A semi-automated and high-throughput approach for the detection of honey bee viruses in bee samples

Nikulin, Sofia Levin; Hesketh-Best, Poppy J.; Mckeown, Dean A.; Spivak, Marla; Schroeder, Declan C.

doi:10.1371/journal.pone.0297623

Cited by 2 publications

(2 citation statements)

References 72 publications

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“…The assays described here can further be improved by their application together with a recently developed semi-automated magnetic-beads-based technology applied to DWVA and DWVB RNA extraction from honeybee samples that allows for the rapid screening of viral loads in 96 samples simultaneously [ 47 ].…”

Section: Discussionmentioning

confidence: 99%

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Selection and Multiplexing of Reverse Transcription–Quantitative PCR Tests Targeting Relevant Honeybee Viral Pathogens

Rossi,

Del Matto,

Ricchiuti

et al. 2024

Microorganisms

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Verifying the inclusivity of molecular detection methods gives indications about the reliability of viral infection diagnosis because of the tendency of viral pathogens to undergo sequence variation. This study was aimed at selecting inclusive probes based on reverse transcription–quantitative PCR (RT-qPCR) assays for the diagnosis of the most widespread and detrimental viruses infecting honeybees, namely the acute bee paralysis virus (ABPV), the black queen cell virus (BQCV), the chronic paralysis bee virus (CBPV), the deformed wing virus variants A (DWVA) and B (DWVB), and the sacbrood virus (SBV). Therefore, previously described detection methods were re-evaluated in silico for their specificity and inclusivity. Based on this evaluation, selected methods were modified, or new ones were designed and tested in duplex RT-qPCR reactions. The limits of detection (LODs), effect of multiplexing on sensitivity and the viral RNA quantification potential in bees and hive debris were assessed. This study made available diagnostic assays able to detect an increased number of virus variants compared with previously described tests and two viral pathogens in a single PCR reaction.

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Section: Discussionmentioning

confidence: 99%

“…Most tests have already been applied in ecological studies and diagnostic screenings [ 4 , 9 , 25 , 43 , 44 , 45 , 46 , 47 , 48 ].…”

Section: Introductionmentioning

confidence: 99%

Selection and Multiplexing of Reverse Transcription–Quantitative PCR Tests Targeting Relevant Honeybee Viral Pathogens

Rossi,

Del Matto,

Ricchiuti

et al. 2024

Microorganisms

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Sacbrood viruses and select Lake Sinai virus variants dominated Apis mellifera colonies symptomatic for European foulbrood

Hesketh-Best,

Fowler,

Odogwu

et al. 2024

Microbiol Spectr

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European foulbrood (EFB) is a prevalent disease in the European honey bee ( Apis mellifera ) in the United States, which can lead to colony decline and collapse. The bacterial components of EFB are well-studied, but the diversity of viral infections within infected colonies has not been explored. In this study, we use meta-transcriptomics sequencing of 12 honey bee hives, symptomatic (+, n = 6) and asymptomatic (–, n = 6) for EFB, to investigate viral infection associated with the disease. We assembled 41 viral genomes, belonging to three families (Iflaviridae, Dicistroviridae, and Sinhaliviridae), all previously reported in honey bees, including Lake Sinai virus, deformed wing virus, sacbrood virus, Black queen cell virus, and Israeli acute paralysis virus. In colonies with severe EFB, we observed a higher occurrence of viral genomes (34 genomes) in contrast to fewer recovered from healthy colonies (seven genomes) and a complete absence of Dicistroviridae genomes.We observed specific Lake Sinai virus clades associated exclusively with EFB + or EFB – colonies, in addition to EFB-afflicted colonies that exhibited an increase in relative abundance of sacbrood viruses. Multivariate analyses highlighted that a combination of site and EFB disease status influenced RNA virome composition, while EFB status alone did not significantly impact it, presenting a challenge for comparisons between colonies kept in different yards. These findings contribute to the understanding of viral dynamics in honey bee colonies compromised by EFB and underscore the need for future investigations to consider viral composition when investigating EFB. IMPORTANCE This study on the viromes of honey bee colonies affected by European foulbrood (EFB) sheds light on the dynamics of viral populations in bee colonies in the context of a prevalent bacterial brood disease. The identification of distinct Lake Sinai virus and sacbrood virus clades associated with colonies affected by severe EFB suggests a potential connection between viral composition and disease status, emphasizing the need for further investigation into the role of viruses during EFB infection. The observed increase in sacbrood viruses during EFB infection suggests a potential viral dysbiosis, with potential implications for honey bee brood health. These findings contribute valuable insights related to beekeeping practices, offering a foundation for future research aimed at understanding and mitigating the impact of bacterial and viral infection in commercial honey bee operations and the management of EFB.

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A semi-automated and high-throughput approach for the detection of honey bee viruses in bee samples

Cited by 2 publications

References 72 publications

Selection and Multiplexing of Reverse Transcription–Quantitative PCR Tests Targeting Relevant Honeybee Viral Pathogens

Selection and Multiplexing of Reverse Transcription–Quantitative PCR Tests Targeting Relevant Honeybee Viral Pathogens

Sacbrood viruses and select Lake Sinai virus variants dominated Apis mellifera colonies symptomatic for European foulbrood

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