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Keywords = honey bee immune response

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16 pages, 1778 KiB  
Article
Synergistic Effects of Amitraz and Dinotefuran on Honey Bee Health: Impacts on Survival, Gene Expression, and Hypopharyngeal Gland Morphology
by Mojtaba Esmaeily, Tekalign Begna, Hyeonjeong Jang, Sunho Kwon and Chuleui Jung
Int. J. Mol. Sci. 2025, 26(14), 6850; https://doi.org/10.3390/ijms26146850 - 17 Jul 2025
Viewed by 258
Abstract
Honey bees (Apis mellifera) are major pollinators, playing a critical role in global food production, biodiversity, and ecosystem stability. However, their populations are increasingly threatened by multiple interacting stressors, including pesticide exposure. Among these, agricultural insecticides and anti-Varroa acaricides such [...] Read more.
Honey bees (Apis mellifera) are major pollinators, playing a critical role in global food production, biodiversity, and ecosystem stability. However, their populations are increasingly threatened by multiple interacting stressors, including pesticide exposure. Among these, agricultural insecticides and anti-Varroa acaricides such as dinotefuran and amitraz can persist in hive matrices, resulting in chronic and combined exposure. This study investigates the low lethal (LC10 and LC30) effects of these compounds, individually and in combination, on honey bee survival, immune function, oxidative stress responses, detoxification pathways, and hypopharyngeal gland morphology. Both pesticides negatively affected honey bee health at low lethal concentrations, with dinotefuran showing higher toxicity. Exposure led to the reduced survival, suppression of vitellogenin expression, and dysregulation of genes related to antioxidant defense, immunity, and detoxification. Additionally, high concentrations of dinotefuran and its combination with amitraz impaired hypopharyngeal gland morphology. Notably, co-exposure resulted in synergistic toxic effects, exacerbating physiological damage beyond individual treatments. These findings emphasize the potential risks of combined exposure to agricultural and beekeeping pesticides. A more comprehensive risk assessment and stricter regulations are urgently needed. Full article
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21 pages, 3620 KiB  
Article
Immune Gene Expression and Locomotor Activity in Response to Vairimorpha ceranae Infection Across Five Honey Bee Subspecies
by Cansu Özge Tozkar and Jay D. Evans
Insects 2025, 16(6), 593; https://doi.org/10.3390/insects16060593 - 5 Jun 2025
Viewed by 3110
Abstract
This study evaluated immune gene expression and locomotor behavior across five Apis mellifera subspecies (Carniolan, Caucasian, Syrian, Muğla ecotype, and Yığılca ecotype) following controlled Vairimorpha ceranae infection. Six days post-infection, Caucasian, Carniolan, and Yığılca bees exhibited a significant upregulation of antimicrobial peptide (AMP) [...] Read more.
This study evaluated immune gene expression and locomotor behavior across five Apis mellifera subspecies (Carniolan, Caucasian, Syrian, Muğla ecotype, and Yığılca ecotype) following controlled Vairimorpha ceranae infection. Six days post-infection, Caucasian, Carniolan, and Yığılca bees exhibited a significant upregulation of antimicrobial peptide (AMP) transcripts—hymenoptaecin, abaecin, defensin, and apidaecin—indicating a robust humoral response. Conversely, Syrian and Muğla bees showed weaker AMP expression and higher V. ceranae mRNA levels, indicating lower immunity and higher susceptibility. Positive correlations among AMP transcripts, especially in Caucasian, Carniolan, and Yığılca bees, suggested a coordinated response. Eater gene expression, critical for cellular immunity, decreased in infected Caucasian and Yığılca bees, coinciding with AMP upregulation. Vitellogenin expression, linked to immunity and longevity, increased in Carniolan and Syrian bees, correlating with higher early locomotor activity. Locomotor analysis revealed subspecies-specific behavioral responses. Syrian bees maintained the highest activity despite elevated V. ceranae mRNA and minimal AMP expression, suggesting unique resilience possibly mediated by vitellogenin. Muğla bees, despite high pathogen loads, exhibited decreased activity. Caucasian bees showed strong immune responses but reduced activity post-infection, reflecting potential physiological trade-offs. Overall, these findings underscore the role of genetic variability in shaping honey bee immune and behavioral responses to Vairimorpha and support subspecies-targeted breeding and disease management strategies to enhance resilience. Full article
(This article belongs to the Special Issue Bee Conservation: Behavior, Health and Pollination Ecology)
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21 pages, 946 KiB  
Review
Immunomodulatory and Anti-Inflammatory Properties of Honey and Bee Products
by Bashar Saad
Immuno 2025, 5(2), 19; https://doi.org/10.3390/immuno5020019 - 30 May 2025
Viewed by 1585
Abstract
Honey and other bee products, including propolis, royal jelly, and bee pollen, are widely recognized for their medicinal properties. Among their numerous biological activities, their anti-inflammatory and immunomodulatory effects have gained significant attention in recent years. Immune and inflammatory disorders contribute significantly to [...] Read more.
Honey and other bee products, including propolis, royal jelly, and bee pollen, are widely recognized for their medicinal properties. Among their numerous biological activities, their anti-inflammatory and immunomodulatory effects have gained significant attention in recent years. Immune and inflammatory disorders contribute significantly to the development of chronic conditions, including cancer and diabetes. Bee-derived products, along with their bioactive compounds such as polyphenols, have shown promising therapeutic effects in modulating inflammatory mediators. Studies indicate that these products help regulate tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6), and interleukin-7 (IL-7) levels while reducing reactive oxygen species (ROS) production. Additionally, both in vitro and in vivo research, along with clinical studies, highlight their role in enhancing immune responses by activating B and T lymphocytes. This review explores the molecular mechanisms underlying these properties, emphasizing the role of bioactive compounds such as flavonoids, phenolic acids, and proteins in modulating immune responses and reducing inflammation. Evidence from in vitro, in vivo, and clinical studies suggests that honey and bee products influence cytokine production, regulate immune cell activity, and mitigate oxidative stress, making them potential therapeutic agents for inflammatory and immune-related disorders. To gather relevant information, databases such as Google Scholar, PubMed, and ScienceDirect were searched using various keyword combinations, including immunomodulatory, anti-inflammatory, bee products, honey, propolis, royal jelly, bee venom, and bee pollen. Given their anti-inflammatory, immune-protective, antioxidant, anti-apoptotic, and antimicrobial properties, bee products remain a subject of interest for further clinical evaluation. Full article
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20 pages, 2253 KiB  
Article
Molecular Assessment of Genes Linked to Honeybee Health Fed with Different Diets in Nuclear Colonies
by Worrel A. Diedrick, Lambert H. B. Kanga, Rachel Mallinger, Manuel Pescador, Islam Elsharkawy and Yanping Zhang
Insects 2025, 16(4), 374; https://doi.org/10.3390/insects16040374 - 2 Apr 2025
Cited by 1 | Viewed by 713
Abstract
Honeybees are of economic importance not only for honey production, but also for crop pollination, which amounts to USD 20 billion per year in the United States. However, the number of honeybee colonies has declined more than 40% during the last few decades. [...] Read more.
Honeybees are of economic importance not only for honey production, but also for crop pollination, which amounts to USD 20 billion per year in the United States. However, the number of honeybee colonies has declined more than 40% during the last few decades. Although this decline is attributed to a combination of factors (parasites, diseases, pesticides, and nutrition), unlike other factors, the effect of nutrition on honeybee health is not well documented. In this study, we assessed the differential expression of seven genes linked to honeybee health under three different diets. These included immune function genes [Cactus, immune deficiency (IMD), Spaetzle)], genes involved in nutrition, cellular defense, longevity, and behavior (Vitellogenin, Malvolio), a gene involved in energy metabolism (Maltase), and a gene associated with locomotory behavior (Single-minded). The diets included (a) commercial pollen patties and sugar syrup, (b) monofloral (anise hyssop), and (c) polyfloral (marigold, anise hyssop, sweet alyssum, and basil). Over the 2.7-month experimental periods, adult bees in controls fed pollen patties and sugar syrup showed upregulated Cactus (involved in Toll pathway) and IMD (signaling pathway controls antibacterial defense) expression, while their counterparts fed monofloral and polyfloral diets downregulated the expression of these genes. Unlike Cactus and IMD, the gene expression profile of Spaetzle (involved in Toll pathway) did not differ across treatments during the experimental period except that it was significantly downregulated on day 63 and day 84 in bees fed polyfloral diets. The Vitellogenin gene indicated that monofloral and polyfloral diets significantly upregulated this gene and enhanced lifespan, foraging behavior, and immunity in adult bees fed with monofloral diets. The expression of Malvolio (involved in sucrose responsiveness and foraging behavior) was upregulated when food reserves (pollen and nectar) were limited in adult bees fed polyfloral diets. Adult bees fed with monofloral diets significantly upregulated the expression of Maltase (involved in energy metabolisms) compared to their counterparts in control diets to the end of the experimental period. Single-Minded Homolog 2 (involved in locomotory behavior) was also upregulated in adult bees fed pollen patties and sugar syrup compared to their counterparts fed monofloral and polyfloral diets. Thus, the food source significantly affected honeybee health and triggered an up- and downregulation of these genes, which correlated with the health and activities of the honeybee colonies. Overall, we found that the companion crops (monofloral and polyfloral) provided higher nutritional benefits to enhance honeybee health than the pollen patty and sugar syrup used currently by beekeepers. Furthermore, while it has been reported that bees require pollen from diverse sources to maintain a healthy physiology and hive, our data on nuclear colonies indicated that a single-species diet (such as anise hyssop) is nutritionally adequate and better or comparable to polyfloral diets. To the best of our knowledge, this is the first report indicating better nutritional benefits from monofloral diets (anise hyssop) over polyfloral diets for honeybee colonies (nucs) in semi-large-scale experimental runs. Thus, we recommend that the landscape of any apiary include highly nutritious food sources, such as anise hyssop, throughout the season to enhance honeybee health. Full article
(This article belongs to the Special Issue Insect Mitogenome, Phylogeny, and Mitochondrial Genome Expression)
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17 pages, 1776 KiB  
Article
Centrality of Hygienic Honey Bee Workers in Colony Social Networks
by Adrian Perez and Brian R. Johnson
Insects 2025, 16(1), 58; https://doi.org/10.3390/insects16010058 - 10 Jan 2025
Viewed by 1055
Abstract
Many social and environmental variables can affect the interactions among individuals in an insect colony that fundamentally structure its social organization. Along with important attributes such as age and caste, immunity-related factors such as the performance of sanitary tasks or exposure to a [...] Read more.
Many social and environmental variables can affect the interactions among individuals in an insect colony that fundamentally structure its social organization. Along with important attributes such as age and caste, immunity-related factors such as the performance of sanitary tasks or exposure to a pathogen can also influence an individual’s social interactions and their place in the resulting social network. Most work on this subject has supported the hypothesis that health-compromised individuals will exhibit altered social or spatial behavior that presumably limits the spread of infection. Here, we test this hypothesis using honey bee workers recently involved in hygienic behavior, an important set of sanitary tasks in which unhealthy brood are uncapped and then removed from the colony. Using static social networks, we quantify the interaction patterns of workers recently involved in hygienic tasks and compare their network centrality to non-hygienic workers. Using dynamic networks, we analyze the capability of hygienic workers to spread a potential infection throughout the colony. We find no substantial differences in how connected hygienic workers are in the network, and we show that hygienic workers would spread a novel infection throughout the colony to the same extent as non-hygienic workers. Our results suggest that experience with certain sanitary tasks may not necessarily produce rapid changes in social behavior. This work highlights the importance of considering the benefits of remaining socially integrated in important information networks and the temporal limitations for how quickly organized immune responses can occur in response to potential infections. Full article
(This article belongs to the Section Social Insects and Apiculture)
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17 pages, 4216 KiB  
Article
Evaluating the Influence of Different Artificial Diets on Apis mellifera L. Using Health Biomarkers and Performance Metrics
by Shams Ul Islam, Muhammad Anjum Aqueel, Muhammad Usman Yousuf, Asim Abbasi, Muhammad Yasin, Rashid Iqbal, Muhammad Fahim Raza, Aqsa Parvaiz and Nazih Y. Rebouh
Insects 2024, 15(11), 905; https://doi.org/10.3390/insects15110905 - 19 Nov 2024
Viewed by 1418
Abstract
The diet of Apis mellifera L. is a crucial factor for managing its colonies particularly during dearth periods. Numerous diets have been developed; however, their global implementation faces challenges due to diverse climatic conditions and some other factors. To address this issue, three [...] Read more.
The diet of Apis mellifera L. is a crucial factor for managing its colonies particularly during dearth periods. Numerous diets have been developed; however, their global implementation faces challenges due to diverse climatic conditions and some other factors. To address this issue, three previously evaluated diets (selected from seven) were tested to assess their effects on A. mellifera using key health biomarkers (immune function, stress response) and performance metrics (foraging activity, honey quality, and social interactions). The experiment was conducted using 12 colonies, including three replications, in The Islamia University of Bahawalpur, Pakistan, from June to September 2023. The results revealed that all the tested parameters were significantly affected by diets. Highest phenol-oxidase activity was recorded in T1 (28.7 U/mg). Heat shock protein (HSP) bands showed that T1 had the fewest (Hsp70), while T0 had more bands (Hsp40, Hsp60, and Hsp70), indicating stress differences. In foraging activity, average number of outgoing bees were highest in T1 (81.8) and lowest in T0 (31.2) and similar trend was followed for returning bees, i.e., T1 (81.8) and T0 (31.2). For pollen-carrying bees, the highest bees were counted in T1 (34.9), and the lowest in T0 (4.10). Honey quality was also significantly affected by diets, pH was highest in T1 (3.85), while moisture was highest in T0 (19.44%). Diastase activity, ash content, and electrical conductivity were best in T1 (13.74 units/g, 0.17%, 0.94 mS/cm, respectively). Mineral content was highest in T1 (406.54 mg/kg), and fructose content also peaked in T1 (396.21 mg/kg). Antioxidant contents, total phenolic content, flavonoid content, and ORAC value were highest in T1 (60.50 mg GAE/100 g, 44.41 mg QE/100 g, 10,237.30 µmol TE/g), while T0 consistently showed the lowest values across all parameters. In social interaction experiments, trophallaxis events were most frequent in T1 (7.38), and T1 also exhibited the longest trophallaxis time (5.51 s). The number of bees per trophallaxis event and antennation frequency followed a similar trend, with the highest recorded in T1 (5.16 bees/event, 10.1 antennation frequency) and the lowest in T0 (2.94 bees/event, 4.18 antennation frequency). Therefore, diet-1 (Watermelon juice 20 mL + Fenugreek powder 2 g + Chickpea flour 20 g + Lupin flour 20 g + Mung bean flour 20 g + Yeast 10 g + Powdered sugar 40 g + vegetable oil 10 mL) is recommended as a suitable substitute for managing A. mellifera colonies during dearth periods. Full article
(This article belongs to the Collection Science of Insect Rearing Systems)
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18 pages, 2338 KiB  
Article
Development and Optimization of Indian Propolis Formulation for Enhanced Immunomodulatory Potential
by Harshad S. Kapare, Harshal Rele, Mayuri Kisanrao Bhosale, Ritesh P. Bhole, Deepak Kulkarni, Vrushali Neve and Sadhana Raut
Immuno 2024, 4(4), 426-443; https://doi.org/10.3390/immuno4040027 - 24 Oct 2024
Cited by 2 | Viewed by 1544
Abstract
Propolis, a complex natural product that honey bees produce by mastication to protect and maintain their hive structures, comprises various bioactive constituents, including phenolic acids, flavonoids, diterpenes, sesquiterpenes, lignans, vitamins, minerals, etc. The objective of the current research was to extract crude propolis [...] Read more.
Propolis, a complex natural product that honey bees produce by mastication to protect and maintain their hive structures, comprises various bioactive constituents, including phenolic acids, flavonoids, diterpenes, sesquiterpenes, lignans, vitamins, minerals, etc. The objective of the current research was to extract crude propolis to enrich the total polyphenolic and flavonoid content, conduct preliminary phytochemical screening, and develop and evaluate dosage form to improve formulation characteristics and immunomodulatory potential. Total balsam, polyphenols, and flavonoids were found to be 46% w/w, 34.82 ± 0.078 mg equivalent of gallic acid/g, and 23.61 ± 0.045 mg equivalent of quercetin/g, respectively. DSC and FTIR studies demonstrated molecular dispersion of the propolis extract. Formulation was optimized with a 32 factorial design, and an optimized batch showed 92.20 ± 1.72% drug release in 1 h, an elevated hypersensitivity (DTH) response (p < 0.0001), increased phagocytic activity (p < 0.01), and a significantly (p < 0.001) higher total leukocyte count ((5.015 ± 0.19) × 103/mm3). The developed formulation showed significantly modulated immune modulatory potential compared with the propolis extract and conventional levamisole. This study can be further extended for clinical evaluations. Full article
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14 pages, 3636 KiB  
Article
Changes in Selected Biochemical Markers of Honey Bees Exposed to Fermented Common Tansy Solution (Tanacetum vulgare L.)
by Natalia Białecka, Klaudia Garbacz, Ewelina Berbeć, Agnieszka Murawska, Beata Madras-Majewska and Paweł Migdał
Animals 2024, 14(19), 2857; https://doi.org/10.3390/ani14192857 - 4 Oct 2024
Cited by 1 | Viewed by 1346
Abstract
Honey bees use pollen and nectar from flowers to produce food. Because they often forage on crops, they are at risk of being exposed to plant protection products (PPPs), both directly and in stored food. Due to the adverse effects of synthetic PPPs [...] Read more.
Honey bees use pollen and nectar from flowers to produce food. Because they often forage on crops, they are at risk of being exposed to plant protection products (PPPs), both directly and in stored food. Due to the adverse effects of synthetic PPPs on pollinators, biopesticides may be a viable alternative. Common tansy extract is used as one of the natural substitutes for synthetic pesticides. In our study, the effect of fermented common tansy extract on aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and gamma-glutamyl transpeptidase (GGTP) activity and the concentration of triglycerides (TGs), total protein (TP), total antioxidant status (TAS), and glucose in honey bee workers’ hemolymph was assessed. These biochemical markers give valuable information about the immunity, detoxification, and nutrition of a bee’s body. Caged bees were given tansy extract added at various concentrations in sugar syrup for 24 h. Then, they were provided with only sugar syrup. After 7 days of the experiment, hemolymph was collected and analyzed. We observed changes in the activity of AST, ALT, GGTP enzymes and TG, TP, and glucose levels, but not all changes were statistically significant. In terms of AST activity, statistically significant differences were found. All groups tested, including the negative control group, showed reduced enzyme activity values compared to the positive control group. In TG concentration, differences were observed between the groups receiving 2% extract and 1% ethanol. Glucose levels differed between the groups receiving 1% extract and 2% extract and between the positive control group and 1% extract. Bee body proper functioning is affected by changes in enzyme activity, especially those responsible for immunity and detoxification, such as AST, ALT, ALP, and GGTP. Despite the short time of bees’ exposure to the agent, the results of study show visible effects. Our results provide a basis for further research on the impact of tansy extract on honey bees. Full article
(This article belongs to the Section Animal Physiology)
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17 pages, 2640 KiB  
Review
Bee-Inspired Healing: Apitherapy in Veterinary Medicine for Maintenance and Improvement Animal Health and Well-Being
by Jevrosima Stevanović, Uroš Glavinić, Marko Ristanić, Vladimira Erjavec, Barış Denk, Slobodan Dolašević and Zoran Stanimirović
Pharmaceuticals 2024, 17(8), 1050; https://doi.org/10.3390/ph17081050 - 9 Aug 2024
Cited by 3 | Viewed by 3958
Abstract
This review aims to present current knowledge on the effects of honey bee products on animals based on in vivo studies, focusing on their application in clinical veterinary practice. Honey’s best-proven effectiveness is in treating wounds, including those infected with antibiotic-resistant microorganisms, as [...] Read more.
This review aims to present current knowledge on the effects of honey bee products on animals based on in vivo studies, focusing on their application in clinical veterinary practice. Honey’s best-proven effectiveness is in treating wounds, including those infected with antibiotic-resistant microorganisms, as evidenced in horses, cats, dogs, mice, and rats. Propolis manifested a healing effect in numerous inflammatory and painful conditions in mice, rats, dogs, and pigs and also helped in oncological cases in mice and rats. Bee venom is best known for its effectiveness in treating neuropathy and arthritis, as shown in dogs, mice, and rats. Besides, bee venom improved reproductive performance, immune response, and general health in rabbits, chickens, and pigs. Pollen was effective in stimulating growth and improving intestinal microflora in chickens. Royal jelly might be used in the management of animal reproduction due to its efficiency in improving fertility, as shown in rats, rabbits, and mice. Drone larvae are primarily valued for their androgenic effects and stimulation of reproductive function, as evidenced in sheep, chickens, pigs, and rats. Further research is warranted to determine the dose and method of application of honey bee products in animals. Full article
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22 pages, 5003 KiB  
Article
Regulatory Roles of Long Non-Coding RNAs Relevant to Antioxidant Enzymes and Immune Responses of Apis cerana Larvae Following Ascosphaera apis Invasion
by Rui Guo, Siyi Wang, Sijia Guo, Xiaoxue Fan, He Zang, Xuze Gao, Xin Jing, Zhitan Liu, Zhihao Na, Peiyuan Zou and Dafu Chen
Int. J. Mol. Sci. 2023, 24(18), 14175; https://doi.org/10.3390/ijms241814175 - 16 Sep 2023
Cited by 4 | Viewed by 1545
Abstract
Long non-coding RNAs (lncRNAs) play an essential part in controlling gene expression and a variety of biological processes such as immune defense and stress-response. However, whether and how lncRNAs regulate responses of Apis cerana larvae to Ascosphaera apis invasion has remained unclear until [...] Read more.
Long non-coding RNAs (lncRNAs) play an essential part in controlling gene expression and a variety of biological processes such as immune defense and stress-response. However, whether and how lncRNAs regulate responses of Apis cerana larvae to Ascosphaera apis invasion has remained unclear until now. Here, the identification and structural analysis of lncRNAs in the guts of A. cerana worker larvae were conducted, and the expression profile of larval lncRNAs during the A. apis infection process was then analyzed, followed by an investigation of the regulatory roles of differentially expressed lncRNAs (DElncRNAs) in the host response. In total, 76 sense lncRNAs, 836 antisense lncRNAs, 184 intron lncRNAs, 362 bidirectional lncRNAs, and 2181 intron lncRNAs were discovered in the larval guts. Additionally, 30 known and 9 novel lncRNAs were potential precursors for 36 and 11 miRNAs, respectively. In the three comparison groups, 386, 351, and 272 DElncRNAs were respectively identified, indicating the change in the overall expression pattern of host lncRNAs following the A. apis invasion. Analysis of cis-acting effect showed that DElncRNAs in the 4-, 5-, and 6-day-old comparison groups putatively regulated 55, 30, and 20 up- and down-stream genes, respectively, which were involved in a series of crucial functional terms and pathways, such as MAPK signaling pathway, and cell process. Analysis showed that 31, 8, and 11 DElncRNAs as potential antisense lncRNAs may interact with 26, 8, and 9 sense-strand mRNAs. Moreover, investigation of the competing endogenous RNA (ceRNA) network indicated that 148, 283, and 257 DElncRNAs were putatively regulated. The expression of target genes by targeting corresponding DEmiRNAs included those associated with antioxidant enzymes and immune responses. These results suggested that DElncRNAs played a potential part in the larval guts responding to the A. apis infection through a cis-acting manner and ceRNA mechanisms. Our findings deepen our understanding of interactions between A. cerana larvae and A. apis and offer a basis for clarifying the DElncRNA-mediated mechanisms underlying the host response to fungal invasion. Full article
(This article belongs to the Section Molecular Immunology)
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23 pages, 1245 KiB  
Article
Transcriptomic Responses Underlying the High Virulence of Black Queen Cell Virus and Sacbrood Virus following a Change in Their Mode of Transmission in Honey Bees (Apis mellifera)
by Yahya Al Naggar, Hassan Shafiey and Robert J. Paxton
Viruses 2023, 15(6), 1284; https://doi.org/10.3390/v15061284 - 30 May 2023
Cited by 5 | Viewed by 3361
Abstract
Background: Over the last two decades, honey bees (Apis mellifera) have suffered high rates of colony losses that have been attributed to a variety of factors, chief among which are viral pathogens, such as deformed wing virus (DWV), whose virulence has [...] Read more.
Background: Over the last two decades, honey bees (Apis mellifera) have suffered high rates of colony losses that have been attributed to a variety of factors, chief among which are viral pathogens, such as deformed wing virus (DWV), whose virulence has increased because of vector-based transmission by the invasive, ectoparasitic varroa mite (Varroa destructor). A shift in the experimental mode of transmission of the black queen cell virus (BQCV) and sacbrood virus (SBV) from fecal/food–oral (direct horizontal) to vector-mediated (indirect horizontal) transmission also results in high virulence and viral titers in pupal and adult honey bees. Agricultural pesticides represent another factor that acts independently or in interaction with pathogens, and they are also thought to cause colony loss. Understanding the molecular mechanisms underlying the higher virulence following a vector-based mode of transmission provides deeper insight into honey bee colony losses, as does determining whether or not host–pathogen interactions are modulated by exposure to pesticides. Methods: Through an experimental design with controlled laboratory, we investigated the effects of the modes of transmission of BQCV and SBV (feeding vs. vector-mediated via injection) alone or in combination with chronic exposure to sublethal and field-realistic concentrations of flupyradifurone (FPF), a novel agricultural insecticide, on honey bee survival and transcription responses by using high-throughput RNA sequencing (RNA-seq) analysis. Results: Co-exposure to viruses via feeding (VF) or injection (VI) and FPF insecticide had no statistically significant interactive effect on their survival compared to, respectively, VF or VI treatments alone. Transcriptomic analysis revealed a distinct difference in the gene expression profiles of bees inoculated with viruses via injection (VI) and exposed to FPF insecticide (VI+FPF). The number of differentially expressed genes (DEGs) at log2 (fold-change) > 2.0 in VI bees (136 genes) or/and VI+FPF insecticide (282 genes) was very high compared to that of VF bees (8 genes) or the VF+FPF insecticide treatment (15 genes). Of these DEGs, the expression in VI and VI+FPF bees of some immune-related genes, such as those for antimicrobial peptides, Ago2, and Dicer, was induced. In short, several genes encoding odorant binding proteins, chemosensory proteins, odor receptors, honey bee venom peptides, and vitellogenin were downregulated in VI and VI+FPF bees. Conclusions: Given the importance of these suppressed genes in honey bees’ innate immunity, eicosanoid biosynthesis, and olfactory associative function, their inhibition because of the change in the mode of infection with BQCV and SBV to vector-mediated transmission (injection into haemocoel) could explain the high virulence observed in these viruses when they were experimentally injected into hosts. These changes may help explain why other viruses, such as DWV, represent such a threat to colony survival when transmitted by varroa mites. Full article
(This article belongs to the Special Issue Virus-Vector Interactions 2023)
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20 pages, 1288 KiB  
Review
Viral Co-Infections and Antiviral Immunity in Honey Bees
by Alice Mélusine Durand, Anne Bonjour-Dalmon and Eric Dubois
Viruses 2023, 15(5), 1217; https://doi.org/10.3390/v15051217 - 22 May 2023
Cited by 10 | Viewed by 4081
Abstract
Over the past few decades, honey bees have been facing an increasing number of stressors. Beyond individual stress factors, the synergies between them have been identified as a key factor in the observed increase in colony mortality. However, these interactions are numerous and [...] Read more.
Over the past few decades, honey bees have been facing an increasing number of stressors. Beyond individual stress factors, the synergies between them have been identified as a key factor in the observed increase in colony mortality. However, these interactions are numerous and complex and call for further research. Here, in line with our need for a systemic understanding of the threats that they pose to bee health, we review the interactions between honey bee viruses. As viruses are obligate parasites, the interactions between them not only depend on the viruses themselves but also on the immune responses of honey bees. Thus, we first summarise our current knowledge of the antiviral immunity of honey bees. We then review the interactions between specific pathogenic viruses and their interactions with their host. Finally, we draw hypotheses from the current literature and suggest directions for future research. Full article
(This article belongs to the Special Issue Molecular Virus-Insect Interactions)
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14 pages, 899 KiB  
Article
The Fungus Nosema ceranae and a Sublethal Dose of the Neonicotinoid Insecticide Thiamethoxam Differentially Affected the Health and Immunity of Africanized Honey Bees
by Alvaro De la Mora, Nuria Morfin, José C. Tapia-Rivera, José O. Macías-Macías, José M. Tapia-González, Francisca Contreras-Escareño, Tatiana Petukhova and Ernesto Guzman-Novoa
Microorganisms 2023, 11(5), 1258; https://doi.org/10.3390/microorganisms11051258 - 10 May 2023
Cited by 7 | Viewed by 2270
Abstract
Honey bees (Apis mellifera L.) are affected by different biotic and abiotic stressors, such as the fungus Nosema ceranae and neonicotinoid insecticides, that negatively impact their health. However, most studies so far conducted have focused on the effect of these stressors separately [...] Read more.
Honey bees (Apis mellifera L.) are affected by different biotic and abiotic stressors, such as the fungus Nosema ceranae and neonicotinoid insecticides, that negatively impact their health. However, most studies so far conducted have focused on the effect of these stressors separately and in European honey bees. Therefore, this study was conducted to analyze the impact of both stressors, singly and in combination, on honey bees of African descent that have demonstrated resistance to parasites and pesticides. Africanized honey bees (AHBs, Apis mellifera scutellata Lepeletier) were inoculated with N. ceranae (1 × 105 spores/bee) and/or chronically exposed for 18 days to a sublethal dose of thiamethoxam (0.025 ng/bee) to evaluate their single and combined effects on food consumption, survivorship, N. ceranae infection, and immunity at the cellular and humoral levels. No significant effects by any of the stressors were found for food consumption. However, thiamethoxam was the main stressor associated to a significant decrease in AHB survivorship, whereas N. ceranae was the main stressor affecting their humoral immune response by upregulating the expression of the gene AmHym-1. Additionally, both stressors, separately and combined, significantly decreased the concentration of haemocytes in the haemolymph of the bees. These findings indicate that N. ceranae and thiamethoxam differentially affect the lifespan and immunity of AHBs and do not seem to have synergistic effects when AHBs are simultaneously exposed to both stressors. Full article
(This article belongs to the Section Parasitology)
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17 pages, 2135 KiB  
Article
Expression of Molecular Markers of Resilience against Varroa destructor and Bee Viruses in Ethiopian Honey Bees (Apis mellifera simensis) Focussing on Olfactory Sensing and the RNA Interference Machinery
by Haftom Gebremedhn, David Claeys Bouuaert, Michel Asperges, Bezabeh Amssalu, Lina De Smet and Dirk C. de Graaf
Insects 2023, 14(5), 436; https://doi.org/10.3390/insects14050436 - 3 May 2023
Cited by 8 | Viewed by 3498
Abstract
Varroa destructor mites and the viruses it vectors are two major factors leading to high losses of honey bees (Apis mellifera) colonies worldwide. However, honey bees in some African countries show resilience to varroa infestation and/or virus infections, although little is [...] Read more.
Varroa destructor mites and the viruses it vectors are two major factors leading to high losses of honey bees (Apis mellifera) colonies worldwide. However, honey bees in some African countries show resilience to varroa infestation and/or virus infections, although little is known about the mechanisms underlying this resilience. In this study, we investigated the expression profiles of some key molecular markers involved in olfactory sensing and RNA interference, as these processes may contribute to the bees’ resilience to varroa infestation and virus infection, respectively. We found significantly higher gene expression of the odorant binding protein, OBP14, in the antennae of Ethiopian bees compared to Belgian bees. This result suggests the potential of OBP14 as a molecular marker of resilience to mite infestation. Scanning electron microscopy showed no significant differences in the antennal sensilla occurrence and distribution, suggesting that resilience arises from molecular processes rather than morphological adaptations. In addition, seven RNAi genes were upregulated in the Ethiopian honey bees and three of them—Dicer-Drosha, Argonaute 2, and TRBP2—were positively correlated with the viral load. We can conclude that the antiviral immune response was triggered when bees were experiencing severe viral infection and that this might contribute to the bees’ resilience to viruses. Full article
(This article belongs to the Section Insect Physiology, Reproduction and Development)
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18 pages, 12969 KiB  
Article
Systematic Characterization and Regulatory Role of lncRNAs in Asian Honey Bees Responding to Microsporidian Infestation
by Zixin Wang, Siyi Wang, Xiaoxue Fan, Kaiyao Zhang, Jiaxin Zhang, Haodong Zhao, Xuze Gao, Yiqiong Zhang, Sijia Guo, Dingding Zhou, Qiming Li, Zhihao Na, Dafu Chen and Rui Guo
Int. J. Mol. Sci. 2023, 24(6), 5886; https://doi.org/10.3390/ijms24065886 - 20 Mar 2023
Cited by 6 | Viewed by 2433
Abstract
Long noncoding RNAs (lncRNAs) are pivotal regulators in gene expression and diverse biological processes, such as immune defense and host–pathogen interactions. However, little is known about the roles of lncRNAs in the response of the Asian honey bee (Apis cerana) to [...] Read more.
Long noncoding RNAs (lncRNAs) are pivotal regulators in gene expression and diverse biological processes, such as immune defense and host–pathogen interactions. However, little is known about the roles of lncRNAs in the response of the Asian honey bee (Apis cerana) to microsporidian infestation. Based on our previously obtained high-quality transcriptome datasets from the midgut tissues of Apis cerana cerana workers at 7 days post inoculation (dpi) and 10 dpi with Nosema ceranae (AcT7 and AcT10 groups) and the corresponding un-inoculated midgut tissues (AcCK7 and AcCK10 groups), the transcriptome-wide identification and structural characterization of lncRNAs were conducted, and the differential expression pattern of lncRNAs was then analyzed, followed by investigation of the regulatory roles of differentially expressed lncRNAs (DElncRNAs) in host response. Here, 2365, 2322, 2487, and 1986 lncRNAs were, respectively, identified in the AcCK7, AcT7, AcCK7, and AcT10 groups. After removing redundant ones, a total of 3496 A. c. cerana lncRNAs were identified, which shared similar structural characteristics with those discovered in other animals and plants, such as shorter exons and introns than mRNAs. Additionally, 79 and 73 DElncRNAs were screened from the workers’ midguts at 7 dpi and 10 dpi, respectively, indicating the alteration of the overall expression pattern of lncRNAs in host midguts after N. ceranae infestation. These DElncRNAs could, respectively, regulate 87 and 73 upstream and downstream genes, involving a suite of functional terms and pathways, such as metabolic process and Hippo signaling pathway. Additionally, 235 and 209 genes co-expressed with DElncRNAs were found to enrich in 29 and 27 terms, as well as 112 and 123 pathways, such as ABC transporters and the cAMP signaling pathway. Further, it was detected that 79 (73) DElncRNAs in the host midguts at 7 (10) dpi could target 321 (313) DEmiRNAs and further target 3631 (3130) DEmRNAs. TCONS_00024312 and XR_001765805.1 were potential precursors for ame-miR-315 and ame-miR-927, while TCONS_00006120 was the putative precursor for both ame-miR-87-1 and ame-miR-87-2. These results together suggested that DElncRNAs are likely to play regulatory roles in the host response to N. ceranae infestation through the regulation of neighboring genes via a cis-acting effect, modulation of co-expressed mRNAs via trans-acting effect, and control of downstream target genes’ expression via competing endogenous RNA networks. Our findings provide a basis for disclosing the mechanism underlying DElncRNA-mediated host N. ceranae response and a new perspective into the interaction between A. c. cerana and N. ceranae. Full article
(This article belongs to the Special Issue Molecular Biology of RNA: Recent Progress)
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