Rhinoceros beetle horn development reveals deep parallels with dung beetles.

Beetle horns are attractive models for studying the evolution of novel traits, as they display diverse shapes, sizes, and numbers among closely related species within the family Scarabaeidae. Horns radiated prolifically and independently in two distant subfamilies of scarabs, the dung beetles (Scarabaeinae), and the rhinoceros beetles (Dynastinae). However, current knowledge of the mechanisms underlying horn diversification remains limited to a single genus of dung beetles, Onthophagus. Here we unveil 11 horn formation genes in a rhinoceros beetle, Trypoxylus dichotomus. These 11 genes are mostly categorized as larval head- and appendage-patterning genes that also are involved in Onthophagus horn formation, suggesting the same suite of genes was recruited in each lineage during horn evolution. Although our RNAi analyses reveal interesting differences in the functions of a few of these genes, the overwhelming conclusion is that both head and thoracic horns develop similarly in Trypoxylus and Onthophagus, originating in the same developmental regions and deploying similar portions of appendage patterning networks during their growth. Our findings highlight deep parallels in the development of rhinoceros and dung beetle horns, suggesting either that both horn types arose in the common ancestor of all scarabs, a surprising reconstruction of horn evolution that would mean the majority of scarab species (~35,000) actively repress horn growth, or that parallel origins of these extravagant structures resulted from repeated co-option of the same underlying developmental processes.

Evaluation of morphological traits and physiological variables of several Chinese goat breeds and their crosses.

The current study was undertaken to evaluate some morphological traits of the goat breeds raised in Southwest China. The field experimentation and data collection were from 434 animals presenting seven breeds of the Dazu black goat (DBG; n = 203), Saanen milk goat (SMG; n = 50), Black Boer × Dazu black goat (BXC; n = 28), Hechuan white goat (HWG; n = 49), Inner Mongolia Cashmere goat (IMCG; n = 25), IMCG × DBG (F1; n = 57) and F1 × F1 (F2; n = 22). All studied animals were adult and selected to be at the same age (36.50 ± 0.75 months). After editing, more than 20 morphological and production traits like body condition score (BCS), testicle measurements, coat colour, fibre traits, skin colour, horn colour, horn shape, horn orientation, wattles, front hair, beard, ear shape, ear size, rump angle, hind leg angulation and physiological variables were analysed. BXC and DBG had dark coat colour, whilst SMG, HWG and IMCG had light colour, whilst F1 and F2 ranged from light to dark coat colour. Concerning BCS, the breeds BXC and DBG were characterized as fat goats, whilst SMG, HWG, F1 and F2 were average, whilst IMCG was thin. The maximum values for testis measurements were recorded for BXC, SMG and DBG. For fibre traits, IMCG, F1 and F2 were the most superior. BXC and DBG males have good fertility parameters. The highest values for rectal temperature, skin temperature and breath rate were recorded for SMG. These findings revealed the presence of a wide range of morphological differences among studied goat breeds. Such diversity in the performance of goat breeds raised in Southwest China is crucial to implement reliable selection strategies for breeding goats in this area.

Morphological divergence in the West African shorthorn Lagune cattle populations from Benin.

This study explored the potential role of agro-ecology in shaping the morphology of Lagune cattle population of Benin. A total of 708 adult Lagune cattle were sampled randomly from nine provenances in two agro-ecological zones (AEZs) and were assessed for eight qualitative and twelve linear body measurements. Data were analyzed using generalized linear model procedures (PROC GLM) followed by the multiple comparison of least square means (LSMEAN) according to the Tukey-Kramer method and multivariate analytical methods, including canonical discrimination analysis (CDA) and hierarchical ascendant classification. Irrespective of AEZ and sex, the body length (102.3 ± 9.31 cm) was greater than the wither height (93.1 ± 7.39 cm) and the body index smaller than 0.85. However, there were significant differences between the two AEZs for most of the measured morphometric and qualitative traits. Moreover, a male-biased sexual size dimorphism was recorded. The CDA based on only four basic body measurements (rump height, body length, heart girth, and ear length) and the calculated Mahalanobis distances suggest that the populations from the two AEZs are distinct and could be further considered ecotypes. Nevertheless, the overall moderate classification rate (70%) of the individual animals into their group of origin indicates interbreeding between the two populations. The pairwise Mahalanobis distances between provenances in the same AEZ were also significant. Together, these results provide supporting evidence for the existence of subdivisions in the Lagune cattle populations from South Benin. The high morphological diversity in the Lagune cattle recorded in the present study could serve as a starting point for the development of efficient selection and sound subpopulation management strategies but also for further phenotypic and genetic characterizations.

Patterns of divergence in the morphology of ceratopsian dinosaurs: sympatry is not a driver of ornament evolution.

Establishing the origin and function of unusual traits in fossil taxa provides a crucial tool in understanding macroevolutionary patterns over long periods of time. Ceratopsian dinosaurs are known for their exaggerated and often elaborate horns and frills, which vary considerably between species. Many explanations have been proposed for the origin and evolution of these 'ornamental' traits, from predator defence to socio-sexual dominance signalling and, more recently, species recognition. A key prediction of the species recognition hypothesis is that two or more species possessing divergent ornamental traits should have been at least partially sympatric. For the first time to our knowledge, we test this hypothesis in ceratopsians by conducting a comparison of the morphological characters of 46 species. A total of 350 ceratopsian cladistic characters were categorized as either 'internal', 'display' (i.e. ornamental) or 'non display'. Patterns of diversity of these characters were evaluated across 1035 unique species pairs. Display characters were found to diverge rapidly overall, but sympatric species were not found to differ significantly in their ornamental disparity from non-sympatric species, regardless of phylogenetic distance. The prediction of the species recognition hypothesis, and thus the idea that ornamentation evolved as a species recognition mechanism, has no statistical support among known ceratopsians.

Evolutionary allometry reveals a shift in selection pressure on male horn size.

How selection pressures acting within species interact with developmental constraints to shape macro-evolutionary patterns of species divergence is still poorly understood. In particular, whether or not sexual selection affects evolutionary allometry, the increase in trait size with body size across species, of secondary sexual characters, remains largely unknown. In this context, bovid horn size is an especially relevant trait to study because horns are present in both sexes, but the intensity of sexual selection acting on them is expected to vary both among species and between sexes. Using a unique data set of sex-specific horn size and body mass including 91 species of bovids, we compared the evolutionary allometry between horn size and body mass between sexes while accounting for both the intensity of sexual selection and phylogenetic relationship among species. We found a nonlinear evolutionary allometry where the allometric slope decreased with increasing species body mass. This pattern, much more pronounced in males than in females, suggests either that horn size is limited by some constraints in the largest bovids or is no longer the direct target of sexual selection in very large species.

Structural adaptations to diverse fighting styles in sexually selected weapons.

The shapes of sexually selected weapons differ widely among species, but the drivers of this diversity remain poorly understood. Existing explanations suggest weapon shapes reflect structural adaptations to different fighting styles, yet explicit tests of this hypothesis are lacking. We constructed finite element models of the horns of different rhinoceros beetle species to test whether functional specializations for increased performance under species-specific fighting styles could have contributed to the diversification of weapon form. We find that horns are both stronger and stiffer in response to species-typical fighting loads and that they perform more poorly under atypical fighting loads, which suggests weapons are structurally adapted to meet the functional demands of fighting. Our research establishes a critical link between weapon form and function, revealing one way male-male competition can drive the diversification of animal weapons.

Changes in horn size of Stone's sheep over four decades correlate with trophy hunting pressure.

Selective harvest may lead to rapid evolutionary change. For large herbivores, trophy hunting removes males with large horns. That artificial selection, operating in opposition to sexual selection, can lead to undesirable consequences for management and conservation. There have been no comparisons of long-term changes in trophy size under contrasting harvest pressures. We analyzed horn measurements of Stone's rams (Ovis dalli stonei) harvested over 37 years in two large regions of British Columbia, Canada, with marked differences in hunting pressure to identify when selective hunting may cause a long-term decrease in horn growth. Under strong selective harvest, horn growth early in life and the number of males harvested declined by 12% and 45%, respectively, over the study period. Horn shape also changed over time: horn length became shorter for a given base circumference, likely because horn base is not a direct target of hunter selection. In contrast, under relatively lower hunting pressure, there were no detectable temporal trends in early horn growth, number of males harvested, or horn length relative to base circumference. Trophy hunting is an important recreational activity and can generate substantial revenues for conservation. By providing a reproductive advantage to males with smaller horns and reducing the availability of desirable trophies, however, excessive harvest may have the undesirable long-term consequences of reducing both the harvest and the horn size of rams. These consequences can be avoided by limiting offtake.

A Late Cretaceous ceratopsian dinosaur from Europe with Asian affinities.

Ceratopsians (horned dinosaurs) represent a highly diverse and abundant radiation of non-avian dinosaurs known primarily from the Cretaceous period (65-145 million years ago). This radiation has been considered to be geographically limited to Asia and western North America, with only controversial remains reported from other continents. Here we describe new ceratopsian cranial material from the Late Cretaceous of Iharkút, Hungary, from a coronosaurian ceratopsian, Ajkaceratops kozmai. Ajkaceratops is most similar to 'bagaceratopsids' such as Bagaceratops and Magnirostris, previously known only from Late Cretaceous east Asia. The new material unambiguously demonstrates that ceratopsians occupied Late Cretaceous Europe and, when considered with the recent discovery of possible leptoceratopsid teeth from Sweden, indicates that the clade may have reached Europe on at least two independent occasions. European Late Cretaceous dinosaur faunas have been characterized as consisting of a mix of endemic 'relictual' taxa and 'Gondwanan' taxa, with typical Asian and North American groups largely absent. Ajkaceratops demonstrates that this prevailing biogeographical hypothesis is overly simplified and requires reassessment. Iharkút was part of the western Tethyan archipelago, a tectonically complex series of island chains between Africa and Europe, and the occurrence of a coronosaurian ceratopsian in this locality may represent an early Late Cretaceous 'island-hopping' dispersal across the Tethys Ocean.

Comparing measures of breeding inequality and opportunity for selection with sexual selection on a quantitative character in bighorn rams.

The reliability and consistency of the many measures proposed to quantify sexual selection have been questioned for decades. Realized selection on quantitative characters measured by the selection differential i was approximated by metrics based on variance in breeding success, using either the opportunity for sexual selection Is or indices of inequality. There is no consensus about which metric best approximates realized selection on sexual characters. Recently, the opportunity for selection on character mean OSM was proposed to quantify the maximum potential selection on characters. Using 21 years of data on bighorn sheep (Ovis canadensis), we investigated the correlations between seven indices of inequality, Is , OSM and i on horn length of males. Bighorn sheep are ideal for this comparison because they are highly polygynous and sexually dimorphic, ram horn length is under strong sexual selection, and we have detailed knowledge of individual breeding success. Different metrics provided conflicting information, potentially leading to spurious conclusions about selection patterns. Iδ, an index of breeding inequality, and, to a lesser extent, Is showed the highest correlation with i on horn length, suggesting that these indices document breeding inequality in a selection context. OSM on horn length was strongly correlated with i, Is and indices of inequality. By integrating information on both realized sexual selection and breeding inequality, OSM appeared to be the best proxy of sexual selection and may be best suited to explore its ecological bases.

Intralocus tactical conflict: genetic correlations between fighters and sneakers of the dung beetle Onthophagus taurus.

Males and females differ in their phenotypic optima for many traits, and as the majority of genes are expressed in both sexes, some alleles can be beneficial to one sex but harmful to the other (intralocus sexual conflict; ISC). ISC theory has recently been extended to intrasexual dimorphisms, where certain alleles may have opposite effects on the fitness of males of different morphs that employ alternative reproductive tactics (intralocus tactical conflict; ITC). Here, we use a half-sib breeding design to investigate the genetic basis for ISC and ITC in the dung beetle Onthophagus taurus. We found positive heritabilities and intersexual genetic correlations for almost all traits investigated. Next, we calculated the intrasexual genetic correlation between males of different morphs for horn length, a sexually selected trait, and compared it to intrasexual correlations for naturally selected traits in both sexes. Intrasexual genetic correlations did not differ significantly between the sexes or between naturally and sexually selected traits, failing to support the hypothesis that horns present a reduction of intrasexual genetic correlations due to ITC. We discuss the implications for the idea of developmental reprogramming between male morphs and emphasize the importance of genetic correlations as constraints for the evolution of dimorphisms.

The role of doublesex in the evolution of exaggerated horns in the Japanese rhinoceros beetle.

Male-specific exaggerated horns are an evolutionary novelty and have diverged rapidly via intrasexual selection. Here, we investigated the function of the conserved sex-determination gene doublesex (dsx) in the Japanese rhinoceros beetle (Trypoxylus dichotomus) using RNA interference (RNAi). Our results show that the sex-specific T. dichotomus dsx isoforms have an antagonistic function for head horn formation and only the male isoform has a role for thoracic horn formation. These results indicate that the novel sex-specific regulation of dsx during horn morphogenesis might have been the key evolutionary developmental event at the transition from sexually monomorphic to sexually dimorphic horns.

Deep homology and the origins of evolutionary novelty.

Do new anatomical structures arise de novo, or do they evolve from pre-existing structures? Advances in developmental genetics, palaeontology and evolutionary developmental biology have recently shed light on the origins of some of the structures that most intrigued Charles Darwin, including animal eyes, tetrapod limbs and giant beetle horns. In each case, structures arose by the modification of pre-existing genetic regulatory circuits established in early metazoans. The deep homology of generative processes and cell-type specification mechanisms in animal development has provided the foundation for the independent evolution of a great variety of structures.

Mechanical limits to maximum weapon size in a giant rhinoceros beetle.

The horns of giant rhinoceros beetles are a classic example of the elaborate morphologies that can result from sexual selection. Theory predicts that sexual traits will evolve to be increasingly exaggerated until survival costs balance the reproductive benefits of further trait elaboration. In Trypoxylus dichotomus, long horns confer a competitive advantage to males, yet previous studies have found that they do not incur survival costs. It is therefore unlikely that horn size is limited by the theoretical cost-benefit equilibrium. However, males sometimes fight vigorously enough to break their horns, so mechanical limits may set an upper bound on horn size. Here, I tested this mechanical limit hypothesis by measuring safety factors across the full range of horn sizes. Safety factors were calculated as the ratio between the force required to break a horn and the maximum force exerted on a horn during a typical fight. I found that safety factors decrease with increasing horn length, indicating that the risk of breakage is indeed highest for the longest horns. Structural failure of oversized horns may therefore oppose the continued exaggeration of horn length driven by male-male competition and set a mechanical limit on the maximum size of rhinoceros beetle horns.

Sexually antagonistic association between paternal phenotype and offspring viability reinforces total selection on a sexually selected trait.

The evolution of conspicuous sexually selected traits, such as horns or antlers, has fascinated biologists for more than a century. Elaborate traits can only evolve if they substantially increase reproduction, because they probably incur survival costs to the bearer. Total selection on these traits, however, includes sexual selection on sires and viability selection on offspring and can be influenced by changes in each of these components. Non-random associations between paternal phenotype and offspring viability may thus affect total selection on sexually selected traits. Long-term data on wild bighorn sheep (Ovis canadensis) provide the first evidence in nature that association between paternal phenotype and lamb viability strengthens total selection on horn size of adult rams, a sexually selected trait. The association of paternal horn length and offspring viability was sexually antagonistic: long-horned males sired sons with high viability but daughters of low viability. These results shed new light on the evolutionary dynamics of an iconic sexually selected trait and have important implications for sustainable wildlife management.

A new species of Shaanxispira (Bovidae, Artiodactyla) from the upper Miocene of China.

A new species of the bovid Shaanxispira, from the upper Miocene deposits of the Linxia Basin, Gansu Province, China, is described here. Shaanxispira is endemic to Northern China and was previously known only from the Lantian area, Shaanxi Province, by two species, S. chowi and S. baheensis. The new species, S. linxiaensis nov. sp., is of early Bahean in age, slightly older than the species from the Lantian area. The horn-cores of the new species are more derived, with large wing-shaped antero-medial keels, suggesting the occurrence of a different lineage of Shaanxispira in the Linxia Basin. Although Shaanxispira has homonymously twisted horn-cores, it is not closely related to other late Miocene bovids with homonymously twisted horn-cores, like Oioceros and Samotragus. Its phylogenetic status is still in debate, but might be more closely related to the late Miocene "ovibovines." 

The morphology of the interfacial tissue between bighorn sheep horn and bony horncore increases contact surface to enhance strength and facilitate load transfer from the horn to the horncore.

The horns of bighorn sheep rams are permanent cranial appendages used for high energy head-to-head impacts during interspecific combat. The horns attach to the underlying bony horncore by a layer of interfacial tissue that facilitates load transfer between the impacted horn and underlying horncore, which has been shown to absorb substantial energy during head impact. However, the morphology and mechanical properties of the interfacial tissue were previously unknown. Histomorphometry was used to quantify the interfacial tissue composition and morphology and lap-shear testing was used to quantify its mechanical properties. Histological analyses revealed the interfacial tissue is a complex network of collagen and keratin fibers, with collagen being the most abundant protein. Sharpey's fibers provide strong attachment between the interfacial tissue and horncore bone. The inner horn surface displayed microscopic porosity and branching digitations which increased the contact surface with the interfacial tissue by approximately 3-fold. Horn-horncore samples tested by lap-shear loading failed primarily at the horn surface, and the interfacial tissue displayed non-linear strain hardening behavior similar to other soft tissues. The elastic properties of the interfacial tissue (i.e., low- and high-strain shear moduli) were comparable to previously measured values for the equine laminar junction. The interfacial tissue contact surface was positively correlated with the interfacial tissue shear strength (1.23 ± 0.21 MPa), high-strain shear modulus (4.5 ± 0.7 MPa), and strain energy density (0.38 ± 0.07 MJ/m3). STATEMENT OF SIGNIFICANCE: The bony horncore in bighorn sheep rams absorbs energy to reduce brain cavity accelerations and mitigate brain injury during head butting. The interfacial zone between the horn and horncore transfers energy from the impacted horn to the energy absorbing horncore but has been largely neglected in previous models of bighorn sheep ramming since interfacial tissue properties were previously unknown. This study quantified the morphology and mechanical properties of the horn-horncore interfacial tissue to better understand structure-property relationships that contribute to energy transfer during ramming. Results from this study will improve models of bighorn sheep ramming used to study mechanisms of brain injury mitigation and may inspire novel materials and structures for brain injury prevention in humans.

Gene expression supports a single origin of horns and antlers in hoofed mammals.

Horns, antlers, and other bony cranial appendages of even-toed hoofed mammals (ruminant artiodactyls) challenge traditional morphological homology assessments. Cranial appendages all share a permanent bone portion with family-specific integument coverings, but homology determination depends on whether the integument covering is an essential component or a secondary elaboration of each structure. To enhance morphological homology assessments, we tested whether juvenile cattle horn bud transcriptomes share homologous gene expression patterns with deer antlers relative to pig outgroup tissues, treating the integument covering as a secondary elaboration. We uncovered differentially expressed genes that support horn and antler homology, potentially distinguish them from non-cranial-appendage bone and other tissues, and highlight the importance of phylogenetic outgroups in homology assessments. Furthermore, we found differentially expressed genes that could support a shared cranial neural crest origin for horns and antlers and expression patterns that refine our understanding of the timing of horn and antler differentiation.

Evolution of horn length and lifting strength in the Japanese rhinoceros beetle Trypoxylus dichotomus.

What limits the size of nature's most extreme structures? For weapons like beetle horns, one possibility is a tradeoff associated with mechanical levers: as the output arm of the lever system-the beetle horn-gets longer, it also gets weaker. This "paradox of the weakening combatant" could offset reproductive advantages of additional increases in weapon size. However, in contemporary populations of most heavily weaponed species, males with the longest weapons also tend to be the strongest, presumably because selection drove the evolution of compensatory changes to these lever systems that ameliorated the force reductions of increased weapon size. Therefore, we test for biomechanical limits by reconstructing the stages of weapon evolution, exploring whether initial increases in weapon length first led to reductions in weapon force generation that were later ameliorated through the evolution of mechanisms of mechanical compensation. We describe phylogeographic relationships among populations of a rhinoceros beetle and show that the "pitchfork" shaped head horn likely increased in length independently in the northern and southern radiations of beetles. Both increases in horn length were associated with dramatic reductions to horn lifting strength-compelling evidence for the paradox of the weakening combatant-and these initial reductions to horn strength were later ameliorated in some populations through reductions to horn length or through increases in head height (the input arm for the horn lever system). Our results reveal an exciting geographic mosaic of weapon size, weapon force, and mechanical compensation, shedding light on larger questions pertaining to the evolution of extreme structures.

A new African fossil caprin and a combined molecular and morphological Bayesian phylogenetic analysis of caprini (Mammalia: Bovidae).

Given that most species that have ever existed on Earth are extinct, no evolutionary history can ever be complete without the inclusion of fossil taxa. Bovids (antelopes and relatives) are one of the most diverse clades of large mammals alive today, with over a hundred living species and hundreds of documented fossil species. With the advent of molecular phylogenetics, major advances have been made in the phylogeny of this clade; however, there has been little attempt to integrate the fossil record into the developing phylogenetic picture. We here describe a new large fossil caprin species from ca. 1.9-Ma deposits from the Middle Awash, Ethiopia. To place the new species phylogenetically, we perform a Bayesian analysis of a combined molecular (cytochrome b) and morphological (osteological) character supermatrix. We include all living species of Caprini, the new fossil species, a fossil takin from the Pliocene of Ethiopia (Budorcas churcheri), and the insular subfossil Myotragus balearicus. The combined analysis demonstrates successful incorporation of both living and fossil species within a single phylogeny based on both molecular and morphological evidence. Analysis of the combined supermatrix produces superior resolution than with either the molecular or morphological data sets considered alone. Parsimony and Bayesian analyses of the data set are also compared and shown to produce similar results. The combined phylogenetic analysis indicates that the new fossil species is nested within Capra, making it one of the earliest representatives of this clade, with implications for molecular clock calibration. Geographical optimization indicates no less than four independent dispersals into Africa by caprins since the Pliocene.