Animals may be used only if the researcher’s best efforts to find a non-animal replacement or a method that does not interfere with the animal (e.g., a non-invasive method such as camera traps, scat collection, or use of tracks) to obtain the required information have failed. Where the use of animals is necessary, consideration must be given to the substitution of non-cephalopod invertebrates for vertebrates or cephalopods. It is recognized that where the aim of a scientific activity in the field is to understand the ecology, ecophysiology, or behaviour of wildlife, replacement by a non-animal-based method, or even replacement of one species with another, may not be an option (Griffin and Gauthier, 2004). However, there may be opportunities to minimize contact with animals to obtain the data required for the scientific activity (e.g., environmental DNA (eDNA) (Thomsen and Willerslev, 2015; Neice and McRae, 2021), genetic non-invasive sampling (gNIS) (Schultz et al., 2021), hair traps, remote imaging, and data banks). In addition, research involving endangered or threatened species may be necessary to support the species’ conservation. While replacement of a rare or threatened species with a more common species or a species of lesser conservation concern (Curzer et al., 2013) may be more desirable in terms of conservation impacts, it does not address the Three Rs principle of replacement.