Mellifera Populations Research Articles

Rapid identification of Africanized honey bee Apis mellifera (hymenoptera: apidae) using high-resolution melting analysis DNA from honey

Genetic characterization of honey bee populations, particularly the identification of Africanized honey bees, has traditionally been based on different molecular methods that involve the amplification and analysis of different regions of the mitochondrial or nuclear genome which often take a long time. We introduce high-resolution melting analysis (HRM) as a novel approach for rapid and cost-effective identification of the African lineage in Apis mellifera populations from mitochondrial DNA extracted from honey. Samples were collected from 111 apiaries in 13 Argentinean provinces between 2012 and 2018. Total DNA was extracted from 10 g honey sediment and subjected to quantitative PCR (qPCR) using specific primers (APIS-F/Afr207R) amplifying a 207 bp product. Positive controls (Africanized/Non-Africanized) were used for mitotype validation and a subset of qPCR products were sequenced to compare different haplotypes. HRM analysis enabled the quick detection of sequence polymorphisms in short amplicons generated by qPCR. Among the analyzed apiaries, 26 located above 35° S in Misiones, La Rioja, Catamarca, Formosa, Corrientes, Córdoba, and Santa Fé exhibited the Africanized mitotypes while the remaining apiaries below this latitude, mainly in Buenos Aires, Rio Negro, Neuquén, and Entre Ríos, showed the European mitotypes. These results align with previous characterizations in the region. HRM analysis demonstrated high sensitivity and effectiveness, requiring minimal DNA input, and reducing costs and sampling efforts. This approach provides a rapid and reliable method for identifying the African lineage, facilitating honey bee breeding programs, conservation efforts, and management strategies in regions with Africanized honey bee populations.

Recapping behavior in Apis cerana: does it contribute to resistance against Varroa spp.?

The invasion of the ectoparasitic mite Varroa destructor into European honey bee, Apis mellifera, populations has contributed to the collapse of most wild populations and economic losses in beekeeping operations. Understanding how some A. mellifera populations survive infestation by this parasite is of great fundamental and practical interest and has led to numerous studies of potential resistance mechanisms. One such mechanism is the uncapping and recapping of comb cells containing infested brood by nurse bees. Recapping has been observed in most surviving populations, but its link to V. destructor resistance remains unclear. Investigating the occurrence of recapping in the Eastern honey bee, Apis cerana, the original host of the parasite, could provide a better understanding of the evolution and function of this behaviour in the Apis genus. Here, we determined the frequency of recapping in two A. cerana populations in China and Thailand at different stages of brood development and compared them with a sympatric A. mellifera population in China. The species, which differ in their susceptibility to infestation, did not show significant differences in recapping frequency. A specific association between recapping and resistance to Varroa spp. in A. cerana is therefore not supported. We discuss possible functions and evolutionary scenarios for this behavior.

Thermal Tolerance of Honeybees (Apis mellifera L.) Changes Across an Elevation Gradient in the Mexican Transition Zone

The objective of this study was to determine the critical thermal minimum [CTmin], critical thermal maximum [CTmax], and thermal tolerance range of A. mellifera at three different elevations located in the Mexican Transition Zone: 11; 1,324, and 3,304 m.a.s.l. In general, we found that the CTmin of A. mellifera was lower at the site with the highest elevation (i.e., they tolerate colder temperatures). At the same time, the CTmax remained constant across the three studied elevations, revealing higher plasticity for cold tolerance rather than heat. Moreover, we did not find evidence that the body mass of the individuals was associated with their thermal tolerance at any of the three sampled elevations. Our findings suggest processes of local adaptation of A. mellifera populations in environmentally contrasting sites, allowing them to expand their range of distribution, which could be useful in predicting responses to future environmental change.

Genetic Diversity of West African Honey Bee (Apis ‎mellifera adansonii Latreille, 1804) from Rural and Urban ‎Areas of ‎Kwara State, North-Central Nigeria

Over one third of the world’s crops– including fruits, vegetables, nuts, spices, and oilseed–‎require insect pollination, and human reliance on ‎pollination services by honey bees (Apis ‎mellifera) to promote these crops continues to rise due to increasing demands from growing ‎human ‎populations. Identifying the effects of urbanization on genetic diversity on this ‎pollinating insect is important in the field of bioscience. This study aimed to investigate genetic diversity of A. mellifera in Kwara State, Nigeria, using the random amplified polymorphic DNA (RAPD) marker. ‎Thirty honey bees ‎were simultaneously collected from both rural and urban regions in ‎Kwara state, Nigeria. Samples were morphologically identified using ‎standard methods, ‎genomic DNA isolated and amplified using five RAPD primers. Data collected were ‎analysed using PyElph, ‎ARLEQUIN, and GeneAlEx version 6.501 software. The results ‎showed that the DNA fragment sizes produced per primer varied from 200 to ‎‎3000 bp. Percentages of polymorphic loci amplified by each primer varied from 17.33 to 33.33%. ‎Analysis of unbiased Nei genetic ‎distance values showed that Agbede (rural) and Adewole ‎‎(urban) showed the highest value of unbiased genetic distance (0.073), while ‎Amoyo ‎‎(rural) to Idofian (urban) exhibited the lowest value (0.027). Dendrogram analysis revealed ‎genetically close relationships among the sampled ‎A. mellifera‎ populations. The low level of genetic ‎polymorphisms observed among the honey bee populations in the two ‎regions ‎indicated that there is genetic relatedness among them. This study concluded that RAPD ‎marker is a useful method for ‎understanding population genetic structure of the African honey ‎bees. These results can be used as baseline information for future genetic ‎diversity ‎assessment of honey bees in Nigeria with larger samples. It is therefore recommended that ‎there is a need to safeguard the genetic ‎diversity of A. mellifera‎ to prevent extinction or ‎gradual loss of diversity‎‎‎.

Genetic Diversity of Kırklareli Honey Bee (Apis mellifera L.) Populations in Thrace Region of Turkey: Identification of Mitochondrial COI and ND5 Gene Regions

The main goal of genetic resource conservation is to keep as much genetic diversity as possible within each species. In this respect, some difficulties in the protection of honey bee gene resources make it necessary to reveal the genetic structures of the subspecies and the genetic relationships between the subspecies. In this study, Kırklareli honey bee populations which were officially registered as an ecotype of Turkey’s honey bee (Apis mellifera L.) gene resources by the Republic of Turkey, Ministry of Agriculture and Forestry, were examined in the COI and ND5 genes of mitochondrial DNA. The restriction fragment length polymorphism (RFLP) together with the polymerase chain reaction (PCR) was used to define Apis mellifera populations. A total of 117 worker bee samples were used which were collected from mostly the Kırklareli province. A newly found single nucleotide polymorphism (SNP), G→A transition in the COI gene region formed a novel NcoI restriction site resulting in a new haplotype. This new haplotype has been abbreviated as haplotype C. As a result of the COI/SspI digestion, the previously reported C haplotype was determined. No restriction was found with the treatment of COI/StyI enzyme. On the other hand, as a result of ND5/AluI restriction, 2 restriction site and previously reported haplotype C was obtained in all of the studied samples. No restriction was screened with ND5/FokI and ND5/HincII enzymes in the whole samples, only a reported uncut B haplotype was observed. Within this study, novel genetic information has been revealed for the Kırklareli honey bee ecotype registered as the Thrace honey bee of Turkey’s honey bee gene resources. Moreover, detailed studies with larger sample sizes should be conducted to characterize the origin and the subspecies of Kırklareli honey bees in detail. It is thought that this study will be useful in the identification and registration of the Kırklareli honey bees to be carried out in the future, and also in the creation of a database.

Signatures of adaptive decreased virulence of deformed wing virus in an isolated population of wild honeybees (Apis mellifera).

Understanding the ecological and evolutionary processes that drive host-pathogen interactions is critical for combating epidemics and conserving species. The Varroa destructor mite and deformed wing virus (DWV) are two synergistic threats to Western honeybee (Apis mellifera) populations across the globe. Distinct honeybee populations have been found to self-sustain despite Varroa infestations, including colonies within the Arnot Forest outside Ithaca, NY, USA. We hypothesized that in these bee populations, DWV has been selected to produce an avirulent infection phenotype, allowing for the persistence of both host and disease-causing agents. To investigate this, we assessed the titre of viruses in bees from the Arnot Forest and managed apiaries, and assessed genomic variation and virulence differences between DWV isolates. Across groups, we found viral abundance was similar, but DWV genotypes were distinct. We also found that infections with isolates from the Arnot Forest resulted in higher survival and lower rates of symptomatic deformed wings, compared to analogous isolates from managed colonies, providing preliminary evidence to support the hypothesis of adaptive decreased viral virulence. Overall, this multi-level investigation of virus genotype and phenotype indicates that host ecological context can be a significant driver of viral evolution and host-pathogen interactions in honeybees.

Haplotypes of the tRNAleu-COII mtDNA Region in Russian Apis mellifera Populations.

Analysis of the mtDNA tRNAleu-COII locus is a widely used tool to establish belonging to a particular evolutionary lineage of Apis mellifera L. (lineages A, M, C, O, and Y). In Russia, most of the area was once inhabited by Apis mellifera mellifera from the M evolutionary lineage, but the introduction of bee subspecies from the southern regions of Russia (A. m. caucasica, A. m. carnica) and from abroad (A. m. carnica, A. m. ligustica) led to fragmentation of their native range. In this study, the results of assessing the haplotype number for the tRNAleu-COII locus of mtDNA in Russian Apis mellifera populations were presented. We analyzed 269 colonies from 19 regions of Russia. As a result, two evolutionary lineages were identified: the East European lineage C (26.4%) and the Northwestern European lineage M (73.6%). A total of 29 haplotypes were identified, 8 of them were already reported, and 21 were found to be novel. From the C lineage, haplotypes C1, C2, C2c, C2j, and C3 were predominant. All M lineage samples from Russia belong to the M17 and M4' haplogroups but have only minor variations in the form of nucleotide substitutions. An analysis of publications devoted to the tRNAleu-COII locus haplotypes, as well as an analysis of the available tRNAleu-COII sequences in GenBank, showed that there is still a problem with the haplotype nomenclature.

Delivery mechanism can enhance probiotic activity against honey bee pathogens

Managed honey bee (Apis mellifera) populations play a crucial role in supporting pollination of food crops but are facing unsustainable colony losses, largely due to rampant disease spread within agricultural environments. While mounting evidence suggests that select lactobacilli strains (some being natural symbionts of honey bees) can protect against multiple infections, there has been limited validation at the field-level and few methods exist for applying viable microorganisms to the hive. Here, we compare how two different delivery systems—standard pollen patty infusion and a novel spray-based formulation—affect supplementation of a three-strain lactobacilli consortium (LX3). Hives in a pathogen-dense region of California are supplemented for 4 weeks and then monitored over a 20-week period for health outcomes. Results show both delivery methods facilitate viable uptake of LX3 in adult bees, although the strains do not colonize long-term. Despite this, LX3 treatments induce transcriptional immune responses leading to sustained decreases in many opportunistic bacterial and fungal pathogens, as well as selective enrichment of core symbionts including Bombilactobacillus, Bifidobacterium, Lactobacillus, and Bartonella spp. These changes are ultimately associated with greater brood production and colony growth relative to vehicle controls, and with no apparent trade-offs in ectoparasitic Varroa mite burdens. Furthermore, spray-LX3 exerts potent activities against Ascosphaera apis (a deadly brood pathogen) likely stemming from in-hive dispersal differences, whereas patty-LX3 promotes synergistic brood development via unique nutritional benefits. These findings provide a foundational basis for spray-based probiotic application in apiculture and collectively highlight the importance of considering delivery method in disease management strategies.

Microplastics incorporated by honeybees from food are transferred to honey, wax and larvae

Microplastics (MP) are ubiquitous in the environment, and there is little information available on their impact on terrestrial organisms. Their effect on insects and particularly on honeybees is relevant, given the prevalence of these organisms in the environment and the fact that they provide key ecosystem services. We conducted a field study to assess (1) the fate of these ingested MP within the hive, and (2) MP effect on Apis mellifera population growth during chronic exposure. We aimed to determine if MP ingested by honeybees are incorporated into hive matrices, including honey, and their effect on colony development and honey reserves. We fed beehives with sucrose solutions treated or untreated with 50 mg of Polyester microfibers/L for one month. Microplastic fibers (MF) from treated syrup were incorporated by adult worker bees, remaining in their cuticle, digestive tract, larvae, honey, and wax. Most of the MF were accumulated in wax showing that honey remains as a safe food. At the end of the experiment, no differences in honey reserves or bee population were observed. This is the first study to evaluate in the field the effects and dynamics of MP inside honeybee hives. Our results showed that bees can incorporate MP from the environment and deliver them into the different matrices of the hive. Concentration of MF found in honey of treated hives was like that found in commercial honey, suggesting that honeybees might be exposed to similar MP contamination levels in the environment compared to our experiment. Finally, our results highlight a way in which MP might enter the food chain, with direct implication for human health.

Nosema disease is an actual problem of beekeeping

The article presents an information review on the infectious disease of bees – nosematosis. The features of the structure of the spore, the life cycle of development, transmission of infection are described, as well as methods of diagnosis, treatment, and prevention are displayed. Alternative methods of treatment and their effectiveness are shown. A decrease in the population of the honey bee (Apis mellifera) was noted. One of the reasons is an opportunistic infection of bees caused by microsporidia of the genus Nozema. Nosematosis occurs in most countries of the world. At the moment, three species of this parasite have been described in honey bees: N. aris, N. ceranae and N. neumanni. The most common are N. aris, N. ceranae. In 2011, Gisder et al. a model of tissue cell culture was developed, which made it possible to see the intracellular life cycle of these microsporidia. It is important for the preservation of the Apis mellifera population to study up-to-date information and look for new ways to combat this disease. Honey bees play an important role for humanity and are an integral part of biological diversity. Its work as a pollinator of agricultural crops and other plants cannot be overestimated.

A genotyping by sequencing approach can disclose Apis mellifera population genomic information contained in honey environmental DNA

Awareness has been raised over the last years on the genetic integrity of autochthonous honey bee subspecies. Genomic tools available in Apis mellifera can make it possible to measure this information by targeting individual honey bee DNA. Honey contains DNA traces from all organisms that contributed or were involved in its production steps, including the honey bees of the colony. In this study, we designed and tested a genotyping by sequencing (GBS) assay to analyse single nucleotide polymorphisms (SNPs) of A. mellifera nuclear genome using environmental DNA extracted from honey. A total of 121 SNPs (97 SNPs informative for honey bee subspecies identification and 24 SNPs associated with relevant traits of the colonies) were used in the assay to genotype honey DNA, which derives from thousands of honey bees. Results were integrated with information derived from previous studies and whole genome resequencing datasets. This GBS method is highly reliable in estimating honey bee SNP allele frequencies of the whole colony from which the honey derived. This assay can be used to identify the honey bee subspecies of the colony that produced the honey and, in turn, to authenticate the entomological origin of the honey.

Varroa destructor in Portugal: an exploratory assessment of pyrethroids resistance status

The drastic and abrupt loss of honey bee colonies around the world has long unsettled the beekeeping community, and although the phenomenon is far from being solved, there is a relative consensus regarding the detrimental effect the mite Varroa destructor exerts over Apis mellifera populations. In Portugal, this parasite was first recorded in the late 1980s and since then, the beekeepers’ method of choice for controlling the mite are synthetic acaricides, most notably the pyrethroids tau-fluvalinate and flumethrin. Unsurprisingly, resistance to these active ingredients quickly surfaced. Here, we report the first PCR-RFLP resistance assessment in Portuguese apiaries, where 96 V. destructor mites, from different geographic regions, were genotyped between April and August of 2019. The results showed that 47% of the sampled mites exhibit the mutation correlated with resistance. These findings stress the need for a current characterization of the acaricides being used and a description of how the treatments are been applied.

Exposure of Larvae to Sublethal Thiacloprid Delays Bee Development and Affects Transcriptional Responses of Newly Emerged Honey Bees.

Understanding the cause of honey bee (Apis mellifera) population decline has attracted immense attention worldwide in recent years. Exposure to neonicotinoid pesticides is considered one of the most probable factors due to the physiological and behavioral damage they cause to honey bees. However, the influence of thiacloprid, a relatively less toxic cyanogen-substituted form of neonicotinoid, on honey bee (Apis mellifera L.) development is not well studied. The toxicity of sublethal thiacloprid to larvae, pupae, and emerging honey bees was assessed under laboratory conditions. We found that thiacloprid reduced the survival rate of larvae and pupae, and delayed the development of bees which led to lower bodyweight and size. Furthermore, we identified differentially expressed genes involved in metabolism and immunity though RNA-sequencing of newly-emerged adult bees. GO enrichment analysis identified genes involved in metabolism, catalytic activity, and transporter activity. KEGG pathway analysis indicated that thiacloprid induced up-regulation of genes related to glutathione metabolism and Toll-like receptor signaling pathway. Overall, our results suggest that chronic sublethal thiacloprid can affect honey bee colonies by reducing survival and delaying bee development.

Dynamics of the Genetic Structure of Apis mellifera Populations in the Southern Urals

Dynamics of the Genetic Structure of Apis mellifera Populations in the Southern Urals

Malate Dehydrogenase allele frequencies in the commercial honey bee (Apis mellifera) population in Thailand reflect those in source populations

Malate Dehydrogenase allele frequencies in the commercial honey bee (Apis mellifera) population in Thailand reflect those in source populations

Dissipation kinetics of imidacloprid in cotton flower, nectariferous tissue, pollen and Apis mellifera products using QuEChERS method.

Exposure of Apis mellifera to neonicotinoid insecticides is one of the factors attributed to the recent decline in A. mellifera populations resulting in economic and ecological losses due to loss of pollination services. Honey bees can get exposed to neonicotinoids like imidacloprid directly in the field at the time of application as well as during consumption of pollen and nectar from treated plants. So, the fate of imidacloprid in commodities to which honey bees get exposed needs to be overhauled. Residue of imidacloprid was investigated following imidacloprid application as seed treatment (2.4 and 4.8 g a.i. kg-1 seed) at the time of sowing and as foliar spray (17.8 and 35.6 g a.i. ha-1 ) at 70 days after sowing when the crop was at full bloom stage. The imidacloprid residue was below limit of quantification (LOQ) in flowers, necatariferous tissue, pollen from seed-treated cotton plants and honey collected from hives kept in plots with seed-treated cotton plants. However, average initial imidacloprid residue (2 h after spray) was 1.84 and 1.95 mg kg-1 in flowers; 0.22 and 0.24 mg kg-1 in nectariferous tissue; 0.88 and 0.96 mg kg-1 in pollen collected from plants sprayed with imidacloprid @ 17.8 g a.i. ha-1 at locations Faridkot and Bathinda, respectively. The average initial imidacloprid residue (21 days after spray) in honey collected from hives was 0.01 mg kg-1 . The residue in different substrates sampled from seed treated cotton plants was below LOQ. However, its foliar spray at bloom time resulted in imidacloprid residue in flower, nectariferous tissue, pollen and honey sampled from hives placed in plots. © 2021 Society of Chemical Industry.

The Current State of the Protected Apis mellifera mellifera Population in Russia: Hybridization and Nosematosis.

Simple SummaryThe Southern Ural Mountains are a habitat for one of the remaining populations of the dark forest bees Apis mellifera mellifera. Using molecular genetic methods, we have established that there are processes of hybridization of this population with subspecies from the evolutionary lineage C. In addition, some colonies are affected by nosematosis. Therefore, it is necessary to take urgent measures to preserve this population.The Southern Urals of Russia are the habitat of one of the surviving populations of the dark forest bee—the Burzyan population of Apis mellifera mellifera. In this study, we present the results of the subspecies identification of bee colonies in the Altyn-Solok Nature Reserve in the Southern Ural Mountains using the intergenic mtDNA COI-COII locus and the assessment of the prevalence of nosematosis. Analysis of the mtDNA COI-COII intergenic locus in the studied sample showed that 30.4% of the colonies belong to the lineage C. The PCR diagnostics of nosematosis in 92 colonies selected from different sectors of the Altyn-Solok Nature Reserve showed that about half of the analyzed colonies were infected with Nosema apis. Nosema ceranae was found in eight colonies. Both of these factors can lead to the extinction of this population of the dark forest bee.

Honey bee viruses are highly prevalent but at low intensities in wild pollinators of cucurbit agroecosystems

Managed and wild bee populations are in decline around the globe due to several biotic and abiotic stressors. Pathogenic viruses associated with the Western honey bee (Apis mellifera) have been identified as key contributors to losses of managed honey bee colonies, and are known to be transmitted to wild bee populations through shared floral resources. However, little is known about the prevalence and intensity of these viruses in wild bee populations, or how bee visitation to flowers impacts viral transmission in agroecosystems. This study surveyed honey bee, bumble bee (Bombus impatiens) and wild squash bee (Eucera (Peponapis) pruinosa) populations in Cucurbita agroecosystems across Pennsylvania (USA) for the prevalence and intensity of five honey bee viruses: acute bee paralysis virus (ABPV), deformed wing virus (DWV), Israeli acute paralysis virus (IAPV), Kashmir bee virus (KBV), and slow bee paralysis virus (SBPV). We investigated the potential role of bee visitation rate to flowers on DWV intensity among species in the pollinator community, with the expectation that increased bee visitation to flowers would increase the opportunity for transmission events between host species. We found that honey bee viruses are highly prevalent but in lower titers in wild E. pruinosa and B. impatiens than in A. mellifera populations throughout Pennsylvania (USA). DWV was detected in 88% of B. impatiens, 48% of E. pruinosa, and 95% of A. mellifera. IAPV was detected in 5% of B. impatiens and 4% of E. pruinosa, compared to 9% in A. mellifera. KBV was detected in 1% of B. impatiens and 5% of E. pruinosa, compared to 32% in A. mellifera. Our results indicate that DWV titers are not correlated with bee visitation in Cucurbita fields. The potential fitness impacts of these low viral titers detected in E. pruinosa remain to be investigated.

Validation of QuEChERS Method for Estimation of Imidacloprid and its Metabolites in Cotton Flower, Pollen, Nectariferous Tissue, and Honey.

Exposure of Apis mellifera to neonicotinoid insecticides is one of the factors attributed to the recent decline in A. mellifera populations resulting in economic and ecological losses due to loss of pollination services. Honeybees can get exposed to neonicotinoids like imidacloprid directly in the field at the time of application as well as during consumption of pollen and nectar from treated plants. In addition, some metabolites of imidacloprid are more toxic than the parent compound. So, the fate of imidacloprid and its metabolites in commodities to which honeybees get exposed needs to be overhauled. To validate QuEChERS method for estimation of imidacloprid and its metabolites in cotton flower, pollen, nectariferous tissue, and honey using HPLC. The QuEChERS method was validated in terms of selectivity, linearity, LOD, LOQ, matrix match, accuracy, and precision. The estimation of residues was done by HPLC. Recoveries of imidacloprid and its metabolites for cotton flowers, nectariferous tissue, pollen and honey samples were in the range of 80.42-99. 83%. LOQ for imidacloprid and its metabolites was 0.01 µg/g. Acceptable precision (RSD < 20%) was obtained. The method allows simple and fast extraction of imidacloprid and its metabolites from cotton flower, pollen, nectariferous tissue, and honey. An accurate, simple, and sensitive analytical method was validated for imidacloprid and its metabolites. The method was validated according to the SANTE/12682/2019 guidelines.

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

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