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Honey bees (Apis mellifera) are an agriculturally important pollinator species that live in easily managed social groups (i.e., colonies). Unfortunately, annual losses of honey bee colonies in many parts of the world have reached unsustainable levels. Multiple abiotic and biotic stressors, including viruses, are associated with individual honey bee and colony mortality. Honey bees have evolved several antiviral defense mechanisms including conserved immune pathways (e.g., Toll, Imd, JAK/STAT) and dsRNA-triggered responses including RNA interference and a non-sequence specific dsRNA-mediated response. In addition, transcriptome analyses of virus-infected honey bees implicate an antiviral role of stress response pathways, including the heat shock response. Herein, we demonstrate that the heat shock response is antiviral in honey bees. Specifically, heat-shocked honey bees (i.e., 42 °C for 4 h) had reduced levels of the model virus, Sindbis-GFP, compared with bees maintained at a constant temperature. Virus-infection and/or heat shock resulted in differential expression of six heat shock protein encoding genes and three immune genes, many of which are positively correlated. The heat shock protein encoding and immune gene transcriptional responses observed in virus-infected bees were not completely recapitulated by administration of double stranded RNA (dsRNA), a virus-associated molecular pattern, indicating that additional virus–host interactions are involved in triggering antiviral stress response pathways.
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A honey bee-associated virus remains infectious and quantifiable in postmortem hosts
Corpse-mediated pathogen transmission is a viable route through which naïve hosts can become infected, but its likelihood for honey bee-associated viruses is largely unknown. While these viruses can be easily detected in deceased bees, it remains unclear if they stay infectious within postmortem hosts or if enough viral RNA degradation-and subsequently virus inactivation-occurs post-host death to render these viruses inviable. This knowledge gap has important implications for how researchers perform honey bee virus studies and for our general understanding of honey bee virus transmission. To better understand the resiliency of honey bee-associated viruses within deceased hosts, we first tested the hypothesis that postmortem specimens, stored in colony-normal temperature and humidity conditions, can be reliably used to quantify virus abundance. To determine this, we experimentally-infected adult honey bees with Israeli acute paralysis virus (IAPV) and then measured the virus levels of individuals sampled live or at different postmortem time points (4, 12, 24, and 48 hours post-death) using RT-qPCR and a standard curve absolute quantification method. We found no significant differences based on when bees were sampled, indicating that postmortem honey bees are statistically comparable to using live-sampled bees and can be reliably used to quantify absolute IAPV abundance. We then performed a follow-up experiment that determined whether or not the IAPV detected in postmortem bees remained infectious over time. We found that IAPV extracted from postmortem bees remained highly infectious for at least 48 hours post-death, indicating that any viral RNA degradation that may have occurred during the postmortem interval did not adversely affect IAPV's overall infectivity. The results from this study suggest that IAPV is more resilient to degradation than previously assumed, support the use of postmortem bees for downstream IAPV analyses, and indicate that postmortem hosts can act as sources of IAPV infection for susceptible individuals.
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- Award ID(s):
- 2022049
- PAR ID:
- 10628782
- Publisher / Repository:
- Elsevier Inc.
- Date Published:
- Journal Name:
- Journal of Invertebrate Pathology
- Volume:
- 209
- Issue:
- C
- ISSN:
- 0022-2011
- Page Range / eLocation ID:
- 108258
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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- Free Publicly Accessible Full Text
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Accepted Manuscript
- Journal Article:
- https://doi.org/10.1016/j.jip.2024.108258
