Efficacy of anesthetics in an echinoderm based on multifaceted stress biomarkers

Animal care committees remain ambiguous on the need for anesthetics during experimental procedures on invertebrate taxa due to long-standing questioning of their sentience and pain perception. When used, anesthetizing procedures for invertebrates have commonly been adapted from those developed for vertebrates, under the largely unverified assumption that they afford the same benefits. The present study formally tested the efficacy of four common anesthetics of aquatic invertebrates (ethanol, MgCl₂, clove oil, MS-222) using behavioural (reaction to physical contact and presence of a predator), physiological (respiration rate), cellular (coelomocytes), and hormonal (cortisol) biomarkers in the holothuroid Cucumaria frondosa (Echinodermata). While subjects recovered from exposures to all anesthetics tested, their responses differed markedly. Ethanol did not immobilize the individuals and concurrently increased their respiration rate, and cellular and hormonal stress markers. MgCl₂ and clove oil reduced the behavioural and physiological responses, and decreased the cellular markers, but increased the cortisol levels. Only MS-222 fully immobilized the treated individuals and decreased their respiration rate, both during exposure and throughout ulterior interactions with a predator, while keeping coelomocyte counts and cortisol concentrations at baseline levels. MS-222 thus appears to induce the loss of sensation, representing a promising anesthetic and sedative in soft-bodied aquatic invertebrates.