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Apidologie

, Volume 49, Issue 4, pp 459–461 | Cite as

A scientific note on improved isolation methods for Melissococcus plutonius from diseased Apis mellifera larvae

  • Yuka Nakai
  • Michika Ishihara
  • Rie Arai
  • Daisuke Takamatsu
Scientific note
  • 88 Downloads

European foulbrood (EFB) caused by Melissococcus plutonius is a significant infectious disease of honey bee brood. EFB is typically diagnosed based on the visual inspection of brood combs and detection of diseased larvae; however, since symptoms of EFB may easily be confused with other diseases or abnormalities in larvae, verification of the presence of M. plutonius in diseased larvae is required for a reliable diagnosis of EFB in many cases. The isolation and storage of M. plutonius strains are also important for future EFB studies. However, difficulties are sometimes associated with the isolation of M. plutonius from diseased larvae by culture methods due to the growth of secondary bacteria associated with EFB.

In an inspection of foulbrood on May 16, 2017, an abnormal Apis melliferacolony was detected in an apiary in Saitama prefecture, Japan. The colony showed irregular capping, and dead larvae were observed in uncapped cells. Gram-positive cocci were detected in dead larvae by...

Keywords

European foulbrood Melissococcus plutonius Apis mellifera Enterococcus faecalis royal jelly 

Notes

Authors’ contributions

YN and MI: Diagnosed the EFB case described in this study and performed experiments; RA: Designed the first experiment; DT: Designed the second experiment and contributed to writing the manuscript. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

13592_2018_572_MOESM1_ESM.pdf (972 kb)
ESM 1 (PDF 972 kb)

References

  1. Arai, R., Tominaga, K., Wu, M., Okura, M., Ito, K., Okamura, N., Onishi, H., Osaki, M., Sugimura, Y., Yoshiyama, M., Takamatsu, D. (2012) Diversity of Melissococcus plutonius from honeybee larvae in Japan and experimental reproduction of European foulbrood with cultured atypical isolates. PLoS One 7 (3), e33708CrossRefPubMedPubMedCentralGoogle Scholar
  2. Arai, R., Miyoshi-Akiyama, T., Okumura, K., Morinaga, Y., Wu, M., Sugimura, Y., Yoshiyama, M., Okura, M., Kirikae, T., Takamatsu, D. (2014) Development of duplex PCR assay for detection and differentiation of typical and atypical Melissococcus plutonius strains. J. Vet. Med. Sci. 76 (4), 491–498CrossRefPubMedGoogle Scholar
  3. Budge, G. E., Shirley, M. D. F., Jones, B., Quill, E., Tomkies, V., Feil, E. J., Brown, M. A., Haynes, E. G. (2014) Molecular epidemiology and population structure of the honey bee brood pathogen Melissococcus plutonius. ISME J. 8 (8), 1588–1597CrossRefPubMedPubMedCentralGoogle Scholar
  4. Forsgren, E., Budge, G. E., Charrière J.-D., Hornitzky, M. A. Z. (2013) Standard methods for European foulbrood research. J. Apic. Res. 52 (1), 1–14CrossRefGoogle Scholar
  5. Haynes, E., Helgason, T., Young, J. P. W., Thwaites, R., Budge, G. E. (2013) A typing scheme for the honeybee pathogen Melissococcus plutonius allows detection of disease transmission events and a study of the distribution of variants. Environ. Microbiol. Rep. 5 (4), 525–529CrossRefPubMedGoogle Scholar
  6. Hornitzky, M. A. Z., Smith, L. (1998) Procedures for the culture of Melissococcus pluton from diseased brood and bulk honey samples. J. Apic. Res. 37 (4): 293–294CrossRefGoogle Scholar
  7. Takamatsu, D., Morinishi, K., Arai, R., Sakamoto, A., Okura, M., Osaki, M. (2014) Typing of Melissococcus plutonius isolated from European and Japanese honeybees suggests spread of sequence types across borders and between different Apis species. Vet. Microbiol. 171 (1–2), 221–226CrossRefPubMedGoogle Scholar
  8. Takamatsu, D., Osawa, A., Nakamura, K., Yoshiyama, M., Okura, M. (2017) High-level resistance of Melissococcus plutonius clonal complex 3 strains to antimicrobial activity of royal jelly. Environ. Microbiol. Rep. 9 (5), 562–570CrossRefPubMedGoogle Scholar

Copyright information

© INRA, DIB and Springer-Verlag France SAS, part of Springer Nature 2018

Authors and Affiliations

  • Yuka Nakai
    • 1
  • Michika Ishihara
    • 1
  • Rie Arai
    • 2
  • Daisuke Takamatsu
    • 3
    • 4
  1. 1.Saitama Prefectural Chuo Livestock Hygiene Service CenterSaitamaJapan
  2. 2.Livestock Safety Division, Department of Agriculture and Forestry, Saitama Prefectural GovernmentSaitamaJapan
  3. 3.Division of Bacterial and Parasitic Disease, National Institute of Animal Health, National Agriculture and Food Research OrganizationTsukubaJapan
  4. 4.The United Graduate School of Veterinary SciencesGifu UniversityGifuJapan

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