In vitro neurotoxicity of Chinese Krait (Bungarus multicinctus) venom and neutralization by antivenoms

<i>Bungarus multicinctus</i>, the Chinese krait, is a highly venomous elapid snake which causes considerable morbidity and mortality in southern China. <i>B. multicinctus</i> venom contains pre-synaptic PLA2 neurotoxins (i.e., β-bungarotoxins) and post-synaptic neurotoxins (i.e., α-bungarotoxins). We examined the in vitro neurotoxicity of <i>B. multicinctus</i> venom, and the efficacy of specific monovalent Chinese <i>B. multicinctus</i> antivenom, and Australian polyvalent elapid snake antivenom, against venom-induced neurotoxicity. <i>B. multicinctus</i> venom (1–10 μg/mL) abolished indirect twitches in the chick biventer cervicis nerve-muscle preparation as well as attenuating contractile responses to exogenous ACh and CCh, but not KCl. This indicates a post-synaptic neurotoxic action but myotoxicity was not evident. Given that post-synaptic α-neurotoxins have a more rapid onset than pre-synaptic neurotoxins, the activity of the latter in the whole venom will be masked. The prior addition of Chinese <i>B. multicinctus</i> antivenom (12 U/mL) or Australian polyvalent snake antivenom (15 U/mL), markedly attenuated the neurotoxic actions of <i>B. multicinctus</i> venom (3 μg/mL) and prevented the inhibition of contractile responses to ACh and CCh. The addition of <i>B. multicinctus</i> antivenom (60 U/mL), or Australian polyvalent snake antivenom (50 U/mL), at the t₉₀ time point after the addition of <i>B. multicinctus</i> venom (3 μg/mL), did not restore the twitch height over 180 min. The earlier addition of <i>B. multicinctus</i> antivenom (60 U/mL), at the t₂₀ or t₅₀ time points, also failed to prevent the neurotoxic effects of the venom but did delay the time to abolish twitches based on a comparison of t₉₀ values. Repeated washing of the preparation with physiological salt solution, commencing at the t₂₀ time point, failed to reverse the neurotoxic effects of venom or delay the time to abolish twitches. This study showed that <i>B. multicinctus</i> venom displays marked in vitro neurotoxicity in a skeletal muscle preparation which is not reversed by antivenom. This does not appear to be related to antivenom efficacy, but due to the irreversible/pseudo-irreversible nature of the neurotoxins.