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1.
PLoS Pathog ; 20(7): e1012337, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38959190

ABSTRACT

The worldwide dispersal of the ectoparasitic mite Varroa destructor from its Asian origins has fundamentally transformed the relationship of the honey bee (Apis mellifera) with several of its viruses, via changes in transmission and/or host immunosuppression. The extent to which honey bee-virus relationships change after Varroa invasion is poorly understood for most viruses, in part because there are few places in the world with several geographically close but completely isolated honey bee populations that either have, or have not, been exposed long-term to Varroa, allowing for separate ecological, epidemiological, and adaptive relationships to develop between honey bees and their viruses, in relation to the mite's presence or absence. The Azores is one such place, as it contains islands with and without the mite. Here, we combined qPCR with meta-amplicon deep sequencing to uncover the relationship between Varroa presence, and the prevalence, load, diversity, and phylogeographic structure of eight honey bee viruses screened across the archipelago. Four viruses were not detected on any island (ABPV-Acute bee paralysis virus, KBV-Kashmir bee virus, IAPV-Israeli acute bee paralysis virus, BeeMLV-Bee macula-like virus); one (SBV-Sacbrood virus) was detected only on mite-infested islands; one (CBPV-Chronic bee paralysis virus) occurred on some islands, and two (BQCV-Black queen cell virus, LSV-Lake Sinai virus,) were present on every single island. This multi-virus screening builds upon a parallel survey of Deformed wing virus (DWV) strains that uncovered a remarkably heterogeneous viral landscape featuring Varroa-infested islands dominated by DWV-A and -B, Varroa-free islands naïve to DWV, and a refuge of the rare DWV-C dominating the easternmost Varroa-free islands. While all four detected viruses investigated here were affected by Varroa for one or two parameters (usually prevalence and/or the Richness component of ASV diversity), the strongest effect was observed for the multi-strain LSV. Varroa unambiguously led to elevated prevalence, load, and diversity (Richness and Shannon Index) of LSV, with these results largely shaped by LSV-2, a major LSV strain. Unprecedented insights into the mite-virus relationship were further gained from implementing a phylogeographic approach. In addition to enabling the identification of a novel LSV strain that dominated the unique viral landscape of the easternmost islands, this approach, in combination with the recovered diversity patterns, strongly suggests that Varroa is driving the evolutionary change of LSV in the Azores. This study greatly advances the current understanding of the effect of Varroa on the epidemiology and adaptive evolution of these less-studied viruses, whose relationship with Varroa has thus far been poorly defined.


Subject(s)
Varroidae , Animals , Bees/virology , Bees/parasitology , Varroidae/virology , Azores , Insect Viruses/genetics , Insect Viruses/isolation & purification , Insect Viruses/classification , RNA Viruses/genetics , RNA Viruses/isolation & purification , RNA Viruses/classification
2.
Proteomics ; 24(9): e2300312, 2024 May.
Article in English | MEDLINE | ID: mdl-38446070

ABSTRACT

The ectoparasitic mite Varroa destructor transmits and triggers viral infections that have deleterious effects on honey bee colonies worldwide. We performed a manipulative experiment in which worker bees collected at emergence were exposed to Varroa for 72 h, and their proteomes were compared with those of untreated control bees. Label-free quantitative proteomics identified 77 differentially expressed A. mellifera proteins (DEPs). In addition, viral proteins were identified by orthogonal analysis, and most importantly, Deformed wing virus (DWV) was found at high levels/intensity in Varroa-exposed bees. Pathway enrichment analysis suggested that the main pathways affected included peroxisomal metabolism, cyto-/exoskeleton reorganization, and cuticular proteins. Detailed examination of individual DEPs revealed that additional changes in DEPs were associated with peroxisomal function. In addition, the proteome data support the importance of TGF-ß signaling in Varroa-DWV interaction and the involvement of the mTORC1 and Hippo pathways. These results suggest that the effect of DWV on bees associated with Varroa feeding results in aberrant autophagy. In particular, autophagy is selectively modulated by peroxisomes, to which the observed proteome changes strongly corresponded. This study complements previous research with different study designs and suggests the importance of the peroxisome, which plays a key role in viral infections.


Subject(s)
Peroxisomes , RNA Viruses , Varroidae , Animals , Bees/virology , Bees/parasitology , Varroidae/virology , Peroxisomes/metabolism , Peroxisomes/virology , RNA Viruses/physiology , Proteomics/methods , Proteome/metabolism , Proteome/analysis , Insect Proteins/metabolism , Signal Transduction , Host-Parasite Interactions
3.
J Virol ; 97(12): e0114923, 2023 Dec 21.
Article in English | MEDLINE | ID: mdl-37966226

ABSTRACT

IMPORTANCE: The parasitic mite Varroa destructor is a significant driver of worldwide colony losses of our most important commercial pollinator, the Western honey bee Apis mellifera. Declines in honey bee health are frequently attributed to the viruses that mites vector to honey bees, yet whether mites passively transmit viruses as a mechanical vector or actively participate in viral amplification and facilitate replication of honey bee viruses is debated. Our work investigating the antiviral RNA interference response in V. destructor demonstrates that key viruses associated with honey bee declines actively replicate in mites, indicating that they are biological vectors, and the host range of bee-associated viruses extends to their parasites, which could impact virus evolution, pathogenicity, and spread.


Subject(s)
Bees , Disease Vectors , Host Specificity , Parasites , Varroidae , Virus Replication , Viruses , Animals , Bees/parasitology , Bees/virology , Parasites/physiology , Parasites/virology , Varroidae/physiology , Varroidae/virology , Viruses/growth & development , Viruses/pathogenicity , RNA Interference
4.
Virol J ; 21(1): 147, 2024 Jun 28.
Article in English | MEDLINE | ID: mdl-38943139

ABSTRACT

Vertical transmission, the transfer of pathogens across generations, is a critical mechanism for the persistence of plant viruses. The transmission mechanisms are diverse, involving direct invasion through the suspensor and virus entry into developing gametes before achieving symplastic isolation. Despite the progress in understanding vertical virus transmission, the environmental factors influencing this process remain largely unexplored. We investigated the complex interplay between vertical transmission of plant viruses and pollination dynamics, focusing on common bean (Phaseolus vulgaris). The intricate relationship between plants and pollinators, especially bees, is essential for global ecosystems and crop productivity. We explored the impact of virus infection on seed transmission rates, with a particular emphasis on bean common mosaic virus (BCMV), bean common mosaic necrosis virus (BCMNV), and cucumber mosaic virus (CMV). Under controlled growth conditions, BCMNV exhibited the highest seed transmission rate, followed by BCMV and CMV. Notably, in the field, bee-pollinated BCMV-infected plants showed a reduced transmission rate compared to self-pollinated plants. This highlights the influence of pollinators on virus transmission dynamics. The findings demonstrate the virus-specific nature of seed transmission and underscore the importance of considering environmental factors, such as pollination, in understanding and managing plant virus spread.


Subject(s)
Phaseolus , Plant Diseases , Pollination , Animals , Plant Diseases/virology , Bees/virology , Phaseolus/virology , Seeds/virology , Infectious Disease Transmission, Vertical , Cucumovirus/physiology , Potyvirus/physiology
5.
J Exp Biol ; 227(13)2024 Jul 01.
Article in English | MEDLINE | ID: mdl-38894668

ABSTRACT

Viral infections can be detrimental to the foraging ability of the western honey bee, Apis mellifera. The deformed wing virus (DWV) is the most common honey bee virus and has been proposed as a possible cause of learning and memory impairment. However, evidence for this phenomenon so far has come from artificially infected bees, while less is known about the implications of natural infections with the virus. Using the proboscis extension reflex (PER), we uncovered no significant association between a simple associative learning task and natural DWV load. However, when assessed through a reversal associative learning assay, bees with higher DWV load performed better in the reversal learning phase. DWV is able to replicate in the honey bee mushroom bodies, where the GABAergic signalling pathway has an antagonistic effect on associative learning but is crucial for reversal learning. Hence, we assessed the pattern of expression of several GABA-related genes in bees with different learning responses. Intriguingly, mushroom body expression of selected genes was positively correlated with DWV load, but only for bees with good reversal learning performance. We hypothesise that DWV might improve olfactory learning performance by enhancing the GABAergic inhibition of responses to unrewarded stimuli, which is consistent with the behavioural patterns that we observed. However, at higher disease burdens, which might be induced by an artificial infection or by a severe, natural Varroa infestation, this DWV-associated increase in GABA signalling could impair associative learning as previously reported by other studies.


Subject(s)
Mushroom Bodies , RNA Viruses , Animals , Bees/virology , Bees/physiology , Mushroom Bodies/virology , Mushroom Bodies/physiology , RNA Viruses/physiology , Signal Transduction , Reversal Learning/physiology , Association Learning/physiology
6.
Vet Res ; 55(1): 118, 2024 Sep 27.
Article in English | MEDLINE | ID: mdl-39334245

ABSTRACT

Honey bees are rapidly declining, which poses a significant threat to our environment and agriculture industry. These vital insects face a disease complex believed to be caused by a combination of parasites, viruses, pesticides, and nutritional deficiencies. However, the real aetiology is still enigmatic. Due to the conventional analysis methods, we still lack complete insights into the honey bee virome and the presence of pathogenic bacteria. To fill this knowledge gap, we employed third-generation nanopore metagenomic sequencing on honey bee haemolymph to monitor the presence of pathogens over almost a year. This study provides valuable insights into the changes in bacterial and viral loads within honey bee colonies. We identified different pathogens in the honey bee haemolymph, which are not included in honey bee screenings. These pathogens comprise the Apis mellifera filamentous virus, Apis rhabdoviruses, and various bacteria such as Frischella sp. and Arsenophonus sp. Furthermore, a sharp contrast was observed between young and old bees. Our research proposes that transgenerational immune priming may play a role in shaping infection patterns in honey bees. We observed a significant increase in pathogen loads in the spring, followed by a notable decrease in pathogen presence during the summer and autumn months. However, certain pathogens seem to be able to evade this priming effect, making them particularly intriguing as potential factors contributing to mortality. In the future, we aim to expand our research on honey bee transgenerational immune priming and investigate its potential in natural settings. This knowledge will ultimately enhance honey bee health and decrease colony mortality.


Subject(s)
Hemolymph , Seasons , Animals , Bees/virology , Bees/microbiology , Hemolymph/virology , Hemolymph/microbiology , Nanopore Sequencing/methods , Nanopore Sequencing/veterinary , Bacteria/genetics , Bacteria/isolation & purification , Bacteria/classification , Virome
7.
Biol Lett ; 20(5): 20230600, 2024 05.
Article in English | MEDLINE | ID: mdl-38715462

ABSTRACT

Novel transmission routes change pathogen landscapes and may facilitate disease emergence. The varroa mite is a virus vector that switched to western honeybees at the beginning of the last century, leading to hive mortality, particularly in combination with RNA viruses. A recent invasion of varroa on the French island of Ushant introduced vector-mediated transmission to one of the last varroa-naive native honeybee populations and caused rapid changes in the honeybee viral community. These changes were characterized by a drastic increase in deformed wing virus type B prevalence and titre in honeybees, as well as knock-on effects in bumblebees, particularly in the year following the invasion. Slow bee paralysis virus also appeared in honeybees and bumblebees, with a 1 year delay, while black queen cell virus declined in honeybees. This study highlights the rapid and far-reaching effects of vector-borne transmission that can extend beyond the directly affected host species, and that the direction of the effect depends on the pathogen's virulence.


Subject(s)
RNA Viruses , Varroidae , Animals , Bees/virology , Varroidae/virology , Varroidae/physiology , RNA Viruses/physiology , RNA Viruses/genetics , France/epidemiology , Introduced Species , Dicistroviridae/genetics , Dicistroviridae/physiology , Prevalence
8.
Arch Virol ; 169(9): 173, 2024 Aug 06.
Article in English | MEDLINE | ID: mdl-39105883

ABSTRACT

In this study, seven bee viruses of significant importance for bee health in Türkiye were investigated using one-step RT-PCR. For this purpose, larvae from 1183 hives and adult bees from 1196 hives were sampled from 400 apiaries in 40 provinces. The prevalence of viral infections in hives was as follows: acute bee paralysis virus (ABPV), 6.4%; black queen cell virus (BQCV), 77%; chronic bee paralysis virus (CBPV), 3.2%; deformed wing virus (DWV), 63.8%; Israel acute bee paralysis virus (IAPV), 7%; Kashmir bee virus (KBV), 2.7%; sacbrood virus (SBV), 49.7%. Moreover, 50 different combinations of viral infections were identified in the hives. While dual infections (36.1%) were the most common in hives, triple infections with BQCV, DWV, and SBV were found to have the highest prevalence (22.1%). At least one viral infection was detected in all of the apiaries tested. Phylogenetic analysis showed that the isolates from this study generally exhibited the highest similarity to previously reported Turkish isolates. When similarity ratios and the locations and types of amino acid mutations were analyzed, it was observed that the isolates from our study exhibited high similarity to isolates from various countries, including China, the United Kingdom, Syria, and Germany.


Subject(s)
Insect Viruses , Phylogeny , RNA Viruses , Animals , Bees/virology , Insect Viruses/genetics , Insect Viruses/isolation & purification , Insect Viruses/classification , Prevalence , RNA Viruses/genetics , RNA Viruses/isolation & purification , RNA Viruses/classification , Larva/virology , Coinfection/virology , Coinfection/epidemiology , Dicistroviridae/genetics , Dicistroviridae/isolation & purification , Dicistroviridae/classification
9.
Phytopathology ; 114(9): 2182-2191, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38842916

ABSTRACT

Plant defenses are conserved among closely related species, but domestication can alter host genotypes through artificial selection with potential losses in host defenses. Therefore, both domestication and host phylogenetic structure may influence plant virus infection outcomes. Here, we examined the association of phylogeny and domestication with the fitness of infected plants. We inoculated three pairs of domesticated and wild/noncultivated squash (Cucurbita spp.) with a combination of two viruses commonly found to coinfect cucurbits, zucchini yellow mosaic virus and squash mosaic virus, and recorded fitness traits related to flowers, pollination, fruit, and seed viability in the field over 2 separate years. In an additional field experiment, we recorded the relative abundance of both viruses via RT-qPCR. We found a gradient of susceptibility across the six tested lineages, and phylogenetic structure, but not domestication, contributed to differences in infection outcomes and impacts on several fitness traits, including fruit number, fruit weight, and germination. Plant virus infection also impacted the quantity and quality of floral rewards and visitation rates of specialist bee pollinators. There were no detectable differences in viral load between the six host taxa for either virus individually or the ratio of zucchini yellow mosaic virus to squash mosaic virus. Our results highlight the importance of phylogenetic structure in predicting host susceptibility to disease across wild and domesticated plants and the ability of several hosts to maintain fitness in the field despite infection. Broader consequences of plant pathogens for beneficial insects, such as pollinators, should also be considered in future research.


Subject(s)
Cucurbita , Domestication , Phylogeny , Plant Diseases , Pollination , Potyvirus , Cucurbita/virology , Plant Diseases/virology , Potyvirus/physiology , Potyvirus/genetics , Animals , Flowers/virology , Fruit/virology , Bees/virology , Plant Viruses/physiology , Plant Viruses/genetics , Seeds/virology
10.
J Invertebr Pathol ; 206: 108171, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39084550

ABSTRACT

Honey bees are economically important insects. However, they face multiple biotic and abiotic stresses, such as diseases, pesticides, climate change, and pests, which cause the loss of honey bee colonies worldwide. Among these factors, viruses have been identified as the major cause of colony loss. Research on honey bee viruses in Uzbekistan is limited. This study investigated the viruses affecting honey bees in Uzbekistan. Virome analysis was conducted for each sample using high-throughput sequencing and bioinformatics. Nine honey bee viruses have been identified: the acute bee paralysis virus, aphid lethal paralysis virus, Apis rhabdovirus 1 and 2, black queen cell virus, deformed wing virus, Lake Sinai virus 10, sacbrood virus, and Hubei partiti-like virus 34. Additionally, 15 plant viruses were identified, 7 of which were novel. This study is the first virome analysis of Uzbekistan honey bees and provides a foundation for understanding the viruses affecting honey bees and plants in Uzbekistan.


Subject(s)
Insect Viruses , Virome , Bees/virology , Animals , Uzbekistan , Insect Viruses/genetics , Insect Viruses/classification , Insect Viruses/isolation & purification , Metagenomics
11.
J Invertebr Pathol ; 205: 108124, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38729295

ABSTRACT

The most common viral diseases affecting honey bees (Apis mellifera) in Israel include deformed wing viruses (DWV-A and DWV-B) and acute paralysis viruses (ABPV and IAPV). These viruses are transmitted within and between colonies, both horizontally and vertically. All members of the colony contribute to this transmission, on the other hand individual and social immunity, particularly hygienic behaviour, may affect the outcome of the process. In this study, we evaluated the ontogeny of natural infections of DWV-A, DWV-B, ABPV and IAPV, their prevalence and loads, in workers and drones from high (H) and low (L) hygienic colonies. In parallel, we evaluated the expression of two immune genes: peptidoglycan recognition protein S2(PGRP-S2) and hymenoptaecin. The prevalence of DWV-B and IAPV increased with age and was higher in workers than in drones. ABPV was not detected in drones. The expression of both immune genes was significantly affected by age and sex. Drones from H colonies had higher expression of these genes. The increased expression of immune genes with drones' age, particularly in hygienic colonies, suggest additional value of honey bee breeding for hygienic behaviour for sustainable beekeeping.


Subject(s)
Insect Proteins , Bees/virology , Bees/immunology , Animals , Insect Proteins/genetics , Dicistroviridae , RNA Viruses , Carrier Proteins/genetics , Female , Antimicrobial Cationic Peptides , Male
12.
J Invertebr Pathol ; 204: 108125, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38705353

ABSTRACT

In La Réunion, the established honeybee subspecies Apis mellifera unicolor, an endemic subspecies of African lineage, is facing considerable challenges. Since the introduction of the Varroa destructor mite in 2017 high colony losses have been recorded. We investigated the dynamics of V. destructor and two viruses, the Deformed Wing Virus (DWV), known to be transmitted by the mite, and the Chronic Bee Paralysis Virus (CBPV), in A. m. unicolor. Colonies from two apiaries located at 300 and 900 m a.s.l were monitored twice for one year without any acaricide treatment. The brood area, V. destructor infestation rates, DWV and CBPV prevalence and load were recorded monthly. A. m. unicolor maintained brood rearing throughout the year. Varroa destructor infestation resulted in high colony mortality (up to 85 %) and high phoretic mite rates (up to 52 mites per hundred bees). The establishment of DWV in colonies occurred after that of V. destructor and the mite infestation rate had a significant effect on the virus prevalence and load. CBPV appeared only transiently throughout the surveys. The data showed that, in tropical colonies with permanent brood rearing, V. destructor and DWV can reach high levels, but are still subject to seasonal variations that appear to be influenced by environmental conditions. This suggests that beekeeping practices could be adapted by favouring sites and periods for transhumance or acaricide treatment.


Subject(s)
RNA Viruses , Varroidae , Animals , Bees/virology , Bees/parasitology , Varroidae/virology , Varroidae/physiology , Mite Infestations/veterinary , Mite Infestations/parasitology , Insect Viruses , Introduced Species , Host-Parasite Interactions , Islands , Dicistroviridae/physiology
13.
Ecotoxicol Environ Saf ; 282: 116706, 2024 Sep 01.
Article in English | MEDLINE | ID: mdl-38996647

ABSTRACT

Antibiotics are frequently employed to control bacterial diseases in honeybees, but their broad-spectrum action can disrupt the delicate balance of the gut microbiome, leading to dysbiosis. This imbalance in the gut microbiota of honeybees adversely affects their physiological health and weakens their resistance to pathogens, including viruses that significantly threaten honeybee health. In this study, we investigated whether tetracycline-induced gut microbiome dysbiosis promotes the replication of Israeli acute paralysis virus (IAPV), a key virus associated with colony losses and whether IAPV infection exacerbates gut microbiome dysbiosis. Our results demonstrated that tetracycline-induced gut microbiome dysbiosis increases the susceptibility of honeybees to IAPV infection. The viral titer in worker bees with antibiotic-induced gut microbiome dysbiosis prior to IAPV inoculation was significantly higher than in those merely inoculated with IAPV. Furthermore, we observed a synergistic effect between tetracycline and IAPV on the disruption of the honeybee gut microbiome balance. The progression of IAPV replication could, in turn, exacerbate antibiotic-induced gut microbiome dysbiosis in honeybees. Our research provides novel insights into the role of the gut microbiota in host-virus interactions, emphasizing the complex interplay between antibiotic use, gut microbiome health, and viral susceptibility in honeybees. We highlight the crucial role of a balanced gut microbiota in honey bees for their immune response against pathogens and emphasize the importance of careful, safe antibiotic use in beekeeping to protect these beneficial microbes.


Subject(s)
Anti-Bacterial Agents , Dicistroviridae , Dysbiosis , Gastrointestinal Microbiome , Tetracycline , Animals , Bees/virology , Bees/microbiology , Bees/drug effects , Gastrointestinal Microbiome/drug effects , Dysbiosis/chemically induced , Dysbiosis/virology , Tetracycline/pharmacology , Tetracycline/toxicity , Dicistroviridae/drug effects , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/toxicity
14.
J Insect Sci ; 24(3)2024 May 01.
Article in English | MEDLINE | ID: mdl-38805657

ABSTRACT

Despite the use of various integrated pest management strategies to control the honey bee mite, Varroa destructor, varroosis remains the most important threat to honey bee colony health in many countries. In Canada, ineffective varroa control is linked to high winter colony losses and new treatment options, such as a summer treatment, are greatly needed. In this study, a total of 135 colonies located in 6 apiaries were submitted to one of these 3 varroa treatment strategies: (i) an Apivar® fall treatment followed by an oxalic acid (OA) treatment by dripping method; (ii) same as in (i) with a summer treatment consisting of formic acid (Formic Pro™); and (iii) same as in (i) with a summer treatment consisting of slow-release OA/glycerin pads (total of 27 g of OA/colony). Treatment efficacy and their effects on colony performance, mortality, varroa population, and the abundance of 6 viruses (acute bee paralysis virus [ABPV], black queen cell virus [BQCV], deformed wing virus variant A [DWV-A], deformed wing virus variant B [DWV-B], Israeli acute paralysis virus [IAPV], and Kashmir bee virus [KBV]) were assessed. We show that a strategy with a Formic Pro summer treatment tended to reduce the varroa infestation rate to below the economic fall threshold of 15 daily varroa drop, which reduced colony mortality significantly but did not reduce the prevalence or viral load of the 6 tested viruses at the colony level. A strategy with glycerin/OA pads reduced hive weight gain and the varroa infestation rate, but not below the fall threshold. A high prevalence of DWV-B was measured in all groups, which could be related to colony mortality.


Subject(s)
Beekeeping , Seasons , Varroidae , Viral Load , Animals , Varroidae/physiology , Bees/parasitology , Bees/virology , Beekeeping/methods , Acaricides , Formates/pharmacology , Canada
15.
Mol Ecol ; 32(14): 3859-3871, 2023 07.
Article in English | MEDLINE | ID: mdl-37194687

ABSTRACT

Domesticated honeybees and wild bees are some of the most important beneficial insects for human and environmental health, but infectious diseases pose a serious risk to these pollinators, particularly following the emergence of the ectoparasitic mite Varroa destructor as a viral vector. The acquisition of this novel viral vector from the Asian honeybee Apis ceranae has fundamentally changed viral epidemiology in its new host, the western honeybee A. mellifera. While the recently discovered Lake Sinai Viruses (LSV) have been associated with weak honeybee colonies, they have not been associated with vector-borne transmission. By combining a large-scale multi-year survey of LSV in Chinese A. mellifera and A. cerana honeybee colonies with globally available LSV-sequence data, we investigate the global epidemiology of this virus. We find that globally distributed LSV is a highly diverse multi-strain virus, which is predominantly associated with the western honeybee A. mellifera. In contrast to the vector-borne deformed wing virus, LSV is not an emerging disease. Instead, demographic reconstruction and strong global and local population structure indicates that it is a highly variable multi-strain virus in a stable association with its main host, the western honeybee. Prevalence patterns in China suggest a potential role for migratory beekeeping in the spread of this pathogen, demonstrating the potential for disease transmission with the man-made transport of beneficial insects.


Subject(s)
Bees , RNA Viruses , Varroidae , Animals , Humans , Bees/parasitology , Bees/virology , China/epidemiology , RNA Viruses/genetics , Varroidae/virology , Viruses
16.
Proc Natl Acad Sci U S A ; 117(13): 7355-7362, 2020 03 31.
Article in English | MEDLINE | ID: mdl-32179689

ABSTRACT

The honey bee gut microbiota influences bee health and has become an important model to study the ecology and evolution of microbiota-host interactions. Yet, little is known about the phage community associated with the bee gut, despite its potential to modulate bacterial diversity or to govern important symbiotic functions. Here we analyzed two metagenomes derived from virus-like particles, analyzed the prevalence of the identified phages across 73 bacterial metagenomes from individual bees, and tested the host range of isolated phages. Our results show that the honey bee gut virome is composed of at least 118 distinct clusters corresponding to both temperate and lytic phages and representing novel genera with a large repertoire of unknown gene functions. We find that the phage community is prevalent in honey bees across space and time and targets the core members of the bee gut microbiota. The large number and high genetic diversity of the viral clusters seems to mirror the high extent of strain-level diversity in the bee gut microbiota. We isolated eight lytic phages that target the core microbiota member Bifidobacterium asteroides, but that exhibited different host ranges at the strain level, resulting in a nested interaction network of coexisting phages and bacterial strains. Collectively, our results show that the honey bee gut virome consists of a complex and diverse phage community that likely plays an important role in regulating strain-level diversity in the bee gut and that holds promise as an experimental model to study bacteria-phage dynamics in natural microbial communities.


Subject(s)
Bees/microbiology , Bees/virology , Animals , Bacteria/genetics , Bacteriophages/genetics , Bees/genetics , Bifidobacterium/isolation & purification , Bifidobacterium/virology , Gastrointestinal Microbiome , Metagenome , Microbiota , Symbiosis/physiology
17.
Proc Natl Acad Sci U S A ; 117(19): 10406-10413, 2020 05 12.
Article in English | MEDLINE | ID: mdl-32341145

ABSTRACT

Anthropogenic changes create evolutionarily novel environments that present opportunities for emerging diseases, potentially changing the balance between host and pathogen. Honey bees provide essential pollination services, but intensification and globalization of honey bee management has coincided with increased pathogen pressure, primarily due to a parasitic mite/virus complex. Here, we investigated how honey bee individual and group phenotypes are altered by a virus of concern, Israeli acute paralysis virus (IAPV). Using automated and manual behavioral monitoring of IAPV-inoculated individuals, we find evidence for pathogen manipulation of worker behavior by IAPV, and reveal that this effect depends on social context; that is, within versus between colony interactions. Experimental inoculation reduced social contacts between honey bee colony members, suggesting an adaptive host social immune response to diminish transmission. Parallel analyses with double-stranded RNA (dsRNA)-immunostimulated bees revealed these behaviors are part of a generalized social immune defensive response. Conversely, inoculated bees presented to groups of bees from other colonies experienced reduced aggression compared with dsRNA-immunostimulated bees, facilitating entry into susceptible colonies. This reduction was associated with a shift in cuticular hydrocarbons, the chemical signatures used by bees to discriminate colony members from intruders. These responses were specific to IAPV infection, suggestive of pathogen manipulation of the host. Emerging bee pathogens may thus shape host phenotypes to increase transmission, a strategy especially well-suited to the unnaturally high colony densities of modern apiculture. These findings demonstrate how anthropogenic changes could affect arms races between human-managed hosts and their pathogens to potentially affect global food security.


Subject(s)
Bees/virology , Dicistroviridae/metabolism , Host-Pathogen Interactions/physiology , Animals , Beekeeping/methods , Bees/genetics , Behavior, Animal , Colony Collapse/epidemiology , DNA Viruses/genetics , DNA Viruses/metabolism , Dicistroviridae/genetics , Dicistroviridae/pathogenicity , Disease Transmission, Infectious/veterinary , Mites/genetics , Pollination , RNA, Double-Stranded , Social Behavior , Virulence
18.
Proc Natl Acad Sci U S A ; 117(19): 10511-10519, 2020 05 12.
Article in English | MEDLINE | ID: mdl-32341166

ABSTRACT

Honey bees (Apis mellifera) produce an enormous economic value through their pollination activities and play a central role in the biodiversity of entire ecosystems. Recent efforts have revealed the substantial influence that the gut microbiota exert on bee development, food digestion, and homeostasis in general. In this study, deep sequencing was used to characterize prokaryotic viral communities associated with honey bees, which was a blind spot in research up until now. The vast majority of the prokaryotic viral populations are novel at the genus level, and most of the encoded proteins comprise unknown functions. Nevertheless, genomes of bacteriophages were predicted to infect nearly every major bee-gut bacterium, and functional annotation and auxiliary metabolic gene discovery imply the potential to influence microbial metabolism. Furthermore, undiscovered genes involved in the synthesis of secondary metabolic biosynthetic gene clusters reflect a wealth of previously untapped enzymatic resources hidden in the bee bacteriophage community.


Subject(s)
Bacteriophages/genetics , Bees/metabolism , Bees/virology , Animals , Bacteria/genetics , Bacteriophages/metabolism , Bees/genetics , Biodiversity , Ecosystem , Gastrointestinal Microbiome/genetics , High-Throughput Nucleotide Sequencing/methods , Metagenomics/methods , Phylogeny , Pollination/genetics , Symbiosis/genetics
19.
PLoS Biol ; 17(10): e3000502, 2019 10.
Article in English | MEDLINE | ID: mdl-31600204

ABSTRACT

The impacts of invertebrate RNA virus population dynamics on virulence and infection outcomes are poorly understood. Deformed wing virus (DWV), the main viral pathogen of honey bees, negatively impacts bee health, which can lead to colony death. Despite previous reports on the reduction of DWV diversity following the arrival of the parasitic mite Varroa destructor, the key DWV vector, we found high genetic diversity of DWV in infested United States honey bee colonies. Phylogenetic analysis showed that divergent US DWV genotypes are of monophyletic origin and were likely generated as a result of diversification after a genetic bottleneck. To investigate the population dynamics of this divergent DWV, we designed a series of novel infectious cDNA clones corresponding to coexisting DWV genotypes, thereby devising a reverse-genetics system for an invertebrate RNA virus quasispecies. Equal replication rates were observed for all clone-derived DWV variants in single infections. Surprisingly, individual clones replicated to the same high levels as their mixtures and even the parental highly diverse natural DWV population, suggesting that complementation between genotypes was not required to replicate to high levels. Mixed clone-derived infections showed a lack of strong competitive exclusion, suggesting that the DWV genotypes were adapted to coexist. Mutational and recombination events were observed across clone progeny, providing new insights into the forces that drive and constrain virus diversification. Accordingly, our results suggest that Varroa influences DWV dynamics by causing an initial selective sweep, which is followed by virus diversification fueled by negative frequency-dependent selection for new genotypes. We suggest that this selection might reflect the ability of rare lineages to evade host defenses, specifically antiviral RNA interference (RNAi). In support of this hypothesis, we show that RNAi induced against one DWV strain is less effective against an alternate strain from the same population.


Subject(s)
Arachnid Vectors/virology , Bees/virology , Immune Evasion/genetics , RNA Viruses/genetics , Varroidae/virology , Animals , Bees/genetics , Bees/immunology , Bees/parasitology , Clone Cells , Gene Library , Genetic Variation , Genotype , Mutation , Phylogeny , RNA Interference/immunology , RNA Viruses/classification , RNA Viruses/immunology , RNA Viruses/pathogenicity , Recombination, Genetic , Reverse Genetics/methods , Selection, Genetic , Virulence , Virus Replication
20.
J Insect Sci ; 22(1)2022 Jan 01.
Article in English | MEDLINE | ID: mdl-35137131

ABSTRACT

Transgenerational immune priming is the process of increased resistance to infection in offspring due to parental pathogen exposure. Honey bees (Apis mellifera L. (Hymenoptera: Apidae)) are hosts to multiple pathogens, and this complex immune function could help protect against overwhelming infection. Honey bees have demonstrated transgenerational immune priming for the bacterial pathogen Paenibacillus larvae; however, evidence for viral transgenerational immune priming is lacking across insects in general. Here we test for the presence of transgenerational immune priming in honey bees with Deformed wing virus (DWV) by injecting pupae from DWV-exposed queens and measuring virus titer and immune gene expression. Our data suggest that there is evidence for viral transgenerational immune priming in honey bees, but it is highly context-dependent based on route of maternal exposure and potentially host genetics or epigenetic factors.


Subject(s)
Bees , Insect Viruses , RNA Viruses , Animals , Bees/immunology , Bees/virology , Female , Maternal Exposure , Pupa , Viral Load
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