Evaluation of Euthanasia Techniques for an Invertebrate Species, Land Snails (Succinea putris)

The euthanasia of invertebrates used in scientific investigations poses unanswered questions regarding the rapid induction of unconsciousness with minimal distress and pain. Relative to vertebrates, invertebrates' sensory experience of pain, nociception, and physiologic response to aversive stimuli are poorly characterized. The scientific communities in the European Union, Canada, United States, Australia, and New Zealand join in consensus regarding the need to address alleviation of pain and distress in cephalopods (octopus, squid, and so forth), which have the best-characterized nervous system among invertebrates. In the current study, we evaluated various euthanasia techniques in a terrestrial gastropod species, with priority on animal wellbeing, scientific variability, feasibility in both field and laboratory settings, and acceptability by personnel. In addition, we demonstrated that the 2-step method of euthanasia described in the AVMA Guidelines as acceptable for aquatic invertebrates is effective for terrestrial snails and meets all welfare and scientific requirements. This 2-step method first induces anesthesia by immersion in 5% ethanol (laboratory-grade ethanol or beer) followed by immersion in a euthanizing and tissue-preserving solution of 70% to 95% ethanol or 10% neutral buffered formalin. Furthermore, alternative methods of euthanasia for terrestrial snails commonly used in field research, such as live immersion in concentrated ethanol or formalin, were shown to be unacceptable.