scholarly journals Metatranscriptome Analysis of Sympatric Bee Species Identifies Bee Virus Variants and a New Virus, Andrena-Associated Bee Virus-1

Viruses ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 291 ◽  
Author(s):  
Katie F. Daughenbaugh ◽  
Idan Kahnonitch ◽  
Charles C. Carey ◽  
Alexander J. McMenamin ◽  
Tanner Wiegand ◽  
...  

Bees are important plant pollinators in agricultural and natural ecosystems. High average annual losses of honey bee (Apis mellifera) colonies in some parts of the world, and regional population declines of some mining bee species (Andrena spp.), are attributed to multiple factors including habitat loss, lack of quality forage, insecticide exposure, and pathogens, including viruses. While research has primarily focused on viruses in honey bees, many of these viruses have a broad host range. It is therefore important to apply a community level approach in studying the epidemiology of bee viruses. We utilized high-throughput sequencing to evaluate viral diversity and viral sharing in sympatric, co-foraging bees in the context of habitat type. Variants of four common viruses (i.e., black queen cell virus, deformed wing virus, Lake Sinai virus 2, and Lake Sinai virus NE) were identified in honey bee and mining bee samples, and the high degree of nucleotide identity in the virus consensus sequences obtained from both taxa indicates virus sharing. We discovered a unique bipartite + ssRNA Tombo-like virus, Andrena-associated bee virus-1 (AnBV-1). AnBV-1 infects mining bees, honey bees, and primary honey bee pupal cells maintained in culture. AnBV-1 prevalence and abundance was greater in mining bees than in honey bees. Statistical modeling that examined the roles of ecological factors, including floral diversity and abundance, indicated that AnBV-1 infection prevalence in honey bees was greater in habitats with low floral diversity and abundance, and that interspecific virus transmission is strongly modulated by the floral community in the habitat. These results suggest that land management strategies that aim to enhance floral diversity and abundance may reduce AnBV-1 spread between co-foraging bees.

Viruses ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 575 ◽  
Author(s):  
John M. K. Roberts ◽  
Nelson Simbiken ◽  
Chris Dale ◽  
Joel Armstrong ◽  
Denis L. Anderson

The global spread of the parasitic mite Varroa destructor has emphasized the significance of viruses as pathogens of honey bee (Apis mellifera) populations. In particular, the association of deformed wing virus (DWV) with V. destructor and its devastating effect on honey bee colonies has led to that virus now becoming one of the most well-studied insect viruses. However, there has been no opportunity to examine the effects of Varroa mites without the influence of DWV. In Papua New Guinea (PNG), the sister species, V. jacobsoni, has emerged through a host-shift to reproduce on the local A. mellifera population. After initial colony losses, beekeepers have maintained colonies without chemicals for more than a decade, suggesting that this bee population has an unknown mite tolerance mechanism. Using high throughput sequencing (HTS) and target PCR detection, we investigated whether the viral landscape of the PNG honey bee population is the underlying factor responsible for mite tolerance. We found A. mellifera and A. cerana from PNG and nearby Solomon Islands were predominantly infected by sacbrood virus (SBV), black queen cell virus (BQCV) and Lake Sinai viruses (LSV), with no evidence for any DWV strains. V. jacobsoni was infected by several viral homologs to recently discovered V. destructor viruses, but Varroa jacobsoni rhabdovirus-1 (ARV-1 homolog) was the only virus detected in both mites and honey bees. We conclude from these findings that A. mellifera in PNG may tolerate V. jacobsoni because the damage from parasitism is significantly reduced without DWV. This study also provides further evidence that DWV does not exist as a covert infection in all honey bee populations, and remaining free of this serious viral pathogen can have important implications for bee health outcomes in the face of Varroa.


Pathogens ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 372
Author(s):  
Antonio Nanetti ◽  
James D. Ellis ◽  
Ilaria Cardaio ◽  
Giovanni Cilia

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.


Author(s):  
J. L. Kevill ◽  
K. C. Stainton ◽  
D. C. Schroeder ◽  
S. J. Martin

AbstractDeformed wing virus (DWV) has been linked to the global decline of honey bees. DWV exists as three master variants (DWV-A, DWV-B, and DWV-C), each with differing outcomes for the honey bee host. Research in the USA showed a shift from DWV-A to DWV-B between 2010 to 2016 in honey bee colonies. Likewise, in the UK, a small study in 2007 found only DWV-A, whereas in 2016, DWV-B was the most prevalent variant. This suggests a shift from DWV-A to DWV-B might have occurred in the UK between 2007 and 2016. To investigate this further, data from samples collected in 2009/10 (n = 46) were compared to existing data from 2016 (n = 42). These samples also allowed a comparison of DWV variants between Varroa-untreated (feral) and Varroa-treated (managed) colonies. The results revealed that, in the UK, DWV-A was far more prevalent in 2009/10 (87%) than in 2016 (43%). In contrast, DWV-B was less prevalent in 2009/10 (76%) than in 2016 (93%). Regardless if colonies had been treated for Varroa (managed) or not (feral), the same trend from DWV-A to DWV-B occurred. Overall, the results reveal a decrease in DWV-A and an increase in DWV-B in UK colonies.


2021 ◽  
Author(s):  
Jay D. Evans ◽  
Olubukola Banmeke ◽  
Evan C. Palmer-Young ◽  
Yanping Chen ◽  
Eugene V. Ryabov

ABSTRACTHoney bees face numerous pests and pathogens but arguably none are as devastating as Deformed wing virus (DWV). Development of antiviral therapeutics and virus-resistant honey bee lines to control DWV in honey bees is slowed by the lack of a cost-effective high-throughput screening of DWV infection. Currently, analysis of virus infection and screening for antiviral treatments in bees and their colonies is tedious, requiring a well-equipped molecular biology laboratory and the use of hazardous chemicals. Here we utilize a cDNA clone of DWV tagged with green fluorescent protein (GFP) to develop the Beeporter assay, a method for detection and quantification of DWV infection in live honey bees. The assay involves infection of honey bee pupae by injecting a standardized DWV-GFP inoculum, followed by incubation for up to 44 hours. GFP fluorescence is recorded at intervals via commonly available long-wave UV light sources and a smartphone camera or a standard ultraviolet transilluminator gel imaging system. Nonlethal DWV monitoring allows high-throughput screening of antiviral candidates and a direct breeding tool for identifying honey bee parents with increased antivirus resistance. For even more rapid drug screening, we also describe a method for screening bees using 96-well trays and a spectrophotometer.


2021 ◽  
Author(s):  
◽  
Evan Brenton-Rule

<p>Biological invasions are one of the major causes of biodiversity decline on the planet. The key driver of the global movement of invasive species is international trade. As a response to trade driven invasive species risk, international and domestic regulations have been promulgated with the goal of managing the spread and impact of non-native species. My aims in this thesis were twofold. First, my goal was to review a subset of international and domestic regulations with a view to commenting on their fitness for purpose and suggesting potential improvements. Second, I used the example of non-native and invasive Hymenoptera, as well as their pathogens, to illustrate the risks posed by invasive species and gaps in their management.   In order to assess international and domestic regulations, I reviewed the World Trade Organization’s (WTO) Agreement on Sanitary and Phytosanitary Measures, as well as associated disputes. I argue that the WTO’s regulatory system does, for the most part, allow domestic regulators to manage invasive species risk as they see fit. Subsequently, the focus of the thesis narrows to investigate New Zealand’s pre- and post-border regime managing invasive species. I argue that New Zealand’s pre-border approach represents international best practice, but the post-border management of species is fragmented. The power to manage invasive species has been delegated to sub-national and regional bodies, which typically approach invasive species management in different ways. This variation has led to regulatory inconsistencies in pests managed and funding allocated. There appears to be a substantial lack of planning in some spaces, such as the risk of aquatic invasions. I make recommendations to ameliorate these inconsistencies.   My second aim involved the study of non-native and invasive Hymenoptera in New Zealand, as well as the pathogens they carry, in order to illustrate the risks posed by invasive species and gaps in their management. I show that the globally widespread invasive Argentine ant (Linepithema humile) may play a role in the pathogen dynamics and mortality of honey bee hives where the species occur sympatrically. Hives in the presence of Argentine ants suffered significantly higher mortality rates relative to hives without ants and always had higher levels of a honey bee pathogen Deformed wing virus. I demonstrate that honey bee pathogens are found in a range of invasive Hymenoptera in New Zealand. I amplify entire genomes of the honey bee virus Kashmir bee virus (KBV) from three species of non-native or invasive Hymenoptera (Argentine ants, common wasps and honey bees). I show that there is KBV strain variability within and between regions, but more between regions. Further, I demonstrate the result that as sampled KBV sequence length increases, so too does sampled diversity. These results highlight how ‘an’ invasive species is typically not alone: they carry a range of diseases that are almost always not considered in international and regional management plans.   Patterns of non-native Hymenoptera carrying honey bee diseases were not restricted to New Zealand. I used mitochondrial DNA to find the likely origin of invasive populations of the globally distributed invasive German wasp. I demonstrate that German wasps show reduced genetic diversity in the invaded range compared to the native range. Populations in the introduced range are likely to have arrived from different source populations. In some regions there were likely multiple introductions. Other regions are genetically homogenous and represent potential areas for use of gene drive technologies. All four different honey bee pathogens assayed for were found in German wasp populations worldwide. These results highlight how the introduction of one exotic species likely brings a range of pathogens. This example of pathogens in Hymenoptera is likely to be true for nearly all non-native introductions.  Many of the impacts of biological invasions, such as predation and competition, are relatively obvious and are frequently studied. However some, such as the impact of pathogens, are unseen and poorly understood. Legal regulation is often a post-hoc response implemented once a problem has already arisen. At a global level regulatory regimes operate relatively effectively. As the focus becomes more granular, such as the case of pathogens of Hymenoptera, fewer controls exists. This thesis helps to reduce uncertainty in this area as well as makes recommendations as to how these risks may be managed.</p>


2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Desiderato Annoscia ◽  
Gennaro Di Prisco ◽  
Andrea Becchimanzi ◽  
Emilio Caprio ◽  
Davide Frizzera ◽  
...  

AbstractThe neonicotinoid Clothianidin has a negative impact on NF-κB signaling and on immune responses controlled by this transcription factor, which can boost the proliferation of honey bee parasites and pathogens. This effect has been well documented for the replication of deformed wing virus (DWV) induced by Clothianidin in honey bees bearing an asymptomatic infection. Here, we conduct infestation experiments of treated bees to show that the immune-suppression exerted by Clothianidin is associated with an enhanced fertility of the parasitic mite Varroa destructor, as a possible consequence of a higher feeding efficiency. A conceptual model is proposed to describe the synergistic interactions among different stress agents acting on honey bees.


Proceedings ◽  
2020 ◽  
Vol 50 (1) ◽  
pp. 16
Author(s):  
Katie F. Daughenbaugh ◽  
Alex J. McMenamin ◽  
Laura M. Brutscher ◽  
Fenali Parekh ◽  
Michelle L. Flenniken

Honey bee colony losses are influenced by multiple abiotic and biotic factors, including viruses. To investigate the effects of RNA viruses on honey bees, we infected bees with a model virus (Sindbis-GFP) in the presence or absence of double-stranded RNA (dsRNA). In honey bees, dsRNA is the substrate for sequence-specific RNA interference (RNAi)-mediated antiviral defense and is a trigger of sequence-independent\antiviral responses. Transcriptome sequencing identified more than 200 differentially expressed genes, including genes in the RNAi, Toll, Imd, JAK-STAT, and heat shock response pathways, and many uncharacterized genes. To confirm the virus limiting role of two genes (i.e., dicer and mf116383) in honey bees, we utilized RNAi to reduce their expression in vivo and determined that the virus abundance increased. To evaluate the role of the heat shock stress response in antiviral defense, bees were heat stressed post-virus infection and the virus abundance and gene expression were assessed. Heat-stressed bees had reduced virus levels and a greater expression of several heat shock protein encoding genes (hsps) compared to the controls. To determine if these genes are universally associated with antiviral defense, bees were infected with another model virus, Flock House virus (FHV), or deformed wing virus and the gene expression was assessed. The expression of dicer was greater in bees infected with either FHV or Sindbis-GFP compared to the mock-infected bees, but not in the deformed wing virus-infected bees. To further investigate honey bee antiviral defense mechanisms and elucidate the function of key genes (dicer, ago-2, mf116383, and hsps) at the cellular level, primary honey bee larval hemocytes were transfected with dsRNA or infected with the Lake Sinai virus 2 (LSV2). These studies indicate that mf116383 and hsps mediate dsRNA detection and that MF116383 is involved in limiting LSV2 infection. Together, these results further our understanding of honey bee antiviral defense, particularly dsRNA-mediated antiviral responses, at both the individual bee and cellular levels.


Insects ◽  
2020 ◽  
Vol 11 (4) ◽  
pp. 239 ◽  
Author(s):  
Alexis Beaurepaire ◽  
Niels Piot ◽  
Vincent Doublet ◽  
Karina Antunez ◽  
Ewan Campbell ◽  
...  

In the past centuries, viruses have benefited from globalization to spread across the globe, infecting new host species and populations. A growing number of viruses have been documented in the western honey bee, Apis mellifera. Several of these contribute significantly to honey bee colony losses. This review synthetizes the knowledge of the diversity and distribution of honey-bee-infecting viruses, including recent data from high-throughput sequencing (HTS). After presenting the diversity of viruses and their corresponding symptoms, we surveyed the scientific literature for the prevalence of these pathogens across the globe. The geographical distribution shows that the most prevalent viruses (deformed wing virus, sacbrood virus, black queen cell virus and acute paralysis complex) are also the most widely distributed. We discuss the ecological drivers that influence the distribution of these pathogens in worldwide honey bee populations. Besides the natural transmission routes and the resulting temporal dynamics, global trade contributes to their dissemination. As recent evidence shows that these viruses are often multihost pathogens, their spread is a risk for both the beekeeping industry and the pollination services provided by managed and wild pollinators.


2019 ◽  
Vol 56 (4) ◽  
pp. 636-641 ◽  
Author(s):  
Roman V. Koziy ◽  
Sarah C. Wood ◽  
Ivanna V. Kozii ◽  
Claire Janse van Rensburg ◽  
Igor Moshynskyy ◽  
...  

Deformed wing virus (DWV) is a single-stranded RNA virus of honey bees ( Apis mellifera L.) transmitted by the parasitic mite Varroa destructor. Although DWV represents a major threat to honey bee health worldwide, the pathological basis of DWV infection is not well documented. The objective of this study was to investigate clinicopathological and histological aspects of natural DWV infection in honey bee workers. Emergence of worker honey bees was observed in 5 colonies that were clinically affected with DWV and the newly emerged bees were collected for histopathology. DWV-affected bees were 2 times slower to emerge and had 30% higher mortality compared to clinically normal bees. Hypopharyngeal glands in bees with DWV were hypoplastic, with fewer intracytoplasmic secretory vesicles; cells affected by apoptosis were observed more frequently. Mandibular glands were hypoplastic and were lined by cuboidal epithelium in severely affected bees compared to tall columnar epithelium in nonaffected bees. The DWV load was on average 1.7 × 106 times higher ( P < .001) in the severely affected workers compared to aged-matched sister honey bee workers that were not affected by deformed wing disease based on gross examination. Thus, DWV infection is associated with prolonged emergence, increased mortality during emergence, and hypoplasia of hypopharyngeal and mandibular glands in newly emerged worker honey bees in addition to previously reported deformed wing abnormalities.


2017 ◽  
Vol 91 (16) ◽  
Author(s):  
Emily J. Remnant ◽  
Mang Shi ◽  
Gabriele Buchmann ◽  
Tjeerd Blacquière ◽  
Edward C. Holmes ◽  
...  

ABSTRACT Understanding the diversity and consequences of viruses present in honey bees is critical for maintaining pollinator health and managing the spread of disease. The viral landscape of honey bees (Apis mellifera) has changed dramatically since the emergence of the parasitic mite Varroa destructor, which increased the spread of virulent variants of viruses such as deformed wing virus. Previous genomic studies have focused on colonies suffering from infections by Varroa and virulent viruses, which could mask other viral species present in honey bees, resulting in a distorted view of viral diversity. To capture the viral diversity within colonies that are exposed to mites but do not suffer the ultimate consequences of the infestation, we examined populations of honey bees that have evolved naturally or have been selected for resistance to Varroa. This analysis revealed seven novel viruses isolated from honey bees sampled globally, including the first identification of negative-sense RNA viruses in honey bees. Notably, two rhabdoviruses were present in three geographically diverse locations and were also present in Varroa mites parasitizing the bees. To characterize the antiviral response, we performed deep sequencing of small RNA populations in honey bees and mites. This provided evidence of a Dicer-mediated immune response in honey bees, while the viral small RNA profile in Varroa mites was novel and distinct from the response observed in bees. Overall, we show that viral diversity in honey bee colonies is greater than previously thought, which encourages additional studies of the bee virome on a global scale and which may ultimately improve disease management. IMPORTANCE Honey bee populations have become increasingly susceptible to colony losses due to pathogenic viruses spread by parasitic Varroa mites. To date, 24 viruses have been described in honey bees, with most belonging to the order Picornavirales. Collapsing Varroa-infected colonies are often overwhelmed with high levels of picornaviruses. To examine the underlying viral diversity in honey bees, we employed viral metatranscriptomics analyses on three geographically diverse Varroa-resistant populations from Europe, Africa, and the Pacific. We describe seven novel viruses from a range of diverse viral families, including two viruses that are present in all three locations. In honey bees, small RNA sequences indicate that these viruses are processed by Dicer and the RNA interference pathway, whereas Varroa mites produce strikingly novel small RNA patterns. This work increases the number and diversity of known honey bee viruses and will ultimately contribute to improved disease management in our most important agricultural pollinator.


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