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doi:10.1006/nlme.2000.3995 
Copyright © 2001 Academic Press. All rights reserved.
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Contrasting Effects of Imidacloprid on Habituation in 7- and 8-Day-Old Honeybees (Apis mellifera)*1
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David Guezb, a, f2, Séverine Suchaila, Monique Gauthierc, Ryszard Maleszkab and Luc P. Belzuncesa, f2
Available online 26 February 2002.
Abstract
We examined the effects of sublethal doses (0.1, 1, and 10 ng per animal) of a new neonicotinoid insecticide, Imidacloprid, on habituation of the proboscis extension reflex (PER) in honeybees (Apis mellifera) reared under laboratory conditions. In untreated honeybees, the habituation of the proboscis extension reflex is age-dependent and there is a significant increase in the number of trials required for habituation in older bees (8–10 days old) as compared to very young bees (4–7 days old). Imidacloprid alters the number of trials needed to habituate the honeybee response to multiple sucrose stimulation. In 7-day-old bees, treatment with Imidacloprid leads to an increase in the number of trials necessary to abolish the response, whereas in 8-day-old bees, it leads to a reduction in the number of trials for habituation (15 min and 1 h after treatment), and to an increase 4 h after treatment. The temporal effects of Imidacloprid in both 7- and 8-day-old bees suggest that 4h after treatment the observed effects are due to a metabolite of Imidacloprid, rather than to Imidacloprid itself. Our results suggest the existence of two distinct subtypes of nicotinic receptors in the honeybee that have different affinities to Imidacloprid and are differentially expressed in 7- and 8-day-old individuals.
Author Keywords: neonicotinoid; chloronicotinyl; nicotinic acetylcholine receptor; Apis mellifera; nonassociative learning; behavior; Hymenoptera.
*1 This study was conducted in the honeybee indoor facilities of the Research School of Biological Sciences, The Australian National University in Canberra. D.G was supported in part by CIFRE agreement between INRA and Bayer AG. We thank Paul Helliwell, Carol Conway, and Nick Potter for their friendly help.
f2 Address correspondence and reprint requests to Luc P. Belzunces or David Guez, Laboratoire de Toxicologie Environnementale, INRA, Unité de Zoologie, Site Agroparc, 84914 Avignon Cedex 9, France. Fax: +33 (0)432 72 2602. E-mail: belzunce@avignon.inra.fr; guez@rsbs.anu.edu.au.
