Phylogenomics of Scorpions Reveal Contemporaneous Diversification of Scorpion Mammalian Predators and Mammal-Active Sodium Channel Toxins.

Scorpions constitute a charismatic lineage of arthropods and comprise more than 2500 described species. Found throughout various tropical and temperate habitats, these predatory arachnids have a long evolutionary history, with a fossil record that began in the Silurian. While all scorpions are venomous, the asymmetrically diverse family Buthidae harbors nearly half the diversity of extant scorpions, and all but one of the 58 species that are medically significant to humans. However, the lack of a densely sampled scorpion phylogeny has hindered broader inferences of the diversification dynamics of scorpion toxins. To redress this gap, we assembled a phylogenomic data set of 100 scorpion venom gland transcriptomes and genomes, emphasizing the sampling of highly toxic buthid genera. To infer divergence times of venom gene families, we applied a phylogenomic node dating approach for the species tree in tandem with phylostratigraphic bracketing to estimate the minimum ages of mammal-specific toxins. Our analyses establish a robustly supported phylogeny of scorpions, particularly with regard to relationships between medically significant taxa. Analysis of venom gene families shows that mammal-active sodium channel toxins (NaTx) have independently evolved in five lineages within Buthidae. Temporal windows of mammal-targeting toxin origins are correlated with the basal diversification of major scorpion mammal predators such as shrews, bats, and rodents. These results suggest an evolutionary model of relatively recent diversification of buthid NaTx homologs in response to the diversification of scorpion predators. [Adaptation; arachnids; phylogenomic dating; phylostratigraphy; venom.].

Phylogenomics facilitates stable scorpion systematics: Reassessing the relationships of Vaejovidae and a new higher-level classification of Scorpiones (Arachnida).

The Neartic family Vaejovidae (Scorpiones: Chactoidea) has long been treated as a diverse and systematically cohesive group of scorpions, but its monophyly and relationship to other scorpion families have historically been questioned. Morphological data have supported its monophyly and a variety of phylogenetic placements within the superfamily Chactoidea. Recent phylogenomic analyses have instead recovered vaejovids as polyphyletic (albeit with minimal taxonomic sampling) and Chactoidea as paraphyletic. Here, we reexamined the monophyly and phylogenetic placement of the family Vaejovidae, sampling 17 new vaejovid libraries using high throughput transcriptomic sequencing. Our phylogenomic analyses revealed a previous misplacement of Smeringurus mesaensis. Regardless, we recovered Vaejovidae as diphyletic due to the placement of the enigmatic genus Uroctonus. The remaining vaejovids formed a clade that was strongly supported as the sister group of the superfamily Scorpionoidea, a placement insensitive to matrix completeness or concatenation vs. species tree approaches to inferring the tree topology. Chactoidea was invariably recovered as a paraphyletic group due to the nested placement of Scorpionoidea. As first steps to resolving the paraphyly of Chactoidea, we take the following systematic actions: (1) we establish the superfamily Superstitionoidea (new superfamily) to accommodate Superstitioniidae; (2) we restore Vaejovoidea (status revalidated) as a valid superfamily that excludes Uroctonus; and (3) we treat the families Caraboctonidae, Troglotayosicidae, and the subfamily Uroctoninae as incertae sedis with respect to superfamilial placement. Our systematic actions thus establish the monophyly of the presently redefined Chactoidea and Vaejovoidea.

The male sexual apparatus in the order Scorpiones (Arachnida): a comparative study of functional morphology as a tool to define hypotheses of homology.

BACKGROUND: Insemination in scorpions is carried out by means of a partly sclerotized structure, the spermatophore, which is composed of two separate halves, the hemispermatophores. In most genera these reproductive structures can be used to differentiate species. However, many taxa such as the genus Euscorpius and the family Diplocentridae lack the morphological diversity observed in the copulatory organs of many other arthropods, rendering them useless for species level taxonomy. Such structural stasis, however, suggests that hemispermatophores have evolved relatively slowly and may thus provide a stronger phylogenetic signal for recognizing supra-generic ranks than previously thought. Based on the postulate that the phenotypic stability observed in some groups is the consequence of functional constraint, the most comprehensive comparative study of the male sexual apparatus to date was conducted for a complete reassessment of the morphology, phylogenetic value and hypotheses of homology of these structures. RESULTS: Hemispermatophores, pre- and post-insemination spermatophores, as well as the inherent mechanisms of insemination, were studied across the whole order, allowing the recognition and description of a series of five basic bauplans for the capsular region. For the most part, these patterns appear to be consistent within each major taxonomic group, but several cases of incongruence between spermatophore morphology and taxonomy raises questions about the monophyly of some clades. The Bothriuridae are traditionally regarded as a basal scorpionoid family. However, except for the genus Lisposoma, bothriurid hemispermatophores and spermatophores are morphologically more similar to those of the Chactoidea than to those of scorpionoids. On the other hand, the male copulatory structures of the hormurid clade (Hormiops (Hormurus + Liocheles)) are more akin to those of Diplocentridae and Heteroscorpionidae than to those of other hormurids. CONCLUSIONS: Spermatophore capsular patterns appears to be congruent with a recent phylogeny of the order Scorpiones based on phylogenomic data that placed Bothriuridae outside of Scorpionoidea and Liocheles outside of Hormuridae, in contradicton with earlier phylogenetic reconstructions based on morphology. This raises questions about the potential use of functionally constrained traits to assess the reliability of contradicting phylogenetic hypotheses and emphasizes the need for a thorough reassessment of the scorpion phylogenetic relationships.

Evidence of duplicated Hox genes in the most recent common ancestor of extant scorpions.

Scorpions (order Scorpiones) are unusual among arthropods, both for the extreme heteronomy of their bauplan and for the high gene family turnover exhibited in their genomes. These phenomena appear to be correlated, as two scorpion species have been shown to possess nearly twice the number of Hox genes present in most arthropods. Segmentally offset anterior expression boundaries of a subset of Hox paralogs have been shown to correspond to transitions in segmental identities in the scorpion posterior tagmata, suggesting that posterior heteronomy in scorpions may have been achieved by neofunctionalization of Hox paralogs. However, both the first scorpion genome sequenced and the developmental genetic data are based on exemplars of Buthidae, one of 19 families of scorpions. It is therefore not known whether Hox paralogy is limited to Buthidae or widespread among scorpions. We surveyed 24 high throughput transcriptomes and the single whole genome available for scorpions, in order to test the prediction that Hox gene duplications are common to the order. We used gene tree parsimony to infer whether the paralogy was consistent with a duplication event in the scorpion common ancestor. Here we show that duplicated Hox genes in non-buthid scorpions occur in six of the ten Hox classes. Gene tree topologies and parsimony-based reconciliation of the gene trees are consistent with a duplication event in the most recent common ancestor of scorpions. These results suggest that a Hox paralogy, and by extension the model of posterior patterning established in a buthid, can be extended to non-Buthidae scorpions.

Phylogenomic resolution of scorpions reveals multilevel discordance with morphological phylogenetic signal.

Scorpions represent an iconic lineage of arthropods, historically renowned for their unique bauplan, ancient fossil record and venom potency. Yet, higher level relationships of scorpions, based exclusively on morphology, remain virtually untested, and no multilocus molecular phylogeny has been deployed heretofore towards assessing the basal tree topology. We applied a phylogenomic assessment to resolve scorpion phylogeny, for the first time, to our knowledge, sampling extensive molecular sequence data from all superfamilies and examining basal relationships with up to 5025 genes. Analyses of supermatrices as well as species tree approaches converged upon a robust basal topology of scorpions that is entirely at odds with traditional systematics and controverts previous understanding of scorpion evolutionary history. All analyses unanimously support a single origin of katoikogenic development, a form of parental investment wherein embryos are nurtured by direct connections to the parent's digestive system. Based on the phylogeny obtained herein, we propose the following systematic emendations: Caraboctonidae is transferred to Chactoidea new superfamilial assignment: ; superfamily Bothriuroidea revalidated: is resurrected and Bothriuridae transferred therein; and Chaerilida and Pseudochactida are synonymized with Buthida new parvordinal synonymies: .

Phylogeny of the North American vaejovid scorpion subfamily Syntropinae Kraepelin, 1905, based on morphology, mitochondrial and nuclear DNA.

The first rigorous analysis of the phylogeny of the North American vaejovid scorpion subfamily Syntropinae is presented. The analysis is based on 250 morphological characters and 4221 aligned DNA nucleotides from three mitochondrial and two nuclear gene markers, for 145 terminal taxa, representing 47 species in 11 ingroup genera, and 15 species in eight outgroup genera. The monophyly and composition of Syntropinae and its component genera, as proposed by Soleglad and Fet, are tested. The following taxa are demonstrated to be para- or polyphyletic: Smeringurinae; Syntropinae; Vaejovinae; Stahnkeini; Syntropini; Syntropina; Thorelliina; Hoffmannius; Kochius; and Thorellius. The spinose (hooked or toothed) margin of the distal barb of the sclerotized hemi-mating plug is demonstrated to be a unique, unambiguous synapomorphy for Syntropinae, uniting taxa previously assigned to different subfamilies. Results of the analysis demonstrate a novel phylogenetic relationship for the subfamily, comprising six major clades and 11 genera, justify the establishment of six new genera, and they offer new insights about the systematics and historical biogeography of the subfamily, and the information content of morphological character systems.

Characterization of the fluorescence intensity and color tonality in the exoskeleton of scorpions.

Scorpions are a mesodiverse and nocturnal group of arachnids inhabiting most biomes worldwide. Different species of scorpions have divergent adaptations to the substrate they live in, but most of them share an intriguing characteristic: their exoskeleton contains fluorophores that emit blue-greenish fluorescence under ultraviolet radiation. Although there are some reports in the literature on the study of fluorescence in scorpions, the biological functionality of this light emission is currently unknown and is under debate. In this work, the properties of emission from the scorpion's exoskeleton are studied by means of digitally processed photographs taken of living specimens under ultraviolet illumination and complemented with standard spectroscopic measurements of emission and excitation spectra. With the aim of identifying possible correlations between the fluorescence, the characteristics color of the exoskeleton and the biology of the scorpions, 4 families, 9 genera and 24 species were studied. Our results suggest that the intensity of fluorescence is heterogeneous throughout the scorpion's exoskeleton studied here in such a way that pedipalps and metasomal segments fluoresce more intensely than the mesosomal segments. The spectrum of fluorescence across species is practically identical, suggesting that the same fluorophores are present in their exoskeletons. However, the fluorescence intensity emitted by each species varies according to their characteristic color (associated with the exoskeleton optical reflectance). Since the coloration of the exoskeleton is determined by the concentration of melanin and other pigments according to the substrate where scorpions live in, we conclude by suggesting that fluorescence may correlate directly to the ecomorphotype of the scorpions.

A new species of Centruroides (Scorpiones, Buthidae) from Colima, Mexico.

As part of an ongoing survey of scorpion diversity in Colima, Mexico, the isolated mountain Cerro Grande, part of the Biosphere Reserve Sierra de Manantlán, was investigated. Centruroides possanii sp. nov., the fifth species of the genus from the state, was discovered during fieldwork in the massif and is described in the present paper. Physiographical and climatic features of Cerro Grande may restrict the range of this new species; thus, we hypothesized that it may be a microendemic species that requires priority conservation. The new species is not assigned to any Centruroides species group recognized because some of its morphological features do not fit the current diagnosis of any of these groups, and these different groups are non-monophyletic and consequently ill-diagnosed. The new species is profusely illustrated, particularly the hemispermatophore. A distribution map is presented along with the other two more common species distributed in Colima. Because only indirect data on the potency of its venom is available, the medical importance of this new species described here is yet to be known.

North American scorpion species of public health importance with a reappraisal of historical epidemiology.

Scorpionism is a severe threat to public health in North America. Historically, few species of Centruroides have been considered to be the offending taxa, but we know now that their diversity is greater and our knowledge incomplete. Current distribution maps are inadequate for some species. Epidemiologic studies are sporadic and local, and a complete synthesis for North America is missing. We analyze historical and recent knowledge about the identity, distribution and epidemiology of species of medical importance in North America. PubMed, Google Scholar, the National Collection of Arachnids, and results of recent field work were consulted in the preparation of our analysis. We recognized 21 species and one subspecies of medically important scorpions in need of precise geographical delimitation. All these species are found in Mexico, which is clearly a hotspot for scorpionism. Although mortality has been steadily decreasing, deaths still occur, and morbidity remains high. Mortality is most common at age classes of 0-10 years and >50. Morbidity is highest in age class 15-50 years, including the most economically active segment of the population. The season of the highest incidence of scorpion sting peaks between spring and summer but there appears to be a second, lower peak at the end of the summer. Although the systematics of the genus Centruroides has advanced considerably, our knowledge of its diversity remains fragmentary. There is a disconnection between the actual distribution of the scorpions and the incidence maps constructed from scorpion sting records. Despite a historically robust knowledge of the distribution of well-known species, most recently described species are known from only a few localities. Some of the epidemiological parameters are consistent among studies reported herein.

A new species of Diplocentrus (Scorpiones: Diplocentridae) with punctate pedipalp surfaces, a diagnostic character within the "mexicanus" group.

Diplocentrus duende n. sp. is described based on adult males collected from a locality in the Tehuacán-Cuicatlán Valley, Mexico. This species has punctate pedipalp surfaces, a condition present only in four other species of this specious genus. As suggested here, this condition has evolved independently in these species within the "mexicanus" group of Diplocentrus from the rest of the diplocentrids.

The male of Megacormusgranosus (Gervais, 1844) with comments on its hemispermatophore (Scorpiones, Euscorpiidae).

The male of Megacormusgranosus is described for the first time and the female redescribed. A homology scheme proposed recently is applied to hemispermatophore structures. The specimens were collected in an oak forest from Pico de Orizaba Volcano at an average altitude of 2340 m. All adult males were collected by pitfall traps, whereas all adult females and both sex immatures were collected using Berlese funnels, suggesting that males are comparatively more mobile within the leaf litter layer, probably due to mating season.

New Fossil Scorpion from the Chiapas Amber Lagerstätte.

A new species of scorpion is described based on a rare entire adult male preserved in a cloudy amber from Miocene rocks in the Chiapas Highlands, south of Mexico. The amber-bearing beds in Chiapas constitute a Conservation Lagerstätte with outstanding organic preservation inside plant resin. The new species is diagnosed as having putative characters that largely correspond with the genus Tityus Koch, 1836 (Scorpiones, Buthidae). Accordingly, it is now referred to as Tityus apozonalli sp. nov. Its previously unclear phylogenetic relationship among fossil taxa of the family Buthidae from both Dominican and Mexican amber is also examined herein. Preliminarily results indicate a basal condition of T. apozonalli regarding to Tityus geratus Santiago-Blay and Poinar, 1988, Tityus (Brazilotityus) hartkorni Lourenço, 2009, and Tityus azari Lourenço, 2013 from Dominican amber, as was Tityus (Brazilotityus) knodeli Lourenço, 2014 from Mexican amber. Its close relationships with extant Neotropic Tityus-like subclades such as 'Tityus clathratus' and the subgenus Tityus (Archaeotityus) are also discussed. This new taxon adds to the knowledge of New World scorpions from the Miocene that are rarely found trapped in amber.