coevolution may have played a role in algae diversification. More broadly, convergent evolution may facilitate host switching between deeply diverged host lineages.

SEROLOGIC RESPONSE TO CANINE DISTEMPER VACCINATION IN CAPTIVE LINNAEUS'S TWO-TOED SLOTHS ( CHOLOEPUS DIDACTYLUS) AFTER A FATAL CANINE DISTEMPER VIRUS OUTBREAK.

Canine distemper virus (CDV) affects many wild and captive, nondomestic species worldwide but has not been previously reported in Xenarthra. Paucity of information on vaccination safety and efficacy presents challenges for disease prevention in captive collections. CDV infections and subsequent mortalities in five captive Linnaeus's two-toed sloths ( Choloepus didactylus) in eastern Tennessee are reported. Clinical signs included oculonasal discharge, oral ulcerations, and diarrhea, and the diagnosis was confirmed by necropsy, histopathology, immunohistochemistry, virus isolation, and polymerase chain reaction. Viral sequencing identified the strain to be consistent with a new CDV lineage currently affecting domestic dogs and wildlife in Tennessee. Seven sloths were examined and vaccinated using a recombinant CDV vaccine on days 0 and 21. Subsequent blood samples showed increased titers in 3/4 sloths. Based on the outbreak and serologic findings postvaccination without adverse effects, the authors recommend recombinant CDV vaccination in sloths exposed to known carriers of CDV.

SEROSURVEY OF SELECTED ARBOVIRAL PATHOGENS IN FREE-RANGING, TWO-TOED SLOTHS (CHOLOEPUS HOFFMANNI) AND THREE-TOED SLOTHS (BRADYPUS VARIEGATUS) IN COSTA RICA, 2005-07.

We screened for antibodies to 16 arboviruses in four populations of free-ranging sloths in Costa Rica. Blood samples were taken from 16 Hoffman's two-toed sloths (HTSs; Choloepus hoffmanni ) and 26 brown-throated sloths (BTSs; Bradypus variegatus ) over a 3-yr period. We used serologic assays to detect antibodies against 10 arboviruses previously described in sloths (St. Louis encephalitis [SLEV], Changuinola, Venezuelan equine encephalitis, Ilheus [ILHV], Oropouche, Mayaro, Utinga, Murutucu, Punta Toro, and vesicular stomatitis [VSV] viruses) and six arboviruses not described in sloths (Rio Grande, West Nile [WNV], eastern equine encephalitis, Piry, Munguba, and La Crosse viruses). Overall, 80% of sloths had detectable antibodies to SLEV, 67% had antibodies to ILHV, 32% to Punta Toro virus, 30% to Changuinola virus, 15% to WNV, 14% to VSV, 11% to Venezuelan equine encephalitis virus, and 10% to Rio Grande virus. No samples had detectable antibodies to the remaining eight viruses. We found a significant increase in prevalence of antibody to VSV in HTSs between 2005 and 2007, and for WNV antibody between 2005 and 2006. We found no significant differences in the prevalences of antibodies to the sampled viruses between the two locations. Antibody prevalences were significantly higher in HTSs than in BTSs for SLEV in 2005. Antibody-positive results for ILHV were likely due to cross-reaction with SLEV. The novel finding of antibodies to Rio Grande virus in sloths could be due to cross-reaction with another phlebovirus. These findings might have implications for land management and domestic animal health. Due to the nature of the study, we could not determine whether sloths could represent amplification hosts for these viruses, or whether they were only exposed and could be used as sentinel species. Further studies are needed to fully characterize arboviral exposure in sloths.

Soft Tissue Mineralization in Captive 2-Toed Sloths.

Soft tissue mineralization was diagnosed in 19 captive 2-toed sloths (Choloepus didactylusandCholoepus hoffmanni) ranging from 2 months to 41 years of age. Gross mineralization was evident at necropsy in 6 of 19 sloths and was prominent in the aorta and arteries. Histologically, 11 sloths had arterial mineralization, including mural osseous and chondroid metaplasia and smooth muscle hyperplasia consistent with arteriosclerosis. Visceral mineralization most commonly involved the gastric mucosa (17 sloths), kidneys (17 sloths), and lungs (8 sloths). Eleven sloths ranging in age from 5 to 41 years old had moderate to severe renal disease, which may be an important underlying cause of soft tissue mineralization in adult sloths. However, 5 sloths (juveniles and adults) had severe soft tissue mineralization with histologically normal kidneys or only mild interstitial inflammation or fibrosis, suggesting other causes of calcium and phosphorus imbalance. Degenerative cardiac disease was a common finding in 10 sloths with vascular mineralization and varied from mild to severe with fibrosis and acute noninflammatory myocardial necrosis. Although the prevalence of cardiac disease in adult sloths has not been documented, disease may be exacerbated by hypertension from degenerative arteriosclerosis as noted in this study group. Although renal disease likely contributed substantially to mineralization of tissues in most sloths in this study, nutritional causes of soft tissue mineralization-such as imbalances in dietary vitamin D or calcium and phosphorus-may be an important contributing factor.