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Fatal intoxication of rabbits, sheep and monkeys by the venom of the sea wasp (Chironex fleckeri)
ToxJcae, 1972, Vol. l0, pp . 653-635. Pu~amon Press. Printed in Great ])rifain
SHORT COMMiTNICATION FATAL INTOXICATION OF RABBITS, SHEEP AND MONKEYS BY THE VENOM OF THE SEA WASP (CHIRONEX FLECKERn E. H.
BAXTER, N.
B.
WALDEN,
C. E.
LIEFMAN
and A. G. M. MARK
Commonwealth Serum Laboratories, Parkville, Victoria, Australia (Acceptedjor publlcation 9 Apri11972)
and SOUTHCOTT, 1965, have assembled observations on fatal stingings of human victims by the sea wasp or box jellyfish (Chironex,/feckeri) in Australian waters. Laboratory workers (ENDEAN et al ., 1969 ; FREEMAN and TURNER, 1969, 1971, 1972 ; BAXTER and MARR, 1969 ; KEEN, 1970) have studied experimental envenomations of laboratory species (mice, prawns, rats and rabbits) by injection of venom extracts from the coelenterate . In the course of our studies on the antigenicity (BAXTER et al., 1968) and lethal properties (BAXTER and MARK, 1969) of the venom, as well as studies on antivenene (BAXTER et al., 1970), and toxoid vaccine development (unpublished observations), it has been necessary to fatally envenomate larger species, namely sheep and monkeys, with venom extracts of Chironex,fleckeri. Observation ofthe envenomation syndromes and post-mortem examinations ofthe carcasses of rabbits, sheep and monkeys (Macaca irus) are recorded here. Venom extracts, derived from tentacular nematocysts by the electrical stimulation method of Barnes (BARNES, 1967) and kindly supplied by him, were diluted with physiological saline containing 0~1 per cent peptone to contain in each dose a small number oflethal units appropriate to the species . The number of mouse i .v. LDap in the LDyo doses for each species of animal were : 2500 g rabbit, 10 ; 2500 g monkey, 100 ; 60,000 g sheep, 360. It will be noted that the species' resistance varies (see also BAXTER and MARK, 1969, for other species). Doses, usually in a volume of 0~5 ml, were injected into the marginal ear vein of the rabbit, the jugular vein of the sheep and the femoral vein of the monkey. The animal under observation was released into a suitable enclosure, except for those monkeys on which heartbeat was being monitored. These were suitably restrained on an animal operating table. Our observations derive from experiments on some 50 to 60 rabbits, 20 sheep and 20 to 30 monkeys. Envenomation syndromes for each species did not vary over the range from one to several lethal units for the species, except for slightly longer survival when the dose was marginally lethal. The rabbits remained passive after injection but breathing was deep and laboured . Within two to three min they fell on their sides, kicking their legs and gasping. The head was thrown back, and a spasm occurred, the animal emitting a shrill cry, and respiration ceasing, followed in about one min by cessation of heartbeat. The sheep stood quite still and within two to three min developed deep and laboured breathing and commenced to sway unsteadily . Muscular tremors and spasms developed, the head drooped to one side and the animals collapsed and fell on their sides. A paralysis of CLELAND
TOXICON /972 Vol. 10.
653
654
E. H. BAXTER, N. B. WALDEN, C. E. LIEFMAN and A. G. M. MARR
the tongue occurred, the tongue protruding to one side and becoming cyanotic. Kicking, struggling and gasping developed, there was loss of eye reflexes and following generalized trembling, death ensued within a few min. There was no vocalization . The monkeys very quickly became depressed, inactive, dull, confused and slightly ataxic and uncoordinated . The eyelids drooped and sometimes the eyes were shut, the mouth open and the head weaving (suggesting pain and discomfort) . The heart reaction varied, sometimes becoming fast and irregular, and sometimes slower and weaker. The breathing became laboured, deep and irregular. Within a few min the animals collapsed and lay on their sides. The heartbeat became very spasmodic and irregular and there was obvious cyanosis. Before death a terminal spasm sometimes appeared and copious froth issued from the nostrils. There was no vocalization . Autopsies were carried out on 24 rabbits, 2 sheep and 6 monkeys. The findings are recorded in Table 1 . Observations are reported only for organs in which appearances departed from normal . TABU : I
Rabbit
Organ
Species Sheep
Monkey tCongested blood vessels in subcutaneous tissue
Abdomen Liver
Moderately congested
Congested
Kidney
Markedly congested and slightly swollen
Moderately congested
Congested
"Moderately congested
Spleen
'Moderately congested
Pleural fluid
'Slight straw coloured
Lungs
Markedly congested, oedema
Markedly congested, oedema
Markedly congested, oedema, "blood stained frothy fluid
Heart
Right ventricle filled with incompletely clotted blood
Right side filled with blood
Stopped in systole
Brain
Blood vessels of meninges over surface of cerebral hemispheres engorged
Not examined
Blood vessels of meninges over surface of cerebral hemispheres markedly engorged
" Not always evident
In our observations of laboratory envenomation there is a close similarity to the human situation in both clinical signs and post-mortem findings . It is obvious that all animals show immediate evidence of pain similarly to humans in whom "during the first 15 min pain increased in mounting waves" (BARNES, 1965) . A difference lies in the fact that, in contrast to humans who are frequently reported as screaming or moaning, animals in general remain mute. The exception appears to be the rabbit, which, during its terminal spasms utters a shrill cry. None of the larger species exhibit the skating, jumping, rolling and leaping activity seen in the mouse . 7YJXICON 1971 Vol. /0.
Fatal intoxication of rabbits, sheep and monkeys by the venom of the Sea Wasp (Chironex Jleckeri)
655
REFERENCES BexivES, J. H. (1967) Extraction of cnidarian venom from living tentacle . ln : Anlmal Toxins (RUSSEt.t., F. E. and Snuxnates, P. R., Eds.) . Pergamon Press, Oxford and New York. BARNFS, J. H. (1965) lgjuries to man from marine invertebrates in the Australian region p . 131 . Spec. Rep . Ser. natn . Hlth med. Rcs . Coun., Canberra, No. 12. Bnxt~x, E. H., Mnax, A. G. M, and LwNe, W. R. (1968) Immunity to the venom of the sea wasp Chironex Jieckeri. Toxicon 6, 45 .
Bex't'ea, E. H. and Mwtut, A. G. M . (1969) Sea wasp (Chtroncx Jleckerl) venom: lethal, haemolytic and detmonecrotic properties . Toxicron 7, 195 . H~xa~, E. H., M~tete, A. G. M. and LeNe, W . R . (1970) Sea wasp (Chironex Jleckeri) toxin : experimental immunity . Toxlcon 8, 121. C~.EUtvn, J. B. and Sou~txcol-r, R. V . (1965) Igjuries to man from marine invertebrates in the Australian region . N.H.M.R .C. Spec . Rep . Ser . natn . Hlth . med. Res. Coun., Canberra, No . 12 . Ermruv, R., Ducl~amv, C., McCotre, D. and FxesEa, E. H . (1969) A study of the biological activity of toxic mattrisl derived from nematocysts of thecubomedusan ChironexJleckeri, Toxicon 6, 179. Faee~N, StitxiBV, E. and TURNER, R. l . (1969). A pharmacological study of the toxin of a cnidarian, Chironex~leckeri Southcott. Br . J. Pharnmc . 3S, 510 . FREEMAN, SwRt.EV, E. and TuRNeR, R. J. (1971) Cardiovascular effects of toxins isolated from the cnidarian Chironex~leckeri Southcott. Br. J. Pharmac. 41, 154 . FREEMAN, St~.EV, E. and TURIJER, R. J. (1972) Cardiovascular effects of cnidarian toxins. A comparison of toxins isolated from Chiropsalmus quadrigatus and Chironex fleckeri. Toxicon 10, 31 . KEEN, T. E. B. (1970) Recent investigations on sea-wasp stingings in Australia. Med. J. Aust . 1, 266 .
TOXICON 19i2 VoJ. J0.
SHORT COMMiTNICATION FATAL INTOXICATION OF RABBITS, SHEEP AND MONKEYS BY THE VENOM OF THE SEA WASP (CHIRONEX FLECKERn E. H.
BAXTER, N.
B.
WALDEN,
C. E.
LIEFMAN
and A. G. M. MARK
Commonwealth Serum Laboratories, Parkville, Victoria, Australia (Acceptedjor publlcation 9 Apri11972)
and SOUTHCOTT, 1965, have assembled observations on fatal stingings of human victims by the sea wasp or box jellyfish (Chironex,/feckeri) in Australian waters. Laboratory workers (ENDEAN et al ., 1969 ; FREEMAN and TURNER, 1969, 1971, 1972 ; BAXTER and MARR, 1969 ; KEEN, 1970) have studied experimental envenomations of laboratory species (mice, prawns, rats and rabbits) by injection of venom extracts from the coelenterate . In the course of our studies on the antigenicity (BAXTER et al., 1968) and lethal properties (BAXTER and MARK, 1969) of the venom, as well as studies on antivenene (BAXTER et al., 1970), and toxoid vaccine development (unpublished observations), it has been necessary to fatally envenomate larger species, namely sheep and monkeys, with venom extracts of Chironex,fleckeri. Observation ofthe envenomation syndromes and post-mortem examinations ofthe carcasses of rabbits, sheep and monkeys (Macaca irus) are recorded here. Venom extracts, derived from tentacular nematocysts by the electrical stimulation method of Barnes (BARNES, 1967) and kindly supplied by him, were diluted with physiological saline containing 0~1 per cent peptone to contain in each dose a small number oflethal units appropriate to the species . The number of mouse i .v. LDap in the LDyo doses for each species of animal were : 2500 g rabbit, 10 ; 2500 g monkey, 100 ; 60,000 g sheep, 360. It will be noted that the species' resistance varies (see also BAXTER and MARK, 1969, for other species). Doses, usually in a volume of 0~5 ml, were injected into the marginal ear vein of the rabbit, the jugular vein of the sheep and the femoral vein of the monkey. The animal under observation was released into a suitable enclosure, except for those monkeys on which heartbeat was being monitored. These were suitably restrained on an animal operating table. Our observations derive from experiments on some 50 to 60 rabbits, 20 sheep and 20 to 30 monkeys. Envenomation syndromes for each species did not vary over the range from one to several lethal units for the species, except for slightly longer survival when the dose was marginally lethal. The rabbits remained passive after injection but breathing was deep and laboured . Within two to three min they fell on their sides, kicking their legs and gasping. The head was thrown back, and a spasm occurred, the animal emitting a shrill cry, and respiration ceasing, followed in about one min by cessation of heartbeat. The sheep stood quite still and within two to three min developed deep and laboured breathing and commenced to sway unsteadily . Muscular tremors and spasms developed, the head drooped to one side and the animals collapsed and fell on their sides. A paralysis of CLELAND
TOXICON /972 Vol. 10.
653
654
E. H. BAXTER, N. B. WALDEN, C. E. LIEFMAN and A. G. M. MARR
the tongue occurred, the tongue protruding to one side and becoming cyanotic. Kicking, struggling and gasping developed, there was loss of eye reflexes and following generalized trembling, death ensued within a few min. There was no vocalization . The monkeys very quickly became depressed, inactive, dull, confused and slightly ataxic and uncoordinated . The eyelids drooped and sometimes the eyes were shut, the mouth open and the head weaving (suggesting pain and discomfort) . The heart reaction varied, sometimes becoming fast and irregular, and sometimes slower and weaker. The breathing became laboured, deep and irregular. Within a few min the animals collapsed and lay on their sides. The heartbeat became very spasmodic and irregular and there was obvious cyanosis. Before death a terminal spasm sometimes appeared and copious froth issued from the nostrils. There was no vocalization . Autopsies were carried out on 24 rabbits, 2 sheep and 6 monkeys. The findings are recorded in Table 1 . Observations are reported only for organs in which appearances departed from normal . TABU : I
Rabbit
Organ
Species Sheep
Monkey tCongested blood vessels in subcutaneous tissue
Abdomen Liver
Moderately congested
Congested
Kidney
Markedly congested and slightly swollen
Moderately congested
Congested
"Moderately congested
Spleen
'Moderately congested
Pleural fluid
'Slight straw coloured
Lungs
Markedly congested, oedema
Markedly congested, oedema
Markedly congested, oedema, "blood stained frothy fluid
Heart
Right ventricle filled with incompletely clotted blood
Right side filled with blood
Stopped in systole
Brain
Blood vessels of meninges over surface of cerebral hemispheres engorged
Not examined
Blood vessels of meninges over surface of cerebral hemispheres markedly engorged
" Not always evident
In our observations of laboratory envenomation there is a close similarity to the human situation in both clinical signs and post-mortem findings . It is obvious that all animals show immediate evidence of pain similarly to humans in whom "during the first 15 min pain increased in mounting waves" (BARNES, 1965) . A difference lies in the fact that, in contrast to humans who are frequently reported as screaming or moaning, animals in general remain mute. The exception appears to be the rabbit, which, during its terminal spasms utters a shrill cry. None of the larger species exhibit the skating, jumping, rolling and leaping activity seen in the mouse . 7YJXICON 1971 Vol. /0.
Fatal intoxication of rabbits, sheep and monkeys by the venom of the Sea Wasp (Chironex Jleckeri)
655
REFERENCES BexivES, J. H. (1967) Extraction of cnidarian venom from living tentacle . ln : Anlmal Toxins (RUSSEt.t., F. E. and Snuxnates, P. R., Eds.) . Pergamon Press, Oxford and New York. BARNFS, J. H. (1965) lgjuries to man from marine invertebrates in the Australian region p . 131 . Spec. Rep . Ser. natn . Hlth med. Rcs . Coun., Canberra, No. 12. Bnxt~x, E. H., Mnax, A. G. M, and LwNe, W. R. (1968) Immunity to the venom of the sea wasp Chironex Jieckeri. Toxicon 6, 45 .
Bex't'ea, E. H. and Mwtut, A. G. M . (1969) Sea wasp (Chtroncx Jleckerl) venom: lethal, haemolytic and detmonecrotic properties . Toxicron 7, 195 . H~xa~, E. H., M~tete, A. G. M. and LeNe, W . R . (1970) Sea wasp (Chironex Jleckeri) toxin : experimental immunity . Toxlcon 8, 121. C~.EUtvn, J. B. and Sou~txcol-r, R. V . (1965) Igjuries to man from marine invertebrates in the Australian region . N.H.M.R .C. Spec . Rep . Ser . natn . Hlth . med. Res. Coun., Canberra, No . 12 . Ermruv, R., Ducl~amv, C., McCotre, D. and FxesEa, E. H . (1969) A study of the biological activity of toxic mattrisl derived from nematocysts of thecubomedusan ChironexJleckeri, Toxicon 6, 179. Faee~N, StitxiBV, E. and TURNER, R. l . (1969). A pharmacological study of the toxin of a cnidarian, Chironex~leckeri Southcott. Br . J. Pharnmc . 3S, 510 . FREEMAN, SwRt.EV, E. and TuRNeR, R. J. (1971) Cardiovascular effects of toxins isolated from the cnidarian Chironex~leckeri Southcott. Br. J. Pharmac. 41, 154 . FREEMAN, St~.EV, E. and TURIJER, R. J. (1972) Cardiovascular effects of cnidarian toxins. A comparison of toxins isolated from Chiropsalmus quadrigatus and Chironex fleckeri. Toxicon 10, 31 . KEEN, T. E. B. (1970) Recent investigations on sea-wasp stingings in Australia. Med. J. Aust . 1, 266 .
TOXICON 19i2 VoJ. J0.