Towards rationalisation of antivenom use in funnel-web spider envenoming: enzyme immunoassays for venom concentrations

Abstract Context: Funnel-web spider (Atrax and Hadronyche spp.) envenoming is rare but causes severe neuromuscular, autonomic, and cardiac effects. A rabbit-derived IgG antivenom is available, but venom detection in patients has not been reported. Objective: To use serial venom and antivenom concentrations to better define envenoming and antivenom effectiveness. Materials and methods: Serum was collected from nine patients with suspected funnel-web spider bites and clinical effects were recorded. Venom-specific enzyme immunoassays were developed to measure funnel-web spider venom and antivenom concentrations. Goat anti-rabbit whole serum was coupled to UltraLink resin and added to samples to remove bound venom and measure free venom. Antivenom efficacy was defined as antivenom binding all free venom and antivenom effectiveness as resolution of clinical features. Results: Venom was detectable in samples from six of nine patients. In three patients without venom detected, there were only moderate effects, which did not completely respond to antivenom in all cases and no spider was identified. In five of six cases, a male Atrax spp. (Sydney funnel-web) spider was identified. Three patients had moderate envenoming which responded to antivenom. Three patients had severe envenoming and developed catecholamine-induced myocarditis and acute pulmonary oedema. Although cholinergic and non-specific clinical features appeared to respond to antivenom, myocarditis and pulmonary oedema lasted 2–4 days. Median venom concentration pre-antivenom in five patients with samples was 5.6 ng/ml (3–35 ng/ml), and immediately post-antivenom decreased to a median of 0 ng/ml (0–1.8 ng/ml). Post-antivenom venom concentrations decreased when bound venom was removed; median, 0 ng/ml (0–0.9 ng/ml), indicating that most venom detected post-antivenom was bound. There was recurrence of venom and clinical features in one patient when a pressure bandage was removed. Conclusions: Detection of venom in suspected funnel-web spider bites identified definite cases with characteristic envenoming and a spider was identified. Measurement of venom concentrations pre- and post-antivenom demonstrated that venom was bound by antivenom, but in severe cases cardiac toxicity was not reversed.

[1]  K. Maduwage,et al.  Detection of venom after antivenom administration is largely due to bound venom. , 2015, Toxicon : official journal of the International Society on Toxinology.

[2]  N. Buckley,et al.  Snakebite in Australia: a practical approach to diagnosis and treatment , 2013, The Medical journal of Australia.

[3]  Nicholas A. Buckley,et al.  Clinical Effects and Antivenom Dosing in Brown Snake (Pseudonaja spp.) Envenoming — Australian Snakebite Project (ASP-14) , 2012, PloS one.

[4]  C. Brun-Buisson,et al.  Meta-analysis of controlled studies on immunotherapy in severe scorpion envenomation , 2011, Emergency Medicine Journal.

[5]  W. Hodgson,et al.  Development of a sensitive enzyme immunoassay for measuring taipan venom in serum. , 2010, Toxicon : official journal of the International Society on Toxinology.

[6]  W. Hodgson,et al.  An examination of the activity of expired and mistreated commercial Australian antivenoms. , 2009, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[7]  E. Antunes,et al.  Systemic envenomation caused by the wandering spider Phoneutria nigriventer, with quantification of circulating venom , 2008, Clinical toxicology.

[8]  W. Stoecker,et al.  Diagnosis of loxoscelism in a child confirmed with an enzyme-linked immunosorbent assay and noninvasive tissue sampling. , 2006, Journal of the American Academy of Dermatology.

[9]  B. Currie,et al.  Enzyme immunoassays in brown snake (Pseudonaja spp.) envenoming: detecting venom, antivenom and venom-antivenom complexes. , 2006, Toxicon : official journal of the International Society on Toxinology.

[10]  M. Santoro,et al.  Comparison of the partial proteomes of the venoms of Brazilian spiders of the genus Phoneutria. , 2006, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP.

[11]  B. Stokes,et al.  Funnel‐web spider bite: a systematic review of recorded clinical cases , 2005, The Medical journal of Australia.

[12]  J. Seymour,et al.  The in vivo cardiovascular effects of box jellyfish Chironex fleckeri venom in rats: efficacy of pre-treatment with antivenom, verapamil and magnesium sulphate. , 2004, Toxicon : official journal of the International Society on Toxinology.

[13]  G. Isbister Mouse spider bites (Missulena spp.) and their medical importance , 2004, The Medical journal of Australia.

[14]  G. Isbister,et al.  Bites by Australian mygalomorph spiders (Araneae, Mygalomorphae), including funnel-web spiders (Atracinae) and mouse spiders (Actinopodidae: Missulena spp). , 2004, Toxicon : official journal of the International Society on Toxinology.

[15]  G. Isbister,et al.  Acute Myocardial Injury Caused by Sydney Funnel-web Spider (Atrax robustus) Envenoming , 2003, Anaesthesia and intensive care.

[16]  E. Kalapothakis,et al.  Sandwich-ELISA detection of venom antigens in envenoming by Phoneutria nigriventer spider. , 2001, Toxicon.

[17]  N. Lazar,et al.  Scorpion envenomation and serotherapy in Morocco. , 2000, The American journal of tropical medicine and hygiene.

[18]  D. Gordon,et al.  δ-Atracotoxins from Australian Funnel-web Spiders Compete with Scorpion α-Toxin Binding but Differentially Modulate Alkaloid Toxin Activation of Voltage-gated Sodium Channels* , 1998, The Journal of Biological Chemistry.

[19]  G. Nicholson,et al.  Characterisation of the effects of robustoxin, the lethal neurotoxin from the Sydney funnel-web spider Atrax robustus, on sodium channel activation and inactivation , 1998, Pflügers Archiv.

[20]  D. Gordon,et al.  delta-Atracotoxins from australian funnel-web spiders compete with scorpion alpha-toxin binding but differentially modulate alkaloid toxin activation of voltage-gated sodium channels. , 1998, The Journal of biological chemistry.

[21]  S. Sutherland,et al.  Funnel‐web spider (Atrax robustus) antivenom in the treatment of human envenomation , 1984, The Medical journal of Australia.

[22]  R. Theakston The application of immunoassay techniques, including enzyme-linked immunosorbent assay (ELISA), to snake venom research. , 1983, Toxicon : official journal of the International Society on Toxinology.

[23]  M. Rimsza,et al.  Scorpion envenomation. , 1980, Pediatrics.