Identifying obstacles preventing the uptake of tunnel handling methods for laboratory mice: An international thematic survey Lindsay J. Henderson, Tom V. Smulders, Johnny V. Roughan Published: April 14, 2020 https://doi.org/10.1371/journal.pone.0231454 Abstract Handling of laboratory mice is essential for experiments and husbandry, but handling can increase anxiety in mice, compromising their welfare and potentially reducing replicability between studies. The use of non-aversive handling (e.g., tunnel handling or cupping), rather than the standard method of picking mice up by the tail, has been shown to enhance interaction with a handler, reduce anxiety-like behaviours, and increase exploration and performance in standard behavioural tests. Despite this, some labs continue to use tail handling for routine husbandry, and the extent to which non-aversive methods are being used is currently unknown. Here we conducted an international online survey targeting individuals that work with and/or conduct research using laboratory mice. The survey aimed to identify the handling methods currently being used, and to determine common obstacles that may be preventing the wider uptake of non-aversive handling. We also surveyed opinions concerning the current data in support of non-aversive handling for mouse welfare and scientific outcomes. 390 complete responses were received and analysed quantitatively and thematically. We found that 35% report using tail handling only, and 43% use a combination of tail and non-aversive methods. 18% of respondents reported exclusively using non-aversive methods. The vast majority of participants were convinced that non-aversive handling improves animal welfare and scientific outcomes. However, the survey indicated that researchers were significantly less likely to have heard of non-aversive handling and more likely to use tail handling compared with animal care staff. Thematic analysis revealed there were concerns regarding the time required for non-aversive methods compared with tail handling, and that there was a perceived incompatibility of tunnel handling with restraint, health checks and other routine procedures. Respondents also highlighted a need for additional research into the impact of handling method that is representative of experimental protocols and physiological indicators used in the biomedical fields. This survey highlights where targeted research, outreach, training and funding may have the greatest impact on increasing uptake of non-aversive handling methods for laboratory mice.