Stress indicator gene expression profiles, colony dynamics and tissue development of honey bees exposed to sub-lethal doses of imidacloprid in laboratory and field experiments
[en] In this study, different context-dependent effects of imidacloprid exposure on the honey bee
response were studied. Honey bees were exposed to different concentrations of imidacloprid
during a time period of 40 days. Next to these variables, a laboratory-field comparison
was conducted. The influence of the chronic exposure on gene expression levels was determined
using an in-house developed microarray targeting different immunity-related and
detoxification genes to determine stress-related gene expression changes. Increased levels
of the detoxification genes encoding, CYP9Q3 and CYT P450, were detected in imidacloprid-
exposed honey bees. The different context-dependent effects of imidacloprid exposure
on honey bees were confirmed physiologically by decreased hypopharyngeal gland sizes.
Honey bees exposed to imidacloprid in laboratory cages showed a general immunosuppression
and no detoxification mechanisms were triggered significantly, while honey bees infield
showed a resilient response with an immune stimulation at later time points. However,
the treated colonies had a brood and population decline tendency after the first brood cycle
in the field. In conclusion, this study highlighted the different context-dependent effects of
imidacloprid exposure on the honey bee response. These findings warn for possible pitfalls
concerning the generalization of results based on specific experiments with short exposure
times. The increased levels of CYT P450 and CYP9Q3 combined with an immune response
reaction can be used as markers for bees which are exposed to pesticides in the field.
Disciplines :
Entomology & pest control
Author, co-author :
De Smet, Lina; Université de Gand > Faculté des Sciences, > Laboratoire d'Entomologie Moleculaire et Pathologie des abeilles
Hatjina; Hellenic Agricultural Organisation `DEMETER', Nea Moudania, Greece > Institute of Animal Science > Division of Apiculture
Ioannidis, Pavlos; Hellenic Agricultural Organisation `DEMETER', Nea Moudania, Greece > Institute of Animal Science > Division of Apiculture
Hamamtzoglou, Anna; Hellenic Agricultural Organisation `DEMETER', Nea Moudania, Greece > Institute of Animal Science > Division of Apiculture
Schoonvaere, Karel; Ghent University, Ghent > Faculty of Sciences > Laboratory of Molecular Entomology and Bee Pathology
Francis, Frédéric ; Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Entomologie fonctionnelle et évolutive
Meeus, Ivan; Ghent University > Faculty of Bioscience Engineering > Department of Crop Protection
Smagghe, Guy; Ghent University > Faculty of Bioscience Engineering > Department of Crop Protection
de Graaf, Dirk C.; Ghent University > Faculty of Sciences > Laboratory of Molecular Entomology and Bee Pathology
Language :
English
Title :
Stress indicator gene expression profiles, colony dynamics and tissue development of honey bees exposed to sub-lethal doses of imidacloprid in laboratory and field experiments
Publication date :
09 February 2017
Journal title :
PLoS ONE
eISSN :
1932-6203
Publisher :
Public Library of Science, United States - California
Belzunces LP, Tchamitchian S, Brunet JL. Neural effects of insecticides in the honey bee. Apidologie. 2012; 43(3):348-70.
Krupke CH, Hunt GJ, Eitzer BD, Andino G, Given K. Multiple routes of pesticide exposure for honey bees living near agricultural fields. Plos One. 2012; 7(1).
Cresswell JE, Page CJ, Uygun MB, Holmbergh M, Li Y, Wheeler JG, et al. Differential sensitivity of honey bees and bumble bees to a dietary insecticide (imidacloprid). Zoology (Jena). 2012; 115(6):365-71.
Decourtye A, Devillers J, Cluzeau S, Charreton M, Pham-Delegue MH. Effects of imidacloprid and del-tamethrin on associative learning in honey bees under semi-field and laboratory conditions. Ecotoxicol Environ Saf. 2004; 57(3):410-9. doi: 10.1016/j.ecoenv.2003.08.001 PMID: 15041263
Blacquiere T, Smagghe G, van Gestel CA, Mommaerts V. Neonicotinoids in bees: a review on concentrations, side-effects and risk assessment. Ecotoxicology. 2012; 21(4):973-92. doi: 10.1007/s10646-012-0863-x PMID: 22350105
Sandrock C, Tanadini M, Tanadini LG, Fauser-Misslin A, Potts SG, Neumann P. Impact of chronic neo-nicotinoid exposure on honey bee colony performance and queen supersedure. PLoS One. 2014; 9(8): e103592. doi: 10.1371/journal.pone.0103592 PMID: 25084279
Khoury DS, Myerscough MR, Barron AB. A quantitative model of honey bee colony population dynamics. PLoS One. 2011; 6(4):e18491. doi: 10.1371/journal.pone.0018491 PMID: 21533156
Winston M. The biology of the honey bee. In: Havard University Press, USA. pp. 281.1987.
Wu-Smart J, Spivak M. Sub-lethal effects of dietary neonicotinoid insecticide exposure on honey bee queen fecundity and colony development. Sci Rep. 2016; 6:32108. doi: 10.1038/srep32108 PMID: 27562025
Skerl MIS, Gregorc A. Heat shock proteins and cell death in situ localisation in hypopharyngeal glands of honey bee (Apis mellifera carnica) workers after imidacloprid or coumaphos treatment. Apidologie. 2010; 41(1):73-86.
Hatjina F, Papaefthimiou C, Charistos L, Dogaroglu T, Bouga M, Emmanouil C, et al. Sub-lethal doses of imidacloprid decreased size of hypopharyngeal glands and respiratory rhythm of honey bees in vivo. Apidologie. 2013; 44(4):467-80.
Crailsheim K, Stolberg E. Influence of diet, age and colony condition upon intestinal proteolytic activity and size of the hypopharyngeal glands in the honeybee (Apis mellifera L). J Insect Physiol. 1989; 35(8):595-602.
Boncristiani H, Underwood R, Schwarz R, Evans JD, Pettis J, van Engelsdorp D. Direct effect of acaricides on pathogen loads and gene expression levels in honey bees Apis mellifera. J Insect Physiol. 2012; 58(5):613-20. doi: 10.1016/j.jinsphys.2011.12.011 PMID: 22212860
Garrido PM, Antunez K, Martin M, Porrini MP, Zunino P, Eguaras MJ. Immune-related gene expression in nurse honey bees (Apis mellifera) exposed to synthetic acaricides. J Insect Physiol. 2013; 59(1):113-9. doi: 10.1016/j.jinsphys.2012.10.019 PMID: 23147024
Gregorc A, Evans JD, Scharf M, Ellis JD. Gene expression in honey bee (Apis mellifera) larvae exposed to pesticides and Varroa mites (Varroa destructor). J Insect Physiol. 2012; 58(8):1042-9. doi: 10.1016/j.jinsphys.2012.03.015 PMID: 22497859
Aufauvre J, Misme-Aucouturier B, Vigues B, Texier C, Delbac F, Blot N. Transcriptome analyses of the honeybee response to Nosema ceranae and insecticides. Plos One. 2014; 9(3).
Stork A. Residues of 14C-NTN 33893 (Imidacloprid) in blossoms of sunflowers (Helianthus annuus) after seed dressing. Unpublished report no. MR-550/99.1999.
Bonmatin JM, Moineau I, Charvet R, Fleche C, Colin ME, Bengsch ER. A LC/APCI-MS/MS method for analysis of imidacloprid in soils, in plants, and in pollens. Anal Chem. 2003; 75(9):2027-33. doi: 10.1021/ac020600b PMID: 12720336
Bonmatin JM, Marchand PA, Charvet R, Moineau I, Bengsch ER, Colin ME. Quantification of imidacloprid uptake in maize crops. J Agr Food Chem. 2005; 53(13):5336-41.
Chauzat MP, Martel AC, Cougoule N, Porta P, Lachaize J, Zeggane S, et al. An assessment of honeybee colony matrices, Apis mellifera (Hymenoptera: Apidae) to monitor pesticide presence in continental France. Environ Toxicol Chem. 2011; 30(1):103-11. doi: 10.1002/etc.361 PMID: 20853451
Tapparo A, Giorio C, Marzaro M, Marton D, Solda L, Girolami V. Rapid analysis of neonicotinoid insecticides in guttation drops of corn seedlings obtained from coated seeds. J Environ Monit. 2011; 13(6):1564-8. doi: 10.1039/c1em10085h PMID: 21509402
Costa C, Buchler R, Berg S, Bienkowska M, Bouga M, Bubalo D, et al. A europe-wide experiment for assessing the impact of genotype-environment interactions on the vitality and performance of honey bee colonies: experimental design and trait evaluation. J Apic Sci. 2012; 56(1):147-58.
Delaplane KS, van der Steen J, Guzman-Novoa E. Standard methods for estimating strength parameters of Apis mellifera colonies. J Apicult Res. 2013; 52(1).
Berger B, Carmargo Abdalla F. Programmed cell death during ovarian differentiation in queens of Apis mellifera linne, 1758 (Hymenoptera, Apini). Braz J Morphol Sci. 2005; 22:4.
Rhodes J, Somerrville D. Introduction and early performance of queen bees. Report: Rural Industires Research & Development Corporation, NSW Agriculture. 03.049. 2003.
Johnson RM, Evans JD, Robinson GE, Berenbaum MR. Changes in transcript abundance relating to colony collapse disorder in honey bees (Apis mellifera). Proc Natl Acad Sci USA. 2009; 106(35):14790-5. doi: 10.1073/pnas.0906970106 PMID: 19706391
De Smet L, Ravoet J, Wenseleers T, de Graaf DC. Expression of key components of the RNAi machinery are suppressed in Apis mellifera that suffer a high virus infection. Entomological Science. 2016. Epub september 2016.
Ritchie ME, Phipson B, Wu D, Hu Y, Law CW, Shi W, et al. limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res. 2015; 43(7):e47. doi: 10.1093/nar/gkv007 PMID: 25605792
Smyth GK. Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Stat Appl Genet Mol Biol. 2004; 3:Article3.
Benjamini Y, Hochberg Y. Controlling the False Discovery Rate-a Practical and Powerful Approach to Multiple Testing. J Roy Stat Soc B Met. 1995; 57(1):289-300.
Hellemans J, Mortier G, De Paepe A, Speleman F, Vandesompele J. qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data. Genome Biol. 2007; 8(2):R19. doi: 10.1186/gb-2007-8-2-r19 PMID: 17291332
Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002; 3(7):RESEARCH0034. PMID: 12184808
Guedes RN, Smagghe G, Stark JD, Desneux N. Pesticide-Induced Stress in Arthropod Pests for Optimized Integrated Pest Management Programs. Annu Rev Entomol. 2016; 61:43-62. doi: 10.1146/annurev-ento-010715-023646 PMID: 26473315
Williams GR, Troxler A, Retschnig G, Roth K, Yanez O, Shutler D, et al. Neonicotinoid pesticides severely affect honey bee queens. Sci Rep. 2015; 5:14621. doi: 10.1038/srep14621 PMID: 26459072
Laycock I, Lenthall KM, Barratt AT, Cresswell JE. Effects of imidacloprid, a neonicotinoid pesticide, on reproduction in worker bumble bees (Bombus terrestris). Ecotoxicology. 2012; 21(7):1937-45. doi: 10.1007/s10646-012-0927-y PMID: 22614036
Bloch G, Francoy TM, Wachtel I, Panitz-Cohen N, Fuchs S, Mazar A. Industrial apiculture in the Jordan valley during Biblical times with Anatolian honeybees. Proc Natl Acad Sci USA. 2010; 107(25):11240-4. doi: 10.1073/pnas.1003265107 PMID: 20534519
Cresswell JE, Robert FX, Florance H, Smirnoff N. Clearance of ingested neonicotinoid pesticide (imidacloprid) in honey bees (Apis mellifera) and bumblebees (Bombus terrestris). Pest Manag Sci. 2014; 70(2):332-7. doi: 10.1002/ps.3569 PMID: 23633150
Mommaerts V, Reynders S, Boulet J, Besard L, Sterk G, Smagghe G. Risk assessment for side-effects of neonicotinoids against bumblebees with and without impairing foraging behavior. Ecotoxicology. 2010; 19(1):207-15. doi: 10.1007/s10646-009-0406-2 PMID: 19757031
Williamson SM, Wright GA. Exposure to multiple cholinergic pesticides impairs olfactory learning and memory in honeybees. J Exp Biol. 2013; 216(Pt 10):1799-807. doi: 10.1242/jeb.083931 PMID: 23393272
Wu JY, Smart MD, Anelli CM, Sheppard WS. Honey bees (Apis mellifera) reared in brood combs containing high levels of pesticide residues exhibit increased susceptibility to Nosema (Microsporidia) infection. J Invertebr Pathol. 2012; 109(3):326-9. doi: 10.1016/j.jip.2012.01.005 PMID: 22285445
Pettis JS, Lichtenberg EM, Andree M, Stitzinger J, Rose R, Vanengelsdorp D. Crop Pollination Exposes Honey Bees to Pesticides Which Alters Their Susceptibility to the Gut Pathogen Nosema ceranae. Plos One. 2013; 8(7).
Suryanarayanan S. Balancing Control and Complexity in Field Studies of Neonicotinoids and Honey Bee Health. Insects. 2013; 4(1):153-67. doi: 10.3390/insects4010153 PMID: 26466800
Knecht D, Kaatz HH. Patterns of larval lood-production by hypopharyngeal glands in adult worker honey-bees. Apidologie. 1990; 21(5):457-68.
Lass A, Crailsheim K. Influence of age and caging upon protein metabolism, hypopharyngeal glands and trophallactic behavior in the honey bee (Apis mellifera L). Insect Soc. 1996; 43(4):347-58.
Hrassnigg N, Crailsheim K. The influence of brood on the pollen consumption of worker bees (Apis mellifera L.). J Insect Physiol. 1998; 44(5-6):393-404. PMID: 12770157
Malone LA, Todd JH, Burgess EPJ, Christeller JT. Development of hypopharyngeal glands in adult honey bees fed with a Bt toxin, a biotin-binding protein and a protease inhibitor. Apidologie. 2004; 35(6):655-64.
Deseyn J, Billen J. Age-dependent morphology and ultrastructure of the hypopharyngeal gland of Apis mellifera workers (Ymenoptera, Apidae). Apidologie. 2005; 36(1):49-57.
Wilde J, Frazek R, Siuda M, Bak B, Hatjina F, A M. The influence of sub-lethal doses of imidacloprid on protein content and proteolytic activity in honey bees (Apis mellifera L). Journal of Apicultural Research. 2016.
Henry M, Beguin M, Requier F, Rollin O, Odoux JF, Aupinel P, et al. A common pesticide decreases foraging success and survival in honey bees. Science. 2012; 336(6079):348-50. doi: 10.1126/science.1215039 PMID: 22461498
Boily M, Sarrasin B, DeBlois C, Aras P, Chagnon M. Acetylcholinesterase in honey bees (Apis mellifera) exposed to neonicotinoids, atrazine and glyphosate: laboratory and field experiments. Environ Sci Pollut R. 2013; 20(8):5603-14.
Feyereisen R. Evolution of insect P450. Biochem Soc T. 2006; 34:1252-5.
Feyereisen R. Insect P450 enzymes. Annu Rev Entomol. 1999; 44:507-33. doi: 10.1146/annurev.ento.44.1.507 PMID: 9990722
Li XC, Schuler MA, Berenbaum MR. Molecular mechanisms of metabolic resistance to synthetic and natural xenobiotics. Annu Rev Entomol. 2007; 52:231-53. doi: 10.1146/annurev.ento.51.110104.151104 PMID: 16925478
Mao W, Rupasinghe SG, Johnson RM, Zangerl AR, Schuler MA, Berenbaum MR. Quercetin-metabolizing CYP6AS enzymes of the pollinator Apis mellifera (Hymenoptera: Apidae). Comp Biochem Physiol B Biochem Mol Biol. 2009; 154(4):427-34. doi: 10.1016/j.cbpb.2009.08.008 PMID: 19737624
Johnson RM, Mao WF, Pollock HS, Niu GD, Schuler MA, Berenbaum MR. Ecologically appropriate xenobiotics induce cytochrome P450s in Apis mellifera. Plos One. 2012; 7(2).
Derecka K, Blythe MJ, Malla S, Genereux DP, Guffanti A, Pavan P, et al. Transient exposure to low levels of insecticide affects metabolic networks of honey bee larvae. Plos One. 2013; 8(7).
Mao WF, Schuler MA, Berenbaum MR. CYP9Q-mediated detoxification of acaricides in the honey bee (Apis mellifera). Proc Natl Acad Sci USA. 2011; 108(31):12657-62. doi: 10.1073/pnas.1109535108 PMID: 21775671
Pettis JS, van Engelsdorp D, Johnson J, Dively G. Pesticide exposure in honey bees results in increased levels of the gut pathogen Nosema. Naturwissenschaften. 2012; 99(2):153-8. doi: 10.1007/s00114-011-0881-1 PMID: 22246149
Brandt A, Gorenflo A, Siede R, Meixner M, Buchler R. The neonicotinoids thiacloprid, imidacloprid, and clothianidin affect the immunocompetence of honey bees (Apis mellifera L.). J Insect Physiol. 2016; 86:40-7. doi: 10.1016/j.jinsphys.2016.01.001 PMID: 26776096
