Effects of field-realistic doses of glyphosate on honeybee appetitive behaviour

Glyphosate (GLY) is a broad-spectrum herbicide used for weed control. The sub-lethal impact of GLY on non-target organisms such as insect pollinators has not yet been evaluated. Apis mellifera is the main pollinator in agricultural environments and is a well-known model for behavioural research. Honeybees are also accurate biosensors of environmental pollutants and their appetitive behavioural response is a suitable tool with which to test sub-lethal effects of agrochemicals. We studied the effects of field-realistic doses of GLY on honeybees exposed chronically or acutely to the herbicide. We focused on sucrose sensitivity, elemental and non-elemental associative olfactory conditioning of the proboscis extension response (PER), and foraging-related behaviour. We found a reduced sensitivity to sucrose and learning performance for the groups chronically exposed to GLY concentrations within the range of recommended doses. When olfactory PER conditioning was performed with sucrose reward with the same GLY concentrations (acute exposure), elemental learning and short-term memory retention decreased significantly compared with controls. Non-elemental associative learning was also impaired by an acute exposure to GLY traces. Altogether, these results imply that GLY at concentrations found in agro-ecosystems as a result of standard spraying can reduce sensitivity to nectar reward and impair associative learning in honeybees. However, no effect on foraging-related behaviour was found. Therefore, we speculate that successful forager bees could become a source of constant inflow of nectar with GLY traces that could then be distributed among nestmates, stored in the hive and have long-term negative consequences on colony performance.

[1]  M. Lindauer,et al.  Über die Fluggeschwindigkeit der Bienen und über ihre Richtungsweisung bei Seitenwind , 2004, Naturwissenschaften.

[2]  J. Erber,et al.  The effect of genotype on response thresholds to sucrose and foraging behavior of honey bees (Apis mellifera L.) , 1998, Journal of Comparative Physiology A.

[3]  H. L. Nixon,et al.  Food transmission within the honeybee community , 1952, Proceedings of the Royal Society of London. Series B - Biological Sciences.

[4]  J. Erber,et al.  Tactile learning and the individual evaluation of the reward in honey bees (Apis mellifera L.) , 1999, Journal of Comparative Physiology A.

[5]  R. Menzel,et al.  The flight paths of honeybees recruited by the waggle dance , 2005, Nature.

[6]  N. Koeniger The biology of the honey bee , 1988, Insectes Sociaux.

[7]  John P. Giesy,et al.  Ecotoxicological Risk Assessment for Roundup ® Herbicide , 2000 .

[8]  C. Thom,et al.  The Scent of the Waggle Dance , 2007, PLoS biology.

[9]  O. Rueppell,et al.  Aging without functional senescence in honey bee workers , 2007, Current Biology.

[10]  Robert E. Page,et al.  Levels of behavioral organization and the evolution of division of labor , 2002, Naturwissenschaften.

[11]  R. Morse The Dance Language and Orientation of Bees , 1994 .

[12]  G. H. Lunney,et al.  USING ANALYSIS OF VARIANCE WITH A DICHOTOMOUS DEPENDENT VARIABLE: AN EMPIRICAL STUDY , 1970 .

[13]  S. Williamson,et al.  Exposure to multiple cholinergic pesticides impairs olfactory learning and memory in honeybees , 2013, Journal of Experimental Biology.

[14]  K. Takeda Classical conditioned response in the honey bee , 1961 .

[15]  Cathy Hawes,et al.  Effects on weed and invertebrate abundance and diversity of herbicide management in genetically modified herbicide-tolerant winter-sown oilseed rape , 2005, Proceedings of the Royal Society B: Biological Sciences.

[16]  J. Springett,et al.  Effect of repeated low doses of biocides on the earthworm Aporrectodea caliginosa in laboratory culture , 1992 .

[17]  F. James Rohlf,et al.  Biometry: The Principles and Practice of Statistics in Biological Research , 1969 .

[18]  H. Thompson,et al.  Evaluating exposure and potential effects on honeybee brood (Apis mellifera) development using glyphosate as an example , 2014, Integrated environmental assessment and management.

[19]  Joaquín Goyret,et al.  Non-random nectar unloading interactions between foragers and their receivers in the honeybee hive , 2005, Naturwissenschaften.

[20]  Dean G. Thompson,et al.  Fate of glyphosate in a Canadian forest watershed. 1. Aquatic residues and off-target deposit assessment. , 1990 .

[21]  H. Lachnit,et al.  Configural olfactory learning in honeybees: negative and positive patterning discrimination. , 2001, Learning & memory.

[22]  C. Grüter,et al.  Propagation of olfactory information within the honeybee hive , 2006, Behavioral Ecology and Sociobiology.

[23]  M. Giurfa,et al.  Early olfactory experience induces structural changes in the primary olfactory center of an insect brain , 2012, The European journal of neuroscience.

[24]  V. S. Rao Principles of Weed Science , 2000 .

[25]  J. Lydon,et al.  Oxadiazon Activity is Similar to That of p-Nitro-Diphenyl Ether Herbicides , 1989, Weed Science.

[26]  Christina M Howe,et al.  Toxicity of glyphosate‐based pesticides to four North American frog species , 2004, Environmental toxicology and chemistry.

[27]  W. Kirchner,et al.  Honeybee dance communication , 1988, Naturwissenschaften.

[28]  Elizabeth Phillips,et al.  The Site , 1988, Palestine Is Throwing a Party and the Whole World Is Invited.

[29]  Jia-Jia Ou,et al.  Global pesticide consumption and pollution: with China as a focus , 2011 .

[30]  M. Winston The Biology of the Honey Bee , 1987 .

[31]  R. Menzel,et al.  Revisiting olfactory classical conditioning of the proboscis extension response in honey bees: A step toward standardized procedures , 2012, Journal of Neuroscience Methods.

[32]  W. H. Kirchner 17. Mad-Bee-Disease? Subletale Effekte von Imidacloprid (Gaucho®) auf das Verhalten von Honigbienen , 1999 .

[33]  R. Relyea The Lethal Impacts of Roundup and Predatory Stress on Six Species of North American Tadpoles , 2005, Archives of environmental contamination and toxicology.

[34]  M. Giurfa Behavioral and neural analysis of associative learning in the honeybee: a taste from the magic well , 2007, Journal of Comparative Physiology A.

[35]  M. Giurfa,et al.  Early olfactory experience modifies neural activity in the antennal lobe of a social insect at the adult stage , 2009, The European journal of neuroscience.

[36]  M. Bitterman,et al.  Classical conditioning of proboscis extension in honeybees (Apis mellifera). , 1983, Journal of comparative psychology.

[37]  F. A. Christian,et al.  Effect of Glyphosate on the Development of Pseudosuccinea columella Snails , 1997 .

[38]  L. G. Goldsborough,et al.  Effect of glyphosate (Roundup® formulation) on periphytic algal photosynthesis , 1988, Bulletin of environmental contamination and toxicology.

[39]  Jack T. Trevors,et al.  Glyphosate in the environment , 1988, Water, Air, and Soil Pollution.

[40]  Christoph Grüter,et al.  Past Experiences Affect Interaction Patterns Among Foragers and Hive‐Mates in Honeybees , 2009 .

[41]  Marcelo A. Aizen,et al.  Expansión de la soja y diversidad de la agricultura argentina , 2009 .

[42]  N. Amrhein,et al.  The Site of the Inhibition of the Shikimate Pathway by Glyphosate: II. INTERFERENCE OF GLYPHOSATE WITH CHORISMATE FORMATION IN VIVO AND IN VITRO. , 1980, Plant physiology.

[43]  J. Nieh,et al.  A nicotinic acetylcholine receptor agonist affects honey bee sucrose responsiveness and decreases waggle dancing , 2012, Journal of Experimental Biology.

[44]  M. S. Balbuena,et al.  Honeybee recruitment to scented food sources: correlations between in-hive social interactions and foraging decisions , 2011, Behavioral Ecology and Sociobiology.

[45]  W. Farina,et al.  Associative Learning during Early Adulthood Enhances Later Memory Retention in Honeybees , 2009, PloS one.

[46]  M. Giurfa Cognitive neuroethology: dissecting non-elemental learning in a honeybee brain , 2003, Current Opinion in Neurobiology.

[47]  C. Grüter,et al.  The honeybee waggle dance: can we follow the steps? , 2009, Trends in ecology & evolution.

[48]  W. Farina,et al.  Age and rearing environment interact in the retention of early olfactory memories in honeybees , 2008, Journal of Comparative Physiology A.

[49]  R. Relyea THE IMPACT OF INSECTICIDES AND HERBICIDES ON THE BIODIVERSITY AND PRODUCTIVITY OF AQUATIC COMMUNITIES , 2006 .

[50]  R. Menzel,et al.  Neonicotinoids Interfere with Specific Components of Navigation in Honeybees , 2014, PloS one.

[51]  W. Farina,et al.  Floral scents learned inside the honeybee hive have a long-lasting effect on recruitment , 2012, Animal Behaviour.

[52]  Nestor A. Schmajuk,et al.  Classical conditioning , 2008, Scholarpedia.

[53]  R. Menzel Memory dynamics in the honeybee , 1999, Journal of Comparative Physiology A.