Parasitology Research

, Volume 111, Issue 2, pp 601–607 | Cite as

Nosema ceranae an emergent pathogen of Apis mellifera in Chile

Original Paper


The microsporidian Nosema apis and Nosema ceranae have been associated with colony disorders of Apis mellifera and Apis cerana, respectively. N. apis is endemic in South America. Recently, N. ceranae has been detected in Brazil, Uruguay and Argentina. No report of its presence, distribution and prevalence in Chile is available. Here, we present a real-time PCR-based method that was able to discriminate between N. apis and N. ceranae. The dynamic range of this assay was 100 to 100,000 spores per honeybee. False-negative results were avoided due to the use of ACTIN gene as internal standard. False-positive results were obtained neither in experimentally nor in naturally contaminated samples. Using this method, we screened 240 beehives from the Chilean region where 42% of the total country honey production take places (Región del Biobío). Nosema spp. were detected in the four provinces and in 20 of the 26 communes of the region. Among the samples analysed, 49% were positive for N. ceranae. Their infection level ranged from 200 to more than 100,000 spores per honeybee. N. apis was not detected in this region. Hence, our data show that in Chile N. ceranae is an emergent pathogen that is been replacing N. apis. Also, they support that N. ceranae maybe the actual responsible for nosemosis in A. mellifera in South America.


  1. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410PubMedGoogle Scholar
  2. Anderson DL, Giacon H (1992) Reduced pollen collection by honey-bee (Hymenoptera, Apidae) colonies infected with Nosema apis and sacbrood virus. J Econ Entomol 85:47–51Google Scholar
  3. Botías C, Martín-Hernández R, Meana A, Higes M (2011) Critical aspects of the Nosema spp. diagnostic sampling in honeybee (Apis mellifera L) colonies. Parasitol Res. doi:10.1007/s00436-011-2760-2
  4. Bourgeois AL, Rinderer TE, Beaman LD, Danka RG (2010) Genetic detection and quantification of Nosema apis and N. ceranae in the honey bee. J Invertebr Pathol 103:53–58PubMedCrossRefGoogle Scholar
  5. Calderón RA, Sanchez LA, Yañez O, Fallas N (2008) Presence of Nosema ceranae in Africanized honey bee colonies in Costa Rica. J Apic Res 47:328–329CrossRefGoogle Scholar
  6. Cantwell GE (1970) Standard methods for counting nosema spores. Am Bee J 110:222–223Google Scholar
  7. Chaimanee V, Chen Y, Pettis JS, Scott CR, Chantawannakul P (2011) Phylogenetic analysis of Nosema ceranae isolated from European and Asian honeybees in Northern Thailand. J Invertebr Pathol 107:229–233PubMedCrossRefGoogle Scholar
  8. Chauzat MP, Higes M, Martín-Hernández R, Meana A, Cougoule N, Faucon JP (2007) Presence of Nosema ceranae in French honey bee colonies. J Apic Res 46:127–128CrossRefGoogle Scholar
  9. Chen Y, Evans JD, Smith IB, Pettis JS (2008) Nosema ceranae is a long-present and wide-spread microsporidian infection of the European honey bee (Apis mellifera) in the United States. J Invertebr Pathol 97:186–188PubMedCrossRefGoogle Scholar
  10. Chen YP, Evans JD, Murphy C, Guttel R, Zuker M, Gundensel-Rindal D, Pettis JS (2009) Morphological, molecular, and philogenetic charaterization of Nosema ceranae a microsporidian parasite isolated from the European honeybee, Apis mellifera. J Eukar Microbiol 56:142–147CrossRefGoogle Scholar
  11. de Graaf DC, Raes H, Sabbe G, De Rycke PH, Jacobs FJ (1994) Early development of Nosema apis (Microspora: Nosematidae) in the midgut epithelium of the honeybee (Apis mellifera). J Invertebr Pathol 63:74–81CrossRefGoogle Scholar
  12. Forsgren E, Fries I (2010) Comparative virulence of Nosema ceranae and Nosema apis in individual European honey bees. Vet Parasitol 170:212–217PubMedCrossRefGoogle Scholar
  13. Fries I (1988) Infectivity and multiplication of Nosema apis Z in the ventriculus of the honey bee. Apidologie 19:319–328CrossRefGoogle Scholar
  14. Fries I (1993) Nosema apis—a parasite in the honey bee colony. Bee World 74:5–19Google Scholar
  15. Fries I (2010) Nosema ceranae in European honey bees (Apis mellifera). J Invertebr Pathol 103(Suppl 1):S73–S79PubMedCrossRefGoogle Scholar
  16. Fries I, Feng F, da Silva A, Slemenda SB, Pieniazek NJ (1996) Nosema ceranae n.sp. (Microspora, Nosematidae), morphological and molecular characterization of a microsporidian parasite of the Asian honey bee (Apis cerana) (Himenoptera, Apidae). Eur J Protistol 32:356–365CrossRefGoogle Scholar
  17. Giersch T, Berg T, Galea F, Hornitzky M (2009) Nosema ceranae infects honey bees (Apis melifera) and contaminates honey in Australia. Apidologie 40:117–123CrossRefGoogle Scholar
  18. Guzman-Novoa E, Hamiduzzaman MM, Arechavaleta-Velasco ME, Koleoglu G, Valizadeh P, Correa-Benitez A (2011) Nosema ceranae has parazited Africanized honey bees in Mexico since at least 2004. J Apic Res 50:167–169CrossRefGoogle Scholar
  19. Hamiduzzaman MM, Guzman-Novoa E, Goodwin PH (2010) A multiplex PCR assay to diagnose and quantify Nosema infections in honey bees (Apis mellifera). J Invertebr Pathol 105:151–155PubMedCrossRefGoogle Scholar
  20. Higes M, Martín-Hernández R, Meana A (2006) Nosema ceranae, a new microsporidian parasite in honeybees in Europe. J Invertebr Pathol 92:93–95PubMedCrossRefGoogle Scholar
  21. Higes M, García-Palencia P, Martín-Hernández R, Meana A (2007) Experimental infection of Apis mellifera honeybees with Nosema ceranae (Microsporidia). J Invertebr Pathol 94:211–217PubMedCrossRefGoogle Scholar
  22. Higes M, Martín-Hernández R, Botias C, Bailon EG, Gonzalez-Porto AV, Barrios L, Del Nozal MJ, Bernal JL, Jimenez JJ, Garcia-Palencia P, Meana A (2008) How natural infection by Nosema ceranae causes honeybee colony collapse. Environ Microbiol 10:2659–2669PubMedCrossRefGoogle Scholar
  23. Higes M, Martín-Hernández R, Garrido-Bailón E, Botías C, Meana A (2009) First detection of Nosema ceranae (Microsporidia) in African Honey bees (Apis melifera intermissa). J Apic Res 48:217–219CrossRefGoogle Scholar
  24. Hinojosa A, González D (2004) Prevalencia de parásitos en Apis mellifera L en colmenares del secano costero e interior de la VI Región, Chile. Parasitol Latinoam 59:137–141CrossRefGoogle Scholar
  25. Hornitzky M (2008) Nosema disease. Literature review and three year survey of beekeepers, Part 2. RIRDC Publication 08/006. Rural Industries Research and Development Corporation, Kingstom, Australia. Available at Accessed 25 Jan 2012
  26. Huaiquil SE, Sepúlveda GF, Rebolledo RE (2009) Sanidad apícola en el valle de azapa, región de Arica y Parinacota, Chile. IDESIA (Chile) 27:71–78Google Scholar
  27. Huang WF, Bocquet M, Lee KC, Sung IH, Jiang JH, Chen YW, Wang CH (2008) The comparison of rDNA spacer regions of Nosema ceranae isolates from different hosts and locations. J Invertebr Pathol 97:9–13PubMedCrossRefGoogle Scholar
  28. Invernizzi C, Abud C, Tomasco IH, Harriet J, Ramallo G, Campa 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
  29. Klee J, Besana AM, Genersch E, Gisder S, Nanetti A, Tam DQ, Chinh TX, Puerta F, Ruz JM, Kryger P, Message D, Hatjina F, Korpela S, Fries I, Paxton RJ (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
  30. Martín-Hernández R, Meana A, Prieto L, Salvador AM, Garrido-Bailon E, Higes M (2007) Outcome of colonization of Apis mellifera by Nosema ceranae. Appl Environ Microbiol 73:6331–6338PubMedCrossRefGoogle Scholar
  31. Martín-Hernández R, Botias C, Barrios L, Martinez-Salvador A, Meana A, Mayack C, Higes M (2011a) Comparison of the energetic stress associated with experimental Nosema ceranae and Nosema apis infection of honeybees (Apis mellifera). Parasitol Res 109:605–612PubMedCrossRefGoogle Scholar
  32. Martín-Hernández R, Botias C, Bailon EG, Martinez-Salvador A, Prieto L, Meana A, Higes M (2011b) Microsporidia infecting Apis mellifera: coexistence or competition. Is Nosema ceranae replacing Nosema apis? Environ Microbiol. doi:10.1111/j.1462-2920.2011.02645.x
  33. Medici SK, Sarlo EG, Porrini MP, Braunstein M, Eguaras MJ (2011) Genetic variation and widespread dispersal of Nosema ceranae in Apis mellifera apiaries from Argentina. Parasitol Res. doi:10.1007/s00436-011-2566-2
  34. Oldroyd BP (2007) What’s killing American honey bees? PLoS Biol 5:1195–1199CrossRefGoogle Scholar
  35. Paxton RJ, 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
  36. Porrini MP, Audisio MC, Sabaté DC, Ibarguren C, Medici SK, Sarlo EG, Garrido PM, Eguaras MJ (2010) Effect of bacterial metabolites on microsporidian Nosema ceranae and on its host Apis mellifera. Parasitol Res 107:381–388PubMedCrossRefGoogle Scholar
  37. Ravikumar G, Raje Urs S, Vijaya Prakash NB, Rao CGP, Vardhana KV (2011) Development of a multiplex polymerase chain reaction for the simultaneous detection of microsporidians, nucleopolyhedrovirus, and densovirus infecting silkworms. J Invertebr Pathol 107:193–197PubMedCrossRefGoogle Scholar
  38. Ritter W (2001) Enfermedades de las abejas. In: Acribia SA (ed) Zaragoza, Spain, p 146Google Scholar
  39. Sambrook J, Russel DW (2001) Molecular cloning. A laboratory manual. Cold Springharbor Laboratory, Cold Spring HarborGoogle Scholar
  40. Soares MP, Elias-Neto M, Simoes ZL, Bitondi MM (2007) A cuticle protein gene in the honeybee: expression during development and in relation to the ecdysteroid titer. Insect Biochem Mol Biol 37:1272–1282PubMedCrossRefGoogle Scholar
  41. Stevanovic J, Stanimirovic Z, Genersch E, Kovacevic S, Ljubenkovic J, Radakovic M, Aleksic N (2010) Dominance of Nosema ceranae in honey bees in Balkan countries in the absence of symptoms of colony collapse disorder. Apidologie 42:49–58CrossRefGoogle Scholar
  42. Tapaszti Z, Forgach P, Kovago C, Bekesi L, Bakonyi T, Rusvai M (2009) First detection and dominance of Nosema ceranae in Hungarian honeybee colonies. Acta Vet Hung 57:383–388PubMedCrossRefGoogle Scholar
  43. Traver BE, Fell RD (2011) Prevalence and infection intensity of Nosema in honey bee (Apis mellifera L.) colonies in Virginia. J Invertebr Pathol 107:43–49PubMedCrossRefGoogle Scholar
  44. Weiss LM, Edlind TD, Vossbrinck CR, Hashimoto T (1999) Microsporidian molecular phylogeny: the fungal connection. J Eukariot Microbiol 46:17S–18SCrossRefGoogle Scholar
  45. Whitaker J, Szalanski A, Kence M (2010) Molecular detection of Nosema ceranae and N. apis from Turkish honey bees. Apidologie 42:174–180CrossRefGoogle Scholar
  46. Williams GR, Shafer AB, Rogers RE, Shutler D, Stewart DT (2008a) First detection of Nosema ceranae, a microsporidian parasite of European honey bees (Apis mellifera), in Canada and central USA. J Invertebr Pathol 97:189–192PubMedCrossRefGoogle Scholar
  47. Williams GR, Sampson MA, Shutler D, Rogers RE (2008b) Does fumagillin control the recently detected invasive parasite Nosema ceranae in Western honeybees (Apis mellifera)? J Invertbr Pathol 99:342–344CrossRefGoogle Scholar
  48. Yoshiyama M, Kimura K (2011) Distribution of Nosema ceranae in the European honeybee, Apis mellifera in Japan. J Invertebr Pathol 106:263–267PubMedCrossRefGoogle Scholar
  49. Zander E (1909) Tierische Parasiten als Krankenheitserreger bei der Biene. Munchener Bienenztg 31:196–204Google Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  1. 1.Instituto de CienciasFacultad de Medicina Clínica Alemana Universidad del DesarrolloSantiagoChile
  2. 2.SantiagoChile

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