Identification of deformed wing virus (DWV) in honeybee pupae by RT-PCR

Document Type : Full Research Paper

Authors

1 Department of Honey Bee, Silk Worm and Wildlife Reseach, Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.

2 Islamic Azad University, Karaj Branch, Karaj, Iran and Hemmati, B., Islamic Azad University, Karaj Branch, Karaj, Iran.

Abstract

The virus is widely distributed and prevalent in colonies infested by the mite. Using RT-PCR based assays, the virus infections in honey bees can be detected and quantified in a rapid and accurate manner. Infected honeybees from colonies suffering with symptoms of depopulation, sudden collapse, paralysis, and varroa infestation, used in this study, originated from 23 provinces of Iran. Bee samples were collected between July - September 2011 and 2012. Altogether, samples from 156 apiaries were collected and submitted for virus screening. From each apiary, 100-500 adult worker bees were sampled. RNA extraction and RT-PCR were performed with QIAGEN kits. The primers lead to a fragment of 434 bp. The PCR products were electrophoresed in a 1.2 % agarose gel (stained with ethidium bromide). Following the RT-PCR reaction with the specific primers, out of the 156 apiaries examined, 34 (21.8 %) were infected with DWV. This is the first study of DWV detection in Iranian apiaries. We have looked into the occurrence of the DWV and identified differences in the distribution of the virus in the collected samples from different geographic regions of Iran. The spread of V. destructor has been implicated in a dramatic increase in the prevalence of DWV and it is likely that this virus will become predominant in infested areas. The role of V. destructor in DWV transmission and the appearance of wing deformities in newly emerged bees also deserve more intensive investigation.

Keywords

Main Subjects


1.Allen M. and B. Ball. 1996. The incidence and world distribution of honey bee viruses. Bee World 77,141-162.
2.Anguiano-Baez R., E. Guzman-Novoa, L.G. Espinosa-Montaño and A. Correa-Benítez. 2016. Varroa destructor (Mesostigmata: Varroidae) parasitismand climate differentially influence the prevalence, levels, and overt infections of deformed wing virus in honey bees (Hymenoptera: Apidae). Journal of Insect Science 16.
3.Bailey L., B.V. BALL and J. Perry. 1981. The prevalence of viruses of honey bees in Britain. Annals of Applied Biology 97,109-118.
4.Bailey L., J. Carpenter and R. Woods. 1979. Egypt bee virus and Australian isolates of Kashmir bee virus. Journal of General Virology 43,641-647.
5.Berényi O., T. Bakonyi, I. Derakhshifar, H. Köglberger and N. Nowotny. 2006. Occurrence of six honeybee viruses in diseased Austrian apiaries. Applied and environmental microbiology 72,2414-2420.
6.Bowen-Walker P., S. Martin and A. Gunn. 1999. The Transmission of Deformed Wing Virus between Honeybees (Apis melliferaL.) by the Ectoparasitic Mite Varroa jacobsoni Oud. Journal of Invertebrate Pathology 73,101-106.
7.Brutscher L.M. and M.L. Flenniken. 2015. RNAi and antiviral defense in the honey bee. Journal of Immunology Research 2015.
8.Chen Y., J.S. Pettis, A. Collins and M.F. Feldlaufer. 2006. Prevalence and transmission of honeybee viruses. Applied and Environmental Microbiology 72,606-611.
9.Chen Y., I.B. Smith, A.M. Collins, J.S. Pettis and M.F. Feldlaufer. 2004. Detection of deformed wing virus infection in honey bees, Apis mellifera L., in the United States. American Bee Journal 144,557-559.
10.Francis R.M., S.L. Nielsen and P. Kryger. 2013. Patterns of viral infection in honey bee queens. Journal of General Virology 94,668-676.
11.Gisder S., P. Aumeier and E. Genersch. 2009. Deformed wing virus: replication and viral load in mites (Varroa destructor). Journal of General Virology 90,463-467.
12.Lanzi G., J.R. de Miranda, M.B. Boniotti, C.E. Cameron, A. Lavazza, L. Capucci, S.M. Camazine and C. Rossi. 2006. Molecular and biological characterization of deformed wing virus of honeybees (Apis mellifera L.). Journal of Virology 80,4998-5009.
13.Möckel N., S. Gisder and E. Genersch. 2011. Horizontal transmission of deformed wing virus: pathologicalconsequences in adult bees (Apis mellifera) depend on the transmission route. Journal of General Virology 92,370-377.
14. Nordström S., I. Fries, A. Aarhus, H. Hansen and S. Korpela. 1999. Virus infections in Nordic honey bee colonies with no, low or severe Varroa jacobsoni infestations. Apidologie 30,475-484.
15. Paxton R.J., J. Klee, S. Korpela and I. Fries. 2007. Nosema ceranae has infected Apis mellifera in Europe since at least 1998 and may be more virulent than Nosema apis. Apidologie 38,558-565.
16. Shah K.S. 2009. Localization of Deformed Wing Virus (DWV) in the Brains of Apis mellifera (European Honey Bees).
17. Tentcheva D., L. Gauthier, S. Jouve, L. Canabady-Rochelle, B. Dainat, F. Cousserans, M.E. Colin, B.V. Ball and M. Bergoin. 2004. Polymerase Chain Reaction detection of deformed wing virus (DWV) in Apis mellifera and Varroa destructor. Apidologie 35,431-439.
18. Tentcheva D., L. Gauthier, N. Zappulla, B. Dainat, F. Cousserans, M.E. Colin and M. Bergoin. 2004. Prevalence and seasonal variations of six bee viruses in Apis mellifera L. and Varroa destructor mite populations in France. Applied and Environmental Microbiology 70,7185-7191.
19. Yue C. and E. Genersch. 2005. RT-PCR analysis of Deformed wing virus in honeybees (Apis mellifera) and mites (Varroa destructor). Journal of General Virology 86, 3419-3424.