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Abstract

The spore-forming bacterium is the causative agent of American foulbrood (AFB), a devastating disease of honeybees (). In the present study, we used whole-genome sequencing (WGS) to investigate an extensive outbreak of AFB in northwestern Slovenia in 2019. A total of 59 . isolates underwent WGS, of which 40 originated from a single beekeeping operation, to assess the diversity of within the beekeeping operation, apiary and colony. By applying a case-specific single-linkage threshold of 34 allele differences (AD), whole-genome multilocus sequence typing (wgMLST) identified two outbreak clusters represented by ERIC II-ST11 clones. All isolates from a single beekeeping operation fell within cluster 1 and the median pairwise AD between them was 10 (range=1–22). The median pairwise AD for apiaries of the same beekeeping operation ranged from 8 to 11 (min.=1, max.=22). For colonies of the same apiary and honey samples from these colonies, the median pairwise AD ranged from 8 to 14 (min.=1, max.=20). The maximum within-cluster distance was 33 pairwise AD for cluster 1 and 44 for cluster 2 isolates. The minimum distance between the outbreak-related and non-related isolates was 37 AD, confirming the importance of associated epidemiological data for delineating outbreak clusters. The observed transmission events could be explained by the activities of honeybees and beekeepers. The present study provides insight into the genetic diversity of at different levels and thus provides information for future AFB surveillance.

Funding
This study was supported by the:
  • Slovenian Ministry of Agriculture, Forestry and Food (Award V4-1804)
    • Principle Award Recipient: DarjaKušar
  • Slovenian Research Agency (Award V4-1804)
    • Principle Award Recipient: DarjaKušar
  • Slovenian Research Agency (Award P4-0092)
    • Principle Award Recipient: NotApplicable
  • This is an open-access article distributed under the terms of the Creative Commons Attribution NonCommercial License.
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2021-12-03
2024-05-13
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References

  1. Papić B, Golob M, Zdovc I, Avberšek J, Ocepek M et al. Using whole-genome sequencing to assess the diversity of paenibacillus larvae within an outbreak and a beekeeping operation. Figshare 2021 [View Article]
    [Google Scholar]
  2. OIE Chapter 3.2.2. American Foulbrood of Honey Bees (Infection of Honey Bees with Paenibacillus larvae). In OIE Terrestrial Manual 2018 Paris: World Organisation for Animal Health (OIE); 2018 pp 719–735
    [Google Scholar]
  3. ECOLEX Council Directive 92/65/EEC of 13 July 1992 laying down animal health requirements governing trade in and imports into the Community of animals, semen, ova and embryos not subject to animal health requirements laid down in specific Community rules referred to in Annex A (I) to Directive 90/425/EEC. Off J L 1992; 14:54–72
    [Google Scholar]
  4. Versalovic J, Schneider M, De Bruijn FJ, Lupski JR. Genomic fingerprinting of bacteria using repetitive sequence-based polymerase chain reaction. Methods Mol Cell Biol 1994; 5:25–40
    [Google Scholar]
  5. Genersch E, Otten C. The use of repetitive element PCR fingerprinting (rep-PCR) for genetic subtyping of German field isolates of Paenibacillus larvae subsp. larvae. Apidologie (Celle) 2003; 34:195–206 [View Article]
    [Google Scholar]
  6. Genersch E, Forsgren E, Pentikäinen J, Ashiralieva A, Rauch S et al. Reclassification of Paenibacillus larvae subsp. pulvifaciens and Paenibacillus larvae subsp. larvae as Paenibacillus larvae without subspecies differentiation. Int J Syst Evol Microbiol 2006; 56:501–511 [View Article] [PubMed]
    [Google Scholar]
  7. Beims H, Bunk B, Erler S, Mohr KI, Spröer C et al. Discovery of Paenibacillus larvae ERIC V: phenotypic and genomic comparison to genotypes ERIC I-IV reveal different inventories of virulence factors which correlate with epidemiological prevalences of American Foulbrood. Int J Med Microbiol 2020; 310:151394 [View Article] [PubMed]
    [Google Scholar]
  8. Morrissey BJ, Helgason T, Poppinga L, Fünfhaus A, Genersch E et al. Biogeography of Paenibacillus larvae, the causative agent of American foulbrood, using a new multilocus sequence typing scheme. Environ Microbiol 2015; 17:1414–1424 [View Article] [PubMed]
    [Google Scholar]
  9. Rauch S, Ashiralieva A, Hedtke K, Genersch E. Negative correlation between individual-insect-level virulence and colony-level virulence of Paenibacillus larvae, the etiological agent of american foulbrood of honeybees. Appl Environ Microbiol 2009; 75:3344–3347 [View Article] [PubMed]
    [Google Scholar]
  10. Chen Y, Luo Y, Carleton H, Timme R, Melka D et al. Whole genome and core genome multilocus sequence typing and single nucleotide polymorphism analyses of Listeria monocytogenes isolates associated with an outbreak linked to cheese, united states, 2013. Appl Environ Microbiol 2017; 83:e00633-17 [View Article] [PubMed]
    [Google Scholar]
  11. Pightling AW, Pettengill JB, Luo Y, Baugher JD, Rand H et al. Terpreting whole-genome sequence analyses of foodborne bacteria for regulatory applications and outbreak investigations. Front Microbiol 2018; 9:1482 [View Article] [PubMed]
    [Google Scholar]
  12. Schürch AC, Arredondo-Alonso S, Willems RJL, Goering RV. Whole genome sequencing options for bacterial strain typing and epidemiologic analysis based on single nucleotide polymorphism versus gene-by-gene-based approaches. Clin Microbiol Infect 2018; 24:350–354 [View Article] [PubMed]
    [Google Scholar]
  13. Bertolotti AC, Forsgren E, Schäfer MO, Consortium E, Sircoulomb F et al. Development and evaluation of a core genome multilocus sequence typing scheme for Paenibacillus larvae, the deadly American foulbrood pathogen of honeybees. Environ Microbiol 2021; 23:5042–5051 [View Article] [PubMed]
    [Google Scholar]
  14. Papić B, Diricks M, Kušar D. Analysis of the global population structure of Paenibacillus larvae and outbreak investigation of American foulbrood using a stable wgMLST scheme. Front Vet Sci 2021; 8:582677 [View Article] [PubMed]
    [Google Scholar]
  15. Ågren J, Schäfer MO, Forsgren E. Using whole genome sequencing to study American foulbrood epidemiology in honeybees. PLoS ONE 2017; 12:e0187924 [View Article] [PubMed]
    [Google Scholar]
  16. Žugelj A, Papić B, Zdovc I, Zajc U, Golob M et al. ERIC and WGS typing of Paenibacillus larvae in Slovenia: investigation of ERIC I outbreaks. Insects 2021; 12:362 [View Article] [PubMed]
    [Google Scholar]
  17. Payne M, Octavia S, Luu LDW, Sotomayor-Castillo C, Wang Q et al. Enhancing genomics-based outbreak detection of endemic Salmonella enterica serovar Typhimurium using dynamic thresholds. Microb Genom 2019; 7:000310 [View Article]
    [Google Scholar]
  18. Letunic I, Bork P. Interactive Tree Of Life (iTOL) v4: recent updates and new developments. Nucleic Acids Res 2019; 47:W256–W259 [View Article] [PubMed]
    [Google Scholar]
  19. Jolley KA, Bray JE, Maiden MCJ. Open-access bacterial population genomics: BIGSdb software, the PubMLST.org website and their applications. Wellcome Open Res 2018; 3:124 [View Article] [PubMed]
    [Google Scholar]
  20. Argimón S, Abudahab K, Goater RJE, Fedosejev A, Bhai J et al. Microreact: visualizing and sharing data for genomic epidemiology and phylogeography. Microb Genom 2016; 2:e000093 [View Article]
    [Google Scholar]
  21. Jagadeesan B, Baert L, Wiedmann M, Orsi RH. Comparative analysis of tools and approaches for source tracking Listeria monocytogenes in a food facility using whole-genome sequence data. Front Microbiol 2019; 10:947 [View Article] [PubMed]
    [Google Scholar]
  22. Papić B, Kušar D, Zdovc I, Golob M, Pate M. Retrospective investigation of listeriosis outbreaks in small ruminants using different analytical approaches for whole genome sequencing-based typing of Listeria monocytogenes. Infect Genet Evol 2020; 77:104047 [View Article] [PubMed]
    [Google Scholar]
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