Detection of Three honeybee Viruses in Palestine using RT- PCR

Journal of the Institute of Science and Technology

Viruses infecting honey bees are diseases that cause infection and economic damage in every developmental stage of honey bees all over the world. In this study, a survey was conducted to determine the presence and prevalence of two worldwide common honeybee viruses (deformed wing virus [DWV] and chronic bee paralysis virus [CBPV]). 128 apiaries from 9 different localities in the province of Bingöl were visited to determine viral RNA. Collected 384 honey bee samples were tested molecularly (reverse-transcriptase polymerase chain reaction, RTPCR) using genome-specific primers specific to each virus. In molecular tests, 28 of 128 apiaries (21.87%) gave a positive reaction for DWV, but no CBPV pathogen was detected in any apiary. The 711 bp nucleotide sequence revealed by cloning one of the randomly selected positive samples was registered in the gene bank with the accession number MZ357973. According to the BLASTn analysis in the NCBI database, the nucleotide sequence of the DWV agent ...

Acta Veterinaria, 2014

In this study 55 honey bee colonies from different Serbian regions were monitored for the presence of Deformed Wing Virus (DWV) and Acute Bee Paralysis Virus (ABPV) using TaqMan-based real-time RT-PCR assay. The results revealed the presence of DWV in each sampling location, and ABPV in 10 out of 11 apiaries. High frequency of DWV (76.4%) and ABPV (61.8%) positive samples in asymptomatic colonies can be the consequence of ineffi cient and postponed Varroa treatment concerning the role of this mite in the transmission and activation of honey bee viruses. The real-time RT-PCR technique described in this paper is proved to be the most reliable method for this kind of investigation.

Apidologie, 2014

The prevalence and regional distribution patterns of seven different honeybee viruses, i.e. Acute bee paralysis virus (ABPV), Sacbrood virus (SBV), Black queen cell virus (BQCV), Deformed wing virus (DWV), Kashmir bee virus (KBV), Chronic bee paralysis virus (CBPV), and Israeli acute paralysis virus (IAPV) were investigated using reverse transcription PCR (RT-PCR) analyses. Bee samples were collected in 82 apiaries located in 20 different districts of Croatia, mainly from bee colonies with various health problems. 9.75 % of investigated samples (apiaries) proved to be infected with CBPV, while ABPV, BQCV, SBV, and DWV were found in 10.97 %, 29.26 %, 40.24 %, and 95.12 % of apiaries, respectively. No samples tested positive for KBV and IAPV. Thus, the prevalences of BQCV, ABPV, SBV, and CBPV were generally lower in Croatia compared to other European Union member states, whereas the prevalence of DWV was similarly high. The possible reasons for these differences are discussed. This study provides the first molecular evidence for the presence of honeybee viruses in Croatian honeybees.

Applied and Environmental Microbiology, 2006

Transmission mechanisms of six honeybee viruses, including acute bee paralysis virus (ABPV), black queen cell virus (BQCV), chronic bee paralysis virus (CBPV), deformed wing virus (DWV), Kashmir bee virus (KBV), and sacbrood bee virus (SBV), in honey bee colonies were investigated by reverse transcription-PCR (RT-PCR) methods. The virus status of individual queens was evaluated by examining the presence of viruses in the queens' feces and tissues, including hemolymph, gut, ovaries, spermatheca, head, and eviscerated body. Except for head tissue, all five tissues as well as queen feces were found to be positive for virus infections. When queens in bee colonies were identified as positive for BQCV, DWV, CBPV, KBV, and SBV, the same viruses were detected in their offspring, including eggs, larvae, and adult workers. On the other hand, when queens were found positive for only two viruses, BQCV and DWV, only these two viruses were detected in their offspring. The presence of viruses in the tissue of ovaries and the detection of the same viruses in queens' eggs and young larvae suggest vertical transmission of viruses from queens to offspring. To our knowledge, this is the first evidence of vertical transmission of viruses in honeybee colonies.

Received on 8/3/2011 and Accepted for Publication on 8/8/2012. ABSTRACT The Reverse Transcription-polymerase Chain Reaction (RT-PCR) is an excellent technique for the detection of honeybee viruses. In this study, the presence of Sac Brood Virus (SBV) and Black Queen Cell Virus (BQCV) was demonstrated in 100 samples, collected from Jordanian honeybee colonies by employing RT-PCR. The collected samples represented infected (depopulation, paralysis, or dark coloring), dead, and apparently healthy honeybees of different developmental stages (adult and larvae). SBV was detected in 37% of the samples, whereas BQCV was detected in 5%. Nucleotide sequences of the PCR products from each virus was determined and found to be 433 and 309 nucleotides in SBV and BQCV, respectively. The identities to the Gene Bank were 95% for SBV and 91% for BQCV. This is the first record of BQCV in Jordan.

Apidologie, 2002

A two years survey was undertaken to determine the occurrence of Acute Bee Paralysis Virus (ABPV) in field samples of adult bees and the parasitic mite Varroa destructor. To detect the viral nucleic acid we used polymerase chain reaction following reverse transcription (RT-PCR). We demonstrated the presence of ABPV RNA in 14 from 114 seemingly healthy colony samples collected from Hungarian honey bees. The investigation revealed that two third of the apiaries were infected with ABPV at a 12.2% infection rate. In seven other apiaries out of eight investigated (87.5%) the presence of the virus was detected from colonies following a sudden collapse; these colonies were simultaneously infected with Nosema apis or infested with Varroa destructor. Virus specific nucleic acid was also identified in the mites collected from two apiaries falling into the latter category. The amplicon of RT-PCR was sequenced and the nucleic acid sequence was aligned to the complete ABPV sequence deposited in the GeneBank database revealing a 93% homology.

Jordan Journal of Agricultural Sciences, 2013

The Reverse Transcription-polymerase Chain Reaction (RT-PCR) is an excellent technique for the detection of honeybee viruses. In this study, the presence of Sac Brood Virus (SBV) and Black Queen Cell Virus (BQCV) was demonstrated in 100 samples, collected from Jordanian honeybee colonies by employing RT-PCR. The collected samples represented infected (depopulation, paralysis, or dark coloring), dead, and apparently healthy honeybees of different developmental stages (adult and larvae). SBV was detected in 37% of the samples, whereas BQCV was detected in 5%. Nucleotide sequences of the PCR products from each virus was determined and found to be 433 and 309 nucleotides in SBV and BQCV, respectively. The identities to the Gene Bank were 95% for SBV and 91% for BQCV. This is the first record of BQCV in Jordan.

Insects

Honey bees are essential for crop and wild plant pollination. However, many countries have reported high annual colony losses caused by multiple possible stressors. Diseases, particularly those caused by viruses, are a major cause of colony losses. However, little is known about the prevalence of honey bee pathogens, particularly virus prevalence, in Egyptian honey bees. To address this shortfall, we determined the prevalence of widespread bee viruses in honey bee colonies in Egypt—whether it is affected by geography, the season, or infestation with Varroa destructor (varroa) mites. Honey bee worker samples were collected from 18 geographical regions across Egypt during two seasons: winter and summer of 2021. Three apiaries were chosen in each region, and a pooled sample of 150 worker bees was collected from five colonies in each apiary then screened by qPCR for 10 viral targets: acute bee paralysis virus (ABPV), black queen cell virus (BQCV), chronic bee paralysis virus (CBPV), def...

Apidologie, 2008

Samples of adult honey bees from apiaries with unusually high winter mortality and brood from hives with symptoms of disease were tested for presence of acute bee paralysis virus (ABPV), black queen cell virus (BQCV), chronic bee paralysis virus (CBPV), deformed wing virus (DWV), Kashmir bee virus (KBV) and sacbrood virus (SBV) by RT-PCR. All six viruses were detected, but the frequencies varied significantly: SBV was detected in 78 apiaries, DWV in 55, ABPV in 11, CBPV in 4, BQCV in 1 and KBV in 1. This is the first record of KBV in Denmark. A large majority of the bee samples were infected with one or more viruses. Single, dual and triple infections were observed. Nucleotide sequences of the PCR products from each virus were determined and found to be 98-99% identical to GenBank accessions except CBPV, which was only 88-90% identical to known CBPV sequences.

Journal of Apicultural Science, 2018

The recent alarming loss of honey bee colonies around the world is believed to be related to the presence of viruses. The aim of this study was to detect two major viral diseases, Apis mellifera Filamentous virus (AmFV) and Israeli Acute Paralysis Virus (IAPV) using Reverse Transcription-Polymerase Chain Reaction RT-PCR, in honey bees in Mexico. Adult and larvae honey bee samples were collected from asymptomatic colonies of six major beekeeping regions in the state of Chihuahua, Mexico. Both viruses were detected in both developmental stages of honey bees, IAPV at a higher prevalence (23.5%) as compared to AmFV, only in 0.9% of samples. However, this is the first report on AmFV infection in Mexican apiaries. Further studies are required to understand the AmFV and IAPV impact on colony loss in Mexico and to develop strategies for enhancing the control of viral diseases.