Preparation of Virus-Enriched Inoculum for Oral Infection of Honey Bees (&lt;em&gt;Apis mellifera&lt;/em&gt;)

Hsieh, Edward M.; Carrillo-Tripp, Jimena; Doležal, Adam

doi:10.3791/61725

Cited by 10 publications

(13 citation statements)

References 35 publications

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“…Large quantities of virus particles were produced and amplified inside honey bee pupae using methods similar to those compiled by de Miranda et al [49] and Carrillo-Tripp et al [47] with minor modifications [48]. While virus particles can be derived from adults or pupae, pupal propagation is very efficient at producing large quantities of concentrated virus [48,49]. Pupae were collected using two different methods: (1) pupal excision and (2) larval self-removal.…”

Section: Virus Particle Productionmentioning

confidence: 99%

“…Immediately following the transfer, each cage received a small weigh boat containing either 600 µL of 30% sucrose solution mixed with an IAPV inoculum or an equivalent quantity of sterile but otherwise untreated sucrose solutions (see associated subsections for concentrations of viral inocula employed in specific experiments). The IAPV inoculum used in all subsequent experiments involving a virus inoculation stage was generated according to the protocols by de Miranda et al [49], Carrillo-Tripp et al [47], and Hsieh et al [48] and contained 9.86 × 10 7 viral copies per 100 ng RNA composed of 99.79% IAPV with only trace levels of DWV, BQCV, and sacbrood virus (SBV). After allowing for total consumption of inocula within all cages (approximately 12-14 h), feeder tubes (15 mL Falcon tubes with 18G needle holes poked in the bottom) were inserted into each cage, the contents of which were provided ad libitum and refilled as necessary throughout the course of each experiment (see subsections for experiment-specific feeder solutions).…”

Section: Cage Assays: General Protocolmentioning

confidence: 99%

“…Although it is not quite as ubiquitous as more commonly studied viruses such as DWV, it can produce clearly discernable phenotypes in bees infected as adults, including lethargic movement, reduced reaction to stimulus, and eventual death [45]. Additionally, its effects on survivorship in controlled conditions is relatively well-characterized [9,13,46,47], meaning IAPV can serve as a valuable model virus for testing interactions with other factors, as it allows for high-throughput bioassays with standardized and repeatable laboratory protocols [48].…”

Section: Introductionmentioning

confidence: 99%

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Ameliorative Effects of Phytochemical Ingestion on Viral Infection in Honey Bees

Hsieh

Berenbaum

Doležal

2020

Insects

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Honey bee viruses are capable of causing a wide variety of devastating effects, but effective treatments have yet to be discovered. Phytochemicals represent a broad range of substances that honey bees frequently encounter and consume, many of which have been shown to improve honey bee health. However, their effect on bee viruses is largely unknown. Here, we tested the therapeutic effectiveness of carvacrol, thymol, p-coumaric acid, quercetin, and caffeine on viral infection by measuring their ability to improve survivorship in honey bees inoculated with Israeli acute paralysis virus (IAPV) using high-throughput cage bioassays. Among these candidates, caffeine was the only phytochemical capable of significantly improving survivorship, with initial screening showing that naturally occurring concentrations of caffeine (25 ppm) were sufficient to produce an ameliorative effect on IAPV infection. Consequently, we measured the scope of caffeine effectiveness in bees inoculated and uninoculated with IAPV by performing the same type of high-throughput bioassay across a wider range of caffeine concentrations. Our results indicate that caffeine may provide benefits that scale with concentration, though the exact mechanism by which caffeine ingestion improves survivorship remains uncertain. Caffeine therefore has the potential to act as an accessible and inexpensive method of treating viral infections, while also serving as a tool to further understanding of honey bee–virus interactions at a physiological and molecular level.

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Section: Virus Particle Productionmentioning

confidence: 99%

Section: Cage Assays: General Protocolmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Ameliorative Effects of Phytochemical Ingestion on Viral Infection in Honey Bees

Hsieh

Berenbaum

Doležal

2020

Insects

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“…Further, we also hypothesized that responses to our treatments would differ across seasons. To test these hypotheses, we used Israeli acute paralysis virus (IAPV) as a model virus (48). IAPV can cause a debilitating disease that can be lethal to all honey bee castes and development stages (49)(50)(51)(52), and has been implicated in colony failure (53).…”

Section: Introductionmentioning

confidence: 99%

Butenolide Insecticide Flupyradifurone Affects Honey Bee Worker Antiviral Immunity and Survival

Harwood

Prayugo²,

Doležal³

2022

Front. Insect Sci.

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Honey bees face many environmental stressors, including exposure to pesticides and pathogens. A novel butenolide pesticide, flupyradifurone, was recently introduced to the US and shown to have a bee-friendly toxicity profile. Like the much-scrutinized neonicotinoids that preceded it, flupyradifurone targets the insect nervous system. Some neonicotinoids have been shown to interfere with antiviral immunity, which raised the concern that similar effects may be observed with flupyradifurone. In this study, we investigated how flupyradifurone and a neonicotinoid, clothianidin, affect the ability of honey bee workers to combat an infection of Israeli acute paralysis virus (IAPV). We exposed workers to field-realistic doses of the pesticides either with or without co-exposure with the virus, and then tracked survival and changes in viral titers. We repeated the experiment in the spring and fall to look for any seasonal effects. We found that flupyradifurone caused elevated mortality in the fall, but it did not lead to increased virus-induced mortality. Flupyradifurone also appeared to affect virus clearance, as bees co-exposed to the pesticide and virus tended to have higher viral titers after 48 hours than those exposed to the virus alone. Clothianidin had no effect on viral titers, and it actually appeared to increase resistance to viral infection in spring bees.

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“…Recent development of DWV infectious clones will further our understanding of this virus and its interactions with bee and mite hosts [ 65 , 78 , 99 ]. In the absence of infectious clones, scientists have utilized crude virus preparations obtained from infected honey bees, including at the larval and pupal stages [ 10 , 68 , 86 , 100 , 101 , 102 , 103 , 104 ].…”

Section: Introductionmentioning

confidence: 99%

Investigating Virus–Host Interactions in Cultured Primary Honey Bee Cells

McMenamin

Parekh

Lawrence

et al. 2021

Insects

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Honey bee (Apis mellifera) health is impacted by viral infections at the colony, individual bee, and cellular levels. To investigate honey bee antiviral defense mechanisms at the cellular level we further developed the use of cultured primary cells, derived from either larvae or pupae, and demonstrated that these cells could be infected with a panel of viruses, including common honey bee infecting viruses (i.e., sacbrood virus (SBV) and deformed wing virus (DWV)) and an insect model virus, Flock House virus (FHV). Virus abundances were quantified over the course of infection. The production of infectious virions in cultured honey bee pupal cells was demonstrated by determining that naïve cells became infected after the transfer of deformed wing virus or Flock House virus from infected cell cultures. Initial characterization of the honey bee antiviral immune responses at the cellular level indicated that there were virus-specific responses, which included increased expression of bee antiviral protein-1 (GenBank: MF116383) in SBV-infected pupal cells and increased expression of argonaute-2 and dicer-like in FHV-infected hemocytes and pupal cells. Additional studies are required to further elucidate virus-specific honey bee antiviral defense mechanisms. The continued use of cultured primary honey bee cells for studies that involve multiple viruses will address this knowledge gap.

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Preparation of Virus-Enriched Inoculum for Oral Infection of Honey Bees (<em>Apis mellifera</em>)

Cited by 10 publications

References 35 publications

Ameliorative Effects of Phytochemical Ingestion on Viral Infection in Honey Bees

Ameliorative Effects of Phytochemical Ingestion on Viral Infection in Honey Bees

Butenolide Insecticide Flupyradifurone Affects Honey Bee Worker Antiviral Immunity and Survival

Investigating Virus–Host Interactions in Cultured Primary Honey Bee Cells

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