The harlequin ladybird, Harmonia axyridis, is one of the most successful invasive insect species worldwide. We investigated whether (i) chill coma recovery time (CCRt) changes during the ontogenetic development of this species, (ii) CCRt varies in response to repeated cold shocks, and (iii) CCRt could be a good predictor of winter survival ability in adults. CCRt decreased during larval development, the lowest CCRt values were observed in teneral adults (one and four days old), and significantly higher values were observed for older adults (16 and 32 days old). Repeated cold shocks (two hours at -3 °C), interrupted by short (30 min) warm periods (22 °C) resulted in decreased CCRt after the second cold shock, probably depicting an acclimation response, but then CCRt increased with additional cold shocks, likely revealing the accumulation of chill injuries. The CCRt of pre-overwintering individuals was not correlated with their winter survival. This indicates that CCRt is not a reliable measure of cold tolerance in H. axyridis. However, this result could be partially affected by the experimental setup - the use of laboratory-reared individuals who experienced standardized conditions and thus the variability in CCRt of tested animals could be much lower than the variability present in nature. The substantial variation observed over the ontogenetic development of H. axyridis poses important methodological implications for future studies, as animals of the same stage/age should be compared with each other. The observed U-shaped response to repeated cold shocks indicates that the expectation of linearity between cold exposure and insect response is an oversimplification of real situations.

Department of Ecology Faculty of Environmental Sciences Czech University of Life Sciences Prague Kamýcká 129 Prague Suchdol 16500 Czech Republic

University of Rennes 1 UMR 6553 EcoBio CNRS 263 Avenue du Gal Leclerc 35000 Rennes Cedex France; Institut Universitaire de France 1 Rue Descartes 75231 Paris Cedex 05 France

University of Rennes CNRS ECOBIO UMR 6553 Station Biologique de Paimpont 35380 Paimpont France

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