Since the discovery that honey bee viruses play a role in colony decline, researchers have made major breakthroughs in understanding viral pathology and infection processes in honey bees. Work on virus transmission patterns and virus vectors, such as the mite Varroa destructor, has prompted intense efforts to manage honey bee health. However, little is known about the occurrence of honey bee viruses in bee predators, such as vespids. In this study, we characterized the occurrence of 11 honey bee viruses in five vespid species and one wasp from four provinces in China and two vespid species from four locations in France. The results showed that all the species from China carried certain honey bee viruses, notably Apis mellifera filamentous virus (AmFV), Deformed wing virus (DWV), and Israeli acute paralysis virus (IAPV); furthermore, in some vespid colonies, more than three different viruses were identified. In France, DWV was the most common virus; Sacbrood virus (SBV) and Black queen cell virus (BQCV) were observed in one and two samples, respectively. Phylogenetic analyses of IAPV and BQCV sequences indicated that most of the IAPV sequences belonged to a single group, while the BQCV sequences belonged to several groups. Additionally, our study is the first to detect Lake Sinai virus (LSV) in a hornet from China. Our findings can guide further research into the origin and transmission of honey bee viruses in Vespidae, a taxon of ecological, and potentially epidemiological, relevance.

Honey bees are essential to the functioning of terrestrial ecosystems. However, despite no single factor being blamed for losses of honey bee colonies in Europe and the USA, viruses have been considered as a major driver. Moreover, a virus vector can enhance the titer and virulence of virus such as Varroa destructor can change the virulence of the deformed wing virus. Here, we report molecular evidence for aphid lethal paralysis virus (ALPV) infecting Vespa velutina, which is an important predator of honey bees, especially of Apis cerana. Viral replication and phylogenetic analysis indicated that ALPV can not only replicate in V. velutina and A. cerana, but ALPV from A. cerana (ALPV-Ac) was also significantly associated with that of V. velutina (ALPV-Vv), though distinct from those of Apis mellifera (ALPV-Am). The host state posterior probability displayed that V. velutina is the main viral reservoir between V. velutina and A. cerana. Our results show ALPV had expanded host diversity resulting in potential impacts on the health of pollinators, even on the pollination ecosystem. We suggest further studies should investigate potential risks and impacts on pollinator populations of hornets. These results should have an impact conservation efforts focused on sustaining native pollinator abundance and diversity, and therefore, the crucial ecosystem services that they provide.

Honey bee viruses are associated with honey bee colony decline. Israeli acute paralysis virus (IAPV) is considered to have a strong impact on honey bee survival. Phylogenetic analysis of the viral genomes from several regions of the world showed that various IAPV lineages had substantial differences in virulence. Chronic bee paralysis virus (CBPV), another important honey bee virus, can induce two significantly different symptoms. However, the infection characteristics and pathogenesis of IAPV and CBPV have not been completely elucidated. Here, we constructed infectious clones of IAPV and CBPV using a universal vector to provide a basis for studying their replication and pathogenesis. Infectious IAPV and CBPV were rescued from molecular clones of IAPV and CBPV genomes, respectively, that induced typical paralysis symptoms. The replication levels and expression proteins of IAPV and CBPV in progeny virus production were confirmed by qPCR and Western blot. Our results will allow further dissection of the role of each gene in the context of viral infection while helping to study viral pathogenesis and develop antiviral drugs using reverse genetics systems.

Although it had been reported that Israeli acute paralysis virus (IAPV) can cause systemic infection in honey bees, little is known about how it establishes this infection and results in the typical symptoms, paralysis and trembling. Here, we used our previously constructed IAPV infectious clone to investigate viral loads in different tissues of honey bees and further identify the relation between tissue tropism and paralytic symptoms. Our results showed that tracheae showed a greater concentration of viral abundance than other tissues. The abundance of viral protein in the tracheae was positively associated with viral titers, and was further confirmed by immunological and ultrastructural evidence. Furthermore, higher viral loads in tracheae induced remarkable down-regulation of succinate dehydrogenase and cytochrome c oxidase genes, and progressed to causing respiratory failure of honey bees, resulting in the appearance of typical symptoms, paralysis and body trembling. Our results showed that paralysis symptoms or trembling was actually to mitigate tachypnea induced by IAPV infection due to the impairment of honey bee tracheae, and revealed a direct causal link between paralysis symptoms and tissue tropism. These findings provide new insights into the understanding of the underlying mechanism of paralysis symptoms of honey bees after viral infection and have implications for viral disease prevention and specific therapeutics in practice.

Chinese sacbrood virus (CSBV) is a serious threat to eastern honeybees (Apis cerana), especially larvae. However, the pathological mechanism of this deadly disease is remains unclear. Here, we employed an mRNA-seq and sRNA-seq approach in healthy and CSBV-infected 3 ^rd Apis cerana larval. Gene ontology (GO) and KEGG analysis of 203 differentially expressed genes showed that CSBV infection affected host development by up-regulating the expression of larval cuticle proteins, such as larval cuticle proteins A1A and A3A, resulting in elevated susceptibility to CSBV. In addition, viral infection not only affected the expression of serine protease related to the melanization pathway and but also altered fatty acid metabolism and biosynthesis, thus progressed to disturb host immune response. Interestingly, GO annotation and KEGG analysis on target genes of CSBV-specific siRNA (vsiRNAs) showed that serine/threonine kinase activity and serine-type endopeptidas as well as fatty acid biosynthesis were significantly enriched (P < 0.05). Among these vsiRNAs, vsiRNA-1441 with relatively high expression targeted extracellular serine/threonine protein kinase. This study provides new evidence that CSBV attacks a distinct immune response pathway and mediates the expression of cuticle protein to gain the more chance of proliferation.