Shape analysis of the preorbital bar in caviomorph rodents.

The highly specialised masticatory apparatus of rodents raises interesting questions about how their skull withstands the intensive and sustained forces produced by biting on hard items. In these mammals, major systematics were explored for a long time based on the adductor muscles' architecture and the related bony structures. The infraorbital foramen stands out, where a hypertrophied head of the zygomaticomandibular muscle passes through-in hystricomorphous rodents-as a direct consequence of the lateral and posterior shift of the preorbital bar. Interestingly, this bar moved laterally and backwards-enlarging the foramen-but it never disappeared throughout evolution, even showing morphological convergence among rodents. Previous research proposed this bar as behaving mechanically similar to the postorbital bar in ungulates, i.e., a safety structure against torsion stress while chewing. We analysed its morphology by mathematically modelling it under bending and torsion scenarios (linearly and elliptically shaped, respectively), and as for biting load propagation (catenary curve). Although the preorbital bar primarily seems to be shaped for withstanding torsional stress (as the postorbital bar in ungulates) and as an escaping point for force propagation, these forces are not a consequence of chewing and grinding foods, but preventing the zygomatic arch from failing when the powerful laterally-displaced jaw adductor muscles are pulling the dentary upwards at biting.

Sample pooling for SARS-CoV-2 RT-PCR screening.

OBJECTIVE: To evaluate the efficacy of sample pooling compared to the individual analysis for the diagnosis of coronavirus disease 2019 (COVID-19) by using different commercial platforms for nucleic acid extraction and amplification. METHODS: A total of 3519 nasopharyngeal samples received at nine Spanish clinical microbiology laboratories were processed individually and in pools (342 pools of ten samples and 11 pools of nine samples) according to the existing methodology in place at each centre. RESULTS: We found that 253 pools (2519 samples) were negative and 99 pools (990 samples) were positive; with 241 positive samples (6.85%), our pooling strategy would have saved 2167 PCR tests. For 29 pools (made out of 290 samples), we found discordant results when compared to their correspondent individual samples, as follows: in 22 of 29 pools (28 samples), minor discordances were found; for seven pools (7 samples), we found major discordances. Sensitivity, specificity and positive and negative predictive values for pooling were 97.10% (95% confidence interval (CI), 94.11-98.82), 100%, 100% and 99.79% (95% CI, 99.56-99.90) respectively; accuracy was 99.80% (95% CI, 99.59-99.92), and the kappa concordant coefficient was 0.984. The dilution of samples in our pooling strategy resulted in a median loss of 2.87 (95% CI, 2.46-3.28) cycle threshold (Ct) for E gene, 3.36 (95% CI, 2.89-3.85) Ct for the RdRP gene and 2.99 (95% CI, 2.56-3.43) Ct for the N gene. CONCLUSIONS: We found a high efficiency of pooling strategies for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA testing across different RNA extraction and amplification platforms, with excellent performance in terms of sensitivity, specificity and positive and negative predictive values.

Mandible strength and geometry in relation to bite force: a study in three caviomorph rodents.

The monophyletic group Caviomorpha constitutes the most diverse rodent clade in terms of locomotion, ecology and diet. Caviomorph species show considerable variation in cranio-mandibular morphology that has been linked to the differences in toughness of dietary items and other behaviors, such as chisel-tooth digging. This work assesses the structural strength of the mandible of three caviomorph species that show remarkable differences in ecology, behavior and bite force: Chinchilla lanigera (a surface-dwelling species), Octodon degus (a semi-fossorial species) and Ctenomys talarum (a subterranean species). Finite element (FE) models of the mandibles are used to predict the stresses they withstand during incisor biting; the results are related to in vivo bite forces and interspecific variations in the mandibular geometries. The study concludes that the mandible of C. talarum is better able to withstand strong incisor bites. Its powerful adducting musculature is consistent with the notorious lateral expansion of the angular process and the masseteric crest, and the enhanced cortical bone thickness. Although it has a relatively low bite force, the mandible of O. degus also shows a good performance for mid-to-strong incisor biting, in contrast to that of C. lanigera, which exhibits, from a mechanical point of view, the worst performance. The mandibles of C. talarum and O. degus appear to be better suited to withstand stronger reaction forces from incisor biting, which is consistent with their closer phylogenetic affinity and shared digging behaviors. The contrast between the low in vivo bite force of C. lanigera and the relatively high estimations that result from the models suggests that its adductor musculature could play significant roles in functions other than incisor biting.

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.

Bite it forward bite it better? Incisor procumbency and mechanical advantage in the chisel-tooth and scratch-digger genus Ctenomys (Caviomorpha, Rodentia).

The subterranean genus Ctenomys (∼60 species, ∼100-1000g) constructs its burrows by using both forefeet and teeth throughout a wide range of habitats in South America. They show a high variation in the incisors' angle of attack (procumbency) and a mostly conserved skull morphology, not only amongst their congeners, but within the caviomorph rodents. Traditionally, procumbency has been largely related to tooth-digging. Looking for the possible influence of incisor procumbency on the mechanical advantage (MA) of each of the seven jaw adductor muscles in the genus, we examined 165 skulls representing 24 species. We also evaluated the role of two other relevant traits - i.e. mandibular width and diastema length - in jaw biomechanics and the existence of a relationship between procumbency angle and soil hardness. The in- and out-lever arms (Li and Lo) of the involve muscles were determined based on their insertion's 3D-coordinates and integrated to calculate their MA. Interspecific scaling relationships for skull and muscle measurements were analyzed through reduced major axis regression performed with phylogenetically independent standardized contrasts. Although the procumbency angle ranged between ∼92.5° (C. mendocinus) and ∼107.2 (C. occultus), we found that it was not significantly correlated with the MA of any jaw adductor muscle. This study also showed that the incisor procumbency variation was not associated with the relative rostral length or soil hardness. This result contradicts previous generalizations about a correlation between habitat conditions and the procumbency of the incisors in subterranean rodents. In sum, our results suggest that, within Ctenomys, possessing more procumbent incisors may not represent a biomechanical advantage, but might be beneficial in other aspects related to chisel-tooth digging or food processing behaviors.

Cranial suture complexity in caviomorph rodents (Rodentia; Ctenohystrica).

Due to their flexibility, sutures are regions that experience greater strains than the surrounding rigid cranial bones. Cranial sutures differ in their degree of interdigitation or complexity. There is evidence indicating that a more convoluted suture better enables the absorption of high stresses coming from dynamic masticatory forces, and other functions. The Order Rodentia is an interesting clade to study this because of its taxa with diverse chewing modes. Due to repeated loading resulting from gnawing and grinding, energy absorption by the sutures might be a crucial factor in these mammals. Species within the infraorder Caviomorpha were chosen as a case study because of their ecomorphological and dietary diversity. This study compared five sutures from the rostrum and cranial vault across seven caviomorph families, and assessed their complexity by means of the relative length and fractal dimension. Across these rodents, cranial sutures are morphologically quite diverse. We found that the sutures connecting the rostrum with the vault were relatively more interdigitated than those in the cranial vault itself, especially premaxillofrontal sutures. Suture interdigitation was higher in species that display chisel-tooth digging and burrowing behaviors, especially in the families Ctenomyidae and Octodontidae, than those in families Dasyproctidae and Cuniculidae, which have more gracile masticatory systems. The reconstruction of the ancestral character state, on family and species phylogeny, points toward low suture interdigitation (i.e., low length ratio) as a likely ancestral state for interfrontal, premaxillofrontal and maxillofrontal sutures. Interspecific differences in suture morphology shown here might represent adaptations to different mechanical demands (i.e., soft vs. tough foods) or behaviors (e.g., chisel-tooth digging), which evolved in close association with the diverse environments occupied by caviomorph rodents.

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.

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.

Sexual selection in a polygynous rodent (Ctenomys talarum): an analysis of fighting capacity.

The South American subterranean rodent genus Ctenomys (Caviomorpha: Octodontoidea), which uses both claws and teeth to dig, shows striking morphological adaptations to its specialized mode of life. Among other traits, the genus has evolved a powerful jaw musculature and procumbent incisors that are used for dento-excavation. Behavioral observations indicate that these traits are also used during male aggressive encounters, which characterize the polygynous mating system of one of the species of the genus, Ctenomys talarum. A question emerges about sexual selection: could it have induced further changes in traits primarily evolved as adaptations for digging? To address this issue, we studied functional and morphological attributes of the jaw and incisors in specimens of C. talarum. Incisor bite forces were measured on wild females and males from a local population (Mar de Cobo; Buenos Aires Province) by means of a strain gauge load cell force transducer. Museum specimens coming from the same population were studied to assess anatomical attributes of both sexes. Since this species exhibits dimorphism in body size, the possible effect of body mass on the studied traits was analyzed. Males and females showed significant differences in biting performance and mandibular width, but when size was taken into account these differences disappeared. However, other dimorphic traits can vary with a certain independence with respect to size, particularly the 2nd moment of area of the incisors and, to a lesser extent, incisor procumbency. The former geometrical parameter, which is proportional to the bending strength, was highly dimorphic. This fact suggests that, during aggressive encounters between males, biting would place large bending loads on the incisors.

Scaling and adaptations of incisors and cheek teeth in caviomorph rodents (Rodentia, Hystricognathi).

The South American hystricognath rodents are one of the most diverse mammalian clades considering their occupied habitats, locomotor modes and body sizes. This might have been partly evolved by diversification of their masticatory apparatus' structure and its ecological commitment, for example, chisel-tooth digging. In this phylogeny-based comparative study, we test the relationship between ecological behavior and mechanical features of their incisors and molariforms. In 33 species of nine families of caviomorph rodents, we analyze incisor attributes related to structural stress resistance and molar features related with grinding capacity, for example, second moment of inertia and enamel index (EI) (enamel band length/occlusal surface area), respectively. Most of these variables scaled isometrically to body mass, with a strong phylogenetic effect. A principal component analysis discrimination on the EI clustered the species according to their geographic distribution. We presume that selective pressures in Andean-Patagonian regions, on particular feeding habits and chisel-tooth digging behaviors, have modeled the morphological characteristics of the teeth. Subterranean/burrower ctenomyids, coruros, and plains viscachas showed the highest bending/torsion strength and anchorage values for incisors; a simplified enamel pattern in molariforms would be associated with a better grinding of the more abrasive vegetation present in more open and drier biomes.