Necrotising arachnidism, or skin ulceration due to spider bite, is an unresolved clinical problem in Australia, with both the spiders responsible and disease pathogenesis remaining unclear. We have examined and compared enzymic activity from three species of Australian spiders capable of causing ulceration in humans; the recluse spider (Loxosceles rufescens), the black window spider (Badumna insignis) and the white-tailed spider (Lampona cylindrata). Enzymes which could contribute to skin ulceration, namely hyaluronidase and proteases, were detected in venom extracts of all the three spiders. Significant sphingomyelinase activity was detected in L. rufescens venom and in abdominal extracts from the three spider species, while significantly lower levels of sphingomyelinase activity were detected in abdominal extracts from the non-necrosing red-back spider (Latrodectus hasselti). These results suggest that both venom and gastric enzymes may contribute to the dermonecrotic effects of these spiders bites.