Molecular Assessment of Genes Linked to Immune Response Traits of Honey Bees in Conventional and Organically Managed Apiaries

Honey bees are of great economic importance, not only for honey production but also for crop pollination. However, honey bee populations continue to decline mainly due to exposure to pesticides, pathogens and beekeeping practices. In this study, total soluble protein was measured, total RNA was extracted and first-strand cDNAs were generated. Quantitative PCR was used to assess the relative expression (transcript abundances) of immune function-related genes in honey bees collected from organically and conventionally managed hives. Honey bees collected from conventionally managed hives with 0% Varroa mite infestation levels displayed an upregulated expression of the prophenoloxidase gene (cellular defense). Similarly, honey bees collected from organically managed hives had increased levels of the vitellogenin gene (immune function and longevity). The gene expression for malvolio (sucrose responsiveness) was highest in organically managed hives with 0% Varroa mite infestations. Young adult bees collected from organically managed hives with 5% Varroa mite infestation levels had upregulated expressions of the gene spaetzle, whereas bees from similarly infested, conventionally managed hives did not, suggesting that honey bees from organically managed hives could mount an immune response. In young adult bees collected from organically managed hives only, the expression of the immune deficiency gene (antimicrobial defense) was upregulated. The relative gene expression for superoxide dismutase 1 increased in young adult bees collected from hives with 5% Varroa mite infestation levels as expected. However, for superoxide dismutase 2, there was a high level of gene expression in adult bees from both conventionally managed hives with 0% Varroa mite infestation levels and organically managed hives with 5% Varroa mite infestations. The gene CYP9Q3 (pesticide detoxification) that metabolizes coumaphos and fluvalinate was upregulated in adult bees collected from organically managed bees. Overall, these findings provide useful insights into the genetic response of honey bees to some environmental stressors and could be an important component of best beekeeping practices that intend to enhance honey bee health.

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Esterase Activity is Affected by Genetics, Age, Insecticide Exposure, and Viral Infection in the Honey Bee, Apis mellifera

10.1101/415356 ◽

2018 ◽

Author(s):

Frank D. Rinkevich ◽

Joseph W. Margotta ◽

Michael Simone-Finstrom ◽

Lilia I. de Guzman ◽

Kristen B. Healy

Keyword(s):

Viral Infection ◽

Honey Bee ◽

Honey Bees ◽

Esterase Activity ◽

Pesticide Exposure ◽

Mite Infestation ◽

Activity Levels ◽

Varroa Mite ◽

Genetic Stock ◽

Insecticide Exposure

AbstractNon-target impacts of insecticide treatments are a major public and environmental concern, particularly in contemporary beekeeping. Therefore, it is important to understand the physiological mechanisms contributing to insecticide sensitivity in honey bees. In the present studies, we sought to evaluate the role of esterases as the source of variation in insecticide sensitivity. To address this question, the following objectives were completed: 1) Evaluated esterase activity among honey bee stocks, 2) Assessed the correlation of esterase activity with changes in insecticide sensitivity with honey bee age, 3) Established if esterases can be used as a biomarker of insecticide exposure, and 4) Examined the effects of Varroa mite infestation and viral infection on esterase activity.Results indicated that honey bees have a dynamic esterase capacity that is influenced by genetic stock and age. However, there was no consistent connection of esterase activity with insecticide sensitivity across genetic stocks or with age, suggests other factors are more critical for determining insecticide sensitivity. The trend of increased esterase activity with age in honey bees suggests this physiological transition is consistent with enhanced metabolic rate with age. The esterase inhibition with naled but not phenothrin or clothianidin indicates that reduced esterase activity levels may only be reliable for sublethal doses of organophosphate insecticides. The observation that viral infection, but not Varroa mite infestation, reduced esterase activity shows viruses have extensive physiological impacts. Taken together, these data suggest that honey bee esterase activity toward these model substrates may not correlate well with insecticide sensitivity. Future studies include identification of esterase substrates and inhibitors that are better surrogates of insecticide detoxification in honey bees as well as investigation on the usefulness of esterase activity as a biomarker of pesticide exposure, and viral infection.

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Identification of long noncoding RNAs reveals the effects of dinotefuran on the brain in Apis mellifera (Hymenopptera: Apidae)

BMC Genomics ◽

10.1186/s12864-021-07811-y ◽

2021 ◽

Vol 22 (1) ◽

Author(s):

Minjie Huang ◽

Jie Dong ◽

Haikun Guo ◽

Minghui Xiao ◽

Deqian Wang

Keyword(s):

Immune Response ◽

Honey Bee ◽

Honey Bees ◽

Transforming Growth Factor ◽

Sublethal Effects ◽

Inflammatory Responses ◽

Noncoding Rnas ◽

Enrichment Analysis ◽

Long Noncoding Rnas ◽

Gene Set Enrichment Analysis

Abstract Background Dinotefuran (CAS No. 165252–70-0), a neonicotinoid insecticide, has been used to protect various crops against invertebrate pests and has been associated with numerous negative sublethal effects on honey bees. Long noncoding RNAs (lncRNAs) play important roles in mediating various biological and pathological processes, involving transcriptional and gene regulation. The effects of dinotefuran on lncRNA expression and lncRNA function in the honey bee brain are still obscure. Results Through RNA sequencing, a comprehensive analysis of lncRNAs and mRNAs was performed following exposure to 0.01 mg/L dinotefuran for 1, 5, and 10 d. In total, 312 lncRNAs and 1341 mRNAs, 347 lncRNAs and 1458 mRNAs, and 345 lncRNAs and 1155 mRNAs were found to be differentially expressed (DE) on days 1, 5 and 10, respectively. Gene set enrichment analysis (GSEA) indicated that the dinotefuran-treated group showed enrichment in carbohydrate and protein metabolism and immune-inflammatory responses such as glycine, serine and threonine metabolism, pentose and glucuronate interconversion, and Hippo and transforming growth factor-β (TGF-β) signaling pathways. Moreover, the DE lncRNA TCONS_00086519 was shown by fluorescence in situ hybridization (FISH) to be distributed mainly in the cytoplasm, suggesting that it may serve as a competing endogenous RNA and a regulatory factor in the immune response to dinotefuran. Conclusion This study characterized the expression profile of lncRNAs upon exposure to neonicotinoid insecticides in young adult honey bees and provided a framework for further study of the role of lncRNAs in honey bee growth and the immune response.

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Varroa jacobsoni infestation of adult Africanized and Italian honey bees (Apis mellifera) in mixed colonies in Brazil

Genetics and Molecular Biology ◽

10.1590/s1415-47571999000300006 ◽

1999 ◽

Vol 22 (3) ◽

pp. 321-323 ◽

Cited By ~ 12

Author(s):

Geraldo Moretto ◽

Leonidas João de Mello Jr.

Keyword(s):

Apis Mellifera ◽

Honey Bee ◽

Honey Bees ◽

Mite Infestation ◽

Varroa Jacobsoni ◽

Africanized Honey Bees ◽

African Honey Bees ◽

Africanized Bees ◽

Bee Colonies ◽

Different Levels

Different levels of infestation with the mite Varroa jacobsoni have been observed in the various Apis mellifera races. In general, bees of European races are more susceptible to the mite than African honey bees and their hybrids. In Brazil honey bee colonies are not treated against the mite, though apparently both climate and bee race influence the mite infestation. Six mixed colonies were made with Italian and Africanized honey bees. The percentage infestation by this parasite was found to be significantly lower in adult Africanized (1.69 ± 0.44) than Italian bees (2.79 ± 0.65). This ratio was similar to that found in Mexico, even though the Africanized bees tested there had not been in contact with varroa, compared to more than 20 years of the coexistence in Brazil. However, mean mite infestation in Brazil on both kinds of bees was only about a third of that found in Mexico.

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Challenges associated with the honey bee (Apis mellifera adansonii) colonies establishment in south western Nigeria

African Journal of Food, Agriculture, Nutrition and Development ◽

10.18697/ajfand.57.12175 ◽

2013 ◽

Vol 13 (57) ◽

pp. 7467-7484

Author(s):

KL Akinwande ◽

◽

MA Badejo ◽

SS Ogbogu ◽

◽

...

Keyword(s):

Honey Bees ◽

Aethina Tumida ◽

Ecological Problem ◽

Varroa Mite ◽

Average Percentage ◽

Pests And Diseases ◽

Significant Difference ◽

European Foulbrood ◽

Apis Mellifera Adansonii ◽

Mite Infestations

The southwestern part of Nigeria is a tropical rainforest region having many local beekeepers. The se beekeepers have experienced decline in colony establishment in the recent past . A study carried out in Lagos, Ogun and Osun states between December 2009 and September 2011 examined 14 randomly selected commercial bee farms for problems associated with decline in colony establishment . Sampling and treatments were split equally between each apiary and three colonies were selected in each . All the colonies were housed in Tanzania /local top bar hive s. There were 58.34, 44.84 and 40.61 average percentage declines in colony establishment in Lagos, Ogun and Osun States , respectively. Presence of pests and diseases, pesticide poisoning, poor hive and seasonal management, ecological problem and lack of queen rearing were potential problems identified by the beekeepers . All the apiaries had pests like Crickets, Ants ( Companotus pennsylvanicus ), Small Hive beetle ( Aethina tumida ), Termites ( Macrotermes spp ) and Spider ( Lactrodectus mactan ) . Varroa mite infestations were found in 33 (78.57%) of apiaries sampled. There was no significant difference between the levels of Varroa infestation in all the colonies during the dry and wet seasons at confidence interval of 95 percent ( t = 1.542, df = 13, p = 0.147 ( p > 0.05) . Nosema spores were found in 27 (64.29%) colonies examined. The number of spores range from 16 x 10 3 to 30.4 x 10 3 . T here were no significant differences in the infection from colony to colony, apiary to apiary and between dry and wet seasons ( t = - 0.094 df = 11, P = 0.927 (P > 0.05). Diseases like American and European foulbrood were absent while chalkbrood disease was prevalent. Environmental factors of high temperature, high rainfall and high relative and hive humidity enhanced the spread of pests and disease pathogens. Many insecticides were constantly applied by the farmers on the crops in the surrounding farmland. Two of the commonly used insecticides DDVP (Dichlorvos) and Cyperforce (Cypermetrin) were discovered to have increased mortality on worker honey bees with progressively larger doses. It was obvious that insecticide toxicity had a significant effect on the colony. Hence , ecological and management problems had contributed to the decline in colony establishment in this area.

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

Insects ◽

10.3390/insects9030122 ◽

2018 ◽

Vol 9 (3) ◽

pp. 122 ◽

Cited By ~ 4

Author(s):

Paul Winkler ◽

Frank Sieg ◽

Anja Buttstedt

Keyword(s):

Gene Expression ◽

Apis Mellifera ◽

Juvenile Hormone ◽

Honey Bee ◽

Honey Bees ◽

Transcriptional Control ◽

Royal Jelly ◽

Adult Worker ◽

Gene Expression Dynamics

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

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

Nature Communications ◽

10.1038/s41467-020-19715-8 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Desiderato Annoscia ◽

Gennaro Di Prisco ◽

Andrea Becchimanzi ◽

Emilio Caprio ◽

Davide Frizzera ◽

...

Keyword(s):

Immune Responses ◽

Honey Bee ◽

Honey Bees ◽

Varroa Destructor ◽

Negative Impact ◽

Asymptomatic Infection ◽

Varroa Mite ◽

Deformed Wing Virus ◽

Synergistic Interactions ◽

Parasitic Mite

AbstractThe neonicotinoid Clothianidin has a negative impact on NF-κB signaling and on immune responses controlled by this transcription factor, which can boost the proliferation of honey bee parasites and pathogens. This effect has been well documented for the replication of deformed wing virus (DWV) induced by Clothianidin in honey bees bearing an asymptomatic infection. Here, we conduct infestation experiments of treated bees to show that the immune-suppression exerted by Clothianidin is associated with an enhanced fertility of the parasitic mite Varroa destructor, as a possible consequence of a higher feeding efficiency. A conceptual model is proposed to describe the synergistic interactions among different stress agents acting on honey bees.

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Honey Bee Viruses, Colony Health, and Antiviral Defense

Proceedings ◽

10.3390/proceedings2020050016 ◽

2020 ◽

Vol 50 (1) ◽

pp. 16

Author(s):

Katie F. Daughenbaugh ◽

Alex J. McMenamin ◽

Laura M. Brutscher ◽

Fenali Parekh ◽

Michelle L. Flenniken

Keyword(s):

Gene Expression ◽

Heat Shock ◽

Honey Bee ◽

Honey Bees ◽

Antiviral Defense ◽

Deformed Wing Virus ◽

Antiviral Responses ◽

Model Virus ◽

Virus Abundance

Honey bee colony losses are influenced by multiple abiotic and biotic factors, including viruses. To investigate the effects of RNA viruses on honey bees, we infected bees with a model virus (Sindbis-GFP) in the presence or absence of double-stranded RNA (dsRNA). In honey bees, dsRNA is the substrate for sequence-specific RNA interference (RNAi)-mediated antiviral defense and is a trigger of sequence-independent\antiviral responses. Transcriptome sequencing identified more than 200 differentially expressed genes, including genes in the RNAi, Toll, Imd, JAK-STAT, and heat shock response pathways, and many uncharacterized genes. To confirm the virus limiting role of two genes (i.e., dicer and mf116383) in honey bees, we utilized RNAi to reduce their expression in vivo and determined that the virus abundance increased. To evaluate the role of the heat shock stress response in antiviral defense, bees were heat stressed post-virus infection and the virus abundance and gene expression were assessed. Heat-stressed bees had reduced virus levels and a greater expression of several heat shock protein encoding genes (hsps) compared to the controls. To determine if these genes are universally associated with antiviral defense, bees were infected with another model virus, Flock House virus (FHV), or deformed wing virus and the gene expression was assessed. The expression of dicer was greater in bees infected with either FHV or Sindbis-GFP compared to the mock-infected bees, but not in the deformed wing virus-infected bees. To further investigate honey bee antiviral defense mechanisms and elucidate the function of key genes (dicer, ago-2, mf116383, and hsps) at the cellular level, primary honey bee larval hemocytes were transfected with dsRNA or infected with the Lake Sinai virus 2 (LSV2). These studies indicate that mf116383 and hsps mediate dsRNA detection and that MF116383 is involved in limiting LSV2 infection. Together, these results further our understanding of honey bee antiviral defense, particularly dsRNA-mediated antiviral responses, at both the individual bee and cellular levels.

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

10.21203/rs.3.rs-291062/v1 ◽

2021 ◽

Author(s):

Daniel B. Weaver ◽

Brandi L. Cantarel ◽

Christine Elsik ◽

Dawn L. Lopez ◽

Jay Evans

Keyword(s):

Gene Expression ◽

Honey Bee ◽

Honey Bees ◽

Host Responses ◽

Negative Effects ◽

Pollination Services ◽

Deformed Wing Virus ◽

Mortality And Morbidity ◽

The Individual ◽

Parasitic Mites

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

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Diagnosis of Varroa Mite (Varroa destructor) and Sustainable Control in Honey Bee (Apis mellifera) Colonies—A Review

Diversity ◽

10.3390/d11120243 ◽

2019 ◽

Vol 11 (12) ◽

pp. 243 ◽

Cited By ~ 1

Author(s):

Aleš Gregorc ◽

Blair Sampson

Keyword(s):

Apis Mellifera ◽

Organic Acids ◽

Essential Oils ◽

Honey Bee ◽

Varroa Destructor ◽

Mite Infestation ◽

Varroa Mite ◽

Mite Control ◽

Bee Colonies ◽

Reliable Methods

Determining varroa mite infestation levels in honey bee colonies and the proper method and time to perform a diagnosis are important for efficient mite control. Performing a powdered sugar shake or counting mites that drop from combs and bees onto a hive bottom board are two reliable methods for sampling varroa mite to evaluate the efficacy of an acaricide treatment. This overview summarizes studies that examine the efficacy of organic acids and essential oils, mite monitoring, and brood interruption for integrated varroa mite control in organic beekeeping.

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

Scientific Reports ◽

10.1038/s41598-021-94641-3 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Zih-Ting Chang ◽

Yu-Feng Huang ◽

Yue-Wen Chen ◽

Ming-Ren Yen ◽

Po-Ya Hsu ◽

...

Keyword(s):

Gene Expression ◽

Honey Bee ◽

Larval Stage ◽

Honey Bees ◽

Pupal Stage ◽

Deformed Wing Virus ◽

Expression Levels ◽

The Impact ◽

Honey Bee Larvae ◽

Gene Expression Levels

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

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