Apidologie

, 42:49

Dominance of Nosema ceranae in honey bees in the Balkan countries in the absence of symptoms of colony collapse disorder

  • Jevrosima Stevanovic
  • Zoran Stanimirovic
  • Elke Genersch
  • Sanja R. Kovacevic
  • Jovan Ljubenkovic
  • Milena Radakovic
  • Nevenka Aleksic
Original Article

Abstract

Nosema species were determined in honey bees from Balkan countries. A total of 273 Nosema-positive samples were analysed. Duplex PCR and PCR-RFLP with newly designed primers, nos-16S-fw/rv, were used to differentiate between N. apis and N. ceranae. N. apis was detected in only one sample (collected in 2008 in Serbia) and N. ceranae in all the others (N = 272) including 35 older samples from Serbia collected between 2000 and 2005. No co-infection was detected. The results suggest (1) the dominance of N. ceranae infection in all Balkan countries monitored throughout the last three years; (2) the presence of N. ceranae in Serbia at least since 2000, which means that N. ceranae has not recently displaced N. apis; (3) the higher efficacy of PCR-RFLP with newly designed primers, nos-16S-fw/rv, in comparison with duplex PCR (100%:82%, respectively). The prevalence of N. ceranae in Balkan countries was not associated with an increase in nosemosis or colony losses resembling Colony Collapse Disorder (CCD).

Keywords

Nosema ceranae PCR-RFLP duplex PCR Balkan countries 

References

  1. Ashiralieva A., Genersch E. (2006) Reclassification, genotypes and virulence of Paenibacillus larvae, the etiological agent of American foulbrood in honeybees — a review, Apidologie 37, 411–420.CrossRefGoogle Scholar
  2. Chauzat M.P., Higes M., Martín-Hernández R., Meana A., Cougoule N., Faucon J.P. (2007) Presence of Nosema ceranae in French honeybee colonies, J. Apic. Res. 46, 127–128.CrossRefGoogle Scholar
  3. Chen Y., Evans J.D., Smith I.B., Pettis J.S. (2008) Nosema ceranae is a long-present and widespread microsporidean infection of the European honey bee (Apis mellifera) in the United States, J. Invertebr. Pathol. 97, 186–188.PubMedCrossRefGoogle Scholar
  4. Cox-Foster D.L., Conlan S., Holmes E.C., Palacios G., Evans J.D., Moran N.A., Quan P.L., Briese T., Hornig M., Geiser D.M., Martinson V., van Engelsdorp D., Kalkstein A.L., Drysdale A., Hui J., Zhai J.H., Cui L.W., Hutchison S.K., Simons J.F., Egholm M., Pettis J.S., Lipkin W.I. (2007) A metagenomic survey of microbes in honey bee colony collapse disorder, Science 318, 283–287.PubMedCrossRefGoogle Scholar
  5. De la Rúa P., Jaffé R., Dall’Olio R., Muñoz I., Serrano J.(2009) Biodiversity, conservation and current threats to European honeybees, Apidologie 40, 263–284.CrossRefGoogle Scholar
  6. Franck P., Garnery L., Solignac M., Cornuet J.M. (2000) Molecular confirmation of a fourth lineage in honey bees from the Near East, Apidologie 31, 167–180.CrossRefGoogle Scholar
  7. Fries I. (1997) Protozoa, in: Morse R.A., Flottum K. (Eds.), Honey bee pests, predators and diseases, 3rd ed., A.I. Root Company, Medina, Ohio, USA, pp. 59–76.Google Scholar
  8. Fries I., Feng F., da Silva A., Slemenda S.B., Pieniazek J. (1996) Nosema ceranae n. sp. (Microspora, Nosematidae), morphological and molecular characterization of a microsporidian parasite of the Asian honey bee Apis cerana (Hymenoptera, Apidae), Eur. J. Protistol. 32, 356–365.Google Scholar
  9. Fries I., Martín R., Meana A., García-Palencia P., Higes M. (2006) Natural infections of Nosema ceranae in European honey bees, J. Apic. Res. 45, 230–233.CrossRefGoogle Scholar
  10. Garnery L., Cornuet J.M., Solignac M. (1992) Evolutionary History of the Honey Bee Apis mellifera Inferred from Mitochondrial DNA Analysis, Mol. Ecol. 1, 145–154.PubMedCrossRefGoogle Scholar
  11. Gatehouse H.S., Malone L.A. (1998) The ribosomal RNA gene region of Nosema apis (Microspora): DNA sequence for small and large subunit rRNA genes and evidence of a large tandem repeat unit size, J. Invertebr. Pathol. 71, 97–105.PubMedCrossRefGoogle Scholar
  12. Genersch E., Ashiralieva A., Fries I. (2005) Strainand genotype-specific differences in virulence of Paenibacillus larvae subsp. larvae, a bacterial pathogen causing American foulbrood disease in honeybees, Appl. Environ. Microbiol. 71, 7551–7555.PubMedCrossRefGoogle Scholar
  13. Genersch E., Forsgren E., Pentikäinen J., Ashiralieva A., Rauch S., Kilwinski J., Fries I. (2006) Reclassification of Paenibacillus larvae subsp. pulvifaciens and Paenibacillus larvae subsp. larvae as Paenibacillus larvae without subspecies classification, Int. J. Syst. Evol. Microbiol. 56, 501–511.PubMedCrossRefGoogle Scholar
  14. Giersch T., Berg T., Galea F., Hornitzky M. (2009) Nosema ceranae infects honey bees (Apis mellifera) and contaminates honey in Australia, Apidologie 40, 117–123.CrossRefGoogle Scholar
  15. Higes M., García-Palencia P., Martín-Hernández R., Meana A. (2007) Experimental infection of Apis mellifera honeybees with Nosema ceranae (Microsporidia), J. Invertebr. Pathol. 94, 211–217.PubMedCrossRefGoogle Scholar
  16. Higes M., Martín R., Meana A. (2006) Nosema ceranae, a new microsporidian parasite in honeybees in Europe, J. Invertebr. Pathol. 92, 93–95.PubMedCrossRefGoogle Scholar
  17. Higes M., Martín R., Sanz A., Álvarez N., Sanz A., García-Palencia P., Meana A. (2005) El síndrome de despoblamiento de las colmenas en España. Consideraciones sobre su origen, Vida Apícola 133, 15–21.Google Scholar
  18. Higes M., Martín-Hernández R., Botías C., Garrido-Bailón E., González-Porto A.V., Barrios L., Del Nozal M.J., Bernal J.L., Jiménez J.J., García-Palencia P., Meana A. (2008) How natural infection by Nosema ceranae causes honeybee colony collapse, Environ. Microbiol. 10, 2659–2669.PubMedCrossRefGoogle Scholar
  19. Higes M., Martín-Hernández R., Garrido-Bailón E., Botías C., Meana A. (2009a) The presence of Nosema ceranae (Microsporidia) in North African honey bees (Apis mellifera intermissa), J. Apic. Res. 48, 217–219.CrossRefGoogle Scholar
  20. Higes M., Martín-Hernández R., Garrido-Bailón E., González-Porto A.V., García-Palencia P., Meana A., del Nozal M.J., Mayo R., Bernal J.L. (2009b) Honeybee colony collapse due to Nosema ceranae in professional apiaries, Environ. Microbiol. Rep. 1, 110–113.CrossRefGoogle Scholar
  21. Huang W.F., Jiang J.H., Chen Y.W., Wang C.H. (2005) Complete rRNA sequence of the Nosema ceranae from honeybee (Apis mellifera), https://gra103.aca.ntu.edu.tw/gdoc/F90632004a.pdf.
  22. Huang W.F., Bocquet M., Lee KC, Sung I.H., Jiang J.H., Chen Y.W., Wang C.H. (2008) The comparison of rDNA spacer regions of Nosema ceranae isolates from different hosts and locations, J. Invertebr. Pathol. 97, 9–13.PubMedCrossRefGoogle Scholar
  23. Huang W.F., Jiang J.H., Chen Y.W., Wang C.H. (2007) A Nosema ceranae isolate from the honeybee Apis mellifera, Apidologie 38, 30–37.CrossRefGoogle Scholar
  24. Imdorf A., Charrière J.D., Gallmann P. (2006) Mögliche Ursachen für die Völkerverluste der letzten Jahre, Schweiz. Bienenztg. 129, 6–10Google Scholar
  25. Invernizzi C., Abud C., Tomasco I.H., Harriet J., Ramallo G., Campá J., Katz H., Gardiol G., Mendoza Y. (2009) Presence of Nosema ceranae in honeybees (Apis mellifera) in Uruguay, J. Invertebr. Pathol. 101, 150–153.PubMedCrossRefGoogle Scholar
  26. Johnson R.M., Evans J.D., Robinson G.E., Berenbaum M.R. (2009) Changes in transcript abundance relating to colony collapse disorder in honey bees (Apis mellifera), PNAS 106, 14790–14795.PubMedCrossRefGoogle Scholar
  27. Klee J., Besana A.M., Genersch E., Gisder S., Nanetti A., Tam D.Q., Chinh T.X., Puerta F., Ruz J.M., Kryger P., Message D., Hatjina F., Korpela S., Fries I., Paxton R.J. (2007) Widespread dispersal of the microsporidian Nosema ceranae, an emergent pathogen of the western honey bee, Apis mellifera, J. Invertebr. Pathol. 96, 1–10.PubMedCrossRefGoogle Scholar
  28. Kozmus P., Stevanovic J., Stanimirovic Z., Stojic V., Kulisic Z., Meglic V. (2007) Analysis of mitochondrial DNA in honey bees (Apis mellifera) from Serbia, Acta Vet.-Beograd 57, 465–476.Google Scholar
  29. Martín-Hernández R., Meana A., Prieto L., Martínez Salvador A., Garrido-Bailón E., Higes M. (2007) Outcome of colonization of Apis mellifera by Nosema ceranae, Appl. Environ. Microbiol. 73, 6331–6338.PubMedCrossRefGoogle Scholar
  30. Meixner M.D., Worobik M., Wilde J., Fuchs S., Koeniger N. (2007) Apis mellifera mellifera in eastern Europe — morphometric variation and determination of its range limits, Apidologie 38, 191–197.CrossRefGoogle Scholar
  31. OIE (2008) Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, Chap. 2.2.4., Nosemosis of Honey Bees, http://www.oie.int/fr/normes/mmanual/2008/pdf/2.02.04_NOSEMOSIS.pdf.
  32. Paxton R.J., Klee J., Korpela S., Fries I. (2007) Nosema ceranae has infected Apis mellifera in Europe since at least 1998 and may be more virulent than Nosema apis, Apidologie 38, 558–565.CrossRefGoogle Scholar
  33. Stanimirovic Z., Stevanovic J., Andjelkovic M. (2005) Chromosomal diversity in Apis mellifera carnica from Serbia, Apidologie 36, 31–42.CrossRefGoogle Scholar
  34. Stanimirovic Z., Stevanovic J., Bajic V., Radovic I. (2007) Evaluation of genotoxic effects of fumagillin (dicyclohexylamine) by citogenetic tests in vivo, Mutat. Res. 628, 1–10.PubMedGoogle Scholar
  35. Stanimirovic Z., Stevanovic J., Kulic M., Stojic V. (2006) Frequency of chromosomal aberrations in the evaluation of genotoxic potential of dicyclohexylamine (fumagillin) in vivo, Acta Vet.-Beograd 56, 353–366.Google Scholar
  36. Stevanovic J., Stanimirovic Z., Pejin I.I., Lazarevic M. (2006) Monitoring of mitotic index and frequency of micronuclei in evaluation of genotoxic potential of fumagillin (dicyclohexylamine) in vivo, Acta Vet.-Beograd 56, 437–448.Google Scholar
  37. Stevanovic J., Stanimirovic Z., Radakovic M., Stojic V. (2008) In vitro evaluation of the clastogenicity of fumagillin, Environ. Mol. Mutagen. 49, 594–601.PubMedCrossRefGoogle Scholar
  38. Sušnik S., Kozmus P., Poklukar J., Megliè V. (2004) Molecular characterisation of indigenous Apis mellifera carnica in Slovenia, Apidologie 35, 623–636.CrossRefGoogle Scholar
  39. Tapaszti Z., Forgách P., Kovágó C., Békési L., Bakonyi T., Rusvai M. (2009) First detection and dominance of Nosema ceranae in Hungarian honeybee colonies, Acta Vet. Hung. 57, 383–388.PubMedCrossRefGoogle Scholar
  40. van Engelsdorp D., Evans J.D., Saegerman C., Mullin C., Haubruge E., Nguyen B.K., Frazier M., Frazier J., Cox-Foster D., Chen Y., Underwood R., Tarpy D.R., Pettis J.S. (2009) Colony collapse disorder: a descriptive study, PLoS One 4, e6481.CrossRefGoogle Scholar
  41. Williams G.R., Shafer A.B.A., Rogers R.E.L., Shutler D., Stewart D.T. (2008a) First detection of Nosema ceranae, a microsporidean parasite of European honey bees (Apis mellifera), in Canada and central USA, J. Invertebr. Pathol. 97, 189–192.PubMedCrossRefGoogle Scholar
  42. Williams G.R., Sampson MA, Shutler D., Rogers R.E.L. (2008b) Does fumagillin control the recently detected invasive parasite Nosema ceranae in western honey bees (Apis mellifera)? J. Invertebr. Pathol. 99, 342–344.PubMedCrossRefGoogle Scholar
  43. Zander E. (1909) Tierische Parasiten als Krankheitserreger bei der Biene, Münchener Bienenzeitung 31, 196–204.Google Scholar

Copyright information

© INRA, DIB-AGIB and Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Jevrosima Stevanovic
    • 1
  • Zoran Stanimirovic
    • 1
  • Elke Genersch
    • 2
  • Sanja R. Kovacevic
    • 1
  • Jovan Ljubenkovic
    • 1
  • Milena Radakovic
    • 1
  • Nevenka Aleksic
    • 3
  1. 1.Department of Biology, Faculty of Veterinary MedicineUniversity of BelgradeBelgradeSerbia
  2. 2.Institute for Bee ResearchHohen NeuendorfGermany
  3. 3.Department of Parasitic Diseases, Faculty of Veterinary MedicineUniversity of BelgradeBelgradeSerbia

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