Genetic diversity of acute bee paralysis virus in Slovenian honeybee samples

2013 ◽  
Vol 61 (2) ◽  
pp. 244-256 ◽  
Author(s):  
Urška Jamnikar Ciglenečki ◽  
Ivan Toplak
Keyword(s):  
Genetic Diversity ◽  
Base Pair ◽  
Phylogenetic Trees ◽  
Nucleotide Diversity ◽  
Random Distribution ◽  
Geographical Location ◽  
Rt Pcr ◽  
Orf2 Region ◽  
Acute Bee Paralysis Virus ◽  
Paralysis Virus

The genetic diversity of acute bee paralysis virus (ABPV) in honeybees was studied in Slovenia. A total of 248 honeybee samples obtained from 134 different apiaries in Slovenia were tested for the presence of ABPV by RT-PCR. Specific 398-base pair (bp) products were generated with primers amplifying the ORF2 region and 452-base pair (bp) products with primers amplifying the ORF1 region of the viral genome. To characterise the overall nucleotide diversity among the ABPV sequences, phylogenetic trees with 54 and 29 samples were constructed from 357 nucleotides from ORF2 and 408 nucleotides from ORF1, respectively. The nucleotide comparison of Slovenian ABPV strains revealed two distinct clusters in ORF2 and ORF1, showing 91.2–92.5% and 96.7–97.2% nucleotide identity, respectively. Comparison of data regarding the geographical location of the ABPV-positive samples with the constructed phylogenetic trees revealed the random distribution of the two clusters throughout Slovenia.

scholarly journals A survey of deformed wing virus and acute bee paralysis virus in honey bee colonies from serbia using real-time RT-PCR

2014 ◽  
Vol 64 (1) ◽  
pp. 81-92 ◽  
Author(s):  
Predrag Simeunović ◽  
Jevrosima Stevanović ◽  
Dejan Vidanović ◽  
Jakov Nišavić ◽  
Dejan Radović ◽  
...  
Keyword(s):  
Real Time ◽  
Honey Bee ◽  
Sampling Location ◽  
Rt Pcr ◽  
Deformed Wing Virus ◽  
Bee Colonies ◽  
Acute Bee Paralysis Virus ◽  
Paralysis Virus ◽  
Bee Viruses

Abstract 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 inefficient and postponed Varroa treatment concerning the role of this mite in the transmission and activation of honey bee viruses. The real-time RTPCR technique described in this paper is proved to be the most reliable method for this kind of investigation.

scholarly journals Determination of Genetically Identical Strains of Four Honeybee Viruses in Bumblebee Positive Samples

Viruses ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 1310
Author(s):  
Ivan Toplak ◽  
Laura Šimenc ◽  
Metka Pislak Ocepek ◽  
Danilo Bevk
Keyword(s):  
Direct Sequencing ◽  
Rt Pcr ◽  
Black Queen Cell Virus ◽  
Queen Cell ◽  
Pcr Products ◽  
Acute Bee Paralysis Virus ◽  
Bee Virus ◽  
Paralysis Virus ◽  
High Diversity

In recent years, there has been growing evidence that certain types of honeybee viruses could be transmitted between different pollinators. Within a voluntary monitoring programme, 180 honeybee samples (Apis mellifera carnica) were collected from affected apiaries between 2007 and 2018. Also from August 2017 to August 2018, a total 148 samples of healthy bumblebees (Bombus lapidarius, B. pascuorum, B. terrestris, B. lucorum, B. hortorum, B. sylvarum, B. humilis) were collected at four different locations in Slovenia, and all samples were tested by using RT-PCR methods for six honeybee viruses. Direct sequencing of a total 158 positive samples (acute bee paralysis virus (ABPV n = 33), black queen cell virus (BQCV n = 75), sacbrood bee virus (SBV n = 25) and Lake Sinai virus (LSV n = 25)) was performed from obtained RT-PCR products. The genetic comparison of identified positive samples of bumblebees and detected honeybee field strains of ABPV, BQCV, SBV, and LSV demonstrated 98.74% to 100% nucleotide identity between both species. This study not only provides evidence that honeybees and bumblebees are infected with genetically identical or closely related viral strains of four endemically present honeybee viruses but also detected a high diversity of circulating strains in bumblebees, similar as was observed among honeybees. Important new genetic data for endemic strains circulating in honeybees and bumblebees in Slovenia are presented.

scholarly journals Phylogenetic Analysis of Acute Bee Paralysis Virus Strains

2002 ◽  
Vol 68 (12) ◽  
pp. 6446-6450 ◽  
Author(s):  
Tamás Bakonyi ◽  
Elvira Grabensteiner ◽  
Jolanta Kolodziejek ◽  
Miklós Rusvai ◽  
Gražyna Topolska ◽  
...  
Keyword(s):  
Phylogenetic Trees ◽  
Geographic Origin ◽  
The United States ◽  
Clinical Samples ◽  
Reverse Transcription Pcr ◽  
The United Kingdom ◽  
Pcr Assays ◽  
Acute Bee Paralysis Virus ◽  
Virus Strains ◽  
Paralysis Virus

ABSTRACT Reverse transcription-PCR assays have been established for a quick, sensitive, and specific diagnosis of acute bee paralysis virus (ABPV), a common virus of the honeybee (Apis mellifera), directly from clinical samples. A 3,071-nucleotide fragment of the ABPV genome, which includes the entire capsid polyprotein gene, was amplified from Austrian, German, Polish, and Hungarian ABPV samples and sequenced, and the sequences were compared. The alignment of a smaller fragment with ABPV sequences from the United States and the United Kingdom revealed nucleotide identity rates between 89 and 96%, respectively. Phylogenetic trees which display the molecular relationship between the viruses of different geographic origin were constructed.

scholarly journals Honey bee viruses in Serbian colonies of different strength

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5887 ◽  
Author(s):  
Dragan Cirkovic ◽  
Jevrosima Stevanovic ◽  
Uros Glavinic ◽  
Nevenka Aleksic ◽  
Spomenka Djuric ◽  
...  
Keyword(s):  
Honey Bee ◽  
Infection Intensity ◽  
Single Infection ◽  
Rt Pcr ◽  
Deformed Wing Virus ◽  
Epidemiological Surveys ◽  
The Relationship ◽  
Acute Bee Paralysis Virus ◽  
Paralysis Virus ◽  
Bee Viruses

Protection of honey bees is of great economic importance because of their role in pollination. Crucial steps towards this goal are epidemiological surveys of pathogens connected with honey bee losses. In this study deformed wing virus (DWV), chronic bee paralysis virus (CBPV), acute bee paralysis virus (ABPV) and sacbrood virus (SBV) were investigated in colonies of different strength located in five regions of Serbia. The relationship between colony strength and virus occurrence/infection intensity were assessed as well as the genetic relationship between virus sequences from Serbia and worldwide. Real-time RT-PCR analyses detected at least one virus in 87.33% of colonies. Single infection was found in 28.67% colonies (21.33%, 4.00%, 2.67% and 0.67% in cases of DWV, ABPV, SBV and CBPV, respectively). In the majority of colonies (58.66%) more than one virus was found. The most prevalent was DWV (74%), followed by ABPV, SBV and CBPV (49.30%, 24.00% and 6.70%, respectively). Except for DWV, the prevalence of the remaining three viruses significantly varied between the regions. No significant differences were found between colony strength and either (i) the prevalence of DWV, ABPV, SBV, CBPV and their combinations, or (ii) DWV infection levels. The sequences of honey bee viruses obtained from bees in Serbia were 93–99% identical with those deposited in GenBank.

scholarly journals Detection of Acute Bee Paralysis Virus and Black Queen Cell Virus from Honeybees by Reverse Transcriptase PCR

2001 ◽  
Vol 67 (5) ◽  
pp. 2384-2387 ◽  
Author(s):  
Mongi Benjeddou ◽  
Neil Leat ◽  
Mike Allsopp ◽  
Sean Davison
Keyword(s):  
Reverse Transcriptase ◽  
Pcr Primers ◽  
Rt Pcr ◽  
Black Queen Cell Virus ◽  
Silica Membrane ◽  
Blind Test ◽  
Reverse Transcriptase Pcr ◽  
Queen Cell ◽  
Acute Bee Paralysis Virus ◽  
Paralysis Virus

ABSTRACT A reverse transcriptase PCR (RT-PCR) assay was developed for the detection of acute bee paralysis virus (ABPV) and black queen cell virus (BQCV), two honeybee viruses. Complete genome sequences were used to design unique PCR primers within a 1-kb region from the 3′ end of both genomes to amplify a fragment of 900 bp from ABPV and 700 bp from BQCV. The combined guanidinium thiocyanate and silica membrane method was used to extract total RNA from samples of healthy and laboratory-infected bee pupae. In a blind test, RT-PCR successfully identified the samples containing ABPV and BQCV. Sensitivities were approximately 1,600 genome equivalents of purified ABPV and 130 genome equivalents of BQCV.

Genetic analysis of wild-type Dobrava hantavirus in Slovenia: co-existence of two distinct genetic lineages within the same natural focus

Microbiology ◽  
2000 ◽  
Vol 81 (7) ◽  
pp. 1747-1755 ◽  
Author(s):  
Tatjana Avsic-Zupanc ◽  
Kirill Nemirov ◽  
Miro Petrovec ◽  
Tomi Trilar ◽  
Mario Poljak ◽  
...  
Keyword(s):  
Genetic Analysis ◽  
Phylogenetic Trees ◽  
Nucleotide Diversity ◽  
Wild Type ◽  
Rt Pcr ◽  
Natural Focus ◽  
Genetic Lineages ◽  
Natural Foci ◽  
High Level ◽  
Close Relatedness

Genetic analysis was performed of wild-type (wt) Dobrava hantavirus (DOB) strains from Slovenia, the country where the virus was first discovered and where it was found to cause haemorrhagic fever with renal syndrome (HFRS), with a fatality rate of 12%. Two hundred and sixty mice of the genus Apodemus, trapped in five natural foci of DOB-associated HFRS during 1990–1996, were screened for the presence of anti-hantavirus antibodies and 49 Apodemus flavicollis and four Apodemus agrarius were found to be positive. RT–PCR was used to recover partial sequences of the wt-DOB medium (M) and small (S) genome segments from nine A. flavicollis and one A. agrarius. Sequence comparison and phylogenetic analysis of the Slovenian wt-DOB strains revealed close relatedness of all A. flavicollis-derived virus sequences (nucleotide diversity up to 6% for the M segment and 5% for the S segment) and the geographical clustering of genetic variants. In contrast, the strain harboured by A. agrarius showed a high level of genetic diversity from other Slovenian DOB strains (14%) and clustered together on phylogenetic trees with other DOB strains harboured by A. agrarius from Russia, Estonia and Slovakia. These findings suggest that the DOB variants carried by the two species of Apodemus in Europerepresent two distinct genetic lineages.

scholarly journals Detection of acute bee paralysis virus by RT-PCR in honey bee and Varroa destructor field samples: rapid screening of representative Hungarian apiaries

Apidologie ◽  
2002 ◽  
Vol 33 (1) ◽  
pp. 63-74 ◽  
Author(s):  
Tam�s Bakonyi ◽  
R�bert Farkas ◽  
Andrea Szendr�i ◽  
Mih�ly Dobos-Kov�cs ◽  
Mikl�s Rusvai
Keyword(s):  
Honey Bee ◽  
Varroa Destructor ◽  
Rapid Screening ◽  
Rt Pcr ◽  
Field Samples ◽  
Acute Bee Paralysis Virus ◽  
Paralysis Virus

scholarly journals The First Detection and Genetic Characterization of Four Different Honeybee Viruses in Wild Bumblebees from Croatia

Pathogens ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 808
Author(s):  
Ivana Tlak Gajger ◽  
Laura Šimenc ◽  
Ivan Toplak
Keyword(s):  
Genetic Characterization ◽  
Direct Sequencing ◽  
Rt Pcr ◽  
Black Queen Cell Virus ◽  
Deformed Wing Virus ◽  
Queen Cell ◽  
Acute Bee Paralysis Virus ◽  
Virus Strains ◽  
Paralysis Virus

To determine the presence and the prevalence of four different honeybee viruses (acute bee paralysis virus—ABPV, black queen cell virus—BQCV, chronic bee paralysis virus—CBPV, deformed wing virus—DWV) in wild bumblebees, pooled randomly selected bumblebee samples were collected from twenty-seven different locations in the territory of Croatia. All samples were prepared and examined using the RT-PCR methods for quantification of mentioned honeybee viruses. Determined prevalence (%) of identified positive viruses were in the following decreasing order: BQCV > DWV > ABPV, CBPV. Additionally, direct sequencing of samples positive for BQCV (n = 24) and DWV (n = 2) was performed, as well as a test of molecular phylogeny comparison with those available in GenBank. Selected positive field viruses’ strains showed 95.7 to 100% (BQCV) and 98.09% (DWV) nucleotide identity with previously detected and deposited honeybee virus strains in the geographic areas in Croatia and neighboring Slovenia. In this article, the first detection of four honeybee viruses with genetic characterization of high diversity strains circulating in wild bumblebees in Croatia is presented.

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