Missulena Research Articles

Five new species of mouse spiders in the genus Missulena (Mygalomorphae: Actinopodidae) from national parks and conservation reserves in Western Australia

Five new species of mouse spiders in the genus Missulena (Mygalomorphae: Actinopodidae) from national parks and conservation reserves in Western Australia

Three new species of mouse spider (Araneae: Actinopodidae: Missulena Walckenaer, 1805) from Western Australia, including an assessment of intraspecific variability in a widespread species from the arid biome

Abstract Mouse spiders (genus Missulena Walckenaer, 1805) are a lineage of trapdoor spiders with males of many species having a brightly coloured red cephalic region, an abdomen that is tinged metallic blue, and the habit of wandering during the day in search of a mate. A total of 17 species of Missulena have been described in Australia to date but most descriptions are based exclusively on males and always small numbers of specimens. Here, we describe three new species of Missulena from the Pilbara and Goldfields regions of Western Australia based on morphology and genetic data: Missulena davidisp. nov. (male and female), M. iugumsp. nov. (male) and M. manningensissp. nov. (male). One of them is presently known only from its type locality and another one from a small range based on two specimens but M. davidi sp. nov. has a linear range of almost 300 km and is genetically highly structured. We use genetic data for 75 specimens as a foundation to evaluate morphological variability in this species and note substantial variation in several characters commonly used to identify species such as body size, colouration, rastellum shape and eye distances. This variation does not necessarily relate to phylogeographic structure as inferred from the genetic data, but rather seems to reflect natural variability both within and between localised populations. Overall, our results stress the need to evaluate a large series of specimens for mygalomorph taxonomy and provide an interesting example of intraspecific variability in hard-to-collect species that are usually underrepresented in museum collections.

Phylogenomic reclassification of the world\u2019s most venomous spiders (Mygalomorphae, Atracinae), with implications for venom evolution

Here we show that the most venomous spiders in the world are phylogenetically misplaced. Australian atracine spiders (family Hexathelidae), including the notorious Sydney funnel-web spider Atrax robustus, produce venom peptides that can kill people. Intriguingly, eastern Australian mouse spiders (family Actinopodidae) are also medically dangerous, possessing venom peptides strikingly similar to Atrax hexatoxins. Based on the standing morphology-based classification, mouse spiders are hypothesized distant relatives of atracines, having diverged over 200 million years ago. Using sequence-capture phylogenomics, we instead show convincingly that hexathelids are non-monophyletic, and that atracines are sister to actinopodids. Three new mygalomorph lineages are elevated to the family level, and a revised circumscription of Hexathelidae is presented. Re-writing this phylogenetic story has major implications for how we study venom evolution in these spiders, and potentially genuine consequences for antivenom development and bite treatment research. More generally, our research provides a textbook example of the applied importance of modern phylogenomic research.

A new species of Mouse Spider (Actinopodidae, Missulena) from the Goldfields region of Western Australia

A new species of Mouse Spider (family Actinopodidae Simon, 1892), Missulena harewoodi, is described from near the City of Kalgoorlie-Boulder in the Goldfields region of Western Australia. It differs from all other Missulena species by the unusual light grey colouration of the abdomen in combination with small body size and shiny carapace. A phylogenetic analysis of a fragment (658 bp) of the COI barcoding gene places M. harewoodi sp. n. in a clade with four Missulena species from the Pilbara region of Western Australia, more than 900 km away. Missulena harewoodi sp. n. is one of the many species in this genus that are currently only known from a single, or a very limited number of specimen, highlighting the paucity of fauna collections in many arid regions of Australia and the difficulties in sampling these cryptic spiders.

Four new Mouse Spider species (Araneae, Mygalomorphae, Actinopodidae, Missulena) from Western Australia.

Four new species of the Mouse Spider genus Missulena Walckenaer, 1805 (family Actinopodidae) are described from Western Australia based on morphological features of adult males. Missulena leniae sp. n.(from the Carnarvon and Yalgoo biogeographic regions), Missulena mainae sp. n. (Carnarvon), Missulena melissae sp. n. (Pilbara) and Missulena pinguipes sp. n. (Mallee) represent a broad spectrum of morphological diversity found in this genus and differ from other congeners by details of the male copulatory bulb, colour patterns, eye sizes, leg morphology and leg spination. Two of the species, M. pinguipes sp. n. and M. mainae sp. n., are characterised by swollen metatarsi of the fourth legs in males, a feature not previously recorded in the family. A key to males of all named Missulena species from Australia is presented and allows their identification based on external morphology.

New species of Mouse Spiders (Araneae: Mygalomorphae: Actinopodidae: <i>Missulena</i>) from the Pilbara region, Western Australia

Two new species of Mouse Spiders, genus Missulena, from the Pilbara region in Western Australia are described based on morphological features of males. Missulena faulderi sp. nov. and Missulena langlandsi sp. nov. are currently known from a small area in the southern Pilbara only. Mitochondrial cytochrome c oxidase subunit I (COI) sequence divergence failed in clearly delimiting species in Missulena, but provided a useful, independent line of evidence for taxonomic work in addition to morphology.

Intersexual variations in Northern ( Missulena pruinosa) and Eastern ( M. bradleyi) mouse spider venom

Venoms of both sexes of Australian Northern ( Missulena pruinosa) and Eastern ( Missulena bradleyi) mouse spiders were studied in order to determine intersexual variations in venom yield, composition and bioactivity. Females of both species yielded more venom than males. High-performance liquid chromatography (HPLC) and mass spectrometry data further indicate a substantial degree of intersexual variation in the venom composition of both species. In a cricket ( Acheta domestica) acute toxicity assay, only small intersexual differences were observed, but M. bradleyi venom was found to be considerably more potent than M. pruinosa venom. In the chick biventer cervicis nerve–muscle preparation, male but not female M. bradleyi venom induced large and sustained muscle contractions with fasciculation and decreased twitch height that could be reversed by CSL funnel-web spider antivenom. In contrast, venoms of both sexes of M. pruinosa did not induce significant effects in the chick biventer cervicis nerve–muscle preparation. We therefore conclude that female M. bradleyi venom and venoms from male and female M. pruinosa appear to contain few, if any, orthologs of δ-missulenatoxin-Mb1a, the toxin responsible for the effects of male M. bradleyi venom in vertebrates. These findings are consistent with clinical reports that mouse spiders, particularly species other than male M. bradleyi, do not appear to be a major medical problem in humans.

Mouse spider bites (Missulena spp.) and their medical importance

To determine the clinical significance of definite bites by mouse spiders (Actinopodidae: Missulena spp.) from published case reports/series and museum records. A computerised literature search of MEDLINE and EMBASE was undertaken. All cases reported to major Australian museums and reports from venom researchers working with mouse spiders were also reviewed. Textbooks on clinical toxinology were searched and further reports of cases were located. All cases of definite spider bites where the spider was collected and identified by an expert as a mouse spider were included. All reports were evaluated and the following data extracted: patient demographics (age, sex, geographical location, season), bite site, local and systemic effects, and hospital attendance. Clinical effects were classified into three groups: severe neurotoxic envenoming, local neurotoxic effects or mild systemic effects, and minor local effects. Forty definite bites were identified from around Australia, with only one case of severe envenoming (a 19-month-old child). Local neurotoxic effects occurred in six cases and minor systemic effects in five. There was no evidence of envenoming in most bite victims, and the rate of severe envenoming was 2.5% (95% CI, 0-13%). There were no recorded deaths. In most cases, bites by mouse spiders cause only minor or moderate effects. Severe envenoming is rare and far less common than for funnel-web spider bites. Mouse spider bites do not appear to be a major medical problem.

Bites by Australian mygalomorph spiders (Araneae, Mygalomorphae), including funnel-web spiders (Atracinae) and mouse spiders (Actinopodidae: Missulena spp)

A number of mygalomorph spiders cause bites in Australia, including the funnel-web spiders (Hexathelidae, Atracinae: Hadronyche and Atrax) and mouse spiders (Actinopodidae: Missulena). There is ongoing debate about the significance of bites by mouse spiders and the frequency of severe envenoming by funnel-web spiders. We conducted a prospective cohort study of definite spider bites with expert spider identification and include the analysis of mygalomorph spiders here. Subjects were recruited prospectively from February 1999 to April 2003 from patients presenting to participating hospitals or contacting a state poison information centre. Forty-nine cases of bites by mygalomorph spiders were included: 16 were by funnel-web spiders, 13 by mouse spiders and 20 by other trapdoor spiders (Families Idiopidae and Nemesiidae). Of the 49 bites, 45 (92%) occurred on distal limbs (hands and feet). Local effects included severe pain (53%), puncture marks (61%) and bleeding (27%), local redness (33%). Itchiness did not occur. The following were highly statistically associated with mygalomorph spider bites compared to all other spiders ( p<0.0001): circumstances (gardening at the time (likelihood ratio (LR) 7.9) and distal limb bites (LR 2.0)) and early clinical features (presence of puncture marks OR bleeding (2.3), or severe pain (2.0)). Of 16 funnel-web spider bites, there were 10 cases with minor local effects, four with moderate envenoming (non-specific systemic or local neurotoxicity) and two with severe envenoming requiring antivenom. In addition to local effects, mouse spider bites caused local paraesthesia in three cases, local diaphoresis in one case and non-specific systemic effects in five cases, but not severe envenoming. True trapdoor spider bites caused only minor effects. The data from a mixed species sample of funnel-web spiders confirms previous observations suggesting that only a small proportion of funnel-web bites cause severe effects. Mouse spider bites are unlikely to cause major envenoming but the clinical effects are consistent with neurotoxic venom and are more severe than the trapdoor spiders.

Isolation of δ-missulenatoxin-Mb1a, the major vertebrate-active spider δ-toxin from the venom of Missulena bradleyi (Actinopodidae)

The present study describes the isolation and pharmacological characterisation of the neurotoxin δ-missulenatoxin-Mb1a (δ-MSTX-Mb1a) from the venom of the male Australian eastern mouse spider, Missulena bradleyi. This toxin was isolated using reverse-phase high-performance liquid chromatography and was subsequently shown to cause an increase in resting tension, muscle fasciculation and a decrease in indirect twitch tension in a chick biventer cervicis nerve-muscle bioassay. Interestingly, these effects were neutralised by antivenom raised against the venom of the Sydney funnel-web spider Atrax robustus. Subsequent whole-cell patch-clamp electrophysiology on rat dorsal root ganglion neurones revealed that δ-MSTX-Mb1a caused a reduction in peak tetrodotoxin (TTX)-sensitive sodium current, a slowing of sodium current inactivation and a hyperpolarising shift in the voltage at half-maximal activation. In addition, δ-MSTX-Mb1a failed to affect TTX-resistant sodium currents. Subsequent Edman degradation revealed a 42-residue peptide with unusual N- and C-terminal cysteines and a cysteine triplet (Cys14-16). This toxin was highly homologous to a family of δ-atracotoxins (δ-ACTX) from Australian funnel-web spiders including conservation of all eight cysteine residues. In addition to actions on sodium channel gating and kinetics to δ-ACTX, δ-MSTX-Mb1a caused significant insect toxicity at doses up to 2000 pmol/g. δ-MSTX-Mb1a therefore provides evidence of a highly conserved spider δ-toxin from a phylogenetically distinct spider family that has not undergone significant modification.

Spiders of medical importance in the Asia-Pacific: atracotoxin, latrotoxin and related spider neurotoxins.

1. The spiders of medical importance in the Asia-Pacific region include widow (family Theridiidae) and Australian funnel-web spiders (subfamily Atracinae). In addition, cupboard (family Theridiidae) and Australian mouse spiders (family Actinopodidae) may contain neurotoxins responsible for serious systemic envenomation. Fortunately, there appears to be extensive cross-reactivity of species-specific widow spider antivenom within the family Theridiidae. Moreover, Sydney funnel-web antivenom has been shown to be effective in the treatment of mouse spider envenomation. 2. alpha-Latrotoxin (alpha-LTx) appears to be the main neurotoxin responsible for the envenomation syndrome known as "latrodectism" following bites from widow spiders. This 120 kDa protein binds to distinct receptors (latrophilin 1 and neurexins) to induce neurotransmitter vesicle exocytosis via both Ca2+-dependent and -independent mechanisms, resulting in vesicle depletion. This appears to involve disruption to a process that normally inhibits vesicle fusion in the absence of Ca2+. Precise elucidation of the mechanism of action of alpha-LTx will lead to a major advancement in our understanding of vesicle exocytosis. 3. delta-Atracotoxins (delta-ACTX) are responsible for the primate-specific envenomation syndrome seen following funnel-web spider envenomation. These peptides induce spontaneous repetitive firing and prolongation of action potentials in excitable cells. This results from a hyperpolarizing shift of the voltage-dependence of activation and a slowing of voltage-gated Na+ channel inactivation. This action is due to voltage-dependent binding to neurotoxin receptor site-3 on insect and mammalian voltage-gated Na+ channels in a manner similar, but not identical, to scorpion alpha-toxins and sea anemone toxins. delta-Atracotoxins provide us with highly specific tools to study Na+ channel structure and function 4. omega- and Janus-faced ACTX, from funnel-web spider venom, are novel neurotoxins that show selective toxicity to insects. In particular omega-ACTX define a new insecticide target due to a specific action to block insect voltage-gated Ca2+ channels. Both these ACTX show promise for the development of baculoviral recombinant biopesticides expressing these toxins for the control of insecticide-resistant agricultural pests. In addition, they should provide valuable tools for the pharmacological and structural characterization of insecticide targets.

Clinical effects of bites from formally identified spiders in tropical Northern Territory.

To determine the types of spiders causing bites and the clinical effects of their bites in tropical Northern Territory (north of the town of Katherine). A prospective study of confirmed and suspected spider-bites and a retrospective analysis of data from a standardised, local database of spider- and snake-bites. Confirmed spider-bites were those in which there was a clear history of the bite, and the captured spider was identified by an arachnologist. Emergency department of a teaching hospital. SUBJECTS AND DATA SOURCE: All subjects with confirmed or suspected spider-bite presenting to the Emergency Department or referred from August 1999 to August 2000, or identified from the database. Thirty-four subjects had a confirmed spider-bite from an identified spider: 25 in the prospective group and nine in the retrospective group. The spiders were Sparassidae (huntsman spider) (12 bites), Missulena pruinosa (northern mouse spider) (7), Latrodectus (widow spider) (4), Araneidae (orb-weaver) (4), Salticidae (jumping spider) (4), Nemesidae (trapdoor spider) (1), Conothele (1) and Selenocosmia (whistling spider) (1). Clinical effects were local pain in 97% (severe in 29%), redness in 47% and swelling in 24% of cases. Systemic effects occurred in three victims, two of whom were bitten by M. pruinosa. There were no cases of confirmed necrotic arachnidism. None of the spider-bites resulted in severe effects. Compared with data from other parts of Australia, different species were involved and latrodectism was uncommon. Our study highlighted the importance of correctly identifying the spider species.

Neurotoxic activity of venom from the Australian eastern mouse spider (Missulena bradleyi) involves modulation of sodium channel gating.

Mouse spiders represent a potential cause of serious envenomation in humans. This study examined the activity of Missulena bradleyi venom in several in vitro preparations. Whilst female M. bradleyi venom at doses up to 0.05 microl ml(-1) failed to alter twitch or resting tension in all preparations used, male venom (0.02 and 0.05 microl ml(-1)) produced potent effects on transmitter release in both smooth and skeletal neuromuscular preparations. In the mouse phrenic nerve diaphragm preparation, male M. bradleyi venom (0.02 microl ml(-1)) caused rapid fasciculations and an increase in indirectly evoked twitches. Male venom (0.02 and 0.05 microl ml(-1)) also caused a large contracture and rapid decrease in indirectly evoked twitches in the chick biventer cervicis muscle, however had no effect on responses to exogenous ACh (1 mM) or potassium chloride (40 mM). In the chick preparation, contractile responses to male M. bradleyi venom (0.05 microl ml(-1)) were attenuated by (+)-tubocurarine (100 microM) and by tetrodotoxin (TTX, 1 microM). Both actions of male M. bradleyi venom were blocked by Atrax robustus antivenom (2 units ml(-1)). In the unstimulated rat vas deferens, male venom (0.05 microl ml(-1)) caused contractions which were inhibited by a combination of prazosin (0.3 microM) and P(2X)-receptor desensitization (with alpha,beta-methylene ATP 10 microM). In the rat stimulated vas deferens, male venom (0.05 microl ml(-1)) augmented indirectly evoked twitches. Male venom (0.1 microl ml(-1)) causes a slowing of inactivation of TTX-sensitive sodium currents in acutely dissociated rat dorsal root ganglion neurons. These results suggest that venom from male M. bradleyi contains a potent neurotoxin which facilitates neurotransmitter release by modifying TTX-sensitive sodium channel gating. This action is similar to that of the delta-atracotoxins from Australian funnel-web spiders.