Chironex Fleckeri Research Articles

Is there a role for the use of pressure immobilization bandages in the treatment of jellyfish envenomation in Australia?

The aim of this paper was to review the literature relating to the use of pressure immobilization bandages in the first aid management of jellyfish sting in Australia and to attempt to make a recommendation about their use based on the current literature. A descriptive review of all published cases of jellyfish envenomation in Australia was performed, with specific focus on the discussion of pressure immobilization bandages in the management of such cases. A Medline search was performed using the key words listed for this article. Selected articles were reviewed and further publications were identified from the published reference lists given in the selected articles. The published articles were grouped into three groups: in vitro evidence, case reports and editorial comment (either in journals or book). Fifteen references were identified that discussed the use of pressure immobilization bandages in the management of jellyfish envenomation. Other articles were identified that had significant management issues discussion. Most of the 'jellyfish' literature is in relation to envenomation by Chironex fleckeri. This jellyfish is usually found in tropical Australia and has resulted in the deaths of 67 people in Australia. The last death was near Cairns in 2000. Unfortunately, there are few good data on marine envenomations, with most of the literature being Chironex envenomation case reports. There are minimal data on the effect of pressure immobilization bandages on other jellyfish envenomations. There is no good evidence to support the use of pressure immobilization bandages in the management of jellyfish sting in Australia [corrected].

Toxinological and immunological studies of capillary electrophoresis fractionated Chrysaora quinquecirrha (Desor) fishing tentacle and Chironex fleckeri Southcott nematocyst venoms

Repeated runs of capillary electrophoresis (CE) were used to study partially-purified jellyfish nematocyst venom protein in concentrations sufficient to perform toxinological assays. Nematocyst venoms from Chironex fleckeri ( Cf) and Chysaora quinquecirrha were processed. The CE eluate was divided into quadrants by scanning protein content. The fourth fraction of both jellyfish venoms, contained proteins with the smallest molecular weight components, which were responsible for the highest hemolysins and the humoral and cell-mediated immunological activity. Cytotoxic Cf lethal factor activity against human liver cells was widely dispersed throughout both venoms but more prominent in fraction 4. A V β receptor human T-cell repertoire was not species-specific for either crude or fractionated jellyfish nematocyst venom.

Optimization and preliminary characterization of venom isolated from 3 medically important jellyfish: the box (Chironex fleckeri), Irukandji (Carukia barnesi), and blubber (Catostylus mosaicus) jellyfish

To optimize venom extraction and to undertake preliminary biochemical studies of venom from the box jellyfish (Chironex fleckeri), the Irukandji jellyfish (Carukia barnesi), and the blubber jellyfish (Catostylus mosaicus). Lyophilized crude venoms from box jellyfish tentacles and whole Irukandji jellyfish were prepared in water by homogenization, sonication, and rapid freeze thawing. A second technique, consisting of grinding samples with a glass mortar and pestle and using phosphate-buffered saline, was used to prepare crude venom from isolated nematocysts of the box jellyfish, the bells of Irukandji jellyfish, and the oral lobes of blubber jellyfish. Venoms were compared by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot test. Toxicity of some venoms was determined by intravenous median lethal dose assay in mice. Different venom extraction techniques produced significantly different crude venoms for both box and Irukandji jellyfish. Irukandji and blubber venom SDS-PAGE protein profiles were established for the first time. Analysis of Western blot tests revealed that box jellyfish antivenin reacted specifically with the venom of each jellyfish. Toxicity was found in Irukandji jellyfish venom derived by use of the mortar-and-pestle method, but not in the lyophilized venom. Glass mortar-and-pestle grinding and use of an appropriate buffer was found to be a simple and suitable method for the preparation of venom from each jellyfish species studied. This study contributes to biochemical investigations of jellyfish venoms, particularly the venom of the Irukandji jellyfish, for which there are, to our knowledge, no published studies. It also highlights the importance of optimizing venom extraction as the first step toward understanding the complex biological effects of jellyfish venoms.

Irukandji and Chironex fleckeri jellyfish envenomation in tropical Australia

To compare the temporal distribution of Irukandji and Chironex fleckeri stings, the demographics of victims, the prevailing physical conditions at the time of a sting, and the prevalence of unsuitable first aid strategies. Retrospective assessment of 478 Chironex and 544 Irukandji stings in Queensland and the Northern Territory of Australia. Adolescent and young adult males were the most common victims of Irukandji (median age 21 years) and Chironex stings (median age 16 years). Most Chironex stings occurred on the legs, while Irukandji stings were more common on the arms. Vinegar was correctly used to remove tentacles in 90.5% of Chironex stings, whereas inappropriate treatments were used in the remaining cases. Chironex stings were reported in every month in the Northern Territory, and in all months but June and July in Queensland. The peak prevalence for Chironex stings occurred in January in both areas, while the number of Irukandji stings peaked in December in Queensland and in May in the Northern Territory. Chironex stings were more common on still, cloudy days, whereas Irukandji stings were more common on still, clear days. Irukandji stings were more frequent than Chironex stings on rough days (P = .0005). Chironex and Irukandji stings were similar with respect to tides, moon phases, and rainfall. This study failed to predict exact weather patterns or other contributing factors to reduce the risk of stings to an acceptable level, but did identify several factors that increase the incidence of stings. The "stinger-free" season reported on Chironex warning signs is inaccurate and should be changed to warn bathers that Chironex may be present year round, particularly in the Northern Territory.

Clinical toxicology: a tropical Australian perspective.

Tropical Australia has an amazing diversity of venomous fauna, from "the world's most venomous creature," the multi-tentacled (chirodropid) box jellyfish Chironex fleckeri, to aggressive spiders whose venom remains to be characterized. All genera of highly venomous Australasian elapid snakes are present, except for tiger snakes. Most notable is the taipan (Oxyuranus scutellatus), with the most efficient "snap-release" biting mechanism of any snake and venom components causing the full constellation of clinical envenoming features: coagulopathy from fibrinogen depletion (procoagulant), neurotoxicity (predominantly presynaptic neurotoxin) and rhabdomyolysis (myotoxin). Brown snakes (Pseudonaja textilis and P. nuchalis) now account for most snake bite fatalities in Australia, as a result of severe coagulopathy and a poorly defined early scenario of collapse, postulated to be caused by profound hypotension caused by transient myocardial dysfunction associated with prothrombin activation. Other venomous entities include paralyzing ticks, the blue-ringed octopus, stone fish and other marine animals with venomous spines, paralyzing cone shells, and a wide range of jellyfish including Carukia barnesi and possibly other four-tentacled (carybdeid) box jellyfish causing the Irukandji syndrome.

Effects of verapamil and CSL antivenom on Chironex fleckeri (Box-jellyfish) induced mortality

Intravenous verapamil prolonged survival in mice challenged with intravenous Chironex fleckeri venom prepared from clarified nematocyst suspensions. Ovoid antivenom protected mice who had been challenged but only over a limited dose range. Verapamil also enhanced the beneficial effect of ovine antivenom but again was most active only over a restricted dose range. This data suggests that there may be an, as yet undefined, optimal antivenom dose for humans and that verapamil, used in combination with antivenom is an effective agent.

Partial purification of box jellyfish ( Chironex fleckeri) nematocyst venom isolated at the beachside

Chironex fleckeri, the northern Australian box jellyfish produces one of, if not, the most potent animal venoms. Study of the venom has been hampered by the limits of the animals' range and the venom's thermolability. Using retained lethality and native polyacrylamide gel electrophoresis (NPAGE), we show that lyophilization of autolysis isolated nematocysts is an effective method of transporting the venom. In addition, Sephadex G-200 chromatography, spin concentration, and NPAGE fail to demonstrate the presence of a 600 kDa protein to which the bulk of the lethal activity has been ascribed. Sodium dodecyl sulfate capillary electrophoresis of crude venom yields several protein bands with a molecular weight range of 30–200 kDa. Freeze–thaw studies show a loss of activity and NPAGE bands after two freeze–thaw cycles.

Autonomic neurotoxicity of jellyfish and marine animal venoms.

Venoms and poisons of jellyfish and other marine animals can induce damage to the human nervous and circulatory systems. Clues to the pathogenesis and clinical manifestations of these lesions can be obtained from data of human envenomations and animal experimentation. Because many investigators are unaware that marine animal venoms have autonomic actions, this paper aims to elucidate the broad antagonistic or toxic effects these compounds have on the autonomic nervous system. Marine venoms can affect ion transport of particularly sodium and calcium, induce channels or pores in neural and muscular cellular membranes, alter intracellular membranes of organelles and release mediators of inflammation. The box jellyfish, particularly Chironex fleckeri, in the Indo-Pacific region, is the world's most venomous marine animal and is responsible for autonomic disorders in patients. The symptoms induced by these venoms are vasospasm, cardiac irregularities, peripheral neuropathy, aphonia, ophthalmic abnormalities and parasympathetic dysautonomia. Cases of Irukandji syndrome, caused by the jellyfish Carukia barnesi, have symptoms that mimic excessive catecholamine release. Coelenterate venoms can also target the myocardium, Purkinje fiber, A-V node or aortic ring. Actions on nerves, as well as skeletal, smooth or cardiac muscle occur. Recent studies indicate that the hepatic P-450 enzyme family may be injured by these compounds. The multiplicity of these venom activities means that a thorough understanding of the sting pathogenesis will be essential in devising effective therapies.

The effects of antivenom and verapamil on the haemodynamic actions of Chironex fleckeri (box jellyfish) venom.

The efficacy of antivenom and verapamil against Chironex fleckeri (box jellyfish) venom was investigated in monitored mechanically ventilated piglets. Chironex fleckeri tentacle extract alone, a mixture of tentacle extract with antivenom, and verapamil before tentacle extract were administered intravenously to groups of animals. Tentacle extract caused severe systemic hypotension, cardiac dysrrhythmias, pulmonary hypertension, haemolysis and hyperkalaemia. These effects were prevented by pre-incubation of tentacle extract with antivenom. Verapamil did not prevent any effect of venom, exacerbated cardiovascular collapse and increased mortality. We conclude that antivenom neutralizes the cardiovascular, haemolytic and hyperkalaemic effects of box jellyfish venom. Verapamil does not prevent any of these effects and is contra-indicated for treatment of envenomation.

Marine stingers in far north Queensland.

Two box jellyfish in particular cause problems in tropical Queensland waters. Chironex fleckeri inhabit calm waters close to the shore between November and May. The venom includes three major components: haemolytic dermatonecrotic and myocardial. The dermatonecrotic toxin causes a ladder pattern of whiplash lesions to the skin which ulcerate become necrotic and heal very slowly over months: Neuromuscular paralysis and cardiovascular collapse may be fatal within minutes of envenomation. Emergency treatment comprises inactivation of stinging capsules by vinegar removal of tentacles analgesia, cardiopulmonary resuscitation and the administration of the specific antivenom. Carukia barnesi ('Irukandji') are found in both coastal and open waters. A patch of erythema with papules at the sting site is characteristically followed 30 min later by the onset of a catecholamine mediated syndrome. Headache and severe abdominal and back pain are usual and may be followed by hypertension, tachyarrhythmias and cardiogenic shock.

The mechanism underlying the cardiotoxic effect of the toxin from the jellyfish Chironex fleckeri.

We have investigated the mechanisms underlying the cardiac effects of the toxin from the box jellyfish Chironex fleckeri. Papillary muscles isolated from the hearts of ferrets and ventricular myocytes isolated from the hearts of ferrets and rats were used. Force, intracellular [Ca2+], and membrane potential were monitored in the papillary muscles; contraction, intracellular [Ca2+], intracellular [Na+], and membrane currents were monitored in the isolated myocytes. Application of the toxin to these preparations resulted in a large increase in intracellular [Ca2+] and the adverse symptoms of Ca2+ overload (aftercontractions, spontaneous contractions, a decrease in developed force, and an increase in resting force). The response of papillary muscles to the toxin was not inhibited by blockers of Ca2+ or Na+ channels or by inhibitors of the sarcoplasmic reticulum, Na+/K+ ATPase, or Na+/H+ exchange. The response to the toxin was, however, blocked by prior exposure to a solution which contained no Na+ and by Ni2+. In the isolated myocytes, as well as an increase in intracellular [Ca2+], the toxin also caused an increase in intracellular [Na+] and the appearance of a current which was inward at negative potentials and reversed at about -10 mV. These data can be explained by the toxin increasing Na+ influx into the cell. The increase in intracellular [Na+] will then increase intracellular [Ca2+] via the Na+/Ca2+ exchange mechanism, thus producing the observed Ca2+ overload.

Identification of Chironex fleckeri envenomation by nematocyst recovery from skin

To prospectively compare two methods of nematocyst retrieval from skin for confirmation of Chironex fleckeri jellyfish envenomation. Twenty patients presenting to Royal Darwin Hospital with jellyfish stings. In each, two methods of retrieval of nematocysts from the sting site were tested: scraping the skin with a scalpel blade; and application of transparent sticky tape. Chironex fleckeri nematocysts were identified in 14/20 patients by scalpel blade scraping and in 17/20 by the sticky tape method. In all patients with scalpel blade scrapings positive for nematocysts, nematocysts were also retrieved by the sticky tape method. Only four patients required narcotic analgesia and none required C. fleckeri antivenom. Nematocyst retrieval from skin by a simple sticky tape method is at least as good as scraping with a scalpel blade. Chironex fleckeri causes the majority of jellyfish envenomations presenting to Royal Darwin Hospital.

Swimming, feeding, circulation and vision in the Australian box jellyfish, Chironex fleckeri (Cnidaria:Cubozoa)

Juvenile Chironex fleckeri medusae were maintained in aquaria for several months. One individual was raised over a nine-month period to subadult condition for the first time. The medusae did not feed naturally in most aquaria but they accepted and digested prey items placed by hand onto the manubrium. Medusae maintained in planktonkreisels, however, extended tentacles and captured and ingested live Acetes, large prawns and fish by subumbrellar flexing of the pedalia. Digestion of prey was rapid and food particles were circulated directionally through functional canals and lacunae primarily by contractions of the bell but also by peristaltic contractions of interradial gastrovascular tissues. In the laboratory, medusae visually reacted to dark objects by swimming away from them. Swimming behaviour of medusae in the sea and natural predation by green turtles (Chelonia rnydas) on C. fleckeri are described.

Monoclonal antibodies neutralizing the haemolytic activity of box jellyfish ( Chironex fleckeri) tentacle extracts

1. 1. Three monoclonal antibodies have been produced which neutralize in vitro the haemolytic activity present in tentacle extracts of the box jellyfish ( Chironex fleckeri). 2. 2. Two of these monoclonal antibodies bound specifically to a component of relative molecular mass 50,000 in tentacle extract on Western blots. 3. 3. This binding only occurred when the extracts were electrophoresed under non-reducing conditions. 4. 4. The third monoclonal antibody did not display binding to Western blots of tentacle extract under any of our experimental conditions.

Cubozoan jellyfish envenomation syndromes and their medical treatment in northern Australia

The stings of Chironex fleckeri, Carukia barnesi and two unidentified carybdeids cause severe or fatal symptoms to swimmers in tropical Australia. The cubozoans and their envenomation syndromes are described. First-aid and medical treatment are reviewed, and safety precautions to prevent envenomation are recommended.

Fatal envenomation by Chironex fleckeri, the north Australian box-jellyfish: the continuing search for lethal mechanisms

Fatal envenomation by Chironex fleckeri, the north Australian box-jellyfish: the continuing search for lethal mechanisms

Techniques applicable for purifying Chironex fleckeri (box-jellyfish) venom

A survey of several techniques to isolate a purified lethal factor from the tentacles of Chironex fleckeri was completed. Heterologous band patterns were obtained from specific eluates after gel filtration, ion exchange, immunoaffinity and hydrophobic chromatography. SDS-PAGE revealed a dense band at 24,000 mol. wt in many of these fractions. Isoelectric focusing of the crude venom resulted in considerable loss of activity but indicated significant purification in the fractions having a pI of 5.2-6.8. These fractions were also immunologically active against sera from a convalescing post-evenomation patient. The primary difficulties encountered in jellyfish venom purification are the lack of stability and the tendency of the active toxins to adhere to each other and to various support matrices.

Verapamil potentiation of Chironex (box-jellyfish) antivenom

‘Rescue’ experiments were performed on mice to compare the efficacy of i.v. verapamil, box-jellyfish antivenom, and a combination of both agents to counter the effects of toxic material extracted from the tentacles of Chironex fleckeri. In each of three series of experiments, four sets of mice were injected with doses of Chironex tentacle extracts containing approximately 4 mg/ml protein which were equivalent to twice the ld 50. Three of these groups also received verapamil alone, antivenom alone, or a combination of both treatments. The combination therapy of drug and antivenom was superior to either agent alone. Treatment of verapamil alone appeared to be equivalent to that with only antivenom.

Successful use of Chironex antivenom by members of the Queensland Ambulance Transport Brigade (for editorial comment, see page 610, see also pages 621, 625 and 626)

We report the first administration in Australia by ambulance officers of the Commonwealth Serum Laboratories' Chironex fleckeri antivenom, in two separate, major, potentially life-threatening Chironex envenomations. In both cases, the antivenom was effective in helping to relieve signs and symptoms (including pain). This antivenom still is the most specific part of the treatment of a major Chironex fleckeri sting after stabilization of the airway, breathing and circulation. The additional use of verapamil may contribute to the prevention of some of the cardiac complications that result from the direct effects of the Chironex venom.

Stabilization of lethal and hemolytic activities of box jellyfish (Chironex fleckeri) venom

A. Comis, R. F. Hartwick and M. E. H. Howden. Stabilization of lethal and hemolytic activities of box jellyfish ( Chironex fleckeri) venom. Toxicon 27, 439–447, 1989.—The stability of both the lethal and hemolytic activities of box jellyfish ( Chironex fleckeri) tentacle extract was assessed after various extraction procedures. Both activities were higher when no buffers or water were used during the initial extraction. Also, when the extract was first filtered through a Sep-pak C 18 cartridge, the residual lethal titre, after incubation for 24 hr at room temperature, was increased 16-fold and hemolysis was increased 2.6-fold. Evidence for proteolytic activity in the extract was also obtained and monitored by size exclusion HPLC.