, Volume 44, Issue 5, pp 522–536

Characteristics of Nosema ceranae infection in Serbian honey bee colonies

  • Jevrosima Stevanovic
  • Predrag Simeunovic
  • Bojan Gajic
  • Nada Lakic
  • Dejan Radovic
  • Ingemar Fries
  • Zoran Stanimirovic
Original article


The incidence, symptoms and consequences of Nosema ceranae infection were monitored in 200 honey bee colonies in Serbia over 5 years (2008–2012) to reveal if they display characteristics described for nosemosis type C. Continual high frequency of N. ceranae-positive colonies was recorded within each study year, ranging from 73 to 98 %. A seasonal pattern in N. ceranae incidence was observed over 4 years (2008–2011). Symptoms traditionally attributed to Nosema apis infection were observed in the majority of N. ceranae-infected colonies, both among surviving and those that died. These symptoms could not serve as predictive markers for colony losses, as they were significantly more frequently observed among surviving colonies in 2010, 2011 and 2012. No clear association between N. ceranae infection and colony losses was affirmed neither during the winter nor during the summer season. Thus, N. ceranae infection in Serbian bees do not fit the characteristics for nosemosis type C.


Nosema ceranae Apis mellifera nosemosis type C faeces 


  1. Alaux, C., Brunet, J.L., Dussaubat, C., Mondet, F., Tchamitchan, S., Cousin, M., Brillard, J., Baldy, A., Belzunces, L.P., Le Conte, Y. (2010) Interactions between Nosema microspores and a neonicotinoid weaken honeybees (Apis mellifera). Environ. Microbiol. 12, 774–782PubMedCrossRefGoogle Scholar
  2. Antúnez, K., Martín-Hernández, R., Prieto, L., Meana, A., Zunino, P., Higes, M. (2009) Immune suppression in the honey bee (Apis mellifera) following infection by Nosema ceranae (Microsporidia). Environ. Microbiol. 11, 2284–2290PubMedCrossRefGoogle Scholar
  3. Bacandritsos, N., Granato, A., Budge, G., Papanastasiou, I., Roinioti, E., Caldon, M., Falcaro, C., Gallina, A., Mutinelli, F. (2010) Sudden deaths and colony population decline in Greek honey bee colonies. J. Invertebr. Pathol. 105, 335–340PubMedCrossRefGoogle Scholar
  4. Bailey, L. (1955) The epidemiology and control of Nosema disease of the honey bee. Ann. Appl. Biol. 43, 379–389CrossRefGoogle Scholar
  5. Bailey, L. (1967) Nosema apis and dysentery of the honey bee. J. Apic. Res. 6, 121–125Google Scholar
  6. Botías, C., Martín-Hernández, R., Garrido-Bailón, E., González-Porto, A., Martínez-Salvador, A., De La Rúa, P., Meana, A., Higes, M. (2012) Critical aspects of the Nosema spp. diagnostic sampling in honey bee (Apis mellifera L.) colonies. Parasitol. Res. 110, 2557–2561PubMedCrossRefGoogle Scholar
  7. Bourgeois, A.L., Rinderer, E.T., Beaman, D.L., Danka, G.R. (2010) Genetic detection and quantification of Nosema apis and N. ceranae in the honey bee. J. Invertebr. Pathol. 103, 53–58PubMedCrossRefGoogle Scholar
  8. Bromenshenk, J.J., Henderson, C.B., Wick, C.H., Stanford, M.F., Zulich, A.W., et al. (2010) Iridovirus and microsporidian linked to honey bee colony decline. PLoS One 5, e13181PubMedCrossRefGoogle Scholar
  9. Burgher-MacLellan, K.L., Williams, G.R., Shutler, D., MacKenzie, K., Rogers, R.E.L. (2010) Optimization of duplex real-time PCR with melting-curve analysis for detecting the microsporidian parasites Nosema apis and Nosema ceranae in Apis mellifera. Can. Entomol. 142, 271–283CrossRefGoogle Scholar
  10. Chaimanee, V., Chantawannakul, P., Chen, Y., Evans, J.D., Pettis, J.S. (2012) Differential expression of immune genes of adult honey bee (Apis mellifera) after inoculated by Nosema ceranae. J. Insect Physiol. 58, 1090–1095PubMedCrossRefGoogle Scholar
  11. 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, 127–128CrossRefGoogle Scholar
  12. 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, 186–188PubMedCrossRefGoogle Scholar
  13. Chen, Y.P., Evans, J.D., Murphy, C., Gutell, R., Zuker, M., Gundensen-Rindal, D., Pettis, J.S. (2009) Morphological, molecular, and phylogenetic characterization of Nosema ceranae, a microsporidian parasite isolated from the European honey bee, Apis mellifera. J. Eukaryot. Microbiol. 56, 142–147PubMedCrossRefGoogle Scholar
  14. COLOSS workshop (2009). Conclusions, Proc. Workshop “Nosema disease: lack of knowledge and work standardization” (COST Action FA0803) Guadalajara. http://www.coloss.org/news/nosema-workshop-proceedings-online. Accessed 20 Nov 2009
  15. Cox-Foster, D.L., Conlan, S., Holmes, E.C., Palacios, G., Evans, J.D., et al. (2007) A metagenomic survey of microbes in honey bee colony collapse disorder. Science 318, 283–287PubMedCrossRefGoogle Scholar
  16. Dainat, B., Evans, J.D., Chen, Y.P., Gauthier, L., Neumann, P. (2012) Predictive markers of honey bee colony collapse. PLoS One 7, e32151PubMedCrossRefGoogle Scholar
  17. Ellis, J.D., Evans, J.D., Pettis, J. (2010) Colony losses, managed colony population decline, and colony collapse disorder in the United States. J. Apic. Res. 49, 134–136CrossRefGoogle Scholar
  18. Fernández, J.M., Puerta, F., Cousinou, M., Dios-Palomares, R., Campano, F., Redondo, L. (2012) Asymptomatic presence of Nosema spp. in Spanish commercial apiaries. J. Invertebr. Pathol. 111, 106–110PubMedCrossRefGoogle Scholar
  19. Forsgren, E., Fries, I. (2010) Comparative virulence of Nosema ceranae and Nosema apis in individual European honey bees. Vet. Parasitol. 170, 212–217PubMedCrossRefGoogle Scholar
  20. Fries, I. (2010) Nosema ceranae in European honey bees (Apis mellifera). J. Invertebr. Pathol. 103, S73–S79PubMedCrossRefGoogle Scholar
  21. Fries, I., Forsgren, E. (2008) Undersökning av spridningen av Nosema ceranae i Sverige. Investigation of the distribution of Nosema ceranae in Sweden Bitidningen 107, januari/februari, pp. 26–27 (in Swedish)Google Scholar
  22. Fries, I., Raina, S. (2003) American foulbrood (Paenibacillus larvae larvae) and African honey bees (Apis mellifera scutellata). J. Econ. Entomol. 96, 1641–1646PubMedCrossRefGoogle Scholar
  23. Fries, I., Ekbohm, G., Villumstad, E. (1984) Nosema apis, sampling techniques and honey yield. J. Apic. Res. 23, 102–105Google Scholar
  24. 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, 356–365CrossRefGoogle Scholar
  25. 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, 230–233CrossRefGoogle Scholar
  26. Genersch, E., von der Ohe, W., Kaatz, H., Schroeder, A., Otten, C., et al. (2010) The German bee monitoring project: a long term study to understand periodically high winter losses of honey bee colonies. Apidologie 41, 332–352CrossRefGoogle Scholar
  27. Giersch, T., Berg, T., Galea, F., Hornitzky, M. (2009) Nosema ceranae infects honey bees (Apis mellifera) and contaminates honey in Australia. Apidologie 40, 117–123CrossRefGoogle Scholar
  28. Gisder, S., Hedtke, K., Möckel, N., Frielitz, M.-C., Linde, A., Genersch, E. (2010) Five-year cohort study of Nosema spp. in Germany: does climate shape virulence and assertiveness of Nosema ceranae? Appl. Environ. Microb. 76, 3032–3038CrossRefGoogle Scholar
  29. Hamiduzzaman, M.M., Guzman-Novoa, E., Goodwin, P.H. (2010) A multiplex PCR assay to diagnose and quantify Nosema infections in honey bees (Apis mellifera). J. Invertebr. Pathol. 105, 151–155PubMedCrossRefGoogle Scholar
  30. Hedtke, K., Jensen, P.M., Jensen, A.B., Genersch, E. (2011) Evidence for emerging parasites and pathogens influencing outbreaks of stress-related diseases like chalkbrood. J. Invertebr. Pathol. 108, 167–173PubMedCrossRefGoogle Scholar
  31. Higes, M., Martín, R., Meana, A. (2006) Nosema ceranae, a new microsporidian parasite in honeybees in Europe. J. Invertebr. Pathol. 92, 93–95PubMedCrossRefGoogle Scholar
  32. Higes, M., Martín-Hernández, R., Botías, C., Garrido Bailón, E., González-Porto, A.V., et al. (2008) How natural infection by Nosema ceranae causes honeybee colony collapse. Environ. Microbiol. 10, 2659–2669PubMedCrossRefGoogle Scholar
  33. 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. (2009) Honeybee colony collapse due to Nosema ceranae in professional apiaries. Environ. Microbiol. Rep. 1, 110–113PubMedCrossRefGoogle Scholar
  34. Higes, M., Martín-Hernández, R., Meana, A. (2010a) Nosema ceranae in Europe: an emergent type C nosemosis. Apidologie 41, 375–392CrossRefGoogle Scholar
  35. Higes, M., Martín-Hernández, R., Martínez-Salvador, A., Garrido-Bailón, E., González-Porto, A.V., Meana, A., Bernal, J.L., del Nozal, M.J., Bernal, J. (2010b) A preliminary study of the epidemiological factors related to honey bee colony loss in Spain. Environ. Microbiol. Rep. 2, 243–250PubMedCrossRefGoogle Scholar
  36. Huang, W.F., Jiang, J.H., Chen, Y.W., Wang, C.H. (2007) A Nosema ceranae isolate from the honeybee Apis mellifera. Apidologie 38, 30–37CrossRefGoogle Scholar
  37. 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, 150–153PubMedCrossRefGoogle Scholar
  38. Klee, J., Besana, A.M., Genersch, E., Gisder, S., Nanetti, A., et al. (2007) Widespread dispersal of the microsporidian Nosema ceranae, an emergent pathogen of the western honey bee, Apis mellifera. J. Invertebr. Pathol. 96, 1–10PubMedCrossRefGoogle Scholar
  39. Martín-Hernández, R., Meana, A., Prieto, L., Salvador, A.M., Garrido-Bailón, E., Higes, M. (2007) Outcome of colonization of Apis mellifera by Nosema ceranae. Appl. Environ. Microb. 73, 6331–6338CrossRefGoogle Scholar
  40. Martín-Hernández, R., Meana, A., García-Palencia, P., Marín, P., Botías, C., Garrido-Bailón, E., Barrios, L., Higes, M. (2009) Effect of temperature on the biotic potential of honeybee microsporidia. Appl. Environ. Microb. 75, 2554–2557CrossRefGoogle Scholar
  41. Martín-Hernández, R., Botías, C., Garrido Bailón, E., Martínez-Salvador, A., Prieto, L., Meana, A., Higes, M. (2011a) Microsporidia infecting Apis mellifera: coexistence or competition. Is Nosema ceranae replacing Nosema apis? Environ. Microbiol. 14, 2127–2138PubMedCrossRefGoogle Scholar
  42. Martín-Hernández, R., Botías, C., Barrios, L., Martínez-Salvador, A., Meana, A., Mayack, C., Higes, M. (2011b) Comparison of the energetic stress associated with experimental Nosema ceranae and Nosema apis infection of honeybees (Apis mellifera). Parasitol. Res. 109, 605–612PubMedCrossRefGoogle Scholar
  43. Mayack, C., Naug, D. (2009) Energetic stress in the honeybee Apis mellifera from Nosema ceranae infection. J. Invertebr. Pathol. 100, 185–188PubMedCrossRefGoogle Scholar
  44. Meana, A., Martín-Hernández, R., Higes, M. (2010) The reliability of spore counts to diagnose Nosema ceranae infections in honey bees. J. Apic. Res. 49, 212–214CrossRefGoogle Scholar
  45. Muñoz, I., Stevanovic, J., Stanimirovic, Z., De la Rúa, P. (2012) Genetic variation of Apis mellifera from Serbia inferred from mitochondrial analysis. J. Apic. Sci. 56, 59–69Google Scholar
  46. Nabian, S., Ahmadi, K., Nazem Shirazi, M.H., Gerami Sadeghian, A. (2011) First detection of Nosema ceranae, a microsporidian protozoa of European honeybees (Apis mellifera) in Iran. Iran. J. Parasitol. 6, 89–95PubMedGoogle Scholar
  47. OIE (2008) Office International des Epizooties, Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, Chap. 2.2.4., Nosemosis of Honey Bees. http://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/2.02.04_NOSEMOSIS.pdf
  48. Paxton, R.J. (2010) Does infection by Nosema ceranae cause Colony Collapse Disorder in honey bees (Apis mellifera)? J. Apic. Res. 49, 80–84CrossRefGoogle Scholar
  49. 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, 558–565CrossRefGoogle Scholar
  50. RHMZ, Republic Hydrometeorological Sevice of Serbia. http://www.hidmet.gov.rs/index_eng.php
  51. Runckel, C., Flenniken, M.L., Engel, J.C., Ruby, J.G., Ganem, D., Andino, R., DeRisi, J.L. (2011) Temporal analysis of the honey bee microbiome reveals four novel viruses and seasonal prevalence of known viruses, Nosema, and Crithidia. PLoS One 6, e20656PubMedCrossRefGoogle Scholar
  52. Stanimirovic, Z., Pejovic, D., Stevanovic, J., Vucinic, M., Mirilovic, M. (2002) Investigations of hygienic behaviour and disease resistance in organic beekeeping of two honeybee ecogeographic varieties from Serbia. Acta Vet.-Beograd 52, 169–180CrossRefGoogle Scholar
  53. Stanimirovic, Z., Stevanovic, J., Andjelkovic, M. (2005a) Chromosomal diversity in Apis mellifera carnica from Serbia. Apidologie 36, 31–42CrossRefGoogle Scholar
  54. Stanimirovic, Z., Stevanovic, J., Cirkovic, D. (2005b) Behavioural defenses of the honey bee ecotype from Sjenica—Pester against Varroa destructor. Acta Vet.-Beograd 55, 69–82Google Scholar
  55. Stevanovic, J., Stanimirovic, Z., Radakovic, M., Kovacevic, R.S. (2010) Biogeographic study of the honey bee (Apis mellifera L.) from Serbia, Bosnia and Herzegovina and Republic of Macedonia based on mitochondrial DNA analyses. Russ. J. Genet. 46, 603–609CrossRefGoogle Scholar
  56. Stevanovic, J., Stanimirovic, Z., Genersch, E., Kovacevic, R.S., Ljubenkovic, J., Radakovic, M., Aleksic, N. (2011) Dominance of Nosema ceranae in honey bees in the Balkan countries in the absence of symptoms of colony collapse disorder. Apidologie 41, 49–58CrossRefGoogle Scholar
  57. 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, 383–388PubMedCrossRefGoogle Scholar
  58. Tlak Gajger, I.T., Vugrek, O., Grilec, D., Petrinec, Z. (2010) Prevalence and distribution of Nosema ceranae in Croatian honeybee colonies. Vet. Med.-Czech 55, 457–462Google Scholar
  59. Traver, B.E., Fell, R.D. (2011) Prevalence and infection intensity of Nosema in honey bee (Apis mellifera L.) colonies in Virginia. J. Invertebr. Pathol. 107, 43–49PubMedCrossRefGoogle Scholar
  60. Traver, B.E., Williams, M.R., Fell, R.D. (2012) Comparison of within hive sampling and seasonal activity of Nosema ceranae in honey bee colonies. J. Invertebr. Pathol. 109, 187–193PubMedCrossRefGoogle Scholar
  61. vanEngelsdorp, D., Evans, J.D., Saegerman, C., Mullin, C., Haubruge, E., et al. (2009) Colony collapse disorder: a descriptive study. PLoS One 4, e6481PubMedCrossRefGoogle Scholar
  62. Vejsnæs, F., Nielsen, S.L., Kryger, P. (2010) Factors involved in the recent increase in colony losses in Denmark. J. Apic. Res. 49, 109–110CrossRefGoogle Scholar
  63. Vidau, C., Diogon, M., Aufauvre, J., Fontbonne, R., Viguès, B., et al. (2011) Exposure to sublethal doses of fipronil and thiacloprid highly increases mortality of honeybees previously infected by Nosema ceranae. PLoS One 6, e21550PubMedCrossRefGoogle Scholar
  64. Williams, G.R., Shafer, A.B.A., Rogers, R.E.L., Shutler, D., Stewart, D.T. (2008) First detection of Nosema ceranae, a microsporidean parasite of European honey bees (Apis mellifera), in Canada and central USA. J. Invertebr. Pathol. 97, 189–192PubMedCrossRefGoogle Scholar
  65. Zander, E. (1909) Tierische Parasiten als Krankheitserreger bei der Biene. Münchener Bienenzeitung 31, 196–204Google Scholar

Copyright information

© INRA, DIB and Springer-Verlag France 2013

Authors and Affiliations

  • Jevrosima Stevanovic
    • 1
  • Predrag Simeunovic
    • 1
  • Bojan Gajic
    • 2
  • Nada Lakic
    • 3
  • Dejan Radovic
    • 4
  • Ingemar Fries
    • 5
  • Zoran Stanimirovic
    • 1
  1. 1.Department of Biology, Faculty of Veterinary MedicineUniversity of BelgradeBelgradeSerbia
  2. 2.Department of Parasitology, Faculty of Veterinary MedicineUniversity of BelgradeBelgradeSerbia
  3. 3.Department of Statistics, Faculty of AgricultureUniversity of BelgradeBelgrade-ZemunSerbia
  4. 4.Institute of Zoology, Faculty of BiologyUniversity of BelgradeBelgradeSerbia
  5. 5.Department of EcologySwedish University of Agricultural SciencesUppsalaSweden

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