A comparison of serum antivenom concentrations after intravenous and intramuscular administration of redback (widow) spider antivenom.

AIMS: There are no studies measuring antivenom concentrations following intramuscular administration. This study aimed to compare antivenom concentrations following intravenous and intramuscular administration of redback spider antivenom (RBSAV). METHODS: Twenty patients recruited to a controlled trial comparing intramuscular and intravenous administration of antivenom had serial blood samples collected at 30 min intervals for 2 h after the administration of one or two doses of antivenom. Antivenom concentration was measured using an enzyme immunoassay. RESULTS: Ten patients received intramuscular antivenom but antivenom could not be detected in serum after either one or two vials, at any time point. The median time of the final sample after commencement of antivenom treatment in these patients was 3.2 h (1.8-5 h). Ten patients received intravenous antivenom (three one vial and seven two or more vials) and antivenom was detected in all patients. CONCLUSIONS: RBS AV given by the intramuscular route is unlikely to be effective in the treatment of redback (widow) spider bite.

The in vitro vascular effects of two chirodropid (Chironex fleckeri and Chiropsella bronzie) venoms.

Clinical observations suggest a primary cardiotoxic role in fatal Chironex fleckeri stings. The limited research available indicates that Chiropsella bronzie venom acts in a similar manner although appears to be less potent. The aim of the present study was to elucidate the vascular effects of C. fleckeri and C. bronzie venoms using rat isolated aorta. Both venoms produced a sustained contraction of endothelium-denuded aorta which was not significantly affected by prazosin or box jellyfish antivenom. Felodipine significantly reduced the contractile response to C. fleckeri venom but not C. bronzie venom. Both venoms produced an initial relaxation (Phase 1), followed by a sustained contraction (Phase 2), in pre-contracted endothelium-intact aorta. Removal of the endothelium significantly inhibited both phases of the response. NOLA significantly inhibited Phase 1, but not Phase 2, of the response to both venoms. Atropine, HOE 140 or BQ 123 did not have any significant inhibitory effect on either phase. In conclusion, neither C. fleckeri nor C. bronzie venoms appear to contain components with activity at alpha(1)-adrenoceptors. Antivenom was ineffective in reversing the effects of the venom suggesting it is incapable of completely neutralising nematocyst-derived venom. Determining the mechanism of action of these venoms will allow for the development of better treatment strategies.

The in vivo cardiovascular effects of an Australasian box jellyfish (Chiropsalmus sp.) venom in rats.

Using a new technique to extract venom from the nematocysts of jellyfish, the in vivo cardiovascular effects of Chiropsalmus sp. venom were investigated in anaesthetized rats. Chiropsalmus sp. venom (150 microg/kg, i.v.) produced a transient hypertensive response (44+/-4 mmHg; n=6) followed by hypotension and cardiovascular collapse. Concurrent artificial respiration or pretreatment with Chironex fleckeri antivenom (AV, 3000 U/kg, i.v.) did not have any effect on the venom-induced hypertensive response nor the subsequent cardiovascular collapse. The cardiovascular response of animals receiving venom after the infusion of MgSO4 (50-70 mM @ 0.25 ml/min, i.v.; n=5) alone, or in combination with AV (n=5), was not significantly different from rats receiving venom alone. Prior administration of prazosin (50 microg/kg, i.v.; n=4) or ketanserin (1 mg/kg, i.v.; n=4) did not significantly attenuate the hypertensive response nor prevent the cardiovascular collapse induced by venom (50 microg/kg, i.v.). In contrast to previous work examining C. fleckeri venom, administration of AV alone, or in combination with MgSO4, was not effective in preventing cardiovascular collapse following the administration of Chiropsalmus sp. venom. This indicates that the venom of the two related box jellyfish contain different lethal components and highlights the importance of species identification prior to initiating treatment regimes following jellyfish envenoming.

The in vivo cardiovascular effects of the Irukandji jellyfish (Carukia barnesi) nematocyst venom and a tentacle extract in rats.

Envenoming by Carukia barnesi may produce life-threatening Irukandji syndrome. There is little published on the activity of C. barnesi venom. This is the first study to investigate the in vivo cardiovascular effects of C. barnesi venom and a tentacle extract (devoid of nematocysts). Venom (50 microg/kg or 100 microg/kg, i.v.) produced a pressor response (42+/-3 and 44+/-6 mmHg, respectively; n=4) and increase in heart rate (31+/-5 and 13+/-2 bpm, respectively; n = 4) in anaesthetised rats. These changes were not dose-dependent and were followed by cardiovascular collapse in one of four rats receiving 50 microg/kg and three of four animals receiving 100 microg/kg. Prazosin (50 microg/kg, i.v.) significantly attenuated the venom (50 microg/kg, i.v.)-induced pressor response (-8+/-3 mmHg; P < 0.05; n = 4) and tachycardia (-9+/-4 bpm; P < 0.05; n = 4). Tentacle extract (100 microg/kg; i.v.) produced a pressor response (51+/-12 mmHg; n = 3) and an increase in heart rate (35+/-1 bpm; n = 3) in anaesthetised rats, with no subsequent cardiovascular collapse. The results of this study are consistent with the effects shown by humans envenomed by C. barnesi which are postulated to be a result of catecholamine release. We show, for the first time, that C. barnesi tentacle extract, free of nematocyst material, produces cardiovascular effects which are distinct from those caused by venom derived from isolated nematocysts.

Pharmacologically distinct cardiovascular effects of box jellyfish (Chironex fleckeri) venom and a tentacle-only extract in rats.

Using a recently developed technique to extract jellyfish venom from nematocysts, the present study investigated the in vivo cardiovascular effects of Chironex fleckeri venom and tentacle extract (devoid of nematocysts). In anaesthetised rats, venom (10 microg/kg, i.v.) produced a transient pressor response (23+/-4 mmHg) followed, in two of five animals, by cardiovascular collapse. Tentacle extract (100 microg/kg, i.v.) produced a more prolonged hypertensive effect (31+/-3 mmHg) without cardiovascular collapse. Prazosin (50 microg/kg, i.v.) did not have any significant effect on the cardiovascular effects produced by venom. However, prazosin significantly attenuated the pressor response produced by tentacle extract. Ketanserin (1 mg/kg, i.v.) did not have any significant effect on the cardiovascular response of the anaesthetised rat to venom (10 microg/kg, i.v.; 25+/-1 mmHg). Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed to compare the two jellyfish samples used in the present study. In addition to ensuring reproducibility of future studies and allow comparison with previous research. We show, for the first time, that a pure venom sample extracted from C. fleckeri nematocysts and a tentacle extract have cardiovascular effects in the anaesthetised rat which are different and pharmacologically distinct.

Protein kinase C and the sub-sensitivity and sub-reactivity of the diabetic rat prostate gland to noradrenaline.

Concentration-response curves to noradrenaline (1 nM-100 microM) were obtained in prostates from 6-week streptozotocin diabetic, insulin-treated diabetic or control rats. Compared to the curve obtained in controls, those obtained in prostates from diabetic and insulin-treated diabetic rats were shifted rightward. The alpha1-adrenoceptor antagonist, prazosin (100 nM), caused a rightward shift of the curves in prostates from all groups. In contrast, the uptake 1 inhibitor, nisoxetine (300 nM), only produced a leftward shift of the curves in prostates from control and insulin-treated diabetic rats. However, frequency-response curves obtained in prostates from both control and diabetic rats were shifted leftward by nisoxetine (300 nM). The concentration-response curve to the alpha1-adrenoceptor agonist, methoxamine (10 nM-100 microM), obtained in prostates from diabetic rats was shifted rightward compared with controls. Calphostin C (500 nM), a protein kinase C inhibitor, caused a leftward shift of the curve in prostates from diabetic, but not control, rats. The protein kinase C inhibitor, bisindolylmaleimide I (500 nM), beta-adrenoceptor antagonist, propranolol (500 nM) and muscarinic cholinoceptor antagonist, atropine (300 nM), had no effect on the noradrenaline concentration-response curves of prostates from control or diabetic rats. Our results suggest that diabetes reduces the sensitivity and reactivity of the prostate to noradrenaline-induced stimulation, and this reduction may be due to changes in protein kinase C activity.

The efficacy of two antivenoms against the in vitro myotoxic effects of black snake (Pseudechis) venoms in the chick biventer cervicis nerve-muscle preparation.

Neurotoxicity is rarely seen following human systemic envenoming by Australasian black snakes (genus Pseudechis) with myotoxicity being the most prominent feature following bites by some species. This study investigated the in vitro myotoxicity of venoms from seven species of Australasian Pseudechis and determined the efficacy of CSL black and tiger snake antivenoms. All Pseudechis venoms (10 microg/ml) significantly inhibited direct twitches of the chick biventer cervicis nerve-muscle preparation (p<0.05, one-way ANOVA). Prior addition of black snake antivenom (5 U/ml) prevented the inhibitory effects of all Pseudechis venoms (p<0.05, one-way ANOVA), except Pseudechis butleri. Prior addition of tiger snake antivenom (5 U/ml) prevented the venom-induced reduction in direct twitches to Notechis scutatus venom and all Pseudechis venoms (p<0.05, one-way ANOVA), with the exception of Pseudechis australis and Pseudechis colletti venoms. Black or tiger snake antivenom (5 U/ml) added 1 h after the addition of venom inhibited further reduction of direct twitches by N. scutatus and most Pseudechis venoms, but did not significantly restore twitch height. PLA2 activity was found in all venoms with the following rank order: Pseudechis porphyriacus>P. australis>Pseudechis papuanus>P. butleri>Pseudechis guttatus> or =Pseudechis pailsii>P. colletti>N. scutatus. The results of the present study suggest that Australasian Pseudechis venoms possess variable myotoxic activity. The ability of black or tiger snake antivenom to prevent or inhibit further venom-induced effects varied across the genus.

The in vivo cardiovascular effects of box jellyfish Chironex fleckeri venom in rats: efficacy of pre-treatment with antivenom, verapamil and magnesium sulphate.

Using a new technique to extract venom from the nematocysts, the efficacy of CSL box jellyfish antivenom (AV) and adjunct therapies, verapamil and magnesium sulfate (MgSO(4)), were investigated against the in vivo cardiovascular effects of Chironex fleckeri venom in anaesthetised rats. C. fleckeri venom (30 microg/kg; i.v.) produced a transient hypertensive response followed by hypotension and cardiovascular collapse within 4 min of administration. Prophylactic treatment of anaesthetised rats with CSL box jellyfish AV (3000 U/kg; i.v.) did not have any effect on the venom-induced pressor response, but prevented cardiovascular collapse in four out of 10 animals. Administration of verapamil (20mM@0.25 ml/min; i.v.) either alone or in combination with AV, did not have any effect on the C. fleckeri venom-induced pressor response nor the consequent hypotension or cardiovascular collapse of animals. However, the administration of verapamil negated the partially protective effects of AV. Concurrent artificial respiration of animals with the above treatments did not attenuate the C. fleckeri venom-induced cardiovascular effects. MgSO(4) (0.05-0.07M@0.25 ml/min; i.v.) alone did not have any effect on the venom-induced pressor response nor the consequent cardiovascular collapse of animals. However, although combined AV and MgSO(4) administration could not inhibit the transient pressor effect following the administration of C. fleckeri venom, it prevented cardiovascular collapse in all animals. We show for the first time, the cardiovascular effects of a C. fleckeri venom sample free of tentacular contamination and the potential of MgSO(4) as an adjunct therapy for the treatment of potentially fatal C. fleckeri envenomings.

The in vitro effects of two chirodropid (Chironex fleckeri and Chiropsalmus sp.) venoms: efficacy of box jellyfish antivenom.

The pharmacological and biochemical isolation of cnidarian venoms has been hindered by difficulties with both extracting pure venom from nematocysts and venom stability. The development of a new technique to extract active, pure venom of Chironex fleckeri and Chiropsalmus sp. has enabled identify both neurotoxic and myotoxic activity in their venoms. These activities are similar, but not identical in each species. Venom (50 micro g/ml) from both species significantly inhibited indirect and direct twitches of the chick biventer nerve-muscle preparation. Pre-incubation with 1U/ml box jellyfish antivenom did not have any significant effect on venom-induced reductions of indirect twitches. However, this activity was markedly attenuated by prior addition of 5U/ml antivenom, albeit to a lesser degree for Chiropsalmus sp. In contrast, prior addition of 5U/ml box jellyfish antivenom did not neutralise the myotoxic activity of C. fleckeri venom (50 micro g/ml), although it did inhibit the myotoxicity produced by Chiropsalmus sp. venom (50 micro g/ml). Antivenom (5U/ml) added 1h after the addition of C. fleckeri venom (50 micro g/ml) had no effect on the indirect or direct twitches of the skeletal muscle preparation. However, it partially restored the reduction in indirect twitch height caused by Chiropsalmus sp. venom (50 micro g/ml). Myotoxicity was confirmed in muscle preparations stained with hematoxylin and eosin.Therefore, although antivenom was able to neutralize the neurotoxic effects of both species, and the myotoxic effects of Chiropsalmus sp., when added prior to venom, it was unable to reverse the effects after venom addition. This suggests that antivenom is unlikely to be useful in the treatment of neurotoxic or myotoxic effects in patients, although these effects are rarely seen clinically.

Neurotoxic effects of venoms from seven species of Australasian black snakes (Pseudechis): efficacy of black and tiger snake antivenoms.

1. Pseudechis species (black snakes) are among the most widespread venomous snakes in Australia. Despite this, very little is known about the potency of their venoms or the efficacy of the antivenoms used to treat systemic envenomation by these snakes. The present study investigated the in vitro neurotoxicity of venoms from seven Australasian Pseudechis species and determined the efficacy of black and tiger snake antivenoms against this activity. 2. All venoms (10 microg/mL) significantly inhibited indirect twitches of the chick biventer cervicis nerve-muscle preparation and responses to exogenous acetylcholine (ACh; 1 mmol/L), but not to KCl (40 mmol/L), indicating activity at post-synaptic nicotinic receptors on the skeletal muscle. 3. Prior administration of either black or tiger snake antivenom (5 U/mL) prevented the inhibitory effects of all Pseudechis venoms. 4. Black snake antivenom (5 U/mL) added at t90 (i.e. the time-point at which the original twitch height was reduced by 90%) significantly reversed the effects of P. butleri (28+/-5%), P. guttatus (25+/-8%) and P. porphyriacus (28+/-10%) venoms. Tiger snake antivenom (5 U/mL) added at the t90 time-point significantly reversed the neurotoxic effects of P. guttatus (51+/-4%), P. papuanus (47+/-5%) and P. porphyriacus (20+/-7%) venoms. 5. We show, for the first time, the presence of neurotoxins in the venom of these related snake species and that this activity is differentially affected by either black snake or tiger snake antivenoms.