Quantitative Real-Time Reverse Transcription-PCR Analysis of Deformed Wing Virus Infection in the Honeybee (Apis mellifera L.)

ABSTRACT Deformed wing virus (DWV) can cause wing deformity and premature death in adult honeybees, although like many other bee viruses, DWV generally persists as a latent infection with no apparent symptoms. Using reverse transcription (RT)-PCR and Southern hybridization, we detected DWV in all life stages of honeybees, including adults with and without deformed wings. We also found DWV in the parasitic mite Varroa destructor, suggesting that this mite may be involved in the transmission of DWV. However, the detection of the virus in life stages not normally associated with mite parasitism (i.e., eggs and larvae) suggests that there are other modes of transmission. The levels of DWV in different life stages of bees were investigated by using TaqMan real-time quantitative RT-PCR. The amounts of virus varied significantly in these different stages, and the highest levels occurred in pupae and in adult worker bees with deformed wings. The variability in virus titer may reflect the different abilities of bees to resist DWV infection and replication. The epidemiology of DWV is discussed, and factors such as mite infestation, malnutrition, and climate are also considered.

[1]  J. Pettis,et al.  Transmission of Kashmir bee virus by the ectoparasitic mite Varroa destructor , 2004 .

[2]  Y. P. Chen,et al.  Quantitation of a Glyptapanteles indiensis polydnavirus gene expressed in parasitized host, Lymantria dispar, by real-time quantitative RT-PCR. , 2003, Journal of virological methods.

[3]  R. Farkas,et al.  Detection of acute bee paralysis virus by RT-PCR in honey bee and Varroa destructor field samples: rapid screening of representative Hungarian apiaries , 2002 .

[4]  J. Evans.  Genetic evidence for coinfection of honey bees by acute bee paralysis and Kashmir bee viruses. , 2001, Journal of invertebrate pathology.

[5]  S. Martin.  The role of Varroa and viral pathogens in the collapse of honeybee colonies: a modelling approach , 2001 .

[6]  S. Davison,et al.  Detection of Acute Bee Paralysis Virus and Black Queen Cell Virus from Honeybees by Reverse Transcriptase PCR , 2001, Applied and Environmental Microbiology.

[7]  Wolfgang Ritter,et al.  Sacbrood Virus of the Honeybee (Apis mellifera): Rapid Identification and Phylogenetic Analysis Using Reverse Transcription-PCR , 2001, Clinical Diagnostic Laboratory Immunology.

[8]  M. Pépin,et al.  Detection of chronic honey bee (Apis mellifera L.) paralysis virus infection: application to a field survey , 2000 .

[9]  C. Peng,et al.  Nucleotide sequence variations in Kashmir bee virus isolated from Apis mellifera L. and Varroa jacobsoni Oud. , 2000 .

[10]  A. C. Hung,et al.  A scientific note on the detection of Kashmir bee virus in individual honeybees and Varroa jacobsoni mites , 1999 .

[11]  I. Fries,et al.  Virus infections in Nordic honey bee colonies with no, low or severe Varroa jacobsoni infestations , 1999 .

[12]  Gunn,et al.  The transmission of deformed wing virus between honeybees (Apis mellifera L.) by the ectoparasitic mite varroa jacobsoni Oud , 1999, Journal of invertebrate pathology.

[13]  S. Martin,et al.  A scientific note on Varroa jacobsoni Oudemans and the collapse of Apis mellifera L. colonies in the United Kingdom , 1998 .

[14]  B. Ball,et al.  The incidence and world distribution of honey bee viruses , 1996 .

[15]  B. Ball,et al.  Characterisation and serological relationships of strains of Kashmir bee virus , 1995 .

[16]  G. Sisson,et al.  Molecular diagnosis of Kashmir bee virus infection , 1995 .

[17]  R. Abramson,et al.  Detection of specific polymerase chain reaction product by utilizing the 5'----3' exonuclease activity of Thermus aquaticus DNA polymerase. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[18]  L. Bailey,et al.  2 – THE HONEY BEE , 1991 .

[19]  B. Ball,et al.  Viruses in honey bee colonies infested with Varroa jacobsoni: first findings in Yugoslavia. , 1990 .

[20]  B. Ball,et al.  The prevalence of pathogens in honey bee (Apis mellifera) colonies infested with the parasitic mite Varroa jacobsoni , 1988 .

[21]  J. Antoniw,et al.  The detection of acute paralysis virus in Varroa jacobsoni by the use of a simple indirect ELISA. , 1986 .

[22]  Denis Anderson,et al.  A comparison of serological techniques for detecting and identifying honeybee viruses , 1984 .

[23]  L. Bailey,et al.  Egypt Bee Virus and Australian Isolates of Kashmir Bee Virus , 1979 .

[24]  L. Bailey,et al.  Three previously undescribed viruses from the honey bee. , 1974, The Journal of general virology.

[25]  L. Bailey,et al.  Acute infection of bees with paralysis virus , 1964 .

[26]  A. Gibbs,et al.  TWO VIRUSES FROM ADULT HONEY BEES (APIS MELLIFERA LINNAEUS). , 1963, Virology.