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Rapid diagnosis Development of viral honeybee disease using Multiplex PCR
Ha-Na Jung,Mi-Sun Yoo,Young-Ha Kim,Jin-Hyeong Noh,Nam-Hee Kim,Kondreddy Eswar Reddy,Suk-Chan Jung,Seung-Won Kang 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.10
Virus infections of the honeybee(Apis mellifera) have been increasingly investigated during the last decade. In general, honeybee viruses are widespread and most of them persist as inapparent infections. We screened honeybee colonies for the presence of several bee viruses, including deformed wing virus(DWV), black queen virus(BQCV), Kashmir bee virus(KBV), Israeli acute paralysis virus (IAPV), sacbrood virus(SBV), acute bee paralysis virus(ABPV), using uniplex RT-PCR. Frequently simultaneous infections with different viruses are diagnosed in seemingly healthy bee colonies. Therefore we developed a multiplex RT-PCR assay for the simultaneous detection of multiple bee viruses.
Responses of Varroa-resistant honey bees (Apis mellifera L.) to Deformed wing virus
Kitiphong Khongphinitbunjong,Lilia I. de Guzman,Thomas E. Rinderer,Matthew R. Tarver,Amanda M. Frake,Yanping Chen,Panuwan Chantawannakul 한국응용곤충학회 2016 Journal of Asia-Pacific Entomology Vol.19 No.4
The negative impact of Deformedwing virus (DWV) on European honey bees Apis mellifera is magnified by Varroa destructor parasitism. This study compared the responses of two Varroa-resistant honey bee stocks, pure Russian honey bees (RHB) and out-crossed Varroa Sensitive Hygienic bees, Pol-line (POL) to DWV infection to that of Varroa- susceptible stock, Italian honey bees (IHB). Two-day-old larvae were fed with DWV lysate in different concentrations: undiluted DWV lysate (D1), D1:100, and D1:1000. The unfed larvae served as negative control. Combs containing test larvae were exposed to a common environment during their development using host colonies. Our results showed that only POL displayed variation in DWV levels when fed different DWV concentrations. POL fed highest concentration of DWV inoculum had the highest increase in DWV level than those fed low concentrations and unfed POL. This high increase in DWV level probably contributed to the decrease in the survival and median longevity (LT50) of D1-fed POL. Weights of newly eclosed D1-fed POL were similar to those of the two controls and DWV-fed bees. However, within IHB, D1-fed bees showed significant reductions in weight, days of survival and LT50. Regardless of the concentrations of DWV inoculum, the DWV levels were similarly low within RHB; adult bees had similar weights. Overall, larvae fed D1 had the highest rate of wing deformation. POL and RHB had numerically lower proportions of bees with deformed wings. This study suggests that RHB showed some degree of resistant to DWV as shown by no reduction on weight and numerically lower proportion of wing deformity when compared with the other bee stocks.
Reddy, Kondreddy Eswar,Noh, Jin Hyeong,Yoo, Mi-Sun,Kim, Young-Ha,Kim, Nam-Hee,Doan, Huong Thi Thanh,Ramya, Mummadireddy,Jung, Suk-Chan,Van Quyen, Dong,Kang, Seung-Won Elsevier 2013 Veterinary microbiology Vol.167 No.3
<P><B>Abstract</B></P> <P>Deformed wing virus (DWV) is one of the most common viral infection in honeybees. Phylogenetic trees were constructed for 16 partial nucleotide sequences of the structural polyprotein region and the RNA helicase region of South Korean DWVs. The sequences were compared with 10 previously reported DWV sequences from different countries and the sequences of two closely related viruses, Kakugo virus (KGV) and <I>Varroa destructor</I> virus-1 (VDV-1). The phylogeny based on these two regions, the Korean DWV genomes were highly conserved with 95–100% identity, while they also shared 93–97% similarity with genotypes from other countries, although they formed a separate cluster. To investigate this phenomenon in more detail, the complete DWV genome sequences of Korea-1 and Korea-2 were determined and aligned with six previously reported complete DWV genome sequences from different countries, as well as KGV and VDV-1, and a phylogenetic tree was constructed. The two Korean DWVs shared 96.4% similarity. Interestingly, the Korea-2 genome was more similar to the USA (96.5%) genome than the Korea-1. The Korean genotypes highly conserved with USA (96%) but low similarity with the United Kingdom3 (UK3) genome (89%). The end of the 5′ untranslated region (UTR), the start of the open reading frame (ORF) region, and the 3′ UTR were variable and contained several substitutions/transitions. This phenomenon may be explained by intramolecular recombination between the Korean and other DWV genotypes.</P>
Occurrence of seven honeybee viruses in Korean apiaries
Yong Soo Choi,Myeong Lyeol Lee,Man Young Lee,Hye Kyung Kim,Kwang Gill Lee 한국응용곤충학회 2008 한국응용곤충학회 학술대회논문집 Vol.2008 No.10
Mortality of honeybees is a serious problem that beekeepers have to face periodically in Korea and worldwide. The presence of RNA viruses, in addition to other pathogens may be one of its possible causes. In this work, we detected Deformed wing virus(DWV), Israle Acute Paralysis Virus (IAPV), Black queen cell virus (BQCV), Cloudy wing virus(CWV), Kashmir bee virus(KBV), Sacbrood virus(SBV), Chronic bee paralysis virus(CBPV) in samples of korea honeybees with or without Varroa destructor and Nosema apis. The detection of viruses in all provinces, simultaneous co-infection of colonies by several viruses and the fact that 96.3% of the samples were infected with one or more virus, indicates they are widely spread in the region. Using uniplex and multiplex RT-PCR we screened honey bee colonies for the presence of several bee viruses, including DWV, IAPV, BQCV, KBV, CWV, and described the detection of mixed virus infections in bees from these colonies. Conclusively, investigated disease of the bee, and confirmed new virus that lead to bee disease, this is thought by valuable thing as data for development of beekeeping industry such as CCD(Colony Collapse Disorder)'s cause searching examination.
Joo-Seong Lee,Byoung-Su Yoon 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.10
Deformed wing virus(DWV) is one of infectious disease of honey bee that is caused to wings of immature or mutation and death at last. In this study 4 kinds of polyprotein of DWV are compared and then selected 2 kinds of polyprotein, RNA-dependent RNA polymerase(RdRP) and peptidase C3G, which has relatively higher homology than others. Analysis of both RdRP region and peptidase C3G of DWV deposited in Genbank of NCBI showed 99~100% and 95~97% of homology on genomic level, respectively whereas analysis of CRPV-capsid and RNA helicase showed 86~98% and 89~98% of homology, repectively. According to the result of gene analysis, two kinds of polyprotein are constructed and analyzed the homology, resulting in RdRP and Peptidase -C3G showed about 96% and 97% of homology, respectively. It indicated that both region is able to use for generation of specific antibody for the diagnosis of Deformed Wing Virus (DWV).
Diagnosis of deformed wing virus (DWV) from bumblebee(Bombus terriestris L.) in Korea
Yong Soo Choi,Man Young Lee,Hye Kyung Kim,Kyu-Ho Byeon,Kwang Gill Lee,Hyung Joo Yoon 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.10
Deformed wing virus (DWV) of honeybees (Apis mellifera) is closely associated with characteristic wing deformities, abdominal bloating, paralysis, and rapid mortality of emerging adult bees. Here, we report for the first time the occurrence of DWV-infected bumble bees (Bombus terrestris). For the present study, the detection of DWV virus from the adult bumble bee, death adult bumble bee, mail bumble bee, pupa and larva to the infection cycle was investigated in the same colony. Using specific RT-PCR protocols for the detection of DWV followed by sequencing of the PCR products we could demonstrate that the bumble bees were indeed infected with DWV. The virus was detected from diseased insects, and its partial DWV gene was cloned and sequenced. The partial DWV gene encoding the polyprotein is 711-nt of 235 amino acid residues. The deduced nucleotide sequence of the polyprotein partial gene of DWV showed 96.9%, 96.2%, 96.8%, and 96.5% homology to other structure polyprotein partial gene of DWV, respectively. Phylogenetic analysis further conformed that the deduced nucleotide sequence of the polyprotein partial gene of DWV divided to the outside tree. We describe the first time that presence of Deformed wing virus(DWV) from bumble bee(Bombus terrestris) in korea using RT-PCR.
First detection of deformed wing virus(DWV) from bumblebee (Bombus ignitus)
Yong Soo Choi,Man Young Lee,Hye Kyung Kim,Nam-Suk Kim,In-Pyo Hong,Kyu-Ho Byeon,Kwang Gill Lee,Hyung Joo Yoon 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.10
We report for the first time the occurrence of DWV-infected bumble bees (Bombus ignitus). For the present study, the detection of DWV virus from the female and male bumble bee was investigated in the same colony. The Deformed wing virus (DWV) of honeybee (Apis mellifera) is closely associated with characteristic wing deformities, abdominal bloating, paralysis, and rapid mortality of emerging adult bees. Using specific RT-PCR protocols for the detection of DWV followed by sequencing of the PCR products we could demonstrate that the bumble bees were indeed infected with DWV. The virus was detected from Bombus ignitus, and its partial DWV gene was cloned and sequenced. The partial DWV gene encoding the polyprotein is 711-nt of 235 amino acid residues. The deduced nucleotide sequence of the polyprotein partial gene of DWV showed 96.9%, 96.2%, 96.8%, and 96.5% homology to other structure polyprotein partial gene of DWV from insects, respectively. Phylogenetic analysis further conformed that the deduced nucleotide sequence of the polyprotein partial gene of DWV divided to the outside tree. We describe the first time that presence of Deformed wing virus(DWV) from bumble bee(Bombus terrestris) in korea using RT-PCR.