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ToxSci Advance Access originally published online on May 28, 2003
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Toxicological Sciences 74, 352-360 (2003)
Copyright © 2003 by the Society of Toxicology

IN VITRO TOXICOLOGY AND ALTERNATIVE TESTING

Species-Dependent Variations in the in Vitro Myotoxicity of Death Adder (Acanthophis) Venoms

Janith C. Wickramaratna*, Bryan G. Fry{dagger} and Wayne C. Hodgson*,1

* Monash Venom Group, Department of Pharmacology, Monash University, Victoria 3800, Australia; and {dagger} Australian Venom Research Unit, Department of Pharmacology, University of Melbourne, Victoria 3010, Australia

Based on early studies on Acanthophis antarcticus (common death adder) venom, it has long been thought that death adder snake venoms are devoid of myotoxicity. However, a recent clinical study reported rhabdomyolysis in patients following death adder envenomations, in Papua New Guinea, by a species thought to be different to A. antarcticus. Subsequently, a myotoxic phospholipase A2 component was isolated from A. rugosus (Irian Jayan death adder) venom. The present study examined the venoms of A. praelongus (northern), A. pyrrhus (desert), A. hawkei (Barkly Tableland), A. wellsi (black head), A. rugosus, A. sp. Seram and the regional variants of A. antarcticus for in vitro myotoxicity. Venoms (10–50 µg/ml) were examined for myotoxicity using the chick directly (0.1 Hz, 2 ms, supramaximal V) stimulated biventer cervicis nerve-muscle preparation. A significant contracture of skeletal muscle and/or inhibition of direct twitches were considered signs of myotoxicity. This was confirmed by histological examination. All venoms displayed high phospholipase A2 activity. The venoms (10–50 µg/ml) of A. sp. Seram, A. praelongus, A. rugosus ,and A. wellsi caused a significant inhibition of direct twitches and an increase in baseline tension compared to the vehicle (n = 4–6; two-way ANOVA, p < 0.05). Furthermore, these venoms caused dose-dependent morphological changes in skeletal muscle. In contrast, the venoms (10–50 µg/ml; n = 3–6) of A. hawkei, A. pyrrhus , and regional variants of A. antarcticus were devoid of myotoxicity. Prior incubation (10 min) of CSL death adder antivenom (5 U/ml) prevented the myotoxicity caused by A. sp. Seram, A. praelongus, A. rugosus , and A. wellsi venoms (50 µg/ml; n = 4–7). In conclusion, clinicians may need to be mindful of possible myotoxicity following envenomations by A. praelongus, A. rugosus, A. sp. Seram, and A. wellsi species.

Key Words: Acanthophis; A. antarcticus; antivenom; death adder; myotoxic; phospholipase A2; A. praelongus; rhabdomyolysis; A. rugosus; venom.


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