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Concept: Box jellyfish

200

Chironex fleckeri (Australian box jellyfish) stings can cause acute cardiovascular collapse and death. We developed methods to recover venom with high specific activity, and evaluated the effects of both total venom and constituent porins at doses equivalent to lethal envenomation. Marked potassium release occurred within 5 min and hemolysis within 20 min in human red blood cells (RBC) exposed to venom or purified venom porin. Electron microscopy revealed abundant ∼12-nm transmembrane pores in RBC exposed to purified venom porins. C57BL/6 mice injected with venom showed rapid decline in ejection fraction with progression to electromechanical dissociation and electrocardiographic findings consistent with acute hyperkalemia. Recognizing that porin assembly can be inhibited by zinc, we found that zinc gluconate inhibited potassium efflux from RBC exposed to total venom or purified porin, and prolonged survival time in mice following venom injection. These findings suggest that hyperkalemia is the critical event following Chironex fleckeri envenomation and that rapid administration of zinc could be life saving in human sting victims.

Concepts: Blood, Red blood cell, Cnidaria, Potassium chloride, Box jellyfish, Chironex fleckeri, Cubozoa, Chirodropidae

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The nematocyst is a complex intracellular structure unique to Cnidaria. When triggered to discharge, the nematocyst explosively releases a long spiny, tubule that delivers an often highly venomous mixture of components. The box jellyfish, Chironex fleckeri, produces exceptionally potent and rapid-acting venom and its stings to humans cause severe localized and systemic effects that are potentially life-threatening. In an effort to identify toxins that could be responsible for the serious health effects caused by C. fleckeri and related species, we used a proteomic approach to profile the protein components of C. fleckeri venom. Collectively, 61 proteins were identified, including toxins and proteins important for nematocyte development and nematocyst formation (nematogenesis). The most abundant toxins identified were isoforms of a taxonomically restricted family of potent cnidarian proteins. These toxins are associated with cytolytic, nociceptive, inflammatory, dermonecrotic and lethal properties and expansion of this important protein family goes some way to explaining the destructive and potentially fatal effects of C. fleckeri venom. Venom proteins and their post-translational modifications (PTMs) were further characterized using toxin-specific antibodies and phosphoprotein/glycoprotein-specific stains. Results indicated that glycosylation is a common PTM of the toxin family while a lack of cross-reactivity by toxin-specific antibodies infers there is significant divergence in structure and possibly function among family members. This study provides insight into the depth and diversity of protein toxins produced by harmful box jellyfish and represents the first description of a cubozoan jellyfish venom proteome.

Concepts: Protein, Proteomics, Cnidaria, Jellyfish, Toxin, Cnidocyte, Box jellyfish, Chironex fleckeri

166

Evolutionary constraints which limit the forces produced during bell contractions of medusae affect the overall medusan morphospace such that jet propulsion is limited to only small medusae. Cubomedusae, which often possess large prolate bells and are thought to swim via jet propulsion, appear to violate the theoretical constraints which determine the medusan morphospace. To examine propulsion by cubomedusae, we quantified size related changes in wake dynamics, bell shape, swimming and turning kinematics of two species of cubomedusae, Chironex fleckeri and Chiropsella bronzie. During growth, these cubomedusae transitioned from using jet propulsion at smaller sizes to a rowing-jetting hybrid mode of propulsion at larger sizes. Simple modifications in the flexibility and kinematics of their velarium appeared to be sufficient to alter their propulsive mode. Turning occurs during both bell contraction and expansion and is achieved by generating asymmetric vortex structures during both stages of the swimming cycle. Swimming characteristics were considered in conjunction with the unique foraging strategy used by cubomedusae.

Concepts: Dynamics, Cnidaria, Limited, Swimming, Jet engine, Box jellyfish, Cubozoa, Propulsive efficiency

86

Cnidarian envenomations are the leading cause of severe and lethal human sting injuries from marine life. The total amount of venom discharged into sting-site tissues, sometimes referred to as “venom load”, has been previously shown to correlate with tentacle contact length and sequelae severity. Since <1% of cnidae discharge upon initial tentacle contact, effective and safe removal of adherent tentacles is of paramount importance in the management of life-threatening cubozoan stings. We evaluated whether common rinse solutions or scraping increased venom load as measured in a direct functional assay of venom activity (hemolysis). Scraping significantly increased hemolysis by increasing cnidae discharge. For Alatina alata, increases did not occur if the tentacles were first doused with vinegar or if heat was applied. However, in Chironex fleckeri, vinegar dousing and heat treatment were less effective, and the best outcomes occurred with the use of venom-inhibiting technologies (Sting No More(®) products). Seawater rinsing, considered a "no-harm" alternative, significantly increased venom load. The application of ice severely exacerbated A. alata stings, but had a less pronounced effect on C. fleckeri stings, while heat application markedly reduced hemolysis for both species. Our results do not support scraping or seawater rinsing to remove adherent tentacles.

Concepts: Cnidaria, Jellyfish, Cnidocyte, Box jellyfish, Chironex fleckeri, Chirodropidae

30

Jellyfish (cnidarians) have a worldwide distribution. Despite most being harmless, some species may cause local and also systemic reactions. Treatment of jellyfish envenomation is directed at: alleviating the local effects of venom, preventing further nematocyst discharges and controlling systemic reactions, including shock. In severe cases, the most important step is stabilizing and maintaining vital functions. With some differences between species, there seems to be evidence and consensus on oral/topical analgesics, hot water and ice packs as effective painkillers and on 30 s application of domestic vinegar (4%-6% acetic acid) to prevent further discharge of unfired nematocysts remaining on the skin. Conversely, alcohol, methylated spirits and fresh water should be carefully avoided, since they could massively discharge nematocysts; pressure immobilization bandaging should also be avoided, as laboratory studies show that it stimulates additional venom discharge from nematocysts. Most treatment approaches are presently founded on relatively weak evidence; therefore, further research (especially randomized clinical trials) is strongly recommended. Dissemination of appropriate treatment modalities should be deployed to better inform and educate those at risk. Adequate signage should be placed at beaches to notify tourists of the jellyfish risk. Swimmers in risky areas should wear protective equipment.

Concepts: Clinical trial, Ethanol, Acetic acid, Cnidaria, Jellyfish, Vinegar, Cnidocyte, Box jellyfish

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QUESTION Which treatments are effective for treating jellyfish stings from species in North America and Hawaii? REVIEW SCOPE Included studies compared interventions for pain relief, prevention of nematocyst discharge, or extrusion of venom from true jellyfish (Cnidaria), box jellyfish, or Physalia species found in North American and Hawaiian waters. Outcomes were pain or erythema from envenomation, discharge of nematocysts, and extrusion of venom. REVIEW METHODS MEDLINE, EMBASE Excerpta Medica, CINAHL, Cochrane Reviews, Google Scholar, reviews, and reference lists were searched for English-language studies. 19 studies met inclusion criteria; 6 were randomized controlled trials (RCTs) (n = 390), all measuring pain outcomes, and are reported in this abstract. 2 RCTs had inadequate randomization, 2 had inadequate blinding, and 2 had low follow-up. No RCTs measured discharge of nematocysts or extrusion of venom, and none studied stings from Chiropsalmus quadrumanus or Chrysaora quinquecirrha. MAIN RESULTS In general, hot water or application of heat reduced pain from Physalia species and Carybdea alata stings (Table). CONCLUSION Evidence from randomized controlled trials suggests that heat or hot water reduces pain from Physalia species and Carybdea alata jellyfish stings, but few studies were found.Interventions for reducing pain from jellyfish stings (6 randomized controlled trials)SpeciesnComparisonsResultsPhysalia spp.96Hot water immersion vs ice packHot water reduced pain more than ice pack.54Hot shower vs ice packHot showers reduced time to complete relief and had greater overall pain reduction at 15 minutes.20Vinegar vs Stingose* vs methylated spirits vs saltwaterVinegar and Stingose reduced pain; methylated spirits increased pain.Carybdea alata25Hot water immersion vs control (papain or vinegar)Hot water reduced pain.133Hot pack vs cold pack vs placeboHot pack reduced pain more than placebo and cold pack; cold pack did not differ from placebo.62Adolph’s meat tenderizer (papain) vs freshwater vs Sting-Aid† vs seawaterNo difference in resolution of pain.*Stingose = 20% aluminum sulfate in detergent.†Sting-Aid = aerolized mixture of water, detergent, and aluminum sulfate.

Concepts: Randomized controlled trial, Aluminium, North America, Cnidaria, Jellyfish, Cnidocyte, Box jellyfish, Carybdea alata

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Directional swimming in the box jellyfish Tripedalia cystophora (cubozoa, cnidaria) is controlled by the shape of the velarium, which is a thin muscular sheet that forms the opening of the bell. It was unclear how different patterns of visual stimulation control directional swimming and that is the focus of this study. Jellyfish were tethered inside a small experimental tank, where the four vertical walls formed light panels. All four panels were lit at the start of an experiment. The shape of the opening in the velarium was recorded in response to switching off different combinations of panels. We found that under the experimental conditions the opening in the velarium assumed three distinct shapes during a swim contraction. The opening was (1) centred or it was off-centred and pocketed out either towards (2) a rhopalium or (3) a pedalium. The shape of the opening in the velarium followed the direction of the stimulus as long as the stimulus contained directional information. When the stimulus contained no directional information, the percentage of centred pulses increased and the shape of the off-centred pulses had a random orientation. Removing one rhopalium did not change the directional response of the animals, however, the number of centred pulses increased. When three rhopalia were removed, the percentage of centred pulses increased even further and the animals lost their ability to respond to directional information.

Concepts: Animal, Cnidaria, Jellyfish, Ctenophora, Cnidocyte, Box jellyfish, Chironex fleckeri, Medusa

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Cnidarian venom research has lagged behind other toxinological fields due to technical difficulties in recovery of the complex venom from the microscopic nematocysts. Here we report a newly developed rapid, repeatable and cost effective technique of venom preparation, using ethanol to induce nematocyst discharge and to recover venom contents in one step. Our model species was the Australian box jellyfish (Chironex fleckeri), which has a notable impact on public health. By utilizing scanning electron microscopy and light microscopy, we examined nematocyst external morphology before and after ethanol treatment and verified nematocyst discharge. Further, to investigate nematocyst content or “venom” recovery, we utilized both top-down and bottom-up transcriptomics-proteomics approaches and compared the proteome profile of this new ethanol recovery based method to a previously reported high activity and recovery protocol, based upon density purified intact cnidae and pressure induced disruption. In addition to recovering previously characterized box jellyfish toxins, including CfTX-A/B and CfTX-1, we recovered putative metalloproteases and novel expression of a small serine protease inhibitor. This study not only reveals a much more complex toxin profile of Australian box jellyfish venom but also suggests that ethanol extraction method could augment future cnidarian venom proteomics research efforts.

Concepts: Protein, Serine protease, Protease, Cnidaria, Jellyfish, Cnidocyte, Box jellyfish, Chironex fleckeri

12

A large number of humans are stung by jellyfish all over the world. The stings cause acute pain followed by persistent pain and local inflammation. Harmful jellyfish species typically cause strong pain, whereas harmless jellyfish cause subtle or no pain. Jellyfish sting humans by injecting a tubule, contained in the nematocyst, the stinging organ of jellyfish. The tubule penetrates into the skin leading to venom injection. The detailed morphology of the nematocyst tubule and molecular structure of the venom in the nematocyst has been reported; however, the mechanism responsible for the difference in pain that is caused by harmful and harmless jellyfish sting has not yet been explored or explained. Therefore, we hypothesized that differences in the length of the nematocyst tubule leads to different degrees of epithelial damage. The initial acute pain might be generated by penetration of the tubule, which stimulates pain receptor neurons, whilst persistent pain might be caused by injection of venom into the epithelium. To test this hypothesis we compared the lengths of discharged nematocyst tubules from harmful and harmless jellyfish species and evaluated their ability to penetrate human skin. The results showed that the harmful jellyfish species, Chrysaora pacifica, Carybdea brevipedalia, and Chironex yamaguchii, causing moderate to severe pain, have nematocyst tubules longer than 200 μm, compared with a jellyfish species that cause little or no pain, Aurelia aurita. The majority of the tubules of harmful jellyfishes, C. yamaguchii and C. brevipedalia, were sufficiently long to penetrate the human epidermis and physically stimulate the free nerve endings of Aδ pain receptor fibers around plexuses to cause acute pain and inject the venom into the human skin epithelium to cause persistent pain and inflammation.

Concepts: Pain, Sensory system, Skin, Nociception, Nociceptor, Cnidaria, Jellyfish, Box jellyfish

12

The potentially fatal Irukandji syndrome is relatively common in tropical waters throughout the world. It is caused by the sting of the Irukandji jellyfish, a family of box jellyfish that are almost impossible to detect in the water owing to their small size and transparency. Using collated medical records of stings and local weather conditions, we show that the presence of Irukandji blooms in coastal waters can be forecast on the basis of wind conditions. On the Great Barrier Reef, blooms largely coincide with relaxation of the prevailing southeasterly trade winds, with average conditions corresponding to near zero alongshore wind on the day prior to the sting. These conditions are consistent with hypotheses long held by local communities and provide a basis for designing management interventions that have the potential to eliminate the majority of stings.

Concepts: Jellyfish, Wind, Great Barrier Reef, Box jellyfish, Prevailing winds, Irukandji jellyfish, Irukandji syndrome, Carukia barnesi