Scorpions(Arachnida) from Bocas del Toro archipelago,Panamá

Four species of buthid scorpions (Ananteris platnicki Louren?o, 1993; Centruroides limbatus [Pocock, 1898]; Tityus pachyurus [Pocock, 1897]; and T. ocelote Francke and Stockwell, 1987) are recorded for the first time from some islands and cays of the Bocas del Toro Archipelago, Panama. Morphological variation and ecological data are given for all the species. This is the first Panamanian record for both C. limbatus and T. ocelote. The scorpion fauna of the Bocas de Toro Archipelago is related with the recent fauna of the Atlantic continental lowland region of Costa Rica and Panama and includes Amazonian-Guyanese (genera Ananteris and Tityus) as well as Mexican-North Central American (genus Centruroides) elements.     

Identification and phylogenetic analysis of Tityus pachyurus and Tityus obscurus novel putative Na+-channel scorpion toxins

Background

Colombia and Brazil are affected by severe cases of scorpionism. In Colombia the most dangerous accidents are caused by Tityus pachyurus that is widely distributed around this country. In the Brazilian Amazonian region scorpion stings are a common event caused by Tityus obscurus. The main objective of this work was to perform the molecular cloning of the putative Na⁺-channel scorpion toxins (NaScTxs) from T. pachyurus and T. obscurus venom glands and to analyze their phylogenetic relationship with other known NaScTxs from Tityus species.

Methodology/Principal Findings

cDNA libraries from venom glands of these two species were constructed and five nucleotide sequences from T. pachyurus were identified as putative modulators of Na⁺-channels, and were named Tpa4, Tpa5, Tpa6, Tpa7 and Tpa8; the latter being the first anti-insect excitatory β-class NaScTx in Tityus scorpion venom to be described. Fifteen sequences from T. obscurus were identified as putative NaScTxs, among which three had been previously described, and the others were named To4 to To15. The peptides Tpa4, Tpa5, Tpa6, To6, To7, To9, To10 and To14 are closely related to the α-class NaScTxs, whereas Tpa7, Tpa8, To4, To8, To12 and To15 sequences are more related to the β-class NaScTxs. To5 is possibly an arthropod specific toxin. To11 and To13 share sequence similarities with both α and β NaScTxs. By means of phylogenetic analysis using the Maximum Parsimony method and the known NaScTxs from Tityus species, these toxins were clustered into 14 distinct groups.

Conclusions/Significance

This communication describes new putative NaScTxs from T. pachyurus and T. obscurus and their phylogenetic analysis. The results indicate clear geographic separation between scorpions of Tityus genus inhabiting the Amazonian and Mountain Andes regions and those distributed over the Southern of the Amazonian rainforest. Based on the consensus sequences for the different clusters, a new nomenclature for the NaScTxs is proposed.     

The Life History of Chactas reticulatus Kraepelin, 1912 (Scorpiones, Chactidae), with a Comparative Analysis of the Reproductive Traits of Three Scorpion Lineages in Relation to Habitat

An account is given of the life history, growth, population densities, habitat, reproductive systems, courtship and mating, litter size and sex ratio, iteroparity, molting and development of the scorpion Chactas reticulatus Kraepelin. These same traits are compared in three distinct scorpion lineages, buthid, chactid and ischnurid in relation to habitat characteristics. Correlations are drawn between reproductive and adaptative strategies, and it is suggested that if non-buthid scorpions may present some homogeneity as equilibrium species, buthid scorpions are not necessarily opportunistic species. Many groups can be characterized within the buthids, and these different lineages show variable ecological requirements.     

Proteomic analysis of the venom from the scorpion Tityus stigmurus: biochemical and physiological comparison with other Tityus species

The venom from the Brazilian scorpion Tityus stigmurus was fractionated by high performance liquid chromatography (HPLC) and the corresponding components were used for molecular mass determination using electrospray ion trap mass spectrometry. One hundred distinct components were clearly assigned showing molecular masses from 216.5 to 44,800.0 Da. Fifteen new components were isolated and sequenced, four of them to completion: Tst-3 (similar to Na(+) channel specific scorpion toxins), Tst-17 (a K(+) channel blocking peptide similar to Tc1), Tst beta KTx (a peptide with identical sequence as that of TsTX-K beta toxin earlier described to exist in T. serrulatus venom) and finally a novel proline-rich peptide of unknown function. Among the eleven components partially sequenced were two enzymes: hyaluronidase and lysozyme. The first enzyme has a molecular mass of 44,800.0 Da. This enzyme showed high activity against the substrate hyaluronan in vitro. Amino acid sequence of the second enzyme showed that it is similar to other known lysozymes, with similar molecular mass and sequence to that of bona fide lysozymes reported in public protein data banks. Finally, this communication reports a correlation among HPLC retention times and molecular masses of folded scorpion toxins as well as a comparative structural and physiological analysis of components from the venom of several species of the genus Tityus.     

Molecular characterization of a new scorpion venom lipolysis activating peptide: Evidence for disulfide bridge-mediated functional switch of peptides

Venoms from scorpions contain extremely rich bioactive peptides that often carry diverse functions and are presumably needed to achieve synergistic effects for rapidly immobilizing prey and defending themselves. BotLVP1 is a unique heterodimer protein recently found in the scorpion Buthus occitanus tunetanus venom that is structurally related to scorpion toxins affecting sodium channels (NaScTxs) but exhibits adipocyte lipolysis activity. We have isolated and identified two cDNA clones encoding subunits and β of a BotLVP1-like peptide (named BmLVP1) from the Chinese scorpion Buthus martensii venom gland and determined the first complete gene structure of this subfamily. These results highlight a genetic link between these lipolysis activating peptides and NaScTxs. Comparison of cDNA and genomic sequences combined with protein structural and functional analysis provides evidence supporting the existence of RNA editing mechanism in scorpion venom glands, which could mediate functional switch of BmLVP1 gene, from adipocyte lipolysis to neurotoxicity, by altering the wrapper disulfide bridge (WDB) pattern of the peptides.     

The complete amino acid sequence of toxin TsTX-VI isolated from the venom of the scorpion Tityus serrulatus

The complete sequence of the toxin TsTX-VI from the venom of the scorpionTityus serrulatus Lutz and Mello is presented. The sequence has been determined by automated Edman analysis of the reduced and carboxymethylated protein as well as of the resulting peptides, obtained fromS. aureus protease and tryptic digestions. TsTX-VI is composed of 62 residues and has a calculated molecular weight of 6717. Homology studies with other scorpion toxins show that TsTX-VI is more similar to the Old World than to the North American scorpion toxins. The hydropathic index indicates that TsTX-VI is more hydrophobic than Ts-. Toxicity studies carried out in mice demonstrate that i.v. injection of TsTX-VI is unable to evoke the usual symptoms induced by the typical neurotoxins of this venom, but only a generalized allergic reaction. These properties are important in clarifying the relationship between primary structure and biological function of scorpion toxins.     

New Fossil Scorpion from the Chiapas Amber Lagerst?tte

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.     

The complete amino acid sequence of toxin TsTX-VI isolated from the venom of the scorpionTityus serrulatus

The complete sequence of the toxin TsTX-VI from the venom of the scorpionTityus serrulatus Lutz and Mello is presented. The sequence has been determined by automated Edman analysis of the reduced and carboxymethylated protein as well as of the resulting peptides, obtained fromS. aureus protease and tryptic digestions. TsTX-VI is composed of 62 residues and has a calculated molecular weight of 6717. Homology studies with other scorpion toxins show that TsTX-VI is more similar to the Old World than to the North American scorpion toxins. The hydropathic index indicates that TsTX-VI is more hydrophobic than Ts-γ. Toxicity studies carried out in mice demonstrate that i.v. injection of TsTX-VI is unable to evoke the usual symptoms induced by the typical neurotoxins of this venom, but only a generalized allergic reaction. These properties are important in clarifying the relationship between primary structure and biological function of scorpion toxins.     

Proteomic analysis of the venom and characterization of toxins specific for Na+ - and K+ -channels from the Colombian scorpion Tityus pachyurus

The Colombian scorpion Tityus pachyurus is toxic to humans and is capable of producing fatal accidents, but nothing is known about its venom components. This communication reports the separation of at least 57 fractions from the venom by high performance liquid chromatography. From these, at least 104 distinct molecular weight compounds were identified by mass spectrometry analysis. The complete amino acid sequences of three peptides were determined and the partial sequences of three others were also identified. Electrophysiological experiments conducted with ion-channels expressed heterologously on Sf9 cells showed the presence of a potent Shaker B K(+)-channel blocker. This peptide (trivial name Tpa1) contains 23 amino acid residues closely packed by three disulfide bridges with a molecular mass of 2,457 atomic mass units. It is the third member of the sub-family 13, for which the systematic name is proposed to be alpha-KTx13.3. The mice assay showed clearly the presence of toxic peptides to mammals. One of them named Tpa2, containing 65 amino acid residues with molecular mass of 7,522.5 atomic mass units, is stabilized by four disulfide bridges. It was shown to modify the Na(+)-currents of F-11 and TE671 cells in culture, similar to the beta scorpion toxins. These results demonstrate the presence of toxic peptides in the venom of T. pachyurus and confirm that accidents with this species of scorpion should be considered an important human hazard in Colombia.     

History of study,updated checklist,distribution and key of scorpions (Arachnida：Scorpiones) from China

This review describes the history of taxonomic research on scorpions and provides an updated checklist and key of the scorpions currently known in China. This checklist is based on a thorough review of the extant literatures on scorpion species whose presence has been confirmed in China through field expeditions and examination of scorpion collections, excepting a few members that have no clear distribution or are currently in doubt. Totally, the scorpion fauna of China consists of 53 species and subspecies belonging to 12 genera crossing five families, with 33 species (62.3%) and one genus being recorded as endemic. Additionally, identification key and the distribution of scorpions from China are provided.     

The effect of habitat fragmentation on the scorpion assemblage of a Brazilian Atlantic Forest

Habitat fragmentation is a topic widely studied in ecology; however, its effects on the assemblage of the order Scorpiones is less well understood. Aiming to fill this gap, this study assessed the effect of fragmentation on the assemblage of these arachnids in 12 Brazilian Atlantic forest fragments. Five environmental variables were measured (depth and dry mass of litter, understory density, canopy openness, and diameter at breast height of the trees), and the fragment area, vegetation cover, connectivity and elevation assessed. The animals were collected during the dry season and, identified at night with the use of ultraviolet light lamps. The analyzed scorpion assemblage in the landscape was characterized by the species Tityus pusillus, T. stigmurus, T. neglectus, T. brazilae, and Ananteris mauryi, with a maximum of three species co-occurring per fragment. Only the fragment size and the dry mass of litter showed a positive relationship with the composition of scorpions. These results suggest that the habitat of scorpions responds to environmental attributes and landscape metrics at both higher (fragment size) and lower (leaf litter) scales. Our study was able to expand our knowledge of how scorpions respond to habitat changes in the Atlantic Forest. We conclude that fragmentation and habitat quality are determining factors for the assemblage of these arachnids.     

Expression of murine poly(A) binding protein II gene in testis

Insect defensin refers to a group of antibacterial peptides derived from a variety of insect species as well as from scorpion and possessing a three-dimensional structure highly similar to that of scorpion toxins. A full-length cDNA encoding an insect defensin-like peptide was isolated from the venom gland cDNA library of the Chinese scorpion Buthus martensii Karsch. The precursor, the overall organization of which is similar to that of insect defensins, consists of 61 amino acid residues with a putative signal peptide of 15 residues, a propeptide of 7 residues, and a mature peptide of 39 residues (named BmTXKS2). The positions of six cysteines and a conserved glycine in mature BmTXKS2 are the same as those in LqDef, the first defensin found in scorpions, which suggests these peptides should present a similar cysteine-stabilized alphabetamotif. Phylogenetic analysis further shows that the structure of BmTXKS2 is closer to that of ancient defensins (e.g., LqDef and AaDef, two insect defensins present in the scorpion hemolymph) than to scorpion toxins.     

Molecular cloning and sequencing of two 'short chain' and two 'long chain' K(+) channel-blocking peptides from the Chinese scorpion Buthus martensii Karsch.

Five full-length cDNAs encoding the precursors of two 'short chain' scorpion non-toxic peptides active on Ca(2+)-activated K(+) channels (BmP02 and BmP03) and two novel putative long chain K(+) channel-blocking peptides (named BmTXKbeta and BmTXKbeta2) were first isolated from the venom gland cDNA library of the Chinese scorpion Buthus martensii Karsch (BmK). BmTXKbeta2 showed a high similarity with AaTXKbeta, while BmTXKbeta was completely different in the deduced primary structure from the long chain and short chain scorpion toxins already characterized. Thus, BmTXKbeta expands the scorpion long chain K(+) channel-blocking peptide family. Although little sequence similarity exists between the above two short and two long peptides, they are similar at the positions of six cysteines, suggesting that they should all share a similar scaffold composed of an alpha-helix and a three-stranded beta-sheet.     

A new genus and species of fossil scorpion (?Euscorpiidae) from the Early–Middle Eocene of Pesciara (Bolca,Italy)

Fossil scorpions are among the oldest terrestrial arthropods known from the fossil record. They have a worldwide distribution and a rich fossil record, especially for the Paleozoic. Fossil scorpions from Mesozoic and Cenozoic deposits are usually rare (except in amber-deposits). Here, we describe the only fossil scorpion from the Early to Middle Eocene Pesciara Lagerstätte in Italy. Eoeuscorpius ceratoi gen. et sp. nov. is probably a genus and species within the family Euscorpiidae. This may be the first fossil record of the Euscorpiidae, which are so far only known from four extant genera. Eoeuscorpius ceratoi gen. et sp. nov. was found in the “Lower Part” of the Pesciara Limestone, which is actually dated Late Ypresian stage (between 49.5 and 49.7 Ma). Besides a possible pseudoscorpion, the here-described fossil scorpion is the second arachnid species known from the Bolca Locality.     

Novel structural class of four disulfide-bridged peptides from Tityus serrulatus venom

A new structural class of short peptides folded by four disulfide-bridges was found in the venom of the Brazilian scorpion Tityus serrulatus. Peptides were put on evidence independently by means of two different approaches of structurally guided prospection. First, a cDNA sequence was obtained using a degenerate primer constructed according to the C-terminal sequence of kaliotoxin (KTx2), from the Androctonus australis venom. Second, MALDI-TOF mass spectrometry analyses of toxic fraction FIII from T. serrulatus venom revealed a family of molecules ranging approximately from 2900 to 3000 Da. Three new peptides were isolated and named TsPep1, TsPep2, and TsPep3. Biochemical characterization showed that they are 29 amino acids long, constrained by a new pattern of four disulfide-bridges. These results enable us to classify these new molecules as part of a novel structural class of short peptides from scorpion venoms.