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.

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 The Pathogens Spillover and Incidence Correlation in Bumblebees and Honeybees in Slovenia

Pathogens ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 884
Author(s):  
Metka Pislak Ocepek ◽  
Ivan Toplak ◽  
Urška Zajc ◽  
Danilo Bevk
Keyword(s):  
Pathogen Transmission ◽  
Rt Pcr ◽  
Black Queen Cell Virus ◽  
Deformed Wing Virus ◽  
Nosema Bombi ◽  
Crithidia Bombi ◽  
Queen Cell ◽  
Acute Bee Paralysis Virus ◽  
Bee Virus ◽  
Paralysis Virus

Slovenia has a long tradition of beekeeping and a high density of honeybee colonies, but less is known about bumblebees and their pathogens. Therefore, a study was conducted to define the incidence and prevalence of pathogens in bumblebees and to determine whether there are links between infections in bumblebees and honeybees. In 2017 and 2018, clinically healthy workers of bumblebees (Bombus spp.) and honeybees (Apis mellifera) were collected on flowers at four different locations in Slovenia. In addition, bumblebee queens were also collected in 2018. Several pathogens were detected in the bumblebee workers using PCR and RT-PCR methods: 8.8% on acute bee paralysis virus (ABPV), 58.5% on black queen cell virus (BQCV), 6.8% on deformed wing virus (DWV), 24.5% on sacbrood bee virus (SBV), 15.6% on Lake Sinai virus (LSV), 16.3% on Nosema bombi, 8.2% on Nosema ceranae, 15.0% on Apicystis bombi and 17.0% on Crithidia bombi. In bumblebee queens, only the presence of BQCV, A. bombi and C. bombi was detected with 73.3, 26.3 and 33.3% positive samples, respectively. This study confirmed that several pathogens are regularly detected in both bumblebees and honeybees. Further studies on the pathogen transmission routes are required.

scholarly journals The Pathogen Profile of a Honey Bee Queen Does Not Reflect That of Her Workers

Insects ◽  
2020 ◽  
Vol 11 (6) ◽  
pp. 382 ◽  
Author(s):  
Jessica L. Kevill ◽  
Katie Lee ◽  
Michael Goblirsch ◽  
Erin McDermott ◽  
David R. Tarpy ◽  
...  
Keyword(s):  
Honey Bee ◽  
Horizontal Transmission ◽  
Pathogen Transmission ◽  
Black Queen Cell Virus ◽  
Deformed Wing Virus ◽  
Israeli Acute Paralysis Virus ◽  
Queen Cell ◽  
Honey Bee Queen ◽  
Acute Bee Paralysis Virus ◽  
Paralysis Virus

Throughout a honey bee queen’s lifetime, she is tended to by her worker daughters, who feed and groom her. Such interactions provide possible horizontal transmission routes for pathogens from the workers to the queen, and as such a queen’s pathogen profile may be representative of the workers within a colony. To explore this further, we investigated known honey bee pathogen co-occurrence, as well as pathogen transmission from workers to queens. Queens from 42 colonies were removed from their source hives and exchanged into a second, unrelated foster colony. Worker samples were taken from the source colony on the day of queen exchange and the queens were collected 24 days after introduction. All samples were screened for Nosema spp., Trypanosome spp., acute bee paralysis virus (ABPV), black queen cell virus (BQCV), chronic bee paralysis virus (CBPV), Israeli acute paralysis virus (IAPV), Lake Sinai virus (LSV), and deformed wing virus master variants (DWV-A, B, and C) using RT-qPCR. The data show that LSV, Nosema, and DWV-B were the most abundant pathogens in colonies. All workers (n = 42) were LSV-positive, 88% were Nosema-positive, whilst pathogen loads were low (<1 × 106 genome equivalents per pooled worker sample). All queens (n = 39) were negative for both LSV and Nosema. We found no evidence of DWV transmission occurring from worker to queen when comparing queens to foster colonies, despite DWV being present in both queens and workers. Honey bee pathogen presence and diversity in queens cannot be revealed from screening workers, nor were pathogens successfully transmitted to the queen.

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.

scholarly journals Detection of Lotmaria passim, Crithidia mellificae and Replicative Forms of Deformed Wing Virus and Kashmir Bee Virus in the Small Hive Beetle (Aethina tumida)

Pathogens ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 372
Author(s):  
Antonio Nanetti ◽  
James D. Ellis ◽  
Ilaria Cardaio ◽  
Giovanni Cilia
Keyword(s):  
Honey Bee ◽  
Aethina Tumida ◽  
Small Hive Beetle ◽  
Deformed Wing Virus ◽  
Queen Cell ◽  
Kashmir Bee Virus ◽  
Acute Bee Paralysis Virus ◽  
Bee Virus ◽  
Paralysis Virus ◽  
Lotmaria Passim

Knowledge regarding the honey bee pathogens borne by invasive bee pests remains scarce. This investigation aimed to assess the presence in Aethina tumida (small hive beetle, SHB) adults of honey bee pathogens belonging to the following groups: (i) bacteria (Paenibacillus larvae and Melissococcus plutonius), (ii) trypanosomatids (Lotmaria passim and Crithidia mellificae), and (iii) viruses (black queen cell virus, Kashmir bee virus, deformed wing virus, slow paralysis virus, sacbrood virus, Israeli acute paralysis virus, acute bee paralysis virus, chronic bee paralysis virus). Specimens were collected from free-flying colonies in Gainesville (Florida, U.S.A.) in summer 2017. The results of the molecular analysis show the presence of L. passim, C. mellificae, and replicative forms of deformed wing virus (DWV) and Kashmir bee virus (KBV). Replicative forms of KBV have not previously been reported. These results support the hypothesis of pathogen spillover between managed honey bees and the SHB, and these dynamics require further investigation.

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 Phylogenetic analysis of black queen cell virus and deformed wing virus in honeybee colonies infected by mites in Van, Eastern Turkey

2018 ◽  
Vol 74 (1) ◽  
pp. 5990-2018 ◽  
Author(s):  
ZEYNEP KARAPINAR ◽  
BEKİR OĞUZ ◽  
ENDER DİNÇER ◽  
CİHAT ÖZTÜRK
Keyword(s):  
Phylogenetic Analysis ◽  
Analysis Data ◽  
Parasitic Infections ◽  
Black Queen Cell Virus ◽  
Deformed Wing Virus ◽  
Eastern Turkey ◽  
Geographical Regions ◽  
Queen Cell ◽  
Colony Loss ◽  
Paralysis Virus

This study aimed to determine the presence and prevalence of viral and parasitic infections causing high rates of colony loss in honey bee colonies in Van province, eastern Turkey. Twenty-six different apiaries were collected from five counties in Van province. These samples were tested by Reverse-Transcriptase PCR (RT-PCR) for acute bee paralysis virus (ABPV), chronic bee paralysis virus (CBPV), black queen cell virus (BQCV) and deformed wing virus (DWV). Selected positives were sequenced, phylogenetically analyzed and investigated in terms of Varroa. DWV and BQCV were identified in 69.23% (18/26) and 88.46% (23/26) of the bees respectively whereas ABPV and CBPV were not detected in the sampled apiaries. Results of the phylogenetic analysis of DWV and BQCV sequences showed 94–100% similarity to DWV and BQCV isolates obtained from Genbank. Prevalence of varroasis was 89% (23/26) in Van. The obtained samples were identified as V. Varroa destructor by morphological investigation. The study showed that viral and parasitic agents commonly infect honeybees in Van province, with high prevalence rates for BQCV and DWV. There was also a high degree of conservation of DWV and BQCV sequences distinct from DWV and BQCV isolates from other geographical regions. These findings, including current prevalence and phylogenetic analysis data for DWV, BQCV and varroazis in honeybees, are useful for future studies. .

Sign in / Sign up

Export Citation Format

Share Document