# Mouse aversion to induction with isoflurane using the drop method

## TL;DR

This study finds that lower concentrations of isoflurane (1.7% to 2.7%) using the drop method reduce aversion in mice compared to higher concentrations.

## Contribution

The study identifies an optimal isoflurane concentration range (1.7% to 2.7%) for minimizing mouse aversion during anesthesia induction using the drop method.

## Key findings

- Aversion increased with higher isoflurane concentrations (1.7% to 3.7%) in mice.
- Mice showed the least aversion at 1.7% isoflurane and increased aversion at 2.7% and 3.7%.
- The study provides evidence for using lower isoflurane concentrations to refine mouse handling protocols.

## Abstract

Isoflurane anesthesia prior to carbon dioxide euthanasia is recognized as a refinement by many guidelines. Facilities lacking access to a vaporizer can use the “drop” method, whereby liquid anesthetic is introduced into an induction chamber. Knowing the least aversive concentration of isoflurane is critical. Previous work has demonstrated that isoflurane administered with the drop method at a concentration of 5% is aversive to mice. Other work has shown that lower concentrations (1.7% to 3.7%) of isoflurane can be used to anesthetize mice with the drop method, but aversion to these concentrations has not been tested. We assessed aversion to these lower isoflurane concentrations administered with the drop method, using a conditioned place aversion (CPA) paradigm. Female C57BL/6J (OT-1) mice (n = 28) were randomly allocated to one of three isoflurane concentrations: 1.7%, 2.7%, and 3.7%. Mice were acclimated to a light–dark apparatus. Prior to and following dark (+ isoflurane) and light chamber conditioning sessions, mice underwent an initial and final preference assessment; the change in the duration spent within the dark chamber between the initial and final preference tests was used to calculate a CPA score. Aversion increased with increasing isoflurane concentration: from 1.7% to 2.7% to 3.7% isoflurane, mean ± SE CPA score decreased from 19.6 ± 20.1 s to –25.6 ± 23.2 s, to –116.9 ± 30.6 s (F1,54 = 15.4, p < 0.001). Our results suggest that, when using the drop method to administer isoflurane, concentrations between 1.7% and 2.7% can be used to minimize female mouse aversion to induction.