Investigation of Anesthesia Techniques in Paper Kite Butterflies (Idea Leuconoe)

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Despite the frequency of invertebrates in research, zoological, and personal collections, sedation and anesthesia in these species remains poorly studied. This study assessed anesthetic techniques in paper kite butterflies (Idea leuconoe). Thirty-six recently emerged butterflies were individually placed in 1.9 L air-tight containers and randomly exposed to one of six treatments (n = 6/group): 5% isoflurane via vaporizer (2 L min–1, fraction of inspired oxygen (FiO2) 1.0); (V), 0.5 mL (CB0.5) or 3.0 mL (CB3) liquid isoflurane on a cotton ball (FiO2 0.21); 100% carbon dioxide via flowmeter (2 L min–1) (CO2); cooling at 2.78°C, or no treatment (control) (CT). Three min (V, CB0.5, CB3, CO2) or 10 min (°C) following recumbency, butterflies were removed from exposure, manually manipulated, and monitored for return to upright posture. Times to recumbency, return to upright, and response to manipulation were recorded. Collected data were compared between groups (significance P < 0.05). Except in CT, all butterflies achieved recumbency. Median (range) time to recumbency was 1.3 (0.5–3.6), 2.8 (1.6–4.6), 1.7 (0.9–2.9), 1.2 (0.8–2.2), and 1.4 (1.0–3.5) min in V, CB0.5, CB3, CO2, and °C, respectively (P = 0.106). Median (range) time to upright was 9.3 (0.7–15.2), 13.8 (6.0–24.0), 14.9 (8.2–26.6), 1.3 (0.9–9.0), and 9.6 (3.6–30.0) min in V, CB0.5, CB3, CO2, and °C, respectively (P = 0.019). All butterflies recovered uneventfully. This is the first peer-reviewed evaluation of anesthetic techniques in butterflies.