Assessment of the Artemia salina toxicity assay as a substitute of the mouse lethality assay in the determination of venom-induced toxicity and preclinical efficacy of antivenom

Mice are routinely used in snake venom research but are costly and subject to pain and suffering. The crustacean Artemia salina could be an alternative to mice, but data to support its adoption in snake venom research is limited. The aim of the present study was to evaluate the suitability of A. salina as a surrogate of mice in assessing the toxicity of venoms and the preclinical efficacy of antivenoms. The toxicity of venoms from 22 snakes of medical importance in sub–Saharan Africa was evaluated in mice (intraperitoneally; i.p. and intravenously; i.v.) and in A. salina. Subsequently, the capacity of a commercial antivenom to neutralize the toxicity of these venoms in mice and A. salina was investigated. There was a positive correlation between the i.v. median lethal doses (LD_50s) and the i.p. LD_50s in mice (r = 0.804; p < 0.0001), a moderate correlation between the i.v. LD_50s in mice and the median lethal concentrations (LC_50s) in A. salina (r = 0.606; p = 0.003), and a moderate correlation between the i.p. LD_50s in mice and the LC_50s in A. salina (r = 0.426; p = 0.048). Moreover, there was a strong correlation between the i.p. median effective doses (ED_50s) and the i.v. ED_50s in mice (r = 0.941, p < 0.0001), between the i.p. ED_50s in mice and the ED_50s in A. salina (r = 0.818, p < 0.0001), and between the i.v. ED_50s in mice and the ED_50s in A. salina (r = 0.972, p < 0.0001). These findings present A. salina as a promising candidate for reducing reliance on mice in snake venom research. Future investigations should build upon these findings, addressing potential limitations and expanding the scope of A. salina in venom research and antivenom development.