Search

Varroa destructor and the pathogens they vector have contributed significantly to high annual honey bee colony losses. Deformed wing virus (DWV) is the most prevalent honey bee virus and has been associated with overwintering colony loss. DWV is capable of infecting honey bees at all life stages and castes via horizontal and vertical transmission. Few studies have been conducted to determine the effect that pathogens have on honey bee queen health and performance. Our results suggest that queens may have mechanisms that reduce the establishment of DWV infection following a single exposure event. A queen's exposure to DWV-A prior to colony establishment did not significantly negatively impact colony strength metrics for two of three queen genotypes tested. Additionally, brood showed little subsequent infection. These findings suggest that when queens experience a single oral or venereal inoculation of DWV-A, vertical transmission may not result in significant brood infection and that overall colony strength may be largely unaffected. Increased colony loss experienced by colonies derived from one queen source indicates that differences in susceptibility exist due to factors that need to be investigated more fully. These findings contribute to a greater understanding of host-pathogen interactions in honey bees and suggest a context-dependent variability on the influence of queen exposure to viruses on colony health. [ABSTRACT FROM AUTHOR]

ABSTRACT European foulbrood (EFB) is a prevalent disease in the European honey bee (Apis mellifera) in the United States, which can lead to colony decline and collapse. The bacterial components of EFB are well-studied, but the diversity of viral infections within infected colonies has not been explored. In this study, we use meta-transcriptomics sequencing of 12 honey bee hives, symptomatic (+, n = 6) and asymptomatic (–, n = 6) for EFB, to investigate viral infection associated with the disease. We assembled 41 viral genomes, belonging to three families (Iflaviridae, Dicistroviridae, and Sinhaliviridae), all previously reported in honey bees, including Lake Sinai virus, deformed wing virus, sacbrood virus, Black queen cell virus, and Israeli acute paralysis virus. In colonies with severe EFB, we observed a higher occurrence of viral genomes (34 genomes) in contrast to fewer recovered from healthy colonies (seven genomes) and a complete absence of Dicistroviridae genomes.We observed specific Lake Sinai virus clades associated exclusively with EFB + or EFB – colonies, in addition to EFB-afflicted colonies that exhibited an increase in relative abundance of sacbrood viruses. Multivariate analyses highlighted that a combination of site and EFB disease status influenced RNA virome composition, while EFB status alone did not significantly impact it, presenting a challenge for comparisons between colonies kept in different yards. These findings contribute to the understanding of viral dynamics in honey bee colonies compromised by EFB and underscore the need for future investigations to consider viral composition when investigating EFB.IMPORTANCEThis study on the viromes of honey bee colonies affected by European foulbrood (EFB) sheds light on the dynamics of viral populations in bee colonies in the context of a prevalent bacterial brood disease. The identification of distinct Lake Sinai virus and sacbrood virus clades associated with colonies affected by severe EFB suggests a potential connection between viral composition and disease status, emphasizing the need for further investigation into the role of viruses during EFB infection. The observed increase in sacbrood viruses during EFB infection suggests a potential viral dysbiosis, with potential implications for honey bee brood health. These findings contribute valuable insights related to beekeeping practices, offering a foundation for future research aimed at understanding and mitigating the impact of bacterial and viral infection in commercial honey bee operations and the management of EFB.

The ectoparasitic mite Varroa destructor is the leading threat to the health of Western honey bees (Apis mellifera) primarily through its action as a vector of viruses. However, it is unclear which of the viruses transmitted by V. destructor actively infect and replicate in mites, which could facilitate increased transmission. To better understand the role of V. destructor as a vector, we took advantage of differences between bee and mite antiviral RNA interference pathways to identify the host specificity of replicating viruses. We used small RNA sequencing of individual V. destructor mites to examine viral small interfering RNA (vsiRNA) profiles of deformed wing virus genotypes (DWV-A and DWV-B), associated with colony declines, as well as nine other viruses present in our samples. We found active replication of six V. destructor-associated viruses, including a novel virus, Varroa destructor virus 9 (VDV-9), and replication of two honey bee-associated viruses, including both DWV-A and -B genotypes, confirming that mites are biological vectors for important bee pathogens. We show that the antiviral RNAi response can be used to define the host range of viruses in host-parasite interactions, such as honey bees and their parasites, enabling a better understanding of the role of a vector in the evolution and spread of honey bee pathogens. [ABSTRACT FROM AUTHOR]

ABSTRACTTo date, many viruses have been discovered to infect honey bees. In this study, we used high-throughput sequencing to expand the known virome of the honey bee, Apis mellifera, by identifying several novel DNA viruses. While the majority of previously identified bee viruses are RNA, our study reveals nine new genomes from the Parvoviridae family, tentatively named Bee densoviruses 1 to 9. In addition, we characterized a large DNA virus, Apis mellifera filamentous-like virus (AmFLV), which shares limited protein identities with the known Apis mellifera filamentous virus. The complete sequence of AmFLV, obtained by a combination of laboratory techniques and bioinformatics, spans 152,678 bp. Linear dsDNA genome encodes for 112 proteins, of which 49 are annotated. Another large virus we discovered is Apis mellifera nudivirus, which belongs to a group of Alphanudivirus. The virus has a length of 129,467 bp and a circular dsDNA genome, and has 106 protein encoding genes. The virus contains most of the core genes of the family Nudiviridae. This research demonstrates the effectiveness of viral binning in identifying viruses in honey bee virology, showcasing its initial application in this field.IMPORTANCEHoney bees contribute significantly to food security by providing pollination services. Understanding the virome of honey bees is crucial for the health and conservation of bee populations and also for the stability of the ecosystems and economies for which they are indispensable. This study unveils previously unknown DNA viruses in the honey bee virome, expanding our knowledge of potential threats to bee health. The use of the viral binning approach we employed in this study offers a promising method to uncovering and understanding the vast viral diversity in these essential pollinators.

Honey bee viruses have been shown to negatively affect the vigour and longevity of European honey bees (Apis mellifera L). In the present work, beehive materials were tested for their potential to serve as non-invasive samples for honey bee virus detection.

Multiple viral infections are one of the main drivers of honey bee colony losses and threaten the sustainability of colony health. The Acute bee paralysis virus (ABPV) is one of the common honey bee viruses that can induce overt and/or covert infections in bee colonies. Globally more knowledge is needed about its effects leading to colony losses. ABPV-related pathology includes paralytic, trembling bees, unable to fly and, in some cases, ABPV causes the death of the bee colonies. The molecular phylogenetic of ABPV isolates obtained from apiaries suffering colony losses and/or with CCD-like complaints in Türkiye was performed in this study. The isolates' structural and non-structural gene regions were analyzed phylogenetically and compared according to sampling years and apiaries. According to the results, the nucleotide identity of the RNA-dependent RNA polymerase (RdRp) gene in Turkish ABPV field isolates is more conserved than the capsid gene. The evaluation of molecular differentiation among the ABPV field isolates can be used for determining the origin and relatedness of ABPV strains and periodic monitoring of ABPV infections in apiaries. [ABSTRACT FROM AUTHOR]

The German wasp, Vespula germanica (Hymenoptera: Vespidae), is social insect that honey contributes to pollination and biological control because they feed on agricultural pests. However, V. germanica can also be a pest itself and use honey bee hives as food sources, including predation, robbing and scavenging on dead bees. This situation causes honeybee pathogens to pass and become infected, especially wasps that consume flightless, sick, or dead honey bees. Viruses constitute the majority of pathogens that cause disease and death in bees. In this study, possible viral pathogens of a large number of V. germanica that died massively were investigated. Using primers that partially amplify the genes of the seven most common viruses (deformed wing virus, Israeli acute bee paralysis virus, black queen cell virus, acute bee paralysis virus, kashmir bee virus, sacbrood bee virus, and chronic bee paralysis virus) in honey bees, the virus scanning was done. As a result of the study, it was determined that the black queen cell virus and kashmir virus were found in V. germanica. According to the results of phylogenetic analysis, it was determined that V. germanica Turkey KBV isolate clustered close to Vespula vulgaris New Zealand KBV isolate, and V. germanica Turkey BQCV isolate clustered close to Bombus terrestiris France BQCV isolate. In this study, it was determined for the first time that the black queen bee virus was found in V. germanica. [ABSTRACT FROM AUTHOR]

The hygienic behavior in honey bees is a complex polygenic trait that serves as a natural defense mechanism against bacterial and fungal brood diseases and Varroa destructor mites infesting brood cells. The aim of this study was to evaluate the effect of a dietary amino acids and vitamins supplement “BEEWELL AminoPlus” on hygienic behavior of Apis mellifera colonies combating microsporidial and viral infections. The experiment was performed during a one-year period on 40 colonies alloted to five groups: one supplemented and infected with Nosema ceranae and four viruses (Deformed wing virus - DWV, Acute bee paralysis virus - ABPV, Chronic bee paralysis virus - CBPV and Sacbrood virus – SBV), three not supplemented, but infected with N. ceranae and/ or viruses, and one negative control group. Beside the l isted pathogens, honey bee trypanosomatids were also monitored in all groups.

Simple Summary: Diseases, particularly those caused by viruses, are a major cause of bee colony losses. However, little is known about the prevalence of the pathogens, particularly virus prevalence, of the honey bee in Egypt, one of the most important countries for beekeeping and agricultural production in Africa. To address this shortfall, we determined the prevalence of ten widespread bee viruses in honey bee colonies in Egypt between two major season, winter and summer, in relation to colony infestation by varroa mites. Honey bees are essential for crop and wild plant pollination. However, many countries have reported high annual colony losses caused by multiple possible stressors. Diseases, particularly those caused by viruses, are a major cause of colony losses. However, little is known about the prevalence of honey bee pathogens, particularly virus prevalence, in Egyptian honey bees. To address this shortfall, we determined the prevalence of widespread bee viruses in honey bee colonies in Egypt—whether it is affected by geography, the season, or infestation with Varroa destructor (varroa) mites. Honey bee worker samples were collected from 18 geographical regions across Egypt during two seasons: winter and summer of 2021. Three apiaries were chosen in each region, and a pooled sample of 150 worker bees was collected from five colonies in each apiary then screened by qPCR for 10 viral targets: acute bee paralysis virus (ABPV), black queen cell virus (BQCV), chronic bee paralysis virus (CBPV), deformed wing virus (DWV) genotypes A (DWV-A), B (DWV-B) and D (Egyptian bee virus), Israeli acute paralysis virus (IAPV), Kashmir bee virus (KBV), sacbrood virus (SBV), and slow bee paralysis virus (SBPV). Our results revealed that DWV-A was the most prevalent virus, followed by BQCV and ABPV; the DWV genotype now spreading across the world, DWV-B, was not detected. There was no difference in varroa infestation rates as well as virus prevalence between winter and summer. However, colonies infected with BQCV had a significantly higher varroa count (adjusted p < 0.05) in the winter season, indicating that there is a seasonal association between the intensity of infestation by varroa and the presence of this virus. We provide data on the current virus prevalence in Egypt, which could assist in the protection of Egypt's beekeeping industry. Moreover, our study aids in the systematic assessment of the global honey bee virome by filling a knowledge gap about the prevalence of honey bee viruses in Egypt. [ABSTRACT FROM AUTHOR]

Simple Summary: Honey bee viruses annually cause serious damage to beekeeping worldwide. Therefore, it is important to accumulate more data regarding routes by which viruses spread. This study was carried out to investigate six of the most widely distributed honey bee viruses in pollen samples either collected from different regions in Bulgaria or purchased directly from the trade market. The obtained results showed the presence of deformed wing virus and sacbrood virus with different frequencies within the examined regions. The phylogenetic analyses of Bulgarian isolates were performed considering the most similar worldwide strains available in the GenBank database. Consequently, it has been concluded that pollen is a valuable source for the detection of honey bee viruses, which provides additional information regarding the horizontal way of spreading of these viruses. Among many pathogens and pests, honey bee viruses are known as one of the most common cause of diseases in honey bee colonies. In this study, we demonstrate that pollen grains and bee bread are potential sources of viral DNA. We extracted DNA from 3 types of pollen samples: directly provided by beekeepers (n = 12), purchased from trade markets (n = 5), and obtained from honeycombs (bee bread, n = 10). The extracted DNA was used for molecular detection (RT-PCR analysis) of six of the most widely distributed honey bee viruses: deformed wing virus, sacbrood virus, acute bee paralysis virus, black queen cell virus, Kashmir bee virus, Israeli acute paralysis virus, and chronic bee paralysis virus. We successfully managed to establish only the deformed wing virus (DWV) and the sacbrood virus (SBV), with different distribution frequencies depending on the territory of the country. The phylogenetic analyses of Bulgarian isolates were performed with the most similar sequences available in molecular databases from other countries. Phylogenies of Bulgarian viral strains demonstrated genetically heterogeneous populations of DWV and relatively homogenous populations of SBV. In conclusion, the results obtained from the current study have shown that pollen is a valuable source for molecular detection of honey bee pathogens. This allows epidemiological monitoring of honey bee diseases at a regional and a national level. [ABSTRACT FROM AUTHOR]

After the nationwide, massive winter losses of honey bees in Korea during the winter of 2021, samplings were conducted from live honey bees in colonies and dead honey bees nearby colonies in the same bee-farms in six regions in Korea. Each sample was subjected to virome analysis using high-throughput sequencing technology. The number of viral reads was the lowest in the live honey bee group sample with 370,503 reads and the highest in the dead honey bee group sample with 42,659,622 reads. Viral contigs were matched with the viral genomes of the black queen cell virus, deformed wing virus, Israeli acute paralysis virus, and sacbrood virus, all of which have been previously reported in Korea. However, Apis rhabdovirus 5, bee macula-like virus, Varroa orthomyxovirus-1, Hubei partiti-like virus 34, Lake Sinai virus 2, 3, and 4, and the Ditton virus, were also discovered in this study, which are the first records in Korea. Plant viral sequences resembling those of Arabidopsis latent virus 1, and a novel viral sequence was also discovered. In the present study 55 complete viral genome sequences were identified. This study is the first virome analysis of domestic honey bees and provides the latest information on the diversity of honey bee viruses in Korea.

Honey bees are essential for crop and wild plant pollination. However, many countries have reported high annual colony losses caused by multiple possible stressors. Diseases, particularly those caused by viruses, are a major cause of colony losses. However, little is known about the prevalence of honey bee pathogens, particularly virus prevalence, in Egyptian honey bees. To address this shortfall, we determined the prevalence of widespread bee viruses in honey bee colonies in Egypt—whether it is affected by geography, the season, or infestation with Varroa destructor (varroa) mites. Honey bee worker samples were collected from 18 geographical regions across Egypt during two seasons: winter and summer of 2021. Three apiaries were chosen in each region, and a pooled sample of 150 worker bees was collected from five colonies in each apiary then screened by qPCR for 10 viral targets: acute bee paralysis virus (ABPV), black queen cell virus (BQCV), chronic bee paralysis virus (CBPV), deformed wing virus (DWV) genotypes A (DWV-A), B (DWV-B) and D (Egyptian bee virus), Israeli acute paralysis virus (IAPV), Kashmir bee virus (KBV), sacbrood virus (SBV), and slow bee paralysis virus (SBPV). Our results revealed that DWV-A was the most prevalent virus, followed by BQCV and ABPV; the DWV genotype now spreading across the world, DWV-B, was not detected. There was no difference in varroa infestation rates as well as virus prevalence between winter and summer. However, colonies infected with BQCV had a significantly higher varroa count (adjusted p < 0.05) in the winter season, indicating that there is a seasonal association between the intensity of infestation by varroa and the presence of this virus. We provide data on the current virus prevalence in Egypt, which could assist in the protection of Egypt’s beekeeping industry. Moreover, our study aids in the systematic assessment of the global honey bee virome by filling a knowledge gap about the prevalence of honey bee viruses in Egypt.

Among many pathogens and pests, honey bee viruses are known as one of the most common cause of diseases in honey bee colonies. In this study, we demonstrate that pollen grains and bee bread are potential sources of viral DNA. We extracted DNA from 3 types of pollen samples: directly provided by beekeepers (n = 12), purchased from trade markets (n = 5), and obtained from honeycombs (bee bread, n = 10). The extracted DNA was used for molecular detection (RT-PCR analysis) of six of the most widely distributed honey bee viruses: deformed wing virus, sacbrood virus, acute bee paralysis virus, black queen cell virus, Kashmir bee virus, Israeli acute paralysis virus, and chronic bee paralysis virus. We successfully managed to establish only the deformed wing virus (DWV) and the sacbrood virus (SBV), with different distribution frequencies depending on the territory of the country. The phylogenetic analyses of Bulgarian isolates were performed with the most similar sequences available in molecular databases from other countries. Phylogenies of Bulgarian viral strains demonstrated genetically heterogeneous populations of DWV and relatively homogenous populations of SBV. In conclusion, the results obtained from the current study have shown that pollen is a valuable source for molecular detection of honey bee pathogens. This allows epidemiological monitoring of honey bee diseases at a regional and a national level.

European foulbrood (EFB) is a prevalent disease in the European honey bee ( Apis mellifera ) in the United States, which can lead to colony decline and collapse. The bacterial components of EFB are well-studied, but the diversity of viral infections within infected colonies has not been explored. In this study, we use meta-transcriptomics sequencing of 12 honey bee hives, symptomatic (+, n = 6) and asymptomatic (-, n = 6) for EFB, to investigate viral infection associated with the disease. We assembled 41 viral genomes, belonging to three families (Iflaviridae, Dicistroviridae, and Sinhaliviridae), all previously reported in honey bees, including Lake Sinai virus, deformed wing virus, sacbrood virus, Black queen cell virus, and Israeli acute paralysis virus. In colonies with severe EFB, we observed a higher occurrence of viral genomes (34 genomes) in contrast to fewer recovered from healthy colonies (seven genomes) and a complete absence of Dicistroviridae genomes.We observed specific Lake Sinai virus clades associated exclusively with EFB + or EFB - colonies, in addition to EFB-afflicted colonies that exhibited an increase in relative abundance of sacbrood viruses. Multivariate analyses highlighted that a combination of site and EFB disease status influenced RNA virome composition, while EFB status alone did not significantly impact it, presenting a challenge for comparisons between colonies kept in different yards. These findings contribute to the understanding of viral dynamics in honey bee colonies compromised by EFB and underscore the need for future investigations to consider viral composition when investigating EFB.IMPORTANCEThis study on the viromes of honey bee colonies affected by European foulbrood (EFB) sheds light on the dynamics of viral populations in bee colonies in the context of a prevalent bacterial brood disease. The identification of distinct Lake Sinai virus and sacbrood virus clades associated with colonies affected by severe EFB suggests a potential connection between viral composition and disease status, emphasizing the need for further investigation into the role of viruses during EFB infection. The observed increase in sacbrood viruses during EFB infection suggests a potential viral dysbiosis, with potential implications for honey bee brood health. These findings contribute valuable insights related to beekeeping practices, offering a foundation for future research aimed at understanding and mitigating the impact of bacterial and viral infection in commercial honey bee operations and the management of EFB., Competing Interests: The authors declare no conflict of interest.

Western honey bees (Apis mellifera) are ecologically, agriculturally, and economically important plant pollinators. High average annual losses of honey bee colonies in the US have been partially attributed to agrochemical exposure and virus infections. To examine the potential negative synergistic impacts of agrochemical exposure and virus infection, as well as the potential promise of phytochemicals to ameliorate the impact of pathogenic infections on honey bees, we infected bees with a panel of viruses (i.e., Flock House virus, deformed wing virus, or Sindbis virus) and exposed to one of three chemical compounds. Specifically, honey bees were fed sucrose syrup containing: (1) thyme oil, a phytochemical and putative immune stimulant, (2) fumagillin, a beekeeper applied fungicide, or (3) clothianidin, a grower-applied insecticide. We determined that virus abundance was lower in honey bees fed 0.16 ppm thyme oil augmented sucrose syrup, compared to bees fed sucrose syrup alone. Parallel analysis of honey bee gene expression revealed that honey bees fed thyme oil augmented sucrose syrup had higher expression of key RNAi genes (argonaute-2 and dicer-like), antimicrobial peptide expressing genes (abaecin and hymenoptaecin) , and vitellogenin , a putative honey bee health and age indicator, compared to bees fed only sucrose syrup. Virus abundance was higher in bees fed fumagillin (25 ppm or 75 ppm) or 1 ppb clothianidin containing sucrose syrup relative to levels in bees fed only sucrose syrup. Whereas, honey bees fed 10 ppb clothianidin had lower virus levels, likely because consuming a near lethal dose of insecticide made them poor hosts for virus infection. The negative impact of fumagillin and clothianidin on honey bee health was indicated by the lower expression of argonaute-2 , dicer-like , abaecin , and hymenoptaecin , and vitellogenin. Together, these results indicate that chemical stimulants and stressors impact the outcome of virus infection and immune gene expression in honey bees. [ABSTRACT FROM AUTHOR]

Western honey bees (Apis mellifera) are ecologically, agriculturally, and economically important plant pollinators. High average annual losses of honey bee colonies in the US have been partially attributed to agrochemical exposure and virus infections. To examine the potential negative synergistic impacts of agrochemical exposure and virus infection, as well as the potential promise of phytochemicals to ameliorate the impact of pathogenic infections on honey bees, we infected bees with a panel of viruses (i.e., Flock House virus, deformed wing virus, or Sindbis virus) and exposed to one of three chemical compounds. Specifically, honey bees were fed sucrose syrup containing: (1) thyme oil, a phytochemical and putative immune stimulant, (2) fumagillin, a beekeeper applied fungicide, or (3) clothianidin, a grower-applied insecticide. We determined that virus abundance was lower in honey bees fed 0.16 ppb thyme oil augmented sucrose syrup, compared to bees fed sucrose syrup alone. Parallel analysis of honey bee gene expression revealed that honey bees fed thyme oil augmented sucrose syrup had higher expression of key RNAi genes (argonaute-2 and dicer-like), antimicrobial peptide expressing genes (abaecin and hymenoptaecin), and vitellogenin, a putative honey bee health and age indicator, compared to bees fed only sucrose syrup. Virus abundance was higher in bees fed fumagillin (25 ppm or 75 ppm) or 1 ppb clothianidin containing sucrose syrup relative to levels in bees fed only sucrose syrup. Whereas, honey bees fed 10 ppb clothianidin had lower virus levels, likely because consuming a near lethal dose of insecticide made them poor hosts for virus infection. The negative impact of fumagillin and clothianidin on honey bee health was indicated by the lower expression of argonaute-2, dicer-like, abaecin, and hymenoptaecin, and vitellogenin. Together, these results indicate that chemical stimulants and stressors impact the outcome of virus infection and immune gene expression in honey bees.

The predator Asian hornet (Vespa velutina) represents one of the major threats to honeybee survival. Viral spillover from bee to wasp has been supposed in several studies, and this work aims to identify and study the virome of both insect species living simultaneously in the same foraging area. Transcriptomic analysis was performed on V. velutina and Apis mellifera samples, and replicative form of detected viruses was carried out by strand‐specific RT‐PCR. Overall, 6 and 9 different viral types were reported in V. velutina and A. mellifera, respectively, and five of these viruses were recorded in both hosts. Varroa destructor virus‐1 and Cripavirus NB‐1/2011/HUN (now classified as Triato‐like virus) were the most represented viruses detected in both hosts, also in replicative form. In this investigation, Triato‐like virus, as well as Aphis gossypii virus and Nora virus, was detected for the first time in honeybees. Concerning V. velutina, we report for the first time the recently detected honeybee La Jolla virus. A general high homology rate between genomes of shared viruses between V. velutina and A. mellifera suggests the efficient transmission of the virus from bee to wasp. In conclusion, our findings highlight the presence of several known and newly reported RNA viruses infecting A. mellifera and V. velutina. This confirms the environment role as an important source of infection and indicates the possibility of spillover from prey to predator. [ABSTRACT FROM AUTHOR]

To date, many viruses have been discovered to infect honey bees. In this study, we used high-throughput sequencing to expand the known virome of the honey bee, Apis mellifera , by identifying several novel DNA viruses. While the majority of previously identified bee viruses are RNA, our study reveals nine new genomes from the Parvoviridae family, tentatively named Bee densoviruses 1 to 9. In addition, we characterized a large DNA virus, Apis mellifera filamentous-like virus (AmFLV), which shares limited protein identities with the known Apis mellifera filamentous virus. The complete sequence of AmFLV, obtained by a combination of laboratory techniques and bioinformatics, spans 152,678 bp. Linear dsDNA genome encodes for 112 proteins, of which 49 are annotated. Another large virus we discovered is Apis mellifera nudivirus, which belongs to a group of Alphanudivirus . The virus has a length of 129,467 bp and a circular dsDNA genome, and has 106 protein encoding genes. The virus contains most of the core genes of the family Nudiviridae . This research demonstrates the effectiveness of viral binning in identifying viruses in honey bee virology, showcasing its initial application in this field.IMPORTANCEHoney bees contribute significantly to food security by providing pollination services. Understanding the virome of honey bees is crucial for the health and conservation of bee populations and also for the stability of the ecosystems and economies for which they are indispensable. This study unveils previously unknown DNA viruses in the honey bee virome, expanding our knowledge of potential threats to bee health. The use of the viral binning approach we employed in this study offers a promising method to uncovering and understanding the vast viral diversity in these essential pollinators., Competing Interests: The authors declare no conflict of interest.

Abstract Considering the intensive trading nowadays, the honey from the local market was tested for the presence of the six most common bee viruses. To prove the suitability of honey as a sample for the bee viruses detection, the set of different sample types taken directly from the hives we comparatively tested. The study included 30 samples of domestic and 5 samples of imported honey. Additionally, we tested 40 sets of samples including live bees, dead bees, and the honey taken from four apiaries for the evaluation of honey suitability for the virus detection, Two out of the six most common bee viruses were detected in the samples of honey from the market. Black queen cell virus (BQCV) genome was found in 24 domestic honey samples and Kashmir bee virus (KBV) genome was detected in one sample of imported honey. The nucleotide sequences of 24 BQCV isolates showed the highest identity (86.4%) with strains from Europe at the polyprotein gene, whilst the Serbian isolates between each other showed 98.5% similarity. By comparative testing of the different type of samples, in three out of four apiaries BQCV genome was detected in both bees and honey. Evaluating the suitability of honey for the detection of the viral disease by simultaneous testing of live, dead bees, and honey from the same hive, it was shown that the honey can be successfully used for the detection of BQCV. Since, as of yet, there has been no evidence of KBV circulation in Serbia, after its detection in imported honey, there is a substantial risk of its introduction and consequently the need for its surveillance. Therefore, the programs of bee diseases screening should be included in the regular control procedures for the international trade. In addition to this benefit, honey gives an opportunity to beekeepers for continuous monitoring of bees’ health status.

Honey bee (Apis mellifera) health is impacted by viral infections at the colony, individual bee, and cellular levels. To investigate honey bee antiviral defense mechanisms at the cellular level we further developed the use of cultured primary cells, derived from either larvae or pupae, and demonstrated that these cells could be infected with a panel of viruses, including common honey bee infecting viruses (i.e., sacbrood virus (SBV) and deformed wing virus (DWV)) and an insect model virus, Flock House virus (FHV). Virus abundances were quantified over the course of infection. The production of infectious virions in cultured honey bee pupal cells was demonstrated by determining that naïve cells became infected after the transfer of deformed wing virus or Flock House virus from infected cell cultures. Initial characterization of the honey bee antiviral immune responses at the cellular level indicated that there were virus-specific responses, which included increased expression of bee antiviral protein-1 (GenBank: MF116383) in SBV-infected pupal cells and increased expression of argonaute-2 and dicer-like in FHV-infected hemocytes and pupal cells. Additional studies are required to further elucidate virus-specific honey bee antiviral defense mechanisms. The continued use of cultured primary honey bee cells for studies that involve multiple viruses will address this knowledge gap.

The honey bee queen is the central hub of a colony to produce eggs and release pheromones to maintain social cohesion. Among many environmental stresses, viruses are a major concern to compromise the queen’s health and reproductive vigor. Viruses have evolved numerous strategies to infect queens either via vertical transmission from the queens’ parents or horizontally through the worker and drones with which she is in contact during development, while mating, and in the reproductive period in the colony. Over 30 viruses have been discovered from honey bees but only few studies exist on the pathogenicity and direct impact of viruses on the queen’s phenotype. An apparent lack of virus symptoms and practical problems are partly to blame for the lack of studies, and we hope to stimulate new research and methodological approaches. To illustrate the problems, we describe a study on sublethal effects of Israeli Acute Paralysis Virus (IAPV) that led to inconclusive results. We conclude by discussing the most crucial methodological considerations and novel approaches for studying the interactions between honey bee viruses and their interactions with queen health.

Infection and transmission of honey bee viruses pose a serious threat to the pollination services of crops and wild plants, which plays a vital role in agricultural economy and ecology. RNA interference (RNAi) is an effective defense mechanism against commonly occurring viral infections of animals and plants. However, recent studies indicate that the effects of RNAi on the honey bee can induce additional impacts and might not always be effective in suppressing the virus. Moreover, the RNAi responses differed in relation to the developmental stage of the insect and the target tissue used, even though the same method of delivery was used. These results indicate that further analysis and field experiments should be performed to characterize the varying effectiveness of RNAi-based methods for treating honey bee viral infections. In this review, we provide an overview of the current knowledge and the recent progress in RNAi-based anti-viral treatments for honey bees, focusing in particular highlight the role of the dsRNA-delivery method used and its effect on RNAi efficiency and demonstrate the potential practical value of this tool for controlling the virus. We conclude studying the gene function and disease control of honey bee by RNAi technology requires a complex consideration from physiology, genetics to environment.

As a part of a Pilot Monitoring Program of honey bee health coordinated by the EURL (European Union Reference Laboratory) and according to the criteria established for Spain, 14 apiaries in Castilla-La Mancha were selected at random and sampled during the autumns of 2012-2014 to identify the most prevalent nosogenic agents, potentially those related to the honey bee colony collapse phenomenon. In all the apiaries studied, Nosema ceranae was the most prevalent pathogen detected over the three years, confirming the worldwide spread of this microsporidian, a pathogen that negatively affects honey bee health at an individual and colony level. Trypanosomatids were also very prevalent in honey bee colonies, although the majority of Trypanosomatids detected were not Crithidia mellificae but rather the genetically distinct Lotmaria passim lineage. We also detected Varroa destructor mites, and the particularly high prevalence in 2014 suggests a possible problem regarding mite control in field conditions that requires attention. In agreement with data from other regions, the BQCV and DWV were the most prevalent viruses in honey bee colonies and thus, the Varroa-DVW interaction may be an important cause of bee colony mortality. While there was little evidence of a relationship between the BQCV virus and N. ceranae under field conditions during 2012, this was not the case in 2013 and 2014. Finally, the AKI-complex or LSV-complex was not detected. The information obtained in this study should help orientate future plans for honey bee disease control.

Honey bee colonies suffer from various pathogens, including honey bee viruses. About 24 viruses have been reported so far. However, six of them are considered to cause severe infection which inflicts heavy losses on beekeeping. The aim of this study was to investigate incidence of six honey bee viruses: deformed wing virus (DWV), acute bee paralysis virus (ABPV), chronic bee paralysis virus (CBPV), sacbrood virus (SBV), kashmir bee virus (KBV), and black queen cell virus (BQCV) by a reverse transcription polymerase chain reaction (RT-PCR). A total of 250 adult honey bee samples were obtained from 50 colonies from eight apiaries situated in three different parts of the country (South, North and West Bulgaria). The results showed the highest prevalence of DWV followed by SBV and ABPV, and one case of BQCV. A comparison with homology sequences available in GenBank was performed by phylogenetic analysis, and phylogenetic relationships were discussed in the context of newly described genotypes in the uninvestigated South Eastern region of Europe. In conclusion, the present study has been the first to provide sequencing data and phylogenetics analyses of some honey bee viruses in Bulgaria.

Pollination of flowering plants is an important ecosystem service provided by wild insect pollinators and managed honey bees. Hence, losses and declines of pollinating insect species threaten human food security and are of major concern not only for apiculture or agriculture but for human society in general. Honey bee colony losses and bumblebee declines have attracted intensive research interest over the last decade and although the problem is far from being solved we now know that viruses are among the key players of many of these bee losses and bumblebee declines. With this special issue on bee viruses we, therefore, aimed to collect high quality original papers reflecting the current state of bee virus research. To this end, we focused on newly discovered viruses (Lake Sinai viruses, bee macula-like virus), or a so far neglected virus species (Apis mellifera filamentous virus), and cutting edge technologies (mass spectrometry, RNAi approach) applied in the field.

Twenty managed honey bee colonies, split between 5 apiaries with 4 hives each, were monitored between the summer of 2011 and spring of 2013. Living bees were sampled in July 2011, July 2012, and August 2012. Twenty-five, medium-aged bees, free of varroa mites, were pooled per colony and date, to form one sample. Unlike in France and Belgium, Chronic Bee Paralysis Virus (CBPV) has not been found in Luxembourg. Slow Bee Paralysis Virus (SBPV) and Israeli Acute Paralysis Virus (IAPV) levels were below detection limits. Traces of Kashmir Bee Virus (KBV) were amplified. Black Queen Cell Virus (BQCV), Varroa destructor Virus-1 (VDV-1), and SacBrood Virus (SBV) were detected in all samples and are reported from Luxembourg for the first time. Varroa destructor Macula- Like Virus (VdMLV), Deformed Wing Virus (DWV), and Acute Bee Paralysis Virus (ABPV) were detected at all locations, and in most but not all samples. There was a significant increase in VDV-1 and DWV levels within the observation period. A principal component analysis was unable to separate the bees of colonies that survived the following winter from bees that died, based on their virus contents in summer. The number of dead varroa mites found below colonies was elevated in colonies that died in the following winter. Significant positive relationships were found between the log-transformed virus levels of the bees and the log-transformed number of mites found below the colonies per week, for VDV-1 and DWV. Sacbrood virus levels were independent of varroa levels, suggesting a neutral or competitive relationship between this virus and varroa.

Infection and transmission of honey bee viruses pose a serious threat to the pollination services of crops and wild plants, which plays a vital role in agricultural economy and ecology. RNA interference (RNAi) is an effective defense mechanism against commonly occurring viral infections of animals and plants. However, recent studies indicate that the effects of RNAi on the honey bee can induce additional impacts and might not always be effective in suppressing the virus. Moreover, the RNAi responses differed in relation to the developmental stage of the insect and the target tissue used, even though the same method of delivery was used. These results indicate that further analysis and field experiments should be performed to characterize the varying effectiveness of RNAi-based methods for treating honey bee viral infections. In this review, we provide an overview of the current knowledge and the recent progress in RNAi-based anti-viral treatments for honey bees, focusing in particular highlight the role of the dsRNA-delivery method used and its effect on RNAi efficiency and demonstrate the potential practical value of this tool for controlling the virus. We conclude studying the gene function and disease control of honey bee by RNAi technology requires a complex consideration from physiology, genetics to environment. [ABSTRACT FROM AUTHOR]

Honey bee colonies suffer from various pathogens, including honey bee viruses. About 24 viruses have been reported so far. However, six of them are considered to cause severe infection which inflicts heavy losses on beekeeping. The aim of this study was to investigate incidence of six honey bee viruses: deformed wing virus (DWV), acute bee paralysis virus (ABPV), chronic bee paralysis virus (CBPV), sacbrood virus (SBV), kashmir bee virus (KBV), and black queen cell virus (BQCV) by a reverse transcription polymerase chain reaction (RT-PCR). A total of 250 adult honey bee samples were obtained from 50 colonies from eight apiaries situated in three different parts of the country (South, North and West Bulgaria). The results showed the highest prevalence of DWV followed by SBV and ABPV, and one case of BQCV. A comparison with homology sequences available in GenBank was performed by phylogenetic analysis, and phylogenetic relationships were discussed in the context of newly described genotypes in the uninvestigated South Eastern region of Europe. In conclusion, the present study has been the first to provide sequencing data and phylogenetics analyses of some honey bee viruses in Bulgaria. [ABSTRACT FROM AUTHOR]

The Asian yellow-legged hornet Vespa velutina nigrithorax, a major predator of honeybees, is spreading in Europe in part due to a lack of efficient control methods. In this study, as a first step to identify biological control agents, we characterized viral RNA sequences present in asymptomatic or symptomatic hornets. Among 19 detected viruses, the honey bee virus Deformed wing virus-B was predominant in all the samples, particularly in muscles from the symptomatic hornet, suggesting a putative cause of the deformed wing symptom. Interestingly, two new viruses closely related to Acyrthosiphon pisum virus and Himetobi P virus and viruses typically associated with honey bees, Acute bee paralysis virus and Black queen cell virus, were detected in the brain and muscles, and may correspond to the circulation and possible replication forms of these viruses in the hornet. Aphid lethal paralysis virus, Bee Macula-like virus, and Moku virus, which are known to infect honey bees, were also identified in the gut virus metagenome of hornets. Therefore, our study underlined the urgent need to study the host range of these newly discovered viruses in hornets to determine whether they represent a new threat for honey bees or a hope for the biocontrol of V. velutina.

Queen loss or failure is an important cause of honey bee colony loss. A functional queen is essential to a colony, and the queen is predicted to be well protected by worker bees and other mechanisms of social immunity. Nevertheless, several honey bee pathogens (including viruses) can infect queens. Here, we report a series of experiments to test how virus infection influences queen–worker interactions and the consequences for virus transmission. We used Israeli acute paralysis virus (IAPV) as an experimental pathogen because it is relevant to bee health but is not omnipresent. Queens were observed spending 50% of their time with healthy workers, 32% with infected workers, and 18% without interaction. However, the overall bias toward healthy workers was not statistically significant, and there was considerable individual to individual variability. We found that physical contact between infected workers and queens leads to high queen infection in some cases, suggesting that IAPV infections also spread through close bodily contact. Across experiments, queens exhibited lower IAPV titers than surrounding workers. Thus, our results indicate that honey bee queens are better protected by individual and social immunity, but this protection is insufficient to prevent IAPV infections completely.

Managed insect pollinators are indispensable in modern agriculture. They are used worldwide not only in the open field but also in greenhouses to enhance fruit set, seed production, and crop yield. Managed honey bee ( Apis mellifera , Apis cerana ) colonies provide the majority of commercial pollination although other members of the superfamily Apoidea are also exploited and commercialized as managed pollinators. In the recent past, it became more and more evident that viral diseases play a key role in devastating honey bee colony losses and it was also recognized that many viruses originally thought to be honey bee specific can also be detected in other pollinating insects. However, while research on viruses infecting honey bees started more than 50 years ago and the knowledge on these viruses is growing ever since, little is known on virus diseases of other pollinating bee species. Recent virus surveys suggested that many of the viruses thought to be honey bee specific are actually circulating in the pollinator community and that pollinator management and commercialization of pollinators provide ample opportunity for viral diseases to spread. However, the direction of disease transmission is not always clear and the impact of these viral diseases on the different hosts remains elusive in many cases. With our review we want to provide an up-to-date overview on the viruses detected in different commercialized pollinators in order to encourage research in the field of pollinator virology that goes beyond molecular detection of viruses. A deeper understanding of this field of virology is urgently needed to be able to evaluate the impact of viruses on pollinator health and the role of different pollinators in spreading viral diseases and to be able to decide on appropriate measures to prevent virus-driven pollinator decline. [ABSTRACT FROM AUTHOR]

Honey bees are important pollinators of agricultural crops. Since 2006, US beekeepers have experienced high annual honey bee colony losses, which may be attributed to multiple abiotic and biotic factors, including pathogens. However, the relative importance of these factors has not been fully elucidated. To identify the most prevalent pathogens and investigate the relationship between colony strength and health, we assessed pathogen occurrence, prevalence, and abundance in Western US honey bee colonies involved in almond pollination. The most prevalent pathogens were Black queen cell virus (BQCV), Lake Sinai virus 2 (LSV2), Sacbrood virus (SBV), Nosema ceranae, and trypanosomatids. Our results indicated that pathogen prevalence and abundance were associated with both sampling date and beekeeping operation, that prevalence was highest in honey bee samples obtained immediately after almond pollination, and that weak colonies had a greater mean pathogen prevalence than strong colonies. [ABSTRACT FROM AUTHOR]

After the nationwide, massive winter losses of honey bees in Korea during the winter of 2021, samplings were conducted from live honey bees in colonies and dead honey bees nearby colonies in the same bee-farms in six regions in Korea. Each sample was subjected to virome analysis using high-throughput sequencing technology. The number of viral reads was the lowest in the live honey bee group sample with 370,503 reads and the highest in the dead honey bee group sample with 42,659,622 reads. Viral contigs were matched with the viral genomes of the black queen cell virus, deformed wing virus, Israeli acute paralysis virus, and sacbrood virus, all of which have been previously reported in Korea. However, Apis rhabdovirus 5, bee macula-like virus, Varroa orthomyxovirus-1, Hubei partiti-like virus 34, Lake Sinai virus 2, 3, and 4, and the Ditton virus, were also discovered in this study, which are the first records in Korea. Plant viral sequences resembling those of Arabidopsis latent virus 1, and a novel viral sequence was also discovered. In the present study 55 complete viral genome sequences were identified. This study is the first virome analysis of domestic honey bees and provides the latest information on the diversity of honey bee viruses in Korea., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Kwon, Jung and Kil.)

Pollination of flowering plants is an important ecosystem service provided by wild insect pollinators and managed honey bees. Hence, losses and declines of pollinating insect species threaten human food security and are of major concern not only for apiculture or agriculture but for human society in general. Honey bee colony losses and bumblebee declines have attracted intensive research interest over the last decade and although the problem is far from being solved we now know that viruses are among the key players of many of these bee losses and bumblebee declines. With this special issue on bee viruses we, therefore, aimed to collect high quality original papers reflecting the current state of bee virus research. To this end, we focused on newly discovered viruses (Lake Sinai viruses, bee macula-like virus), or a so far neglected virus species (Apis mellifera filamentous virus), and cutting edge technologies (mass spectrometry, RNAi approach) applied in the field. [ABSTRACT FROM AUTHOR]

For the first time, adults and brood of Africanized and European honey bees ( Apis mellifera ) were compared for relative virus levels over 48 h following Varroa destructor parasitism or injection of V. destructor homogenate. Rates of increase of deformed wing virus (DWV) for Africanized versus European bees were temporarily lowered for 12 h with parasitism and sustainably lowered over the entire experiment (48 h) with homogenate injection in adults. The rates were also temporarily lowered for 24 h with parasitism but were not affected by homogenate injection in brood. Rates of increase of black queen cell virus (BQCV) for Africanized versus European bees were similar with parasitism but sustainably lowered over the entire experiment with homogenate injection in adults and were similar for parasitism and homogenate injection in brood. Analyses of sac brood bee virus and Israeli acute paralysis virus were limited as detection did not occur after both homogenate injection and parasitism treatment, or levels were not significantly higher than those following control buffer injection. Lower rates of replication of DWV and BQCV in Africanized bees shows that they may have greater viral resistance, at least early after treatment. [ABSTRACT FROM AUTHOR]

Honey bees face numerous biotic threats from viruses to bacteria, fungi, protists, and mites. Here we describe a thorough analysis of microbes harbored by worker honey bees collected from field colonies in geographically distinct regions of Turkey. Turkey is one of the World's most important centers of apiculture, harboring five subspecies of Apis mellifera L., approximately 20% of the honey bee subspecies in the world. We use deep ILLUMINA-based RNA sequencing to capture RNA species for the honey bee and a sampling of all non-endogenous species carried by bees. After trimming and mapping these reads to the honey bee genome, approximately 10% of the sequences (9-10 million reads per library) remained. These were then mapped to a curated set of public sequences containing ca. Sixty megabase-pairs of sequence representing known microbial species associated with honey bees. Levels of key honey bee pathogens were confirmed using quantitative PCR screens. We contrast microbial matches across different sites in Turkey, showing new country recordings of Lake Sinai virus, two Spiroplasma bacterium species, symbionts Candidatus Schmidhempelia bombi, Frischella perrara, Snodgrassella alvi, Gilliamella apicola, Lactobacillus spp.), neogregarines, and a trypanosome species. By using metagenomic analysis, this study also reveals deep molecular evidence for the presence of bacterial pathogens (Melissococcus plutonius, Paenibacillus larvae), Varroa destructor-1 virus, Sacbrood virus, and fungi. Despite this effort we did not detect KBV, SBPV, Tobacco ringspot virus, VdMLV (Varroa Macula like virus), Acarapis spp., Tropilaeleps spp. and Apocephalus (phorid fly). We discuss possible impacts of management practices and honey bee subspecies on microbial retinues. The described workflow and curated microbial database will be generally useful for microbial surveys of healthy and declining honey bees. [ABSTRACT FROM AUTHOR]

The seasonal dynamic of four honey bee viruses in 18 honey bee colonies from six selected apiaries were studied using a specific reverse transcriptase-polymerase chain reaction (RT-PCR) method. In each investigated colony, pupae, hive and forager bees were sampled and tested for acute bee paralysis virus (ABPV), chronic bee paralysis virus (CBPV), deformed wing virus (DWV) and black queen cell virus (BQCV) once per month during a 14 month-period. In pupae samples, only BQCV and DWV were found in very low percentages, with no major differences throughout the investigating period. In hive bees ABPV, BQCV, CBPV and DWV were detected, but in lower percentages than in foraging bees where the number of all four viruses was the highest. The BQCV was discovered in 100% of foraging bees and in 94% of hive bees. For the other viruses, seasonal differences were observed in hive bees and foraging bees; however, the highest seasonal variations were detected for ABPV. This study suggests that horizontal transmission of ABPV, BQCV and DWV may occur through contacts between social groups in the direction from forager to hive bees. [ABSTRACT FROM AUTHOR]

We demonstrated that honey bee viruses including Deformed wing virus (DWV), Black queen cell virus (BQCV) and Israeli acute paralysis virus (IAPV) could infect and replicate in the fungal pathogen Ascosphaera apis that causes honey bee chalkbrood disease, revealing a novel biological feature of honey bee viruses. The phylogenetic analysis show that viruses of fungal and honey bee origins form two clusters in the phylogenetic trees distinctly and that host range of honey bee viruses is dynamic. Further studies are warranted to investigate the impact of the viruses on the fitness of their fungal host and phenotypic effects the virus-fungus combination has on honey bee hosts. [ABSTRACT FROM AUTHOR]

The article focuses on the study which analysis the prevalence and distribution of honeybee viruses in apiaries in the Koprivnica-Križevci district in Croatia. It states that 100 percent of samples were infected with deformed wing virus (DWV), while 70 percent affectede with Sacbrood bee virus (SBV), and 20 percent with Black queen virus (BQCV). It mentions that the DWV had highest prevalence and persist in honey bee colonies.

This article offers information on a study which investigated the prevalence of honey bee viruses in the Black Sea region in Turkey. Utilizing the reverse transcriptase-polymerase chain reaction (RT-PCR) method, the study analyzed three virus types namely, acute bee paralysis virus (ABPV), chronic bee paralysis virus (CBPV) and black queen cell virus (BQCV). The Varroa destructor parasitic infestation was also studied in apiaries since the parasite has been known to be associated with viral diseases and bee colony mortality in the country.

Goal: The goal of this review is to summarize viral diseases of honey bees. Observation of indemonstrable colony collapse disorders in several countries considered as a very important problem which may affect the biological equilibrium. Colony losses were initially attributed solely to mite infestation, but observations during the last decades suggested that certain honey bee viruses are responsible for much of the mortality observed in infested colonies. Until now 18 viruses causing infection in honey bees have been determined. The most important are; deformed wing virus, sacbrood virus, black queen cell virus, Kashmir bee virus, chronic bee paralysis virus, acute bee paralysis virus, Thai sacbrood virus, slow paralysis virus and Israeli acute paralysis virus. First six of these agents are most encountered bee viruses in the world and the data about honey bee viruses are acquired by researches conducted on these agents. In this review article general information and recent developments on deformed wing virus, sacbrood virus, Thai sacbrood virus, black queen cell virus and Kashmere bee virus are given. Moreover, disease symptoms and recent developments are described. Discussion and Conclusion: Deformed wing virus cause disease conditions mainly in adult bees while it may also infect other development stages including pupa. Infected adult bees show wrinkles and color loss on the wings and the body of infected bee is getting smaller than the non-infected bees. Virus propagation is very slow and death due to this infection is very rare in pupas. There is a clear relationship demonstrated between deformed wing virus and presence of varroa infestation in the colony. Contrary to deformed wing virus, disease conditions due to sacbrood virus mainly shown in larva and white color of larva is typically turn to yellowish-brown. Combs in the infected colonies are shown as puzzle. Incidence of sacbrood virus infection increases in the spring. It is not clear whether or not this virus transmitted by varroa. Black queen cell virus may infect all life stages of bees, but the incidence of infection is more common in adults. It is transmission is mainly related to the parasite Nosema apis. After infection by black queen cell virus prepupas, pupas and queen of the colony may die and their color will turn to dark-brown. The most virulent agent among honey bee viruses is Kashmere bee virus whish was first demonstrated in Asian honey bees (A.cerena). But later, it is distribution in A. mellifera colonies in many countries have been described. Kashmere bee virus cause trilling, in-coordination and death of infected bees. These symptoms are generally found in young bees because virus leads to persistent infection in adults. [ABSTRACT FROM AUTHOR]