Functional morphology and fiber types of the masseter muscles of two caviomorph rodents with contrasting lifestyles, Ctenomys talarum (Ctenomyidae) and Cavia aperea (Caviidae).

The aim of this work is the analysis of histochemical and morphometric properties of the masseter muscles of Ctenomys talarum and Cavia aperea. The former belongs to a subterranean rodent clade, Ctenomyidae, which has evolved a robust masticatory apparatus adapted to chisel-tooth digging and processing of abrasive grasses; C. aperea belongs to the family Caviidae, with relatively graceful jaws and mandibular musculatures, consistent with less mechanically challenging diets. Adult males were captured, immediately transported to the laboratory, and euthanized in a CO2 chamber. The musculus masseter superficialis and musculus masseter profundus on the left side of the animals were used to analyze the histochemical composition of the fiber types treated with myosin adenosine triphosphatase, succinate dehydrogenase and periodic acid Schiff. The mean fiber diameters, relative areas, and frequencies of each muscle fiber type were calculated. The mm. masseter superficialis and masseter profundus on the right side were used to measure the physiological cross-sectional area (PCSA). Based on this measurement, the internal force (F) was estimated. In the m. masseter profundus of both species and in the m. masseter superficialis of C. aperea intermediate fast oxidative-glycolytic fibers (FOGi) predominated. In the mm. masseter superficialis and masseter profundus of C. talarum the relative area of fast glycolytic (FG) fibers was greater than that of the muscles of C. aperea, whose main muscle fiber component is FOGi fibers. When corrected for body mass differences, PCSA was higher for the mm. masseter superficialis of C. talarum. This and the larger relative area of FG fibers, probably contributes to the exertion of large bite forces in C. talarum, as measured in previous studies.

Functional morphology and identity of the thenar pad in the subterranean genus Ctenomys (Rodentia, Caviomorpha).

As in many other fossorial tetrapods, the most obvious adaptations to scratch-digging in the subterranean tuco-tuco (Rodentia, Ctenomyidae, Ctenomys) are found in the hands, which among other adaptations, present the mesaxonic condition; i.e. the central digits are more developed, and also their claws, which are curved and elongated. The thumb is atrophied and aligned with the rest of the digits, showing a small and flat claw. This configuration of digits and claws seems to be in accordance with what it is expected for rodents: rudimentary movements when handling food items. However, on the palmar side of the hand, tuco-tucos have several pads, the thenar (located under the thumb) being the most developed. In this study, we investigated the functional morphology of the thenar pad through different approaches: musculoskeletal anatomy, histology and functionality. The analysis of radiographs and clarified and double-stained hand samples of Ctenomys talarum and C. australis showed that the thenar pad is supported by a paddle-shaped bone that articulates with a protrusion in the scapholunate bone. This bone, flat and long, continues in a flat cartilaginous structure, with a shape similar to a claw. Dissections showed that the thenar pad has several associated muscles: the m. palmaris longus, the m. abductor pollicis longus, and a massive muscular complex located between the thumb and the thenar pad. By topology it might be inferred that this complex is formed by the m. abductor pollicis brevis, the m. flexor pollicis brevis and the m. adductor pollicis brevis. Longitudinal histological sections of the thenar pad stained with hematoxylin-eosin showed a thick layer of keratin at the distal end, external face. The observation of live specimens of C. talarum foraging on two food items of different size and filmed at 300 fps showed that the thenar pad acts as an opposable thumb, with digit-like movements. Tuco-tucos are able to perform more precise movements than expected, and to grasp and manipulate the food with one hand. In previous studies, it was suggested that the thenar pad was supported by a 'palmar ossicle', or 'prepollex' (= radial sesamoid bone). Our results suggest that this sesamoid underwent a radical change on its morphology, making the thenar pad a part of the food handling system in Ctenomys, so the thenar pad might be considered a 'false thumb', rather than a palmar pad. It is suggested to advance on the description and functional analysis of the thenar pad, redefining the structure, since the terms used so far to define it would not be accurate.

An allometric analysis of sexual dimorphism in Ctenomys australis: integrating classic morphometry and functional performance in vivo.

The South American rodents of the genus Ctenomys (Rodentia, Hystricognathi), which use both forelimbs and incisors to dig, show strong, specialized morphological adaptations to living in the underground niche. In these rodents, the effectiveness of a bite - in this case the potential to inflict physical damage - mostly depends on the strength of the incisors (e.g. bending and torsion stresses) and the power of the masseteric muscle of the jaw. Ctenomys australis (the sand dune tuco-tuco) is a highly territorial subterranean rodent that builds large and exclusive burrow systems in coastal sand dunes found continuously along the Atlantic coast of the Buenos Aires province, Argentina. Incisors in this species are used both in agonistic encounters among congeners (i.e. reproductive or territorial intentions) as well as to dig and increase the size of their burrows daily (e.g. acquisition of plant material as food resources). We first characterized sexual dimorphism in cranial traits involved in aggressive interactions between males, analyzing ontogenetic trajectories of both sexes. We also analyzed some remarkable attributes of sexual dimorphism in subadult and adult individuals, the bending and torsion stresses and the bite performance in vivo, hypothesizing that males are able to apply comparatively stronger bite forces at their incisor tips than females, concordant with a broader mandible for the insertion of a powerful adductor musculature. Bite forces were measured in vivo in wild animals of both sexes using a strain gauge load cell force transducer. These individuals were also used to estimate the incisor area (CA), the 2nd moment of inertia of the incisors (I, or the bending strength to external forces), and the torsional strength of the incisor (J), all traits involved in the production of the force that can be withstood at the incisor tips. Ontogeny explained some sexual differences in the cranial traits of adults, expressed as (i) morphological changes related to body size variation, and (ii) morphological changes associated with differences in the slope of ontogenetic trajectories (regardless of body size variation). Recordings of in vivo bite forces were significantly higher in adult males than in females. These differences in bite forces were not associated with the sex itself, but the sexual dimorphism in body size. Although males did not show significantly higher allometric coefficients for I, J and CA than females, males still showed a strong sexual dimorphism in these traits due to body size variation. On the other hand, measurements of mandibular width - an estimator of the relative size of masseteric muscles - in vivo showed strong differences between sexes, suggesting higher force performance at the incisor tips for males. Overall, we observed that many attributes related to bite performance might have been molded by sexual selection, which implies differences in allometric coefficients from some morphological traits throughout ontogeny.

Analysis of arch-like bones: The rodent mandible as a case study.

Bone strength is determined by the mechanical properties of bone material, and the size and shape of the whole bone, i.e., its architecture. The mandible of vertebrates has been traditionally regarded as a beam oriented in relation to main masticatory loads, i.e., the longer dimension of its cross-section being parallel to the load. Rodents follow this pattern but, in addition, their mandible possesses an intriguing arch-like shape that is apparent when seen in the lateral view. Little attention was given to the structural capacity of this trait. The advantage of an arch is that it can withstand a greater load than a horizontal beam. The objective of this study was to model the rodent mandible like an arch to evaluate its structural strength. The bending moment in an arch-like mandible was 15-25% lower with respect to a beam-like mandible. Further, bending varies with mandible "slenderness" and incisor procumbency, a functionally relevant rodent trait. In the rodent Ctenomys talarum (Caviomorpha; Ctenomyidae), bone stress was substantially reduced when the mandible was modeled as an arch-like structure as compared with a beam-like structure, and safety factors were 15-34% higher. The shape of rodents' mandible might confer a functional advantage to high and repeatedly applied loads resulting from a unique feeding mode: gnawing. J. Morphol. 277:879-887, 2016. © 2016 Wiley Periodicals, Inc.

The role of stimulus complexity, age and experience in the expression of exploratory behaviour in the Chimango Caracara, Milvago chimango.

Exploration represents an important way by which organisms evaluate environment information. The decision of whether or not an animal should investigate environmental changes may influence the extent to which animals learn about their surroundings and cope with habitat modifications. We analysed exploration behaviour in a suburban population of a raptor species, the Chimango Caracara, Milvago chimango, by examining how age, previous experience and object complexity influence novel object exploration. Our findings showed that object complexity did not influence caracaras initial approach and contact with objects, but did influence the degree of engagement during exploratory activities, as measured by total exploration time and number of exploration events. These variables were higher for complex objects than for simple objects. Experience resulted in less exploration of simple objects. It is likely that, for caracaras, simple objects are easier to encode and recall than complex objects, so additional exploration of such objects would not provide further information. Results suggest that exploratory behaviour in this raptor was guided more by the benefits of a greater quantity of information obtained by exploring complex objects, than by the risks associated to this activity. We can conclude that caracaras cope with novel features in their surroundings with a novelty-seeking strategy, characteristic for generalist species in discovering early new resources opportunities, and which might be a determining factor for adaptive responses to environment modification.

Postnatal ontogeny of limb proportions and functional indices in the subterranean rodent Ctenomys talarum (Rodentia: Ctenomyidae).

Burrow construction in the subterranean Ctenomys talarum (Rodentia: Ctenomyidae) primarily occurs by scratch-digging. In this study, we compared the limbs of an ontogenetic series of C. talarum to identify variation in bony elements related to fossorial habits using a morphometrical and biomechanical approach. Diameters and functional lengths of long bones were measured and 10 functional indices were constructed. We found that limb proportions of C. talarum undergo significant changes throughout postnatal ontogeny, and no significant differences between sexes were observed. Five of six forelimb indices and two of four hindlimb indices showed differences between ages. According to discriminant analysis, the indices that contributed most to discrimination among age groups were robustness of the humerus and ulna, relative epicondylar width, crural and brachial indices, and index of fossorial ability (IFA). Particularly, pups could be differentiated from juveniles and adults by more robust humeri and ulnae, wider epicondyles, longer middle limb elements, and a proportionally shorter olecranon. Greater robustness indicated a possible compensation for lower bone stiffness while wider epicondyles may be associated to improved effective forces in those muscles that originate onto them, compensating the lower muscular development. The gradual increase in the IFA suggested a gradual enhancement in the scratch-digging performance due to an improvement in the mechanical advantage of forearm extensors. Middle limb indices were higher in pups than in juveniles-adults, reflecting relatively more gracile limbs in their middle segments, which is in accordance with their incipient fossorial ability. In sum, our results show that in C. talarum some scratch-digging adaptations are already present during early postnatal ontogeny, which suggests that they are prenatally shaped, and other traits develop progressively. The role of early digging behavior as a factor influencing on morphology development is discussed.

Another one bites the dust: bite force and ecology in three caviomorph rodents (Rodentia, Hystricognathi).

Mammals have developed sophisticated strategies adapting to particular locomotor modes, feeding habits, and social interactions. Many rodent species have acquired a fossorial, semi-fossorial, or even subterranean life-style, converging on morphological, anatomical, and ecological features but diverging in the final arrangement. These ecological variations partially depend on the functional morphology of their digging tools. Muscular and mechanical features (e.g., lever arms relationship) of the bite force were analyzed in three caviomorph rodents with similar body size but different habits and ecological demands of the jaws. In vivo forces were measured at incisors' tip using a strain gauge load cell force transducer whereas theoretical maximal performance values, mechanical advantages, and particular contribution of each adductor muscle were estimated from dissections in specimens of Ctenomys australis (subterranean, solitary), Octodon degus (semi-fossorial, social), and Chinchilla laniger (ground-dweller, colonial). Our results showed that C. australis bites stronger than expected given its small size and C. laniger exhibited the opposite outcome, while O. degus is close to the expected value based on mammalian bite force versus body mass regressions; what might be associated to the chisel-tooth digging behavior and social interactions. Our key finding was that no matter how diverse these rodents' skulls were, no difference was found in the mechanical advantage of the main adductor muscles. Therefore, interspecific differences in the bite force might be primarily due to differences in the muscular development and force, as shown for the subterranean, solitary and territorial C. australis versus the more gracile, ground-dweller, and colonial C. laniger.

Evaluation of neophobia and its potential impact upon predator control techniques: a study on two sympatric foxes in southern Patagonia.

An alternative approach to increase the efficiency of predator control and selectivity is to consider the natural behavioural repertoire of the target species and how such behaviours may increase their vulnerability. Neophobia, or the hesitancy to approach a novel food item, object, or place, is an important factor influencing the investigative behaviour of animals, and its incorporation to predator control techniques may help to reduce losses of livestock to predators. In this study, we simultaneously evaluated the existence and intensity of neophobic responses in two sympatric fox species, the Culpeo (Pseudalopex culpaeus) and the Grey (P. griseus) foxes in southern Patagonia, Argentina. For this purpose, we used bait stations to compare fox behavioural responses in the absence (pre-treatment), presence (treatment) and removal (post-treatment) of a novel stimulus, which consisted of an orange PVC-traffic cone. Both fox species showed a neophobic response: bait-station visitation rates decreased (P=0.005 and P=0.048, for Culpeo and Grey foxes, respectively) in the presence of the novel object. The intensity of the response differed between species being higher for Culpeo foxes (approximately 80% of reduction in visitation rate during treatment for Culpeo foxes vs. 10% for Grey foxes). However, the bait-station visitation pattern after novel object removal indicated that animals probably increased exploration of the station. The high level of neophobia achieved by the Culpeo fox, together with an increase in post-treatment site exploration, suggests that behavioural manipulations (reduction of neophobia and its consequent increase in risk taking) could improve selective and efficient fox control in rural areas where livestock production is a major economic activity.

Biting performance and skull biomechanics of a chisel tooth digging rodent (Ctenomys tuconax; Caviomorpha; Octodontoidea).

Biting performance is a key factor in vertebrate groups possessing particular food habits. In subterranean rodents that use the incisors as a digging tool, apart from requirements related to gnawing abrasive diets, the force exerted at the incisors tips must be sufficient to break down soils that are often exceedingly compact. The subterranean genus Ctenomys diversified in the southern portion of South America closely associated with the relatively open environments that characterize that region. This genus is considered a "claw and chisel tooth digger," that is, during the excavation of their galleries, the animals break down the soil with both the fore-claws and the incisors. We report here measurements of in vivo bite force in one of the largest species of the genus, C. tuconax, which occupies highland grasslands with compacted soils. We document the combined use of claws and incisors observed under field conditions, also providing measurements of soil compaction in the habitat occupied by this species. We report estimates of bite force at the level of the incisors and cheek teeth calculated from the physiological cross-sectional area of jaw muscles. To this aim, anatomical and biomechanical analyses of the mandibular apparatus were performed in preserved specimens. We found that C. tuconax bites with a higher force than expected for a mammal of its size. To assess anatomical correlates of biting performance, the morphology of the skull and jaw, and incisor second moment of area were compared with those of other caviomorph rodents with different lifestyle.

Inter-individual and age differences in exploration, neophobia and problem-solving ability in a Neotropical raptor (Milvago chimango).

Animal innovations have far-reaching ecological and evolutionary consequences. The occurrence and persistence of an innovation require several processes, including exploration, social and asocial learning, and low neophobia. In addition, the identity of the innovator may determine how these new behaviours are socially transmitted. Taking into account inter-individual and age differences, we investigated three correlates of animal innovation: object exploration, neophobia level and novel problem-solving ability in an opportunistic generalist raptor, the Chimango Caracara (Milvago chimango). Eighteen individuals (7 adults and 11 juveniles) were caught during the non-breeding period and housed in individual cages in outdoor aviaries. Each bird was given three tests: exploration, neophobia and problem-solving. Individuals differed in their response to novel situations both within and between age groups. Most of the juveniles were more explorative and had a lower neophobic response to a strange object than adult birds, but both age groups were able to solve a novel problem when given a food reward. In juveniles, neophobia level and problem-solving performance were inversely related; however, we found no relationship between these behaviours in adults. Exploration did not correlate with neophobia or problem-solving ability for either age group. This research is one of the few studies exploring the inter-individual and age differences in behavioural innovation and their correlates in a bird of prey. The explorative tendency, low neophobia and ability to innovate showed by M. chimango may be advantageous for this generalist and opportunistic raptor and might be some of the factors underlying its ecological success.

Evolution of brain size in a highly diversifying lineage of subterranean rodent genus Ctenomys (Caviomorpha: Ctenomyidae).

We evaluated brain size evolution in a specialized subterranean mammal, the ctenomyid rodent genus Ctenomys (tuco-tuco) and compared it, within a phylogenetic framework, to other caviomorph rodents differing in body size and modes of life. Although brain size in Ctenomys falls below the regression line obtained for caviomorph rodents, some fossorial species within the sister family Octodontidae, which make use of both above and below ground habitats, have a relatively larger brain size. Ctenomys has experienced a relatively recent and 'explosive' cladogenesis which yielded approximately 60 living species differing almost one order of magnitude in body size. Based on the evolutionary lag hypothesis between body versus brain size evolution, a negative correlation between encephalization quotient and body size would be expected for a group experiencing such a recent diversification in size. This is because a decrease of body size occurring during the emergence of a new species should produce a correspondingly greater encephalization and vice versa. Despite the fact that the allometric coefficient of brain weight versus body weight for 30 living species of Ctenomys was lower than that obtained for families within Caviomorpha, we failed to find any significant relationship between encephalization and body weight. Finally, the relationship between brain size and metabolism was assessed for those species of Ctenomys in which metabolic data are available. Brain size evolution in Ctenomys is discussed in regards to sensory capabilities and behavioral attributes associated with the occupation of a subterranean ecological niche.