Protein toxins (20 kD molecular mass) causing lysis of human erythrocytes were isolated from sea anemones (Heteractis magnifica, Madang, Papua New Guinea, and Entacmaea quadricolor, Red Sea), which host anemonefish (Amphiprion sp.). These toxins are also ichthyotoxic. Freshwater and marine fish exposed to toxin concentrations of 0.5 micrograms/ml water were killed within 2 hr and exhibited extensive pathological alterations of the gill filaments. Amphiprion species, e.g. clarkii and percula, which live in the sea anemones Heteractis crispa and Stichodactyla mertensii, were highly vulnerable to the Heteractis magnifica toxin, whereas A. percula from the sea anemone H. magnifica proved to be toxin resistant. However, another species, A. perideraion also living in H. magnifica, was highly sensitive to the toxin. The two toxins exhibited cross-reactivity: Amphiprion, resistant to H. magnifica toxin, was also resistant to Entacmaea quadricolor toxin; but all fish were killed by other membrane-active substances such as gramicidin, saponin and latrunculin. The results of the study indicate that resistance to toxins secreted by the sea anemone has evolved in some Amphiprion species, but it is not an essential or a major factor in the anemonefish symbiosis. The skin's mucus layer seems to provide protection from nematocyst discharge.