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Jellyfish Sting Newsletters: Number 28 - January 2003

Significant Papers Published 

  1. Alam JM, Qasim R, Ali SA, Jamal Q, Alam SM. Biochemical characterization and pathophysiological properties of two high molecular weight cytolytic proteins from the venom of a coelenterate (jellyfish), Physalia urticulus (Blue Bottle). Pakiston J. Zool, 35:9-17;2002.

    Two high-molecular weight cytolytic proteins from the venom of jellyfish Physalia utriculus were studied. Both were primarily protein in nature with 25-35% polysaccharide content, consisting of two subunits each with molecular weights of 139.00 kDa and 117.00 kDa, respectively. LD50 (i.v.) was determined to be 1.10±0.05 mg/kg and 1.19±0.03 mg/kg, whereas, hemorrhagic, edema-inducing, neurotoxic and hemolytic activities were 48.75±0.73 and 46.37±0.77 lg/mouse (i.d.), 39.55±0.55 and 38.84±0.40 lg/paq (MED) and 331.25±3.70 and 327.50±5.72 lg/kg (i.v.), 9.06±0.01 and 9.46±0.05 total units, respectively. Half LD50 doses induced marked elevation of blood GOT, LDH, CPK, GGT, ALP, amylase and lipase levels in experimental animals. Histopathological studies showed marked cytolytic potency of both toxins with cellular infiltration, disorientation of normal morphological units and necrosis in section of kidneys, liver, lungs in addition to moderate levels of the same in heart and spleen sections.

  2. Edwards LP, Whitter E, Hessinger DA. Apparent membrane pore- formation by Portuguese Man-of-war (Physalia physalis) venom in intact cultured cells. Toxicon 40:1299-1305;2002.

    Intracellular, ratiometric microfluorimetry with fura-2 reveals that low doses of Portuguese Man-of-war (Physalia physalis) venom cause a linear increase in intracellular calcium accumulation by cultured L-929 cells. The influx of calcium is preceded by a lag period that is relatively independent of venom concentration, except at very low concentrations. Electron micrographs of negatively stained preparations of membranes from venom-treated L-929 and GH4C1 cells exhibit 10-80 mm diameter lesions. The number and diameter of these lesions correlate with venom concentration. The venom forms lesions in GH4C1 cells at much lower concentrations than in L-929 cells. Osmotic protectants such as sucrose and polyethylene glycol (PEG), reduce the extent of lactate dehydrogenase (LDH) release from venom-treated cells with higher molecular weight PEG causing a greater inhibition of LDH release than sucrose. These results imply that Man-of-war venom produces pore-like structures in the membranes of target cells, which leads to colloid osmotic swelling with subsequent release of intracellular proteins and cell lysis.

    Our group found a pore forming action of sea nettle venom producing an 80 picosiemen holes. (Dubois JM, Tangay J, Burnett JW. Ionic channels induced by sea nettle toxin in the nodal membrane. Biophysics J 42:199-202; 1983)

  3. Wiltshire CJ, Sutherland SK, Fenner PJ, Young AR. Optimization and preliminary characterization of venom isolated from 3 medically important jellyfish: the box (Chironix flecerki), Irukandji (Carukia barnesi), and blubber (Catostylus mosaicus) jellyfish.Wilderness and Environmental Medicine 11:241-250;2000.

    To optimize venom extraction and to undertake preliminary biochemical studies of venom from the box jellyfish glass mortar-and-pestle grinding and use of an appropriate buffer was found to be a simple suitable method for the preparation of venom (Chironex fleckeri), the Irukandji jellyfish (Carukia barnesi), and the blubber jellyfish (Catosylus mosaicus). Analysis of Western blot tests revealed that box jellyfish antivenin reacted specifically with the venom of each jellyfish. Lethal toxicity was found in Irukandji jellyfish venom derived by use of the mortar-and-pestle method, but not in the lyophilized venom.

  4. Yoshimoto CM, Yanagihara AA. Cnidarian (coelenterate) envenomations in Hawai’i improve following heat application. Transactions of the Royal Society of Tropical Medicine and Hygiene 96:300-303;2002.

    A retrospective review of medical records from 113 patients with cnidarian stings in western O’ahu, Hawai’i, was conducted for the 5-year period 1994-98. The most common clinical feature was acute local pain, but cases of anaphylaxis or anaphylactoid syndrome and a persistent or delayed local cutaneous syndrome were also documented. Six cases resembled the Irukandji syndrome described from northern Australia, characterized by severe pain and signs of catecholamine excess, including muscle cramping, elevated blood pressure, diaphoresis, and tremor. Treatment with heat application, usually by means of a whole-body hot shower, appeared to provide better clinical improvement than parenteral analgesics or tranquilizers, particularly in patients with the Irukandji-like syndrome. The heat sensitivity of one or more of the Carybdea alata venom components might account for the effect of heat treatment. Prospective, randomized, controlled clinical trials should be performed to assess heat treatment for cnidarian envenomation.

    This paper has the disadvantages of being a retrospective medical record investigation and one in which all offending jellyfish are combined not separated by species. A recent past newsletter lists another study with conflicting results.

  5. Fenner P. Marine bites and stings: first aid and medical treatment. Medicine Today 3:26-31;2002.

    A brief overview of multiple causes of marine bites and stings.

  6. La Spada G, Sorrenti G, Soffli A, Montaleone B, Marino A, Musci G. Thiol-induced discharge of acontial nematocysts. Comp Biochem Physio B 132:367-373;2002.

    The discharge of nematocysts, the stinging cells of coelenterata, is a poorly understood phenomenon. In particular, little is known about the chemical stimuli that trigger the discharge. Thiols are able to initiate the nematocyst discharge in isolated nematocysts. Among the thiols tested, reduced glutathione and cysteine were found to be the most effective. The effect of glutathione was likely two-fold: it formed mixed disulfides with membrane thiols, as shown by the ability of the mercapto-blocking reagent iodoacetaminde to abolish its action; and it bound to the membrane through the glutamate moiety, as demonstrated by competitive experiments with free glutamate. Glutathione triggered the discharge at concentrations higher than those sufficient to activate the feeding response of coelenterates.

  7. Freudenthal AR, Barbagallo JS. Ghost anemone dermatitis. J Am Acad Dermatol 47:722-726;2002.

    Haloclava producta, the “ghost anemone”, is a burrowing sea anemone in estuarine sediments of the US East Coast and Gulf of Mexico. It has never been identified as harmful to human beings; however, residents of Long Island, New York develop a pruritic, erythematous, vesiculopapular dermatitis on areas of the body that contact these organisms. H. producta is the apparent causative agent of ghost anemone dermatitis.

  8. Taylor DM, Ashby K, Winkel KD. An analysis of marine animal injuries presenting to emergency departments in Victoria, Australia. Wilderness and Environmental Medicine 13:106-112;2002.

    Two hundred five injuries were identified, and males predominated (71.7%, P<.01). Injuries were most frequent during summer and when jellyfish were most prevalent. Various fish species, stingrays, jellyfish, and sharks were incriminated in 83 (40.5%), 46 (22.4%), 42 (20.5%), and 5 (2.4%) injuries, respectively. Most (65.9%) injuries occurred during leisure or sport, and 72 (35.1%) occurred in a place of recreation. Spikes, spines and barbs caused 82 (40.0%) injuries, and stings caused 54 (26.3%) injuries. Bites were uncommon. Most injuries were to the limbs, with the hands or feet injured in 127 (62.0%) patients. Forty (19.5%) injuries were associated with a retained foreign body. Only 17 (8.3%) patients required admission to the hospital.

  9. Winkel KD, Hawdon GM, Ashby K, Ozanne-Smith J. Eye injury after jellyfish sting in temperate Australia. Wilderness and Environmental Medicine 13:203-205;2002.

    A jellyfish sting-related eye injury, probably caused by the “hair” jellyfish (Cyanea capillata) from southeast Australia is reported. The patient, a 54- year-old man, was stung while swimming without goggles in a jellyfish-infested bay. He experienced severe pain in his right eye, requiring narcotic analgesia, and had decreased visual acuity associated with right-sided facial swelling.

  10. Chung JJ, Ratnapala LA, Cooke IM, Yanagihara AA. Partial purification and characterization of a hemolysin (CAH1) from Hawaiian box jellyfish (Carbdea alata) venom. Toxicon 39:981-990;2002.

    A novel hemolytic protein from the venom of the Hawaiian box jellyfish (Carybdea alata) was isolated and characterized. Hemolysis of sheep red blood cells was used to quantitate hemolytic potency of crude venom extracted from isolated nematocysts and venom after fractionation and purification procedures. Hemolytic activity of crude venom was reduced or lost after exposure to the proteolytic enzymes trypsin, collagenase and papain. The activity exhibited lectin-like properties in that hemolysis was inhibited by D-lactulose and certain other sugars. Activity was irreversibly lost after dialysis of crude venom against divalent-free, 20 mM EDTA buffer; it was optimal in the presence of 10 mM Ca2+ or Mg2+. Two chromatographic purification methods, size fractionation on Sephadex G-200 and anion exchange with quaternary ammonium, provided fractions in which hemolytic activity corresponded to the presence of a protein band with an apparent molecular weight of 42 kDa by SDS-PAGE. The N-terminal sequence of this protein was determined to be: XAADAXSTDIDD/GIIG.

  11. Yanagihara AA, Kuroiwa JMY, Oliver LM, Chung JJ, Kunkel DD. Ultrastructure of a novel eurytele nematocyst of Carybdea alata Reynaud (Cubozoa, Cnidaria). Cell Tissue Res 308:307-318;2002.

    The ultrastructural characteristics of nematocysts from the cubozoan Carybdea alata Reynaud, 1830 (Hawaiian box jellyfish) were examined using light, scanning and transmission electron microscopy. We reclassified the predominant nematocyst in C. alata tentacles as a heterotrichous microbasic eurytele, based on spine, tubule and capsule measurements. These nematocysts exhibited a prominent and singular stylet, herein referred to as the lancet. Discharged nematocysts from fixed tentacle preparations displayed the following structures; a smooth shaft base, lamellae, a hemicircumferential fissure demarking the proximal end of a stratified lancet, and a gradually tapering tubule densely covered with large triangularly shaped spines. The lancet remained partially adjoined to the shaft base in a hinge-like fashion in rapidly fixed, whole-tentacle preparations. In contrast, this structure was not observed in discharged nematocyst preparations which involved multiple transfer steps prior to fixation. Various approaches were designed to detect this structure in the absence of fixative. Detached lancets were located in proximity to discharged tubules in undisturbed coverslip preparations of fresh tentacles. In addition, examination of embedded nematocysts from fresh tentacles laid on polyacrylamide gels revealed still-attached lancets. To examine the function of this structure in prey capture, Artemia sp. laden tentacles were prepared for scanning electron microscopy. While carapace exteriors exhibited structures proximal to the lancet, i.e., the nematocyst capsule and shaft base, neither tubule nor lancet structures were visible. Taken together, the morphological data suggested a series of events involved in the discharge of a novel eurytele from C. alata.

  12. Sakanashi M, Matsuzaki T, Nakasone J, Koyama T, Sakanashi M, Kukita I, Sakanashi M. Effects of diltiazem on in vitro cardiovascular actions of crude venom obtained from Okinawan box-jellyfish (Habu-kurage), Chiropsalmus quadrigatus. Anaesth Intensive Care 30:570-577;2002.

    The venom obtained from Okinawan box-jellyfish (Habu-kurage), Chiropsalmus quadrigatus, produced increases in contractions of isolated rat right atrial preparations in a concentration-dependent manner without changes in a spontaneous beating rate. These increases in contractions were significantly inhibited by diltiazem and did not show tachyphylaxis. The venom also produced increases in contractions of isolated rat aortic ring preparations (endothelium-intact) in a concentration-dependent fashion, which were reproducible with repeated application and were significantly inhibited by diltiazem or heating. These increases in vascular contractions were weakened in endothelim-denuded preparations, and almost abolished in a calcium-free medium. On the other hand, the venom at higher concentrations diminished contractions of both myocardial and vascular preparations and did not show reproducibility. These results suggest that the Habu-kurage venom is heat-labile and may increase contractions of cardiac muscle and aortic smooth muscle by increasing calcium influx into muscle cells, and that the venom at higher concentrations may produce dysfunction of muscle contractile systems due to calcium overload.

    This paper also corroborates our earlier work on verapamil. The diltiazem produced a similar beneficial effect.

  13. Seymour J, Carrette T, Cullen P, Little M, Mulcahy RF, Pereira PL. The use of pressure immobilization bandages in the first aid management of cubozoan evenomings. Toxicon 40:1503-1505;2002.

    This study is aimed to evaluate whether the application of pressure results in additional release of venom from naturally discharged, vinegar soaked nematocysts of the box jellyfish Chironex fleckeri. The results show that large quantities of venom are expressed with the application of pressures similar to that applied by compression pressure immobilization bandages (PIB). The volume of venom expressed by this pressure was similar to the quantity expressed upon initial natural discharge of the nematocysts. The current recommended practice of applying PIB to cubozoan stings might worsen the envenomation. As the existing data now shows that PIB may be detrimental to victims envenomed by cubozoans, we suggest that the current practice of the use of PIB in cubozoan envenomings be discarded until there is direct experimental evidence to support its use.

  14. Haddad V, Lang da Silveira F, Costa Cardoso JL, Morandini AC. A report of 49 cases of cnidarian envenoming from southeastern Brazilian coastal waters. Toxicon 40:1445-1450; 2002.

    Forty-nine accidents caused by jellyfish (Cnidaria) were observed during a period of 5 years on the southeastern coast of Brazil. Most of them involved male patients (65.3%), the injured areas being mainly the legs (71.3%) and the trunk (65.3%). Twenty accidents with Chiropsalmus quadrumanus, four with Physalia physalis and 20 with unidentified jellyfish presented intense pain, linear plaques and systemic symptoms. The five cases with Olindias sambaquiensis caused mild pain, round plaques and no systemic symptoms. There are a few reports on accidents caused by jellyfish, in this country, and scarce clinical or epidemiological data are available up to the present moment.

    Dr. Haddad continues his work in Brazil. His friends have mentioned local cases of Linuche dermatitis to him.

  15. Carrette T, Alderslade P, Seymour J. Nematocyst ratio and prey in two Australian cubomedusans, Chironex fleckeri and Chiropsalmus sp. Toxicon 40:1547-1551;2002.

    This study examined differences in the nematocyst ratios between two species of Australian cubozoans. In Chiropsalmus sp., a species that feeds exclusively on shrimp, no changes in the ratio of the three groups of nematocyst present in the cnidome were detected with size of the individual animals. In Chironex fleckeri, the ratio of different types of nematocysts in the cnidome for small animals (less than 40 mm) was similar to that of Chiropsalmus sp. However, with an increase in body size in C. fleckeri, the nematocyst ratio changed, with mastigophores (nematocysts believed to hold the lethal venom component for prey) increasing in proportion. The change in cnidome ratio is correlated with a change in the prey of C. fleckeri with increased size. Small C. fleckeri appeared to feed exclusively on prawns, medium sized animals fed on fish and prawns and large animals fed predominantly on fish. An increase in the proportion of mastigophores (and presumably the lethal venom component) in the cnidome of C. fleckeri may also be responsible for why this species has caused numerous human fatalities, while the Australian Chiropsalmus sp. has not.

  16. Whiteman L. The blobs of summer. On Earth (summer) 14-19;2002.

    A review of recent toxic jellyfish swarms in Atlantic and Caribbean waters. They describe the appearance Phyllorhiza punctata, and a 60 pound pink medusa, Drymonema dalmatinium, with tentacles over 100 ft. long if extended, in the Gulf of Mexico.

  17. Marques AC, Haddad V, Migotto AE. Envenomation by a benthic Hydrozoa (cnidaria): the case of Nemalecium lighti (Haleciidae). Toxicon 40:213-215;2002.

    A case of envenomation caused by the Nemalecium lighti is described in Brazil. The hydrozoan species lives in many kinds of substrates, being quite common in tropical shallow water. The patient, a marine biologist, had contact with the animal in two different opportunities while snorkeling. Both contacts produced erythematous and highly pruriginous papules in exposed areas of the body. The signs and symptoms persisted for a week and healed without sequellae.

  18. Taylor DM, Pereira P, Seymour J, Winkel KD. A sting from an unknown jellyfish species associated with persistent symptoms and raised troponin I levels.

    A patient stung by an unknown jellyfish species offshore in Far North Queensland was described. The sting caused immediate and severe pain, multiple whip-like skin lesions and constitutional symptoms. The jellyfish tentacular nematocysts were similar to, but distinct from, those of Carukia barnesi, a cause of the ‘Irukandji’ syndrome. The patient’s symptoms largely resolved over seven months and were associated with elevated cardiac troponin levels, in the absence of other evidence of cardiac disease. This case highlights the envenomation risks associated with marine recreation, and the need for critical evaluation of cardiac troponin assays and for further research in marine toxicology.

    This case is unusual because the troponin levels were elevated at the first assay and persisted so a causal relationship was not determined.

Abstracts 

  1. All from the 6th Asia-Pacific Congress on Animal, Plant and Microbial Toxins, Cairns, Australia, July 8-12, 2002.

    1. Little M, Pereira P, Seymour J, Mulcahy R, Cullen P, Carrette T. “Severe Irukandji Syndrome” The epidemiology, management and name change?

      Eleven patients have been known to develop cardiac symptoms with Irukandji syndrome. Cardiac failure developed 1.5-18 hours post-sting. The two patients who received adrenalin as an inotrope improved thus giving doubt to the fact that catecholamines were a cause of the failure.

    2. Seymour J. Mechanisms of envenoming in box jellyfish; could this explain delayed onset of symptoms in Irukandji syndrome victims?

      Jamie Seymour speculates that the delayed onset of Irukandji syndrome may be related to the fact that the nematocyst shaft of the offending animal has only on terminated perforation allowing only a deep intramuscular injection rather than an intradermal capillary injection.

    3. Winkel KD, Tibballs J, Ross-Smith M, Lambert G, Lau C, Wiltshire C, Doube J, Angus JA. In vitro and In vivo analysis of ‘Jimble’ jellyfish (Carybdea rastoni) venom.

      This ‘Jimble’ [Carybdea rastoni] is small, four-tentacled cubozoan jellyfish widespread throughout the Pacific Ocean. Although it usually causes acutely painful stings with local swelling and blistering only, it has also been implicated in some cases of ‘Irukandji-like’ syndrome. As part of our investigations of Irukandji and Irukandji-like syndromes, we examined the effects of C. rastoni venom in vitro and in vivo. The jellyfish were captured in Gulf St Vincent, South Australia, and soluble crude venom extracted (CVE) from the tentacles of pooled specimens using a simple mortar and pestle approach. The in vitro activity of this extract was analysed using rat isolated mesenteric small artery, guinea pig atria and human coronary arteries. The in vivo activity of the CVE was examined in anaesthetized, mechanically ventilated piglets. The CVE (0.1-3μg/ml) contracted the mesenteric small artery. This response was unaffected by ?-conotoxin GVIA (1μM), yohimbine (0.1μM), prazosin (0.1μM), guanethidine (10μM) or tetrodotoxin (0.1μM)(n=6). By contrast, the elicited contraction was reduced up to 50%, but not eliminated, by the L-type voltage-operated calcium channel antagonists (L-VOCC) felodipine (30nM), and nicardipine (0.1μM)(n=2). The positive chronotropic and inotropic responses seen in isolated guinea pig left and right atria treated with the CVE (0.1-3μg/ml) were completely unaffected by propanolol (1μM)(n=2). Concentration-contraction relationships were evident in small (<1mm) and large (4mm) diameter human coronary blood vessels (n=2). CVE (3μg/kg) infused intravenously into the piglets over several minutes caused a small increase in heart rate, systemic arterial pressure and pulmonary artery pressures. These changes were moderate at 10μg/kg and marked at 30μg/kg (n=5). However, in contrast to CVE from Carukia barnesi, peripheral venous blood samples did not demonstrate any significant change in circulating catecholamines at the peak of tachycardia and systemic hypertension. The L-VOCC blocker verapamil (0.1 mg/kg), infused at the peak of these responses, abolished the tachycardia and systemic and pulmonary hypertension. Although the systemic cardiovascular effects of C. rastoni venom appears outwardly similar to that of C. barnesi, a cause of the Irukandji syndrome, the action of the former venom appears predominantly post-junctional. Consequently the ‘Irukandji-like’ syndrome related to jimble stings is not related to hypercatecholaminaemia but is secondary to a direct, Ca2+-dependent action on vascular tissues.

     

Published Letters 

  1. Tibballs J, Hawdon G, Winkel K. Mechanism of cardiac failure in Irukandji syndrome and first aid treatment for stings. Anaesth Intensive Care 29:552;2002.

    A 200-fold increase in serum noradrenaline and a 100-fold increase in adrenaline was observed shortly thereafter injecting piglets with Carukia barnesi venom. This “catecholamine storm” was accompanied by severe systemic and mild pulmonary hypertension persisting for at least two hours. Creatine kinase (MB) enzymes levels did not rise in these piglets in that interval.

Correspondence 

  1. A jellyfish swarm appeared from Daytona Beach to Jacksonville, Florida for 11 days during late July and early August. It was identified by Monty Graham, Ph.D. of the Dauphin Island Marine Laboratory as Chiropsalmus quadrimanus. Lots of a large soccer ball sized medusa which may be a new species were observed off Bonaire by Vicki Carr through Bud Gillan. The overall length of the jellyfish was about three feet. Another medusa with overall length of 0.5M and five banded tentacles was also unidentified. This may have been Carybdea alata var. grandis.

  2. Nidaria Technologies Ltd. organized a group consisting of Drs. Valeh Levy, Arlen Stauffer and Amit Lotan to study the efficacy of Safe Sea lotion versus control on 12 volunteers. Tentacles of C. quadrimanus were laid on their volunteers’ forearms for 10, 20 then 30 second periods until pain was perceived. In 9 instances the lotion protected the subject from acquiring a rash whereas the control arm exhibited a rash.

    This study was cautiously done because of the potential concerns of systemic damage. Unfortunately, there are two disadvantages. Even with controls, the practice of placing a tentacle on intact skin is difficult to reproduce and this system does not mimic the bathing situation in which the swimmer moves in the water thus potentially removing the lotion. We have tried their technique extensively and are familiar with its disadvantages. I cannot think of a better initial step, but further work is needed. Their results corroborate ours in that we can probably conclude that placing jellyfish tentacles on a limited area of skin is a painful but innocuous procedure.

  3. John Serton of the Netherlands writes that on August 8 hundreds of thousand salmon were killed off Scotland by an unidentified species of jellyfish. Also on August 24 the Dutch city of Goes advised against North Sea swimming because of a Gonionemus vertens swarm.

  4. Angel Yanagihara of Hawaii finds long, unsaturated fatty acids which contain ether bonds, and phosphatiyl choline amalogues which could react with exogenous phospholipase A2. This work on C. alata venom is being expanded and should increase our understanding of jellyfish-venom-induced inflammation.

  5. Takatoshi Ishikawa’s daughter forwarded news articles stating the presence of many Stomolphus nomura medusa off the Japan Sea this past summer. In the fall they (umbrella up to 1m, body weight 1150km) appeared through the Tugaru isthmus between Honshu and Hokkaido then down the Pacific coast as far as Sendai. Serious damage to fish, nets, fishing equipment and crabs occurred as these medusa could either swim about or submerge.

Lyophilized crude venoms from box jellyfish tentacles and whole Irunkandji 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 Irunkandji jellyfish, and the oral lobes of blubber jellyfish. Venoms were compared by use of sodim 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.