Biomechanics and energetics in aquatic and semiaquatic mammals: platypus to whale

A variety of mammalian lineages have secondarily invaded the water. To locomote and thermoregulate in the aqueous medium, mammals developed a range of morphological, physiological, and behavioral adaptations. A distinct difference in the suite of adaptations, which affects energetics, is apparent between semiaquatic and fully aquatic mammals. Semiaquatic mammals swim by paddling, which is inefficient compared to the use of oscillating hydrofoils of aquatic mammals. Semiaquatic mammals swim at the water surface and experience a greater resistive force augmented by wave drag than submerged aquatic mammals. A dense, nonwettable fur insulates semiaquatic mammals, whereas aquatic mammals use a layer of blubber. The fur, while providing insulation and positive buoyancy, incurs a high energy demand for maintenance and limits diving depth. Blubber contours the body to reduce drag, is an energy reserve, and suffers no loss in buoyancy with depth. Despite the high energetic costs of a semiaquatic existence, these animals represent modern analogs of evolutionary intermediates between ancestral terrestrial mammals and their fully aquatic descendants. It is these intermediate animals that indicate which potential selection factors and mechanical constraints may have directed the evolution of more derived aquatic forms.     

Swimming performance in semiaquatic and terrestrial Oryzomyine rodents

Semiaquatic and terrestrial mammals frequently have to cross or move along water bodies, both trying to remain on the water surface using one or two pairs of limbs, combining different gaits and stride lengths and frequencies. This is the case of the semiaquatic water rats Nectomys and the cursorial Cerradomys, sister genera of the Oryzomyini tribe, capable of swimming using similar gaits. They provide an opportunity to investigate performance specializations involving the semiaquatic habitat, our objective in this study. Rodents were filmed at 30 frames s⁻¹ in lateral view, swimming in a glass aquarium. Video sequences were analyzed dividing the swimming cycle into power and recovery phases. Differences in swimming performance were detected between species of Nectomys and Cerradomys, but not between species of the same genus. Absolute mean speed did not differ between the semiaquatic and terrestrial groups, but the semiaquatic Nectomys had longer stride lengths with lower stride frequency, whereas the terrestrial Cerradomys had higher stride frequency and relative swimming speed. The widest behavior repertoire of Nectomys allowed more efficient, but not necessarily faster swimming than the terrestrial Cerradomys. Efficient aquatic locomotion in Nectomys is ultimately a result of improved buoyancy by hydrophobic fur and subtle morphological specializations, which allow this genus to perform more efficiently in water than the terrestrial Cerradomys without compromising locomotion in the terrestrial environment.     

Site-specific fatty acid composition in adipose tissues of several northern aquatic and terrestrial mammals

Site-specific differences in fatty acid compositions (by gas-liquid chromatography) were compared in aquatic, semiaquatic and terrestrial mammals: the ringed seals (Phoca hispida hispida and P. h. botnica), otter (Lutra lutra), raccoon dog (Nyctereutes procyonoides), brown bear (Ursus arctos) and grey wolf (Canis lupus). In addition, we briefly discuss our earlier results for the Canadian beaver (Castor canadensis) and muskrat (Ondatra zibethicus). In both aquatic and terrestrial species, large amounts of Δ9-monounsaturated fatty acids (MUFAs) and small amounts of saturated fatty acids and exogenous long-chain MUFAs were found in the cold tissues of the extremities. In seals, the poikilothermic outer blubber had these characteristics and differed from the inner blubber. On the other hand, the subcutaneous and inner fat depots of the coated semiaquatic and terrestrial mammals were uniform. In the bare extremities, however, these mammals also had an excess of A9-MUFAs. The degree of Δ9-desaturation in the outer blubber of the seals was significantly correlated with age. The excess of Δ9-MUFAs in the bare extremities of land mammals increased the overall double bond content of these tissues compared with the inner depots. In contrast, due to the large amounts of dietary polyunsaturated fatty acids, this was not found in the aquatic and semiaquatic species. The observed site-specific differences are discussed as possible inherited evolutionary adaptations to low temperature of the tissues.     

Transitions from Drag-based to Lift-based Propulsion in Mammalian Swimming

The evolution of fully aquatic mammals from quadrupedal, terrestrialmammals was associated with changes in morphology and swimmingmode. Drag is minimized by streamlining body shape and appendages.Improvement in speed, thrust production and efficiency is accomplishedby a change of swimming mode. Terrestrial and semiaquatic mammalsemploy drag-based propulsion with paddling appendages, whereasfully aquatic mammals use lift-based propulsion with oscillatinghydrofoils. Aerobic efficiencies are low for drag-based swimming,but reach a maximum of 30% for lift-based propulsion. Propulsiveefficiency is over 80% for lift-based swimming while only 33%for paddling. In addition to swimming mode, the transition tohigh performance propulsion was associated with a shift fromsurface to submerged swimming providing a reduction in transportcosts. The evolution of aquatic mammals from terrestrial ancestorsrequired increased swimming performance with minimal compromiseto terrestrial movement. Examination of modern analogs to transitionalswimming stages suggests that only slight modification to theneuromotor pattern used for terrestrial locomotion is requiredto allow for a change to lift-based propulsion.     

Examining the Prey Mass of Terrestrial and Aquatic Carnivorous Mammals: Minimum,Maximum and Range

Predator-prey body mass relationships are a vital part of food webs across ecosystems and provide key information for predicting the susceptibility of carnivore populations to extinction. Despite this, there has been limited research on the minimum and maximum prey size of mammalian carnivores. Without information on large-scale patterns of prey mass, we limit our understanding of predation pressure, trophic cascades and susceptibility of carnivores to decreasing prey populations. The majority of studies that examine predator-prey body mass relationships focus on either a single or a subset of mammalian species, which limits the strength of our models as well as their broader application. We examine the relationship between predator body mass and the minimum, maximum and range of their prey''s body mass across 108 mammalian carnivores, from weasels to baleen whales (Carnivora and Cetacea). We test whether mammals show a positive relationship between prey and predator body mass, as in reptiles and birds, as well as examine how environment (aquatic and terrestrial) and phylogenetic relatedness play a role in this relationship. We found that phylogenetic relatedness is a strong driver of predator-prey mass patterns in carnivorous mammals and accounts for a higher proportion of variance compared with the biological drivers of body mass and environment. We show a positive predator-prey body mass pattern for terrestrial mammals as found in reptiles and birds, but no relationship for aquatic mammals. Our results will benefit our understanding of trophic interactions, the susceptibility of carnivores to population declines and the role of carnivores within ecosystems.     

Skeleton weight allometry in aquatic and terrestrial vertebrates

The relation of skeleton weight to body weight with increasing size is compared for aquatic and terrestrial vertebrates. Due to the buoyancy of water, the skeleton weights of aquatic vertebrates (fishes and whales) vary in nearly direct proportion (exponent 1.0) to body weight; while the skeletons of terrestrial vertebrates occupy an increasingly greater proportion of total body weight as size increases (exponent greater than i. i) due to the necessity of supporting their weight on land.     

Head‐turning morphologies: Evolution of shape diversity in the mammalian atlas–axis complex

Mammals flex, extend, and rotate their spines as they perform behaviors critical for survival, such as foraging, consuming prey, locomoting, and interacting with conspecifics or predators. The atlas–axis complex is a mammalian innovation that allows precise head movements during these behaviors. Although morphological variation in other vertebral regions has been linked to ecological differences in mammals, less is known about morphological specialization in the cervical vertebrae, which are developmentally constrained in number but highly variable in size and shape. Here, we present the first phylogenetic comparative study of the atlas–axis complex across mammals. We used spherical harmonics to quantify 3D shape variation of the atlas and axis across a diverse sample of species, and performed phylogenetic analyses to investigate if vertebral shape is associated with body size, locomotion, and diet. We found that differences in atlas and axis shape are partly explained by phylogeny, and that mammalian subclades differ in morphological disparity. Atlas and axis shape diversity is associated with differences in body size and locomotion; large terrestrial mammals have craniocaudally elongated vertebrae, whereas smaller mammals and aquatic mammals have more compressed vertebrae. These results provide a foundation for investigating functional hypotheses underlying the evolution of neck morphologies across mammals.     

Aquatic Adaptation and Swimming Mode Inferred from Skeletal Proportions in the Miocene Desmostylian Desmostylus

Desmostylians are enigmatic, extinct, semiaquatic marine mammals that inhabited coastlines of the northern Pacific Rim during the late Oligocene through middle Miocene. Principal components analysis (PCA) of trunk and limb proportions provides a rational multivariate context for separating living semiaquatic mammals on three orthogonal axes: a size axis (PC-I), a degree of aquatic adaptation axis (PC-II), and a forelimb- versus hind-limb-dominated locomotion axis (PC-III). The necessary skeletal measurements are available for Desmostylus hesperus but not for other desmostylians. Among species similar in size to Desmostylus in the study set, the one most similarly proportioned is the polar bear. Projection of Desmostylus on PC-II shows it to have been more aquatic than a polar bear (indicated by its relatively short ilium and femur, combined with relatively long metapodals and phalanges). Projection of Desmostylus on PC-III suggests that its aquatic locomotion was even more forelimb-dominated than that of a bear (indicated by its relatively long metacarpal III and corresponding proximal phalanx, combined with a relatively short metatarsal III and corresponding proximal phalanx). Desmostylians were different from all living semiaquatic mammals, and desmostylians are properly classified in their own extinct order, but their skeletal proportions suggest that bears provide an appropriate baseline for imagining what desmostylians were like in life.     

Trabecular architecture in the humeral metaphyses of non-avian reptiles (Crocodylia,Squamata and Testudines): Lifestyle,allometry and phylogeny

The lifestyle of extinct tetrapods is often difficult to assess when clear morphological adaptations such as swimming paddles are absent. According to the hypothesis of bone functional adaptation, the architecture of trabecular bone adapts sensitively to physiological loadings. Previous studies have already shown a clear relation between trabecular architecture and locomotor behavior, mainly in mammals and birds. However, a link between trabecular architecture and lifestyle has rarely been examined. Here, we analyzed trabecular architecture of different clades of reptiles characterized by a wide range of lifestyles (aquatic, amphibious, generalist terrestrial, fossorial, and climbing). Humeri of squamates, turtles, and crocodylians have been scanned with microcomputed tomography. We selected spherical volumes of interest centered in the proximal metaphyses and measured trabecular spacing, thickness and number, degree of anisotropy, average branch length, bone volume fraction, bone surface density, and connectivity density. Only bone volume fraction showed a significant phylogenetic signal and its significant difference between squamates and other reptiles could be linked to their physiologies. We found negative allometric relationships for trabecular thickness and spacing, positive allometries for connectivity density and trabecular number and no dependence with size for degree of anisotropy and bone volume fraction. The different lifestyles are well separated in the morphological space using linear discriminant analyses, but a cross-validation procedure indicated a limited predictive ability of the model. The trabecular bone anisotropy has shown a gradient in turtles and in squamates: higher values in amphibious than terrestrial taxa. These allometric scalings, previously emphasized in mammals and birds, seem to be valid for all amniotes. Discriminant analysis has offered, to some extent, a distinction of lifestyles, which however remains difficult to strictly discriminate. Trabecular architecture seems to be a promising tool to infer lifestyle of extinct tetrapods, especially those involved in the terrestrialization.     

Axial allometry in a neutrally buoyant environment: effects of the terrestrial‐aquatic transition on vertebral scaling

Ecological diversification into new environments presents new mechanical challenges for locomotion. An extreme example of this is the transition from a terrestrial to an aquatic lifestyle. Here, we examine the implications of life in a neutrally buoyant environment on adaptations of the axial skeleton to evolutionary increases in body size. On land, mammals must use their thoracolumbar vertebral column for body support against gravity and thus exhibit increasing stabilization of the trunk as body size increases. Conversely, in water, the role of the axial skeleton in body support is reduced, and, in aquatic mammals, the vertebral column functions primarily in locomotion. Therefore, we hypothesize that the allometric stabilization associated with increasing body size in terrestrial mammals will be minimized in secondarily aquatic mammals. We test this by comparing the scaling exponent (slope) of vertebral measures from 57 terrestrial species (23 felids, 34 bovids) to 23 semi‐aquatic species (pinnipeds), using phylogenetically corrected regressions. Terrestrial taxa meet predictions of allometric stabilization, with posterior vertebral column (lumbar region) shortening, increased vertebral height compared to width, and shorter, more disc‐shaped centra. In contrast, pinniped vertebral proportions (e.g. length, width, height) scale with isometry, and in some cases, centra even become more spool‐shaped with increasing size, suggesting increased flexibility. Our results demonstrate that evolution of a secondarily aquatic lifestyle has modified the mechanical constraints associated with evolutionary increases in body size, relative to terrestrial taxa.     

Functional significance of bone ballastin in the evolution of buoyancy control strategies by aquatic tetrapods

The primary function of pachyostosis, pachyosteo‐sclerosis, and osteosclerosis may be to act as ballast, not so much (as previously suggested) to neutralise the buoyancy of existing lungs, but to allow enlargement of the lungs. Enlarged lungs cause an animal to lose buoyancy more rapidly with depth. They also provide a larger oxygen store. These features are useful for slow swimmers and shallow divers, such as feeders on benthic plants and invertebrates. Examples are sirenians, primitive sauropterygians ("not‐hosaurs"), placodonts, and the sea otter Enhydra. These last two show convergent evolution of adaptations to feeding on hard‐shelled invertebrate prey in shallow water. Mesosaurids are problematical. Bone ballast uses body mass and volume less efficiently than other buoyancy control strategies. Theoretical analysis predicts that bone ballast should not occur in semiaquatic forms, fast swimmers or deep divers. It does not usually occur in such organisms. Marine iguanas of the Galápagos, desmostylians, and the aquatic sloth Thalassocnus are all littoral feeders and all lack bone ballast as predicted. Plesiosaurs adopted varied strategies: some used bone ballast, and others used gastroliths. Biomechanical considerations lead to the prediction that a new marine tetrapod clade will typically evolve bone ballast as part of its adaptation to life in water. Slow swimmers and grazers on sessile food, like sirenians and placodonts, develop it more strongly, but active predators like ichthyosaurs and cetaceans secondarily lose this character.     

Temperature‐dependent oxygen limitation and the rise of Bergmann's rule in species with aquatic respiration

Bergmann's rule is the propensity for species‐mean body size to decrease with increasing temperature. Temperature‐dependent oxygen limitation has been hypothesized to help drive temperature–size relationships among ectotherms, including Bergmann's rule, where organisms reduce body size under warm oxygen‐limited conditions, thereby maintaining aerobic scope. Temperature‐dependent oxygen limitation should be most pronounced among aquatic ectotherms that cannot breathe aerially, as oxygen solubility in water decreases with increasing temperature. We use phylogenetically explicit analyses to show that species‐mean adult size of aquatic salamanders with branchial or cutaneous oxygen uptake becomes small in warm environments and large in cool environments, whereas body size of aquatic species with lungs (i.e., that respire aerially), as well as size of semiaquatic and terrestrial species do not decrease with temperature. We argue that oxygen limitation drives the evolution of small size in warm aquatic environments for species with aquatic respiration. More broadly, the stronger decline in size with temperature observed in aquatic versus terrestrial salamander species mirrors the relatively strong plastic declines in size observed previously among aquatic versus terrestrial invertebrates, suggesting that temperature‐dependent oxygen availability can help drive patterns of plasticity, micro‐ and macroevolution.     

Comparative limb bone scaling in turtles: Phylogenetic transitions with changes in functional demands?

Several terrestrial vertebrate clades include lineages that have evolved nearly exclusive use of aquatic habitats. In many cases, such transitions are associated with the evolution of flattened limbs that are used to swim via dorsoventral flapping. Such changes in shape may have been facilitated by changes in limb bone loading in novel aquatic environments. Studies on limb bone loading in turtles found that torsion is high relative to bending loads on land, but reduced compared to bending during aquatic rowing. Release from torsion among rowers could have facilitated the evolution of hydrodynamically advantageous flattened limbs among aquatic species. Because rowing is regarded as an intermediate locomotor stage between walking and flapping, rowing species might show limb bone flattening intermediate between the tubular shapes of walkers and the flattened shapes of flappers. We collected measurements of humeri and femora from specimens representing four functionally divergent turtle clades: sea turtles (marine flappers), softshells (specialized freshwater rowers), emydids (generalist semiaquatic rowers), and tortoises (terrestrial walkers). Patterns of limb bone scaling with size were compared across lineages using phylogenetic comparative methods. Although rowing taxa did not show the intermediate scaling patterns we predicted, our data provide other functional insights. For example, flattening of sea turtle humeri was associated with positive allometry (relative to body mass) for the limb bone diameter perpendicular to the flexion-extension plane of the elbow. Moreover, softshell limb bones exhibit positive allometry of femoral diameters relative to body mass, potentially helping them maintain their typical benthic position in water by providing additional weight to compensate for shell reduction. Tortoise limb bones showed positive allometry of diameters, as well as long humeri, relative to body mass, potentially reflecting specializations for resisting loads associated with digging. Overall, scaling patterns of many turtle lineages appear to correlate with distinctive behaviors or locomotor habits.     

The structural characteristics of the auditory portion of the cerebral cortex in the Baikal seal (Pusa sibirica) and the neuronal reaction to hypoxia]

Methods of light (Nissl, Golgi-Kopsch) and electron microscopy were used for studying the structural organization of the auditory portion of the cerebral cortex of Pusa sibirica. The auditory port of this kind of cortex is characterized by pyramidization of all the layers (layer IV included), thick layer I, two sublayers in layer III which is typical of other semiaquatic and aquatic mammals. Synaptoarchitechtonics of this part of the neocortex may be compared with that of other aquatic, semiaquatic and terrestrial mammals. The investigation of the state of neurons of the auditory portion of the cortex of Pusa sibirica during induced 18-min hypoxia has shown that the alterations in the neurons are of functional and reversible character. The axosomatic synapses remained most intact. The alterations in the auditory portion of the neo-cortex of Pusa sibirica evidences high level of adaptive reactions to hypoxia associated with the adaptation of these mammals to diving and fixed in the evolutionary process.     

The middle ear in the ontogenesis of mammals

The middle ear in mammals is characterized by structural variations and a broad spectrum of adaptive transformations related to peculiarities of species ecology, but it preserves the general basic principle of structure in most mammals. In species remote from a phylogenetic point of view but close in ecologic specialization, features of parallelism are observed concerning the development of separate elements of auditory ossicles as well as the way of their interconnection and attachment to the tympanum. Along the way to the adaptation to the water lifestyle in semi-aquatic and aquatic species, new additional structures, not intrinsic to initial terrestrial forms, have been formed. The use of ecological and morphological approaches to research the peripheral division of the auditory system of mammals with different ecological specialization in the ontogenesis permitted us to reveal that peculiarities of its structure in different groups of mammals are preconditioned by the animals’ adaptation to specific acoustic properties of their environment. Morphological and functional adaptations of the peripheral auditory system aimed at optimizing auditory sensitivity in the environments differing in physical properties are of great importance in evolution. Adaptive specific features in the structure of the middle ear in aquatic species appear at early stages of development in spite of intrauterine growth without the direct influence of environmental conditions.     

Environmental DNA enables detection of terrestrial mammals from forest pond water

Terrestrial animals must have frequent contact with water to survive, implying that environmental DNA (eDNA) originating from those animals should be detectable from places containing water in terrestrial ecosystems. Aiming to detect the presence of terrestrial mammals using forest water samples, we applied a set of universal PCR primers (MiMammal, a modified version of fish universal primers) for metabarcoding mammalian eDNA. The versatility of MiMammal primers was tested in silico and by amplifying DNAs extracted from tissues. The results suggested that MiMammal primers are capable of amplifying and distinguishing a diverse group of mammalian species. In addition, analyses of water samples from zoo cages of mammals with known species composition suggested that MiMammal primers could successfully detect mammalian species from water samples in the field. Then, we performed an experiment to detect mammals from natural ecosystems by collecting five 500‐ml water samples from ponds in two cool‐temperate forests in Hokkaido, northern Japan. MiMammal amplicon libraries were constructed using eDNA extracted from water samples, and sequences generated by Illumina MiSeq were subjected to data processing and taxonomic assignment. We thereby detected multiple species of mammals common to the sampling areas, including deer (Cervus nippon), mouse (Mus musculus), vole (Myodes rufocanus), raccoon (Procyon lotor), rat (Rattus norvegicus) and shrew (Sorex unguiculatus). Many previous applications of the eDNA metabarcoding approach have been limited to aquatic/semiaquatic systems, but the results presented here show that the approach is also promising even for forest mammal biodiversity surveys.     

Energetic benefits and adaptations in mammalian limbs: Scale effects and selective pressures

Differences in limb size and shape are fundamental to mammalian morphological diversity; however, their relevance to locomotor costs has long been subject to debate. In particular, it remains unknown if scale effects in whole limb morphology could partially underlie decreasing mass‐specific locomotor costs with increasing limb length. Whole fore‐ and hindlimb inertial properties reflecting limb size and shape—moment of inertia (MOI), mass, mass distribution, and natural frequency—were regressed against limb length for 44 species of quadrupedal mammals. Limb mass, MOI, and center of mass position are negatively allometric, having a strong potential for lowering mass‐specific locomotor costs in large terrestrial mammals. Negative allometry of limb MOI results in a 40% reduction in MOI relative to isometry's prediction for our largest sampled taxa. However, fitting regression residuals to adaptive diversification models reveals that codiversification of limb mass, limb length, and body mass likely results from selection for differing locomotor modes of running, climbing, digging, and swimming. The observed allometric scaling does not result from selection for energetically beneficial whole limb morphology with increasing size. Instead, our data suggest that it is a consequence of differing morphological adaptations and body size distributions among quadrupedal mammals, highlighting the role of differing limb functions in mammalian evolution.     

Running energetics of the North American river otter: do short legs necessarily reduce efficiency on land?

Semi-aquatic mammals move between two very different media (air and water), and are subject to a greater range of physical forces (gravity, buoyancy, drag) than obligate swimmers or runners. This versatility is associated with morphological compromises that often lead to elevated locomotor energetic costs when compared to fully aquatic or terrestrial species. To understand the basis of these differences in energy expenditure, this study examined the interrelationships between limb morphology, cost of transport and biomechanics of running in a semi-aquatic mammal, the North American river otter. Oxygen consumption, preferred locomotor speeds, and stride characteristics were measured for river otters (body mass=11.1 kg, appendicular/axial length=29%) trained to run on a treadmill. To assess the effects of limb length on performance parameters, kinematic measurements were also made for a terrestrial specialist of comparable stature, the Welsh corgi dog (body mass=12.0 kg, appendicular/axial length=37%). The results were compared to predicted values for long legged terrestrial specialists. As found for other semi-aquatic mammals, the net cost of transport of running river otters (6.63 J kg(-1)min(-1) at 1.43 ms(-1)) was greater than predicted for primarily terrestrial mammals. The otters also showed a marked reduction in gait transition speed and in the range of preferred running speeds in comparison to short dogs and semi-aquatic mammals. As evident from the corgi dogs, short legs did not necessarily compromise running performance. Rather, the ability to incorporate a period of suspension during high speed running was an important compensatory mechanism for short limbs in the dogs. Such an aerial period was not observed in river otters with the result that energetic costs during running were higher and gait transition speeds slower for this versatile mammal compared to locomotor specialists.     

Evolution of morphology and locomotor performance in anurans: relationships with microhabitat diversification

The relationships between morphology, performance, behavior and ecology provide evidence for multiple and complex phenotypic adaptations. The anuran body plan, for example, is evolutionarily conserved and shows clear specializations to jumping performance back at least to the early Jurassic. However, there are instances of more recent adaptation to habit diversity in the post‐cranial skeleton, including relative limb length. The present study tested adaptive models of morphological evolution in anurans associated with the diversity of microhabitat use (semi‐aquatic arboreal, fossorial, torrent, and terrestrial) in species of anuran amphibians from Brazil and Australia. We use phylogenetic comparative methods to determine which evolutionary models, including Brownian motion (BM) and Ornstein‐Uhlenbeck (OU) are consistent with morphological variation observed across anuran species. Furthermore, this study investigated the relationship of maximum distance jumped as a function of components of morphological variables and microhabitat use. We found there are multiple optima of limb lengths associated to different microhabitats with a trend of increasing hindlimbs in torrent, arboreal, semi‐aquatic whereas fossorial and terrestrial species evolve toward optima with shorter hindlimbs. Moreover, arboreal, semi‐aquatic and torrent anurans have higher jumping performance and longer hindlimbs, when compared to terrestrial and fossorial species. We corroborate the hypothesis that evolutionary modifications of overall limb morphology have been important in the diversification of locomotor performance along the anuran phylogeny. Such evolutionary changes converged in different phylogenetic groups adapted to similar microhabitat use in two different zoogeographical regions.     

A biomechanical constraint on body mass in terrestrial mammalian predators

Observations on extant mammals suggest that large body mass is selectively advantageous for a terrestrial predator on large herbivores. Yet, throughout the Cenozoic, some lineages of terrestrial mammalian predators attained greater maximal body masses than others. In order to explain this evolutionary pattern, the following biomechanical constraint on body mass is hypothesized. The stress, set up in the humerus by the bending moment of the peak ground reaction force at maximal running speed, increased with increasing body mass within a given lineage of terrestrial mammalian predators, resulting in a decreasing safety factor for the bone, until a predator could no longer attain the maximal running speed of its smaller relatives. The selective disadvantage of reduced maximal running speed prevented further increase of body mass within the lineage. This hypothesis is tested by examining the scaling of humeral dimensions and estimating maximal body masses in several lineages of terrestrial mammalian predators. Among lineages with otherwise similar postcranial skeletons, those with the more robust humeri at a given body mass attained the greater maximal body masses. Lineages with the longer deltoid ridges/deltopectoral crests of the humeri and/or the more distally located deltoid scars (suggesting the more distal insertions of the humeral flexors) at a given body mass also attained the greater maximal body masses. These results support the existence of the proposed biomechanical constraint, although paleoecological data suggest that some lineages of terrestrial mammalian predators failed to reach the limits, imposed by this constraint, because of the small size of available prey.