In-situ Hybridization for the Detection of Sacbrood Virus in Infected Larvae of the Honey Bee (Apis cerana)

AbstractSince 2016, Apis cerana sacbrood virus (AcSBV) has been recorded in Taiwan. It is epizootic in Apis cerana (Hymenoptera: Apidae) and causing serious loss of A. cerana. Herein, we performed a long-term survey of AcSBV prevalence in the populations of A. cerana in Northern Taiwan from January 2017 to July 2018. The surveillance of AcSBV prevalence in A. mellifera (Hymenoptera: Apidae) populations was starting and further confirmed by sequencing since April 2017; thus, these data were also included in this survey. In our survey, the average prevalence rates of AcSBV were 72 and 53% in A. cerana and A. mellifera, respectively, in 2017, which decreased to 45 and 27% in 2018. For the spatial analysis of AcSBV in two honey bee populations, Hsinchu showed the highest prevalence, followed by New Taipei, Yilan, Taipei, and Keelung, suggesting that AcSBV might have come from the southern part of Taiwan. Interestingly, the AcSBV prevalence rates from A. cerana and A. mellifera cocultured apiaries gradually synchronized. The result of phylogenetic analysis and comparison of the annual AcSBV prevalence in A. cerana-only, A. mellifera-only, and A. cerana/A. mellifera cocultured sample sites indicate cross-infection between A. cerana and A. mellifera; however, AcSBV may lose the advantage of virulence in A. mellifera. The evidence suggested that the transmission of AcSBV might occur among these two honey bee species in the field. Therefore, A. mellifera may serve as a guard species to monitor AcSBV in A. cerana, but the cross-infection still needs to be surveyed.

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In situ hybridization assays for localization of the chronic bee paralysis virus in the honey bee (Apis mellifera) brain

Journal of Virological Methods ◽

10.1016/j.jviromet.2008.06.027 ◽

2008 ◽

Vol 153 (2) ◽

pp. 232-237 ◽

Cited By ~ 12

Author(s):

Violaine Olivier ◽

Isabelle Massou ◽

Olivier Celle ◽

Philippe Blanchard ◽

Frank Schurr ◽

...

Keyword(s):

In Situ Hybridization ◽

Apis Mellifera ◽

Honey Bee ◽

Paralysis Virus ◽

Hybridization Assays

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The high-throughput production of dsRNA against sacbrood virus for use in the honey bee Apis cerana (Hymenoptera: Apidae)

Virus Genes ◽

10.1007/s11262-016-1346-6 ◽

2016 ◽

Vol 52 (5) ◽

pp. 698-705 ◽

Cited By ~ 10

Author(s):

Jianqing Zhang ◽

Yi Zhang ◽

Richou Han

Keyword(s):

Honey Bee ◽

High Throughput ◽

Apis Cerana ◽

Sacbrood Virus

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Transcriptome Profiling Reveals a Novel Mechanism of Antiviral Immunity Upon Sacbrood Virus Infection in Honey Bee Larvae (Apis cerana)

Frontiers in Microbiology ◽

10.3389/fmicb.2021.615893 ◽

2021 ◽

Vol 12 ◽

Author(s):

Yulong Guo ◽

Zhengyi Zhang ◽

Mingsheng Zhuang ◽

Liuhao Wang ◽

Kai Li ◽

...

Keyword(s):

Virus Infection ◽

Molecular Mechanism ◽

Honey Bee ◽

Differentially Expressed Genes ◽

Honey Bees ◽

Apis Cerana ◽

Differentially Expressed ◽

Virus Family ◽

Sacbrood Virus ◽

Honey Bee Larvae

The honey bee is one of the most important pollinators in the agricultural system and is responsible for pollinating a third of all food we eat. Sacbrood virus (SBV) is a member of the virus family Iflaviridae and affects honey bee larvae and causes particularly devastating disease in the Asian honey bees, Apis cerana. Chinese Sacbrood virus (CSBV) is a geographic strain of SBV identified in China and has resulted in mass death of honey bees in China in recent years. However, the molecular mechanism underlying SBV infection in the Asian honey bee has remained unelucidated. In this present study, we employed high throughput next-generation sequencing technology to study the host transcriptional responses to CSBV infection in A. cerana larvae, and were able to identify genome-wide differentially expressed genes associated with the viral infection. Our study identified 2,534 differentially expressed genes (DEGs) involved in host innate immunity including Toll and immune deficiency (IMD) pathways, RNA interference (RNAi) pathway, endocytosis, etc. Notably, the expression of genes encoding antimicrobial peptides (abaecin, apidaecin, hymenoptaecin, and defensin) and core components of RNAi such as Dicer-like and Ago2 were found to be significantly upregulated in CSBV infected larvae. Most importantly, the expression of Sirtuin target genes, a family of signaling proteins involved in metabolic regulation, apoptosis, and intracellular signaling was found to be changed, providing the first evidence of the involvement of Sirtuin signaling pathway in insects’ immune response to a virus infection. The results obtained from this study provide novel insights into the molecular mechanism and immune responses involved in CSBV infection, which in turn will contribute to the development of diagnostics and treatment for the diseases in honey bees.

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DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100122885 ◽

1992 ◽

Vol 50 (1) ◽

pp. 496-497

Author(s):

Barbara Trask ◽

Susan Allen ◽

Anne Bergmann ◽

Mari Christensen ◽

Anne Fertitta ◽

...

Keyword(s):

In Situ Hybridization ◽

Dna Sequence ◽

Dna Sequences ◽

Dual Band ◽

Nick Translation ◽

Metaphase Chromosomes ◽

Band Pass ◽

Texas Red ◽

Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

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Ultrastructural analysis of the spatial distribution of MRNA

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100123167 ◽

1992 ◽

Vol 50 (1) ◽

pp. 552-553

Author(s):

Gary Bassell ◽

Robert H. Singer

Keyword(s):

Electron Microscopy ◽

Spatial Distribution ◽

In Situ Hybridization ◽

High Resolution ◽

Nucleic Acids ◽

Colloidal Gold ◽

Dna Probe ◽

Nucleotide Analogs ◽

Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

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