Sub-zero temperatures and large-scale weather patterns induce tooth damage in Icelandic arctic foxes.

Tooth damage in carnivores can reflect shifts in both diet and feeding habits, and in large carnivores, it is associated with increased bone consumption. Variation in tooth condition in Icelandic arctic foxes, a mesocarnivore, was recorded from 854 individual foxes spanning 29 years. We hypothesized that annual climatic variations, which can influence food abundance and accessibility, will influence tooth condition by causing dietary shifts toward less edible prey. We examined tooth condition in relation to four climatic predictors: mean annual winter temperature, indices of both the El Niño anomaly and North Atlantic subpolar gyre (SPG), and the number of rain-on-snow days (ROS). We found unequivocal evidence for a strong effect of annual climate on tooth condition. Teeth of Icelandic foxes were in better condition when winter temperatures were higher, when the SPG was more positive, and when the number of ROS was low. We also found a substantial subregional effect with foxes from northeastern Iceland having lower tooth damage than those from two western sites. Contradicting our original hypothesis that foxes from northeastern Iceland, where foxes are known to scavenge on large mammal remains (e.g., sheep and horses), would show the highest tooth damage, we suggest that western coastal sites exhibited greater tooth damage because cold winter temperatures lowered the availability of seabirds, causing a shift in diet toward abrasive marine subsidies (e.g., bivalves) and frozen beach wrack. Our study shows that monitoring tooth breakage and wear can be a useful tool for evaluating the impact of climate on carnivore populations and that climate change may influence the condition and fitness of carnivores in complex and potentially conflicting ways.

Teaching Dynamics to Biology Undergraduates: the UCLA Experience.

There is a growing realization that traditional "Calculus for Life Sciences" courses do not show their applicability to the Life Sciences and discourage student interest. There have been calls from the AAAS, the Howard Hughes Medical Institute, the NSF, and the American Association of Medical Colleges for a new kind of math course for biology students, that would focus on dynamics and modeling, to understand positive and negative feedback relations, in the context of important biological applications, not incidental "examples." We designed a new course, LS 30, based on the idea of modeling biological relations as dynamical systems, and then visualizing the dynamical system as a vector field, assigning "change vectors" to every point in a state space. The resulting course, now being given to approximately 1400 students/year at UCLA, has greatly improved student perceptions toward math in biology, reduced minority performance gaps, and increased students' subsequent grades in physics and chemistry courses. This new course can be customized easily for a broad range of institutions. All course materials, including lecture plans, labs, homeworks and exams, are available from the authors; supporting videos are posted online.

Reimagining the Introductory Math Curriculum for Life Sciences Students.

Calculus is typically one of the first college courses encountered by science, technology, engineering, and mathematics (STEM) majors. Calculus often presents major challenges affecting STEM student persistence, particularly for students from groups historically underrepresented in STEM. For life sciences majors, calculus courses may not offer content that is relevant to biological systems or connect with students' interests in biology. We developed a transformative approach to teaching college-level math, using a dynamical systems perspective that focuses first on demonstrating why students need math to understand living systems, followed by providing quantitative and computational skills, including concepts from calculus, that students need to build and analyze mathematical models representing these systems. We found that students who complete these new math courses perform better in subsequent science courses than their counterparts who take traditional calculus courses. We also provide evidence that the new math curriculum positively impacts students' academic performance, with data that show narrowing of the achievement gap, based on students' math grades, between student subgroups in the new math courses. Moreover, our results indicate that students' interest in the concepts and skills critical to the quantitative preparation of 21st-century life sciences majors increases after completing the new contextualized math curriculum.

Naturally-occurring tooth wear, tooth fracture, and cranial injuries in large carnivores from Zambia.

Determining the incidence and causes of craniodental damage in wild carnivores is often constrained by limited access to specimens with associated ecological data, such as prey type and abundance. We assessed dental condition and cranial injuries in lion, leopard, and spotted hyena in relation to prey and predator populations in Zambia's Luangwa Valley, where large prey are more abundant and lion and leopard more numerous, and the Greater Kafue Ecosystem, where smaller prey species are more prevalent and lion and leopard less common. In Luangwa, lions had significantly higher rates of tooth fracture, and blunt trauma injuries attributable to prey-handling, compared to Kafue lions. In contrast, leopards in both regions had similar rates of tooth wear and breakage. Overall, lions showed a significantly higher tooth fracture rate than leopards on a per tooth basis. Spotted hyenas had the highest rates of tooth wear and fracture among all three carnivores, and greatly exceeded previously recorded rates based on historical samples. Despite larger numbers of lion and leopard in Luangwa, there was no difference in incidence of intraspecific injuries between regions. These results are consistent with a greater abundance of large prey species, especially buffalo, in the diets of Luangwa lions, and previous work showing a reliance on smaller prey species in Kafue throughout the large carnivore guild.

Dire wolves were the last of an ancient New World canid lineage.

Dire wolves are considered to be one of the most common and widespread large carnivores in Pleistocene America1, yet relatively little is known about their evolution or extinction. Here, to reconstruct the evolutionary history of dire wolves, we sequenced five genomes from sub-fossil remains dating from 13,000 to more than 50,000 years ago. Our results indicate that although they were similar morphologically to the extant grey wolf, dire wolves were a highly divergent lineage that split from living canids around 5.7 million years ago. In contrast to numerous examples of hybridization across Canidae2,3, there is no evidence for gene flow between dire wolves and either North American grey wolves or coyotes. This suggests that dire wolves evolved in isolation from the Pleistocene ancestors of these species. Our results also support an early New World origin of dire wolves, while the ancestors of grey wolves, coyotes and dholes evolved in Eurasia and colonized North America only relatively recently.

"Mucosal maps" of the canine nasal cavity: Micro-computed tomography and histology.

Nasal turbinals, delicate and complex bones of the nasal cavity that support respiratory or olfactory mucosa (OM), are now easily studied using high resolution micro-computed tomography (µ-CT). Standard µ-CT currently lacks the capacity to identify OM or other mucosa types without additional radio-opaque staining techniques. However, even unstained mucosa is more radio-opaque than air, and thus mucosal thickness can be discerned. Here, we assess mucosal thickness of the nasal fossa using the cranium of a cadaveric adult dog that was µ-CT scanned with an isotropic resolution of 30 µm, and subsequently histologically sectioned and stained. After co-alignment of µ-CT slice planes to that of histology, mucosal thickness was estimated at four locations. Results based on either µ-CT or histology indicate olfactory mucosa is thicker on average compared with non-olfactory mucosa (non-OM). In addition, olfactory mucosa has a lesser degree of variability than the non-OM. Variability in the latter appears to relate mostly to the varying degree of vascularity of the lamina propria. Because of this, in structures with both specialized vascular respiratory mucosa and OM, such as the first ethmoturbinal (ET I), the range of thickness of OM and non-OM may overlap. Future work should assess the utility of diffusible iodine-based contrast enhanced CT techniques, which can differentiate epithelium from the lamina propria, to enhance our ability to differentiate mucosa types on more rostral ethmoturbinals. This is especially critical for structures such as ET I, which have mixed functional roles in many mammals.

The dog-human connection.

This special issue of The Anatomical Record is the end result of a rare convergence of researchers scattered around the globe who came together to explore the mystery of the dog-human connection. Many of the discussions at the 12th International Congress of Vertebrate Morphology in Prague (July 23, 2019) are echoed within this issue. The enigmatic origins of dog domestication (as well as feralized descendants such as the dingo) are discussed, including phases of domestication that we might infer, and our historical knowledge of dog breeding. Emphasized by the morphological and genetic data are the forces of selection, both unintentional and intentional. In our modern life with dogs, we enjoy their companionship and benefit from the utility of many breeds, but we encounter unintended health care issues that are often breed-specific. Dogs are so different in their sensory specializations (especially olfaction), but have uniquely (among other domestic mammals) developed highly sophisticated means of interspecific communication with humans. In sum, the manuscripts within this issue discuss anatomical, paleontological, genetic, and behavioral evidence bearing on the antiquity of the domestic dog, the process of domestication, and the many ways in which dogs continue to affect human life.

The impact of extreme skull morphology in domestic dogs on cribriform plate shape.

Dog owners are often impressed by their dog's sense of smell. Many of these dogs, however, have skulls that are quite altered from those of their closest canid relatives. Housed within these skulls are essential olfactory structures like the cribriform plate that play a role in olfaction and the transmission of olfactory nerve impulses to the olfactory bulb of the brain. With improvements in CT technology and accessibility, we are now able to digitally reconstruct in 3D cribriform plate morphology and study its variation within and among species. In this study, we CT scanned the skulls of 95 dog specimens from 45 different domestic dog breeds and 12 species of wild canid and compared the shape of the cribriform plate among three main groups: domestic dog breeds, wolf-like canids, and fox-like canids. Despite only recent selective pressure for extreme skull morphology, domestic dogs display much more variation in cribriform plate shape than wild canids, indicating that cribriform plate shape is plastic and linked to skull shape. Intense artificial selection on domestic dog skull phenotype in the last 200 years has clear effects on secondary features of the domestic dog skull, implying that selection for overt phenotypes also can impact other anatomical features associated with the skull, like the cribriform plate.

Domesticating olfaction: Dog breeds, including scent hounds, have reduced cribriform plate morphology relative to wolves.

The domestic dog is assumed by nearly everyone to be the consummate smeller. Within the species Canis familiaris individual breeds, such as the bloodhound or beagle, are known as olfactory stars. These are "scent breeds," a grouping variably defined as a genetic clade or breed class commonly used for scent detection tasks. Previous work suggests that the dog has a more robust olfactory anatomy than many mammal species. Now we undertake a closer investigation of the dog's olfactory system, both in relationship to its closest wild relatives, the wolf and coyote, and across individual breeds. First, we seek to resolve whether the dog has lost olfactory capacity through its domestication from the wolf lineage. Second, we test the inertial lore that among dogs, "scent breeds," have a superior olfactory facility. As a measure of relative olfactory capacity, we look to the cribriform plate (CP), a bony cup in the posterior nasal cavity perforated by passageways for all olfactory nerve bundles streaming from the periphery to the brain. Using high-resolution computed tomography (CT) scans and digital quantification, we compare relative CP size in 46 dog breeds, the coyote and gray wolf. Results show the dog has a reduced CP surface area relative to the wolf and coyote. Moreover, we found no significant differences between CP size of "scent" and "non-scent" breeds. Our study suggests that the dog lost olfactory capacity as a result of domestication and this loss was not recovered in particular breed groupings through directed artificial selection for increased olfactory facility.

Olfaction at depth: Cribriform plate size declines with dive depth and duration in aquatic arctoid carnivorans.

It is widely accepted that obligate aquatic mammals, specifically toothed whales, rely relatively little on olfaction. There is less agreement about the importance of smell among aquatic mammals with residual ties to land, such as pinnipeds and sea otters. Field observations of marine carnivorans stress their keen use of smell while on land or pack ice. Yet, one dimension of olfactory ecology is often overlooked: while underwater, aquatic carnivorans forage "noseblind," diving with nares closed, removed from airborne chemical cues. For this reason, we predicted marine carnivorans would have reduced olfactory anatomy relative to closely related terrestrial carnivorans. Moreover, because species that dive deeper and longer forage farther removed from surface scent cues, we predicted further reductions in their olfactory anatomy. To test these hypotheses, we looked to the cribriform plate (CP), a perforated bone in the posterior nasal chamber of mammals that serves as the only passageway for olfactory nerves crossing from the periphery to the olfactory bulb and thus covaries in size with relative olfactory innervation. Using CT scans and digital quantification, we compared CP morphology across Arctoidea, a clade at the interface of terrestrial and aquatic ecologies. We found that aquatic carnivoran species from two lineages that independently reinvaded marine environments (Pinnipedia and Mustelidae), have significantly reduced relative CP than terrestrial species. Furthermore, within these aquatic lineages, diving depth and duration were strongly correlated with CP loss, and the most extreme divers, elephant seals, displayed the greatest reductions. These observations suggest that CP reduction in carnivorans is an adaptive response to shifting selection pressures during secondary invasion of marine environments, particularly to foraging at great depths. Because the CP is fairly well preserved in the fossil record, using methods presented here to quantify CP morphology in extinct species could further clarify evolutionary patterns of olfactory loss across aquatic mammal lineages that have independently committed to life in water.

Iterative evolution of large-bodied hypercarnivory in canids benefits species but not clades.

Ecological specialization has costs and benefits at various scales: traits benefitting an individual may disadvantage its population, species or clade. In particular, large body size and hypercarnivory (diet over 70% meat) have evolved repeatedly in mammals; yet large hypercarnivores are thought to be trapped in a macroevolutionary "ratchet", marching unilaterally toward decline. Here, we weigh the impact of this specialization on extinction risk using the rich fossil record of North American canids (dogs). In two of three canid subfamilies over the past 40 million years, diversification of large-bodied hypercarnivores appears constrained at the clade level, biasing specialized lineages to extinction. However, despite shorter species durations, extinction rates of large hypercarnivores have been mostly similar to those of all other canids. Extinction was size- and carnivory-selective only at the end of the Pleistocene epoch 11,000 years ago, suggesting that large hypercarnivores were not disadvantaged at the species level before anthropogenic influence.

Problems with inferring a lack of competition between Rancho La Brea dire wolves and sabertooth cats based on dental enamel.

Van Valkenburgh et al. challenge the conclusions of a recent study by DeSantis et al. that claimed that sabertooth cats and dire wolves did not compete for similar prey.

Tooth fracture frequency in gray wolves reflects prey availability.

Exceptionally high rates of tooth fracture in large Pleistocene carnivorans imply intensified interspecific competition, given that tooth fracture rises with increased bone consumption, a behavior that likely occurs when prey are difficult to acquire. To assess the link between prey availability and dental attrition, we documented dental fracture rates over decades among three well-studied populations of extant gray wolves that differed in prey:predator ratio and levels of carcass utilization. When prey:predator ratios declined, kills were more fully consumed, and rates of tooth fracture more than doubled. This supports tooth fracture frequency as a relative measure of the difficulty of acquiring prey, and reveals a rapid response to diminished food levels in large carnivores despite risks of infection and reduced fitness due to dental injuries. More broadly, large carnivore tooth fracture frequency likely reflects energetic stress, an aspect of predator success that is challenging to quantify in wild populations.

Distinct Predatory Behaviors in Scimitar- and Dirk-Toothed Sabertooth Cats.

Over the Cenozoic, large cat-like forms have convergently evolved into specialized killers of "megaherbivores" that relied on their large, and laterally compressed (saber-like) canines to rapidly subdue their prey [1-5]. Scimitar- and dirk-toothed sabertooths are distinct ecomorphs that differ in canine tooth length, degree of serration, and postcranial features indicative of dissimilar predatory behavior [6-13]. Despite these differences, it is assumed that they used a similar "canine-shear" bite to kill their prey [14, 15]. We investigated the killing behavior of the scimitar-toothed Homotherium serum and the dirk-toothed Smilodon fatalis using a comparative sample of living carnivores and a new quantitative approach to the analysis of skull function. For the first time, we quantified differences in the relative amount and distribution of cortical and trabecular bone in coronal sections of skulls to assess relative skull stiffness and flexibility [16-19]. We also use finite element analysis to simulate various killing scenarios that load skulls in ways that likely favor distinct proportions of cortical versus trabecular bone across the skull. Our data reveal that S. fatalis had an extremely thick skull and relatively little trabecular bone, consistent with a large investment in cranial strength for a stabbing canine-shear bite. However, H. serum had more trabecular bone and most likely deployed an unusual predatory behavior more similar to the clamp-and-hold technique of the lion than S. fatalis. These data broaden the killing repertoire of sabertooths and highlight the degree of ecological specialization among members of the large carnivore guild during the Late Pleistocene of North America.

Dietary specialization is linked to reduced species durations in North American fossil canids.

How traits influence species persistence is a fundamental question in ecology, evolution and palaeontology. We test the relationship between dietary traits and both species duration and locality coverage over 40 million years in North American canids, a clade with considerable ecomorphological disparity and a dense fossil record. Because ecomorphological generalization-broad resource use-may enable species to withstand disturbance, we predicted that canids of average size and mesocarnivory would exhibit longer durations and wider distributions than specialized larger or smaller species. Second, because locality coverage might reflect dispersal ability and/or survivability in a range of habitats, we predicted that high coverage would correspond with longer durations. We find a nonlinear relationship between species duration and degree of carnivory: species at either end of the carnivory spectrum tend to have shorter durations than mesocarnivores. Locality coverage shows no relationship with size, diet or duration. To test whether generalization (medium size, mesocarnivory) corresponds to an adaptive optimum, we fit trait evolution models to previously generated canid phylogenies. Our analyses identify no single optimum in size or diet. Instead, the primary model of size evolution is a classic Cope's Rule increase over time, while dietary evolution does not conform to a single model.

Olfaction written in bone: cribriform plate size parallels olfactory receptor gene repertoires in Mammalia.

The evolution of mammalian olfaction is manifested in a remarkable diversity of gene repertoires, neuroanatomy and skull morphology across living species. Olfactory receptor genes (ORGs), which initiate the conversion of odorant molecules into odour perceptions and help an animal resolve the olfactory world, range in number from a mere handful to several thousand genes across species. Within the snout, each of these ORGs is exclusively expressed by a discrete population of olfactory sensory neurons (OSNs), suggesting that newly evolved ORGs may be coupled with new OSN populations in the nasal epithelium. Because OSN axon bundles leave high-fidelity perforations (foramina) in the bone as they traverse the cribriform plate (CP) to reach the brain, we predicted that taxa with larger ORG repertoires would have proportionately expanded footprints in the CP foramina. Previous work found a correlation between ORG number and absolute CP size that disappeared after accounting for body size. Using updated, digital measurement data from high-resolution CT scans and re-examining the relationship between CP and body size, we report a striking linear correlation between relative CP area and number of functional ORGs across species from all mammalian superorders. This correlation suggests strong developmental links in the olfactory pathway between genes, neurons and skull morphology. Furthermore, because ORG number is linked to olfactory discriminatory function, this correlation supports relative CP size as a viable metric for inferring olfactory capacity across modern and extinct species. By quantifying CP area from a fossil sabertooth cat (Smilodon fatalis), we predicted a likely ORG repertoire for this extinct felid.

The Influence of Sniffing on Airflow and Odorant Deposition in the Canine Nasal Cavity.

Nasal airflow plays a critical role in olfaction by transporting odorant from the environment to the olfactory epithelium, where chemical detection occurs. Most studies of olfaction neglect the unsteadiness of sniffing and assume that nasal airflow and odorant transport are "quasi-steady," wherein reality most mammals "sniff." Here, we perform computational fluid dynamics simulations of airflow and odorant deposition in an anatomically accurate model of the coyote (Canis latrans) nasal cavity during quiet breathing, a notional quasi-steady sniff, and unsteady sniffing to: quantify the influence of unsteady sniffing, assess the validity of the quasi-steady assumption, and investigate the functional advantages of sniffing compared to breathing. Our results reveal that flow unsteadiness during sniffing does not appreciably influence qualitative (gross airflow and odorant deposition patterns) or quantitative (time-averaged olfactory flow rate and odorant uptake) measures of olfactory function. A quasi-steady approximation is, therefore, justified for simulating time-averaged olfactory function in the canine nose. Simulations of sniffing versus quiet breathing demonstrate that sniffing delivers about 2.5 times more air to the olfactory recess and results in 2.5-3 times more uptake of highly- and moderately-soluble odorants in the sensory region per unit time, suggesting one reason why dogs actively sniff. Simulations also reveal significantly different deposition patterns in the olfactory region during inspiration for different odorants, and that during expiration there is little retronasal odorant deposition in the sensory region. These results significantly improve our understanding of canine olfaction, and have several practical implications regarding computer simulation of olfactory function.

Skeletal trauma reflects hunting behaviour in extinct sabre-tooth cats and dire wolves.

Skeletal-injury frequency and distribution are likely to reflect hunting behaviour in predatory vertebrates and might therefore differ between species with distinct hunting modes. Two Pleistocene predators from the Rancho La Brea asphalt seeps, the sabre-tooth cat, Smilodon fatalis, and dire wolf, Canis dirus, represent ambush and pursuit predators, respectively. On the basis of a collection of over 1,900 pathological elements, the frequency of traumatic injury across skeletal elements in these two species was calculated. Here we show that the frequency of trauma in the sabre-tooth cat exceeds that of the dire wolf (4.3% compared to 2.8%), implying that the killing behaviour of S. fatalis entailed greater risk of injury. The distribution of traumatic injuries also differed between the two species. S. fatalis, an ambush predator, was injured more often than expected across the lumbar vertebrae and shoulders whereas C. dirus, a pursuit predator, had higher than expected levels of injury in the limbs and cervical vertebrae. Spatial analysis was used to quantify differences in the distribution of putative hunting injuries. Analysis of injury locations discriminated true hotspots from injury-dense areas and facilitated interpretation of predatory behaviour, demonstrating the use of spatial analyses in the study of vertebrate behaviour and evolution. These results suggest that differences in trauma distribution reflect distinct hazards of each species' hunting mode.

Comparative Morphology and Histology of the Nasal Fossa in Four Mammals: Gray Squirrel, Bobcat, Coyote, and White-Tailed Deer.

Although the anatomy of the nasal fossa is broadly similar among terrestrial mammals, differences are evident in the intricacies of nasal turbinal architecture, which varies from simple scroll-like to complex branching forms, and in the extent of nonsensory and olfactory epithelium covering the turbinals. In this study, detailed morphological and immunohistochemical examinations and quantitative measurements of the turbinals and epithelial lining of the nasal fossa were conducted in an array of species that include the gray squirrel, bobcat, coyote, and white-tailed deer. Results show that much more of the nose is lined with olfactory epithelium in the smallest species (gray squirrel) than in the larger species. In two species with similar body masses, bobcat and coyote, the foreshortened felid snout influences turbinal size and results in a decrease of olfactory epithelium on the ethmoturbinals relative to the longer canine snout. Ethmoturbinal surface area exceeds that of the maxilloturbinals in all four sampled animals, except the white-tailed deer, in which the two are similar in size. Combining our results with published data from a broader array of mammalian noses, it is apparent that olfactory epithelial surface area is influenced by body mass, but is also affected by aspects of life history, such as diet and habitat, as well as skull morphology, itself a product of multiple compromises between various functions, such as feeding, vision, and cognition. The results of this study warrant further examination of other mammalian noses to broaden our evolutionary understanding of nasal fossa anatomy. Anat Rec, 299:840-852, 2016. © 2016 Wiley Periodicals, Inc.