Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter October 15, 2017

Varroa destructor induces changes in the expression of immunity-related genes during the development of Apis mellifera worker and drone broods

Ewa A. Zaobidna, Krystyna Żółtowska and Elżbieta Łopieńska-Biernat
From the journal Acta Parasitologica


The ectoparasitic mite Varroa destructor has emerged as the major pest of honeybees. Despite extensive research efforts, the pathogenesis of varroosis has not been fully explained. Earlier studies suggested that V. destructor infestation leads to the suppression of the host’s immune system. The aim of this study was to analyze the immune responses of 14 genes in the Toll signal transduction pathways, including effector genes of antimicrobial peptides (AMPs), in developing Apis mellifera workers and drones infested with V. destructor. Four developmental stages (L5 larvae, prepupae, and 2 pupal stages) and newly emerged imagines were analyzed. In workers, the most significant changes were observed in L5 larvae in the initial stages of infestation. A significant increase in the relative expression of 10 of the 14 analyzed genes, including defensin-1 and defensin-2, was observed in infested bees relative to non-infested individuals. The immune response in drones developed at a slower rate. The expression of genes regulating cytoplasmic signal transduction increased in prepupae, whereas the expression of defensin-1 and defensin-2 effector genes increased in P3 pupae with red eyes. The expression of many immunity-related genes was silenced in successive life stages and in imagines, and it was more profound in workers than in drones. The results indicate that V. destructor significantly influences immune responses regulated by the Toll signal transduction pathway in bees. In infested bees, the observed changes in Toll pathway genes varied between life stages and the sexes.


This research was supported by grant No. 2013/11/N/NZ9/00053 from the Polish National Science Centre.


Alaux C., Dantec C., Parrinello H., Le Conte Y. 2011. Nutrigenomics in honey bees: digital gene expression analysis of pollen’s nutritive effects on healthy and varroa-parasitized bees. BMC Genomics, 12, 496. 10.1186/1471-2164-12-496Search in Google Scholar PubMed

Antúnez K., Mendoza Y., Santos E., Invernizzi C. 2013. Differential expression of vitellogenin in honey bees (Apis mellifera) with different degrees of Nosema ceranae infection. Journal of Apicultural Research, 52, 227–234. 10.3896/IBRA. in Google Scholar

Aronstein K.A., Saldivar E., Vega R., Westmiller S., Douglas A.E. 2012. How Varroa parasitism affects the immunological and nutritional status of the honey bee, Apis mellifera. Insects, 3, 601–615. 10.3390/insects3030601Search in Google Scholar PubMed

Boncristiani H., Underwood R., Schwarz R., Evans J.D., Pettis J., vanEngelsdorp D. 2012. Direct effect of acaricides on pathogen loads and gene expression levels of honey bee Apismellifera. Journal of Insect Physiology, 58, 613–620. 10.1016/j.jinsphys.2011.12.011Search in Google Scholar PubMed

Bowen-Walker P.L., Gunn A. 2001. The effect of the ectoparasitic mite, Varroa destructor on adult worker honeybee (Apis mellifera) emergence weights, water, protein, carbohydrate, and lipid levels. Entomologia Experimentalis et Applicata, 101, 207–217. 10.1046/j.1570-7458.2001.00905.xSearch in Google Scholar

Brutscher L.M., Daughenbaugh K.F., Flenniken M.L. 2015. Antiviral defense mechanisms in honey bees. Current Opinion in Insect Science, 10, 71–82. 10.101B/j.cois.2015.04.016.Search in Google Scholar PubMed

Burgett M., Rucker R.R., Thurman W.N. 2004. Economics and honey bee pollination markets. American Bee Journal, 144, 269–271Search in Google Scholar

Chen Y.P., Siede R. 2007. Honey bee viruses. Advances in Virus Research, 70, 33–80. 10.1016/S0065-3527(07)70002-7Search in Google Scholar PubMed

Cytryńska M. 2009. Immunity without antibodies. Postępy Biologii Komórki, 36, 309–324. 10.2478/v10052-009-0003-9Search in Google Scholar

Danihlík J., Aronstein K., Petřivalský M. 2015 Antimicrobial peptides: a key component of honey bee innate immunity. Journal of Apicultural Reearch, 54, 123–136. 10.1080/00218839.2015.1109919Search in Google Scholar

De Jong D. 1997. Mites: Varroa and other parasites of brood. In: (Eds. Morse, R.A., Flottum, K.) Honey Bee Pests, Predators and Diseases. A.I. Root Company, Medina, 278–327.Search in Google Scholar

Dietemann V., Neumann P., Ellis J.D. 2013. The Coloss Beebook volume 1: Standard methods for Apis mellifera research. Journal of Apicultural Research, 52. 10.3896/IBRA. in Google Scholar

Di Prisco G., Annoscia D., Margiotta M., Ferrara R., Varricchio P., Zanni V., et al. 2016. A mutualistic symbiosis between a parasite mite and a pathogenic virus undermines honey bee immunity and health. Proceedings of the National Academy of Sciences 113, 3203–3208. 10.1073/pnas.1523515113Search in Google Scholar PubMed PubMed Central

Ellis J.D. 2001. The future of Varroa control: Integrating current treatments with the latest advancements. American Bee Journal, 141, 127–131Search in Google Scholar

Erler S., Popp M., Lattorff H.M.G. 2011. Dynamics of immune system gene expression upon bacterial challenge and wounding in social insect (Bombus terrestris). PLoS One, 6, e1826. 10.1371/journal.pone.0018126Search in Google Scholar PubMed PubMed Central

Evans J.D., Spivak M. 2010. Socialized medicine: individual and communal disease barriers in honey bees. Journal of Invertebrate Pathology, 103, 62–72. 10.1016/j.jip.2009.06.019Search in Google Scholar PubMed

Evans J.D., Aronstein K., Chen Y.P., Hetru C., Imler J.L., Jiang H., et al. 2006. Immune pathways and defence mechanisms in honey bees Apis mellifera. Insect Molecular Biology, 15, 645–656. 10.1111/j.1365-2583.2006.00682.x.Search in Google Scholar PubMed PubMed Central

Frączek R., Żółtowska K,. Lipiński Z. Dmitryjuk M. 2013. The mutual influence of proteins from Varroa destructor extracts and from honeybee haemolymph on their proteolytic activity – in vitro study. Acta Parasitologica, 58, 317–323. 10.2478/s11686-013-0144-8Search in Google Scholar PubMed

Gätschenberger H., Gimple O., Tautz J., Beier H. 2012. Honey bee drones maintain humoral immune competence throughout all life stages in the absence of vitellogenin production. Journal of Experimental Biology, 215, 1313–1322. 10.1242/jeb.065276Search in Google Scholar PubMed

Gätschenberger H., Gimple O., Tautz J., Beier H. 2012. Honey bee drone maintain humoral immune competence throughout all life stages in the absence of vitellogenin production. Journal Experimental Biology, 215, 1313–1322. 10.1242/jeb.065276Search in Google Scholar

Genersch E. 2010. Honey bee pathology: current threats to honey bees and beekeeping. Applied Microbiology and Biotechnology, 87: 87–97. 10.1007/s00253-010-2573-8Search in Google Scholar PubMed

Garedew A., Schmolz E., Lamrecht I. 2004. The energy and nutrition demand of the parasitic life of the mite Varroa destructor. Apidologie, 35: 419–430. 0.1051/apido:2004032.Search in Google Scholar

Gliński Z., Buczek K. 2003. Response of the Apoidea to fungal infections. Apiacta, 38, 183–189Search in Google Scholar

Goulson D., Nicholls E., Botias C., Rotheray E.L. 2015. Combined stress from parasites, pesticides and lack of flowers drives bee declines. Science, 347, 1255957. 10.1126/science.1255957Search in Google Scholar PubMed

Gregorc A., Evans J.D., Scharf M., Ellis J.D. 2012. Gene expression in honey bee (Apis mellifera) larvae exposed to pesticides and Varroa mites (Varroa destructor). Journal of Insect Physiology, 58, 1042–1049. 10.1016/j.jinsphys.2012.03.015Search in Google Scholar PubMed

Gregory P.G., Evans J.D., Rinderer T.E., De Gruzman L. 2005. Conditional immune-gene suppression of honeybees parasitized by Varroa mites. Journal of Insect Science, 5, 1–5. in Google Scholar

Hamiduzzaman M.M., Sinia A., Guzman-Novoa E., Goodwin P. 2012. Entomopathogenic fungi as potential biocontrol agents of ecto-parasitic mite, Varroa destructor, and their effect on the immune response of honey bees (Apis mellifera L.). Journal of Invertebrate Pathology, 111, 237–243. 10.1016/j.jip.2012.09.001Search in Google Scholar PubMed

Jay C.S. 1963. The development of honeybees in the their cells. Journal Apiculture Researe, 2, 117–134. 10.1080/00218839.1963.11100072Search in Google Scholar

Kanbar G., Engels W. 2005. Communal use of integumental wounds in honey bee (Apis mellifera) pupae multiply infested by the ectoparasitic mite Varroa destructor. Genetics and Molecular Research, 4, 465–472Search in Google Scholar

Khongphinitbunjong K., de Guzman L.I., Tarver M.R., Rinderer T.E., Chen Y., Chantawannakul P. 2015. Differential viral levels and immune gene expression in three stocks of Apis mellifera induced by different numbers of Varroa destructor. Journal of Insect Physiology, 72, 28–34. 10.1016/j.jinsphys.2014.11.005Search in Google Scholar PubMed

Kuster R.D., Boncristiani H.F., Rueppell O. 2014. Immunogene and viral transcript dynamics during parasitic Varroa destructor mite infection of developing honey bee (Apis mellifera) pupae. The Journal of Experimental Biology, 217, 1710–1718. 10.1242/jeb.097766Search in Google Scholar PubMed

Lemaitre B., Hoffmann J.A. 2007. The host defense of Drosophila melanogaster. Annual Review of Immunology, 25, 697–743. 10.1146/annurev.immunol.25.022106.141615Search in Google Scholar PubMed

Li W.F., Ma G.X., Zhou X.X. 2006. Apidaecin-type peptides: biodiversity, structurefunction relationships and mode of action. Peptides, 27, 2350–2359. 10.1016/j.peptides.2006.03.016Search in Google Scholar PubMed

Lourenço A.P., Guidugli – Lazzarini K.R., Freitas F.C., Bitondi M.M., Simoes Z.L. 2013. Bacterial infection activates the immune system response and dysregulates microRNA expression in honey bees. Insect Biochemistry and Molecular Biology, 43, 474–82. 10.1016/j.ibmb.2013.03.001Search in Google Scholar PubMed

Moore P.A., Wilson M.E., Skinner J.A. 2016. Honey Bee Viruses, the Deadly Varroa Mite Associates. Extension Bee Health CoP,–bee-viruses-the-deadly-varroa-mite-associates#.VIcA08ma8UZSearch in Google Scholar

Navajas M., Migeon A., Alaux C., Martin-Magniette M.L., Robinson G.E., Evans J.D., Cros-Arteil S., Crauser D., Le Conte Y. 2008. Differential gene expression of the honey bee Apis mellifera associated with Varroa destructor infection. BMC Genomics, 9, 301. 10.1186/1471-2164-9-301Search in Google Scholar PubMed PubMed Central

Nazzi F., Le Conte Y. 2017. Ecology of Varroa destructort, the major ectoparasite of the Western honey bee, Apis mellifera. Annual Review of Entomology, 61, 417–432. 10.1146/annurevento-010715-023731Search in Google Scholar

Nazzi F., Brown S.P., Annoscia D., Del Piccolo F., Di Prisco G., Varricchio P., Vedova G.D., Cattonaro F., Caprio E., Pennacchio F. 2012. Synergistic parasite–pathogen interactions mediated by host immunity can drive the collapse of honeybee colonies. PLoS Pathogens, 8, e1002735. 10.1371/journal.ppat.1002735Search in Google Scholar PubMed PubMed Central

Otvos L.J., Insung O., Rogers M.E., Consolvo P.J., Condie B.A., Lovas S., Bulet P., Blaszczyk-Thurin M. 2000. Interaction between heat shock proteins and 973 antimicrobial peptides. Biochemistry, 39, 14150–1415910.1021/bi0012843Search in Google Scholar PubMed

Pfaffl M.W. 2001. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Research, 29, e45. 10.1093/nar/29.9.e45.Search in Google Scholar PubMed PubMed Central

Randolt K., Gimple O., Geissendorfer J., Reinders J., Prusko C., Mueller M.J., Albert S., Tautz J., Beier H. 2008. Immune-related proteins induced in the hemolymph after aseptic and septic injury differ in honey bee worker larvae and adults. Archives of Insect Biochemistry and Physiology, 69, 155–167. 10.1002/arch.20269Search in Google Scholar PubMed

Rosenkranz P., Aumeier P., Ziegelmann B. 2010. Biology and control of Varroa destructor. Journal of Invertebrate Pathology, 103, 96–119. 10.1016/j.jip.2009.07.016.Search in Google Scholar PubMed

Sammataro D., Gerson U., Needham G. 2000. Parasitic mites of honey bees: life history, implications and impact. Annual Review of Entomology, 45, 519–548. 10.1146/annurev.ento.45.1.519Search in Google Scholar PubMed

Strachecka A., Borsuk G., Paleolog J., Olszewski K., Chobotow J. 2012. Antipathogenic activity on the body surface of adult workers Apis mellifera. Veterinary Medicine-Science and Practice, 68, 290–292Search in Google Scholar

Strachecka A., Borsuk G., Olszewski K., Paleolog J., Lipiński Z. 2013. Proteolysis on the body surface of pyrethroid-sensitive and resistant Varroa destructor. Acta Parasitologica 58, 64–69. 10.2478/s11686-013-0109-ySearch in Google Scholar PubMed

Yang X., Cox-Foster D.L. 2007. Effects of parasitization by Varroa destructor on survivorship and physiological traits of Apis mellifera in correlation with viral incidence and microbial challenge. Parasitology, 134, 405–412. 10.1017/S0031182006000710Search in Google Scholar PubMed

Yang X., Cox-Foster D.L. 2005. Impact of an ectoparasite on the immunity and pathology of an invertebrate: evidence for host immunosuppression and viral amplification. Proceedings of the National Academy of Sciences, 102, 7470–7475. 10.1073/pnas.0501860102Search in Google Scholar PubMed PubMed Central

Zaobidna E.A., Żółtowska K., Łopieńska-Biernat E. 2015. Expression of the prophenoloxidase gene and phenoloxidase activity, during the development of Apis mellifera brood infected with Varroa destructor. Journal of Apicultural Science, 59, 85–93. 10.1515/JAS-2015-00Search in Google Scholar

Received: 2017-4-12
Revised: 2017-8-2
Accepted: 2017-8-3
Published Online: 2017-10-15
Published in Print: 2017-12-20

© 2017 W. Stefański Institute of Parasitology, PAS