European foulbrood in Czechia after 40 years: application of next-generation sequencing to analyze Melissococcus plutonius transmission and influence on the bacteriome of Apis mellifera
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Abstract
Worker honeybees (Apis mellifera) transmit Melissococcus plutonius between colonies. However, the transmission of M. plutonius, which causes European foulbrood (EFB), is poorly understood. To analyze the first EFB outbreak in 40 years in Czechia, we collected 49 hive worker samples from 18 beehives in two diseased apiaries for bacteriome analysis of the V1-V3 portion of the 16S rRNA gene. When we compared control samples obtained outside of the EFB zone, bees from an EFB apiaries containing colonies without clinical symptoms and bees from colonies with EFB clinical symptoms, there was a 100-fold higher occurrence of M. plutonius in colonies with EFB symptoms. The presence of M. plutonius in controls indicated that this pathogen exists in an enzootic state. EFB influenced the core bacteria in the worker bacteriome because the number of Snodgrassella alvi, Lactobacillus mellis, Lactobacillus melliventris, and Fructobacillus fructosus sequences increased, while Bartonella apis, Frischella perrara, and Commensalibacter intestine sequences decreased. Together, the results of this study suggest worker bees from EFB-diseased apiaries serve as vectors of M. plutonius, and eliminating such colonies is an appropriate method to overcome disease outbreaks. Because M. plutonius exists in honeybee colonies in an enzootic state, there may be similar abundances in control colonies outside the EFB zone to those in asymptomatic colonies from EFB apiaries. High-throughput Illumina next-generation sequencing permitted the quantitative interpretation of M. plutonius within the honeybee worker bacteriome. Future studies focusing on honeybee diseases, colony losses, detection of bacterial pathogens and interactions of bacteriome with pathogenic bacteria will benefit of this study.
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2016. European foulbrood in Czechia after 40 years: application of next-generation sequencing to analyze Melissococcus plutonius transmission and influence on the bacteriome of Apis mellifera. PeerJ Preprints 4:e2618v1 https://doi.org/10.7287/peerj.preprints.2618v1note This preprint is not peer-reviewed. You may wish to reference the subsequent peer-reviewed version of this article.
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Competing Interests
The authors declare that they have no competing interests.
Author Contributions
Tomas Erban conceived and designed the experiments, analyzed the data, contributed reagents/materials/analysis tools, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.
Ondrej Ledvinka analyzed the data, prepared figures and/or tables, reviewed drafts of the paper.
Martin Kamler conceived and designed the experiments, performed the experiments, reviewed drafts of the paper.
Bronislava Hortova performed the experiments, prepared figures and/or tables, reviewed drafts of the paper.
Marta Nesvorna performed the experiments, analyzed the data, reviewed drafts of the paper.
Jan Tyl performed the experiments, reviewed drafts of the paper.
Dalibor Titera conceived and designed the experiments, contributed reagents/materials/analysis tools, reviewed drafts of the paper.
Martin Markovic prepared figures and/or tables, reviewed drafts of the paper.
Jan Hubert conceived and designed the experiments, performed the experiments, analyzed the data, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.
DNA Deposition
The following information was supplied regarding the deposition of DNA sequences:
GenBank as SRA project No. SRP093440
Data Deposition
The following information was supplied regarding data availability:
GenBank as SRA project No. SRP093440
Funding
The study was supported by project No. QJ1310085 of the Ministry of Agriculture of the Czech Republic (http://www.eagri.cz). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
