scholarly journals Multi-tiered analyses of honey bees that resist or succumb to parasitic mites and viruses

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Daniel B. Weaver ◽  
Brandi L. Cantarel ◽  
Christine G. Elsik ◽  
Dawn L. Boncristiani ◽  
Jay D. Evans
Keyword(s):  
Gene Expression ◽  
Honey Bee ◽  
Honey Bees ◽  
Gene Activation ◽  
Constitutive Expression ◽  
Host Responses ◽  
Pollination Services ◽  
Deformed Wing Virus ◽  
Mortality And Morbidity ◽  
Parasitic Mites

Abstract Background Varroa destructor mites, and the numerous viruses they vector to their honey bee hosts, are among the most serious threats to honey bee populations, causing mortality and morbidity to both the individual honey bee and colony, the negative effects of which convey to the pollination services provided by honey bees worldwide. Here we use a combination of targeted assays and deep RNA sequencing to determine host and microbial changes in resistant and susceptible honey bee lineages. We focus on three study sets. The first involves field sampling of sympatric western bees, some derived from resistant stock and some from stock susceptible to mites. The second experiment contrasts three colonies more deeply, two from susceptible stock from the southeastern U.S. and one from mite-resistant bee stock from Eastern Texas. Finally, to decouple the effects of mites from those of the viruses they vector, we experimentally expose honey bees to DWV in the laboratory, measuring viral growth and host responses. Results We find strong differences between resistant and susceptible bees in terms of both viral loads and bee gene expression. Interestingly, lineages of bees with naturally low levels of the mite-vectored Deformed wing virus, also carried lower levels of viruses not vectored by mites. By mapping gene expression results against current ontologies and other studies, we describe the impacts of mite parasitism, as well as viruses on bee health against two genetic backgrounds. We identify numerous genes and processes seen in other studies of stress and disease in honey bee colonies, alongside novel genes and new patterns of expression. Conclusions We provide evidence that honey bees surviving in the face of parasitic mites do so through their abilities to resist the presence of devastating viruses vectored by these mites. In all cases, the most divergence between stocks was seen when bees were exposed to live mites or viruses, suggesting that gene activation, rather than constitutive expression, is key for these interactions. By revealing responses to viral infection and mite parasitism in different lineages, our data identify candidate proteins for the evolution of mite tolerance and virus resistance.

scholarly journals Multi-tiered Analyses of Honey Bees That Resist or Succumb to Parasitic Mites and Viruses

2021 ◽  
Author(s):  
Daniel B. Weaver ◽  
Brandi L. Cantarel ◽  
Christine Elsik ◽  
Dawn L. Lopez ◽  
Jay Evans
Keyword(s):  
Gene Expression ◽  
Honey Bee ◽  
Honey Bees ◽  
Host Responses ◽  
Negative Effects ◽  
Pollination Services ◽  
Deformed Wing Virus ◽  
Mortality And Morbidity ◽  
The Individual ◽  
Parasitic Mites

Abstract Background Varroa destructor mites, and the numerous viruses they vector to their honey bee hosts, are among the most serious threats to honey bee populations, causing mortality and morbidity to both the individual honey bee and colony, the negative effects of which convey to the pollination services provided by honey bees worldwide. Here we use a combination of targeted assays and deep RNA sequencing to determine host and microbial changes in resistant and susceptible honey bee lineages. We focus on three study sets. The first involves field sampling of sympatric western bees, some derived from resistant stock and some from stock susceptible to mites. The second experiment contrasts three colonies more deeply, two from susceptible stock from the southeastern U.S. and one from mite-resistant bee stock from Eastern Texas. Finally, to decouple the effects of mites from those of the viruses they vector, we experimentally expose honey bees to DWV in the laboratory, measuring viral growth and host responses. Results We find strong differences between resistant and susceptible bees in terms of both viral loads and bee gene expression. Interestingly, lineages of bees with naturally low levels of the mite-vectored Deformed wing virus, also carried lower levels of viruses not vectored by mites. By mapping gene expression results against current ontologies and other studies, we describe the impacts of mite parasitism, as well as viruses on bee health against two genetic backgrounds. We identify numerous genes and processes seen in other studies of stress and disease in honey bee colonies, though we find novel genes and new patterns of expression too. Conclusions We provide evidence that honey bees surviving in the face of parasitic mites do so through their abilities to resist the presence of devastating viruses vectored by these mites. By revealing responses to viral infection and mite parasitism in different lineages, our data identify candidate proteins for the evolution of mite tolerance and virus resistance.

scholarly journals Honey Bee Viruses, Colony Health, and Antiviral Defense

Proceedings ◽  
2020 ◽  
Vol 50 (1) ◽  
pp. 16
Author(s):  
Katie F. Daughenbaugh ◽  
Alex J. McMenamin ◽  
Laura M. Brutscher ◽  
Fenali Parekh ◽  
Michelle L. Flenniken
Keyword(s):  
Gene Expression ◽  
Heat Shock ◽  
Honey Bee ◽  
Honey Bees ◽  
Antiviral Defense ◽  
Deformed Wing Virus ◽  
Antiviral Responses ◽  
Model Virus ◽  
Virus Abundance

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.

scholarly journals Transcriptome-level assessment of the impact of deformed wing virus on honey bee larvae

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Zih-Ting Chang ◽  
Yu-Feng Huang ◽  
Yue-Wen Chen ◽  
Ming-Ren Yen ◽  
Po-Ya Hsu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Honey Bee ◽  
Larval Stage ◽  
Honey Bees ◽  
Pupal Stage ◽  
Deformed Wing Virus ◽  
Expression Levels ◽  
The Impact ◽  
Honey Bee Larvae ◽  
Gene Expression Levels

AbstractDeformed wing virus (DWV) prevalence is high in honey bee (Apis mellifera) populations. The virus infects honey bees through vertical and horizontal transmission, leading to behavioural changes, wing deformity, and early mortality. To better understand the impacts of viral infection in the larval stage of honey bees, artificially reared honey bee larvae were infected with DWV (1.55 × 1010 copies/per larva). No significant mortality occurred in infected honey bee larvae, while the survival rates decreased significantly at the pupal stage. Examination of DWV replication revealed that viral replication began at 2 days post inoculation (d.p.i.), increased dramatically to 4 d.p.i., and then continuously increased in the pupal stage. To better understand the impact of DWV on the larval stage, DWV-infected and control groups were subjected to transcriptomic analysis at 4 d.p.i. Two hundred fifty-five differentially expressed genes (DEGs) (fold change ≥ 2 or ≤ -2) were identified. Of these DEGs, 168 genes were downregulated, and 87 genes were upregulated. Gene Ontology (GO) analysis showed that 141 DEGs (55.3%) were categorized into molecular functions, cellular components and biological processes. One hundred eleven genes (38 upregulated and 73 downregulated) were annotated by KO (KEGG Orthology) pathway mapping and involved metabolic pathways, biosynthesis of secondary metabolites and glycine, serine and threonine metabolism pathways. Validation of DEGs was performed, and the related gene expression levels showed a similar tendency to the DEG predictions at 4 d.p.i.; cell wall integrity and stress response component 1 (wsc1), cuticular protein and myo-inositol 2-dehydrogenase (iolG) were significantly upregulated, and small conductance calcium-activated potassium channel protein (SK) was significantly downregulated at 4 d.p.i. Related gene expression levels at different d.p.i. revealed that these DEGs were significantly regulated from the larval stage to the pupal stage, indicating the potential impacts of gene expression levels from the larval to the pupal stages. Taken together, DWV infection in the honey bee larval stage potentially influences the gene expression levels from larvae to pupae and reduces the survival rate of the pupal stage. This information emphasizes the consequences of DWV prevalence in honey bee larvae for apiculture.

Parasitic mites and microsporidians in managed western honey bee colonies on the island of Newfoundland, Canada

2010 ◽  
Vol 142 (6) ◽  
pp. 584-588 ◽  
Author(s):  
Geoffrey R. Williams ◽  
Krista Head ◽  
Karen L. Burgher-MacLellan ◽  
Richard E.L. Rogers ◽  
Dave Shutler
Keyword(s):  
Honey Bee ◽  
Honey Bees ◽  
Varroa Destructor ◽  
Geographic Isolation ◽  
Acarapis Woodi ◽  
Pollination Services ◽  
Chemical Treatments ◽  
Organic Honey ◽  
Parasitic Mites ◽  
Bee Colonies

AbstractWestern honey bees, Apis mellifera L. (Hymenoptera: Apidae), occur in nearly every region inhabited by man because they provide valuable honey, wax, and pollination services. Many commercial honey bee operations are plagued by economically important parasites; however, beekeepers on the island of Newfoundland, Canada, are in a unique position because of the province of Newfoundland and Labrador’s strict import regulations and geographic isolation. We surveyed about 25% of the island’s approximately 100 managed honey bee colonies. The parasitic mites Varroa destructor Anderson and Trueman (Acari: Varroidae) and Acarapis woodi (Rennie) (Acari: Tarsonemidae) were not detected, whereas Nosema spp. microsporidia were detected in two of four beekeeping operations and in 11 of 23 (48%) colonies (intensity = 482 609 ± 1199 489 (mean ± SD); median intensity = 0). Because V. destructor and A. woodi are important pests that typically require chemical treatments, beekeepers on the island of Newfoundland may be uniquely positioned to market organic honey bee products from colonies that could also be a source of mite-naïve bees for research.

scholarly journals Impact of Chronic Exposure to Sublethal Doses of Glyphosate on Honey Bee Immunity, Gut Microbiota and Infection by Pathogens

2021 ◽  
Vol 9 (4) ◽  
pp. 845
Author(s):  
Loreley Castelli ◽  
Sofía Balbuena ◽  
Belén Branchiccela ◽  
Pablo Zunino ◽  
Joanito Liberti ◽  
...  
Keyword(s):  
Gut Microbiota ◽  
Honey Bee ◽  
Chronic Exposure ◽  
Pollination Services ◽  
Deformed Wing Virus ◽  
Immune Genes ◽  
Bee Health ◽  
Honey Bee Health ◽  
Sublethal Doses ◽  
The Impact

Glyphosate is the most used pesticide around the world. Although different studies have evidenced its negative effect on honey bees, including detrimental impacts on behavior, cognitive, sensory and developmental abilities, its use continues to grow. Recent studies have shown that it also alters the composition of the honey bee gut microbiota. In this study we explored the impact of chronic exposure to sublethal doses of glyphosate on the honey bee gut microbiota and its effects on the immune response, infection by Nosema ceranae and Deformed wing virus (DWV) and honey bee survival. Glyphosate combined with N. ceranae infection altered the structure and composition of the honey bee gut microbiota, for example by decreasing the relative abundance of the core members Snodgrassella alvi and Lactobacillus apis. Glyphosate increased the expression of some immune genes, possibly representing a physiological response to mitigate its negative effects. However, this response was not sufficient to maintain honey bee health, as glyphosate promoted the replication of DWV and decreased the expression of vitellogenin, which were accompanied by a reduced life span. Infection by N. ceranae also alters honey bee immunity although no synergistic effect with glyphosate was observed. These results corroborate previous findings suggesting deleterious effects of widespread use of glyphosate on honey bee health, and they contribute to elucidate the physiological mechanisms underlying a global decline of pollination services.

scholarly journals Deformed wing virus variant shift from 2010 to 2016 in managed and feral UK honey bee colonies

2021 ◽  
Author(s):  
J. L. Kevill ◽  
K. C. Stainton ◽  
D. C. Schroeder ◽  
S. J. Martin
Keyword(s):  
Honey Bee ◽  
Honey Bees ◽  
Small Study ◽  
Deformed Wing Virus ◽  
Virus Variant ◽  
The Usa ◽  
The Uk ◽  
Bee Colonies ◽  
Existing Data

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.

Sequential generations of honey bee (Apis mellifera) queens produced using cryopreserved semen

2012 ◽  
Vol 24 (8) ◽  
pp. 1079 ◽  
Author(s):  
Brandon K. Hopkins ◽  
Charles Herr ◽  
Walter S. Sheppard
Keyword(s):  
Apis Mellifera ◽  
Honey Bee ◽  
Honey Bees ◽  
Food Production ◽  
Effective Means ◽  
Eastern Asia ◽  
Pollination Services ◽  
Instrumental Insemination ◽  
Diploid Female ◽  
Frozen Semen

Much of the world’s food production is dependent on honey bees for pollination, and expanding food production will further increase the demand for managed pollination services. Apiculturists outside the native range of the honey bee, in the Americas, Australia and eastern Asia, have used only a few of the 27 described subspecies of honey bees (Apis mellifera) for beekeeping purposes. Within the endemic ranges of a particular subspecies, hybridisation can threaten native subspecies when local beekeepers import and propagate non-native honey bees. For many threatened species, cryopreserved germplasm can provide a resource for the preservation of diversity and recovery of endangered populations. However, although instrumental insemination of queen honey bees is well established, the absence of an effective means to cryopreserve honey bee semen has limited the success of efforts to preserve genetic diversity within the species or to develop repositories of honey bee germplasm for breeding purposes. Herein we report that some queens inseminated with cryopreserved semen were capable of producing a substantial number of fertilised offspring. These diploid female larvae were used to produce two additional sequential generations of new queens, which were then back-crossed to the same stock of frozen semen. Our results demonstrate the ability to produce queens using cryopreserved honey bee spermatozoa and the potential for the establishment of a honey bee genetic repository.

scholarly journals Beeporter: a high-throughput tool for non-invasive analysis of honey bee virus infection

2021 ◽  
Author(s):  
Jay D. Evans ◽  
Olubukola Banmeke ◽  
Evan C. Palmer-Young ◽  
Yanping Chen ◽  
Eugene V. Ryabov
Keyword(s):  
Virus Infection ◽  
Honey Bee ◽  
High Throughput ◽  
Honey Bees ◽  
High Throughput Screening ◽  
Fluorescent Protein ◽  
Imaging System ◽  
Uv Light ◽  
Light Sources ◽  
Deformed Wing Virus

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.

scholarly journals Transcriptional Control of Honey Bee (Apis mellifera) Major Royal Jelly Proteins by 20-Hydroxyecdysone

Insects ◽  
2018 ◽  
Vol 9 (3) ◽  
pp. 122 ◽  
Author(s):  
Paul Winkler ◽  
Frank Sieg ◽  
Anja Buttstedt
Keyword(s):  
Gene Expression ◽  
Apis Mellifera ◽  
Juvenile Hormone ◽  
Honey Bee ◽  
Honey Bees ◽  
Transcriptional Control ◽  
Royal Jelly ◽  
Adult Worker ◽  
Gene Expression Dynamics

One of the first tasks of worker honey bees (Apis mellifera) during their lifetime is to feed the larval offspring. In brief, young workers (nurse bees) secrete a special food jelly that contains a large amount of unique major royal jelly proteins (MRJPs). The regulation of mrjp gene expression is not well understood, but the large upregulation in well-fed nurse bees suggests a tight repression until, or a massive induction upon, hatching of the adult worker bees. The lipoprotein vitellogenin, the synthesis of which is regulated by the two systemic hormones 20-hydroxyecdysone and juvenile hormone, is thought to be a precursor for the production of MRJPs. Thus, the regulation of mrjp expression by the said systemic hormones is likely. This study focusses on the role of 20-hydroxyecdysone by elucidating its effect on mrjp gene expression dynamics. Specifically, we tested whether 20-hydroxyecdysone displayed differential effects on various mrjps. We found that the expression of the mrjps (mrjp1–3) that were finally secreted in large amounts into the food jelly, in particular, were down regulated by 20-hydroxyecdysone treatment, with mrjp3 showing the highest repression value.

scholarly journals Neonicotinoid Clothianidin reduces honey bee immune response and contributes to Varroa mite proliferation

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Desiderato Annoscia ◽  
Gennaro Di Prisco ◽  
Andrea Becchimanzi ◽  
Emilio Caprio ◽  
Davide Frizzera ◽  
...  
Keyword(s):  
Immune Responses ◽  
Honey Bee ◽  
Honey Bees ◽  
Varroa Destructor ◽  
Negative Impact ◽  
Asymptomatic Infection ◽  
Varroa Mite ◽  
Deformed Wing Virus ◽  
Synergistic Interactions ◽  
Parasitic Mite

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.

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