Detection and Identification of a Novel Lactic Acid Bacterial Flora Within the Honey Stomach of the Honeybee Apis mellifera


This investigation concerned the question of whether honeybees collect bacteria that are beneficial for humans from the flowers that contribute to formation of their honey. Bacteria originating from the types of flowers involved, and found in different anatomic parts of the bees, in larvae, and in honey of different types, were sampled during a 2-year period. 16S rRNA sequencing of isolates and clones was employed. A novel bacterial flora composed of lactic acid bacteria (LAB) of the genera Lactobacillus and Bifidobacterium, which originated in the honey stomach of the honeybee, was discovered. It varied with the sources of nectar and the presence of other bacterial genera within the honeybee and ended up eventually in the honey. It appeared that honeybees and the novel LAB flora may have evolved in mutual dependence on one another. It was suggested that honey be considered a fermented food product because of the LAB involved in honey production. The findings are seen as having clear implications for future research in the area, as providing a better understanding the health of honeybees and of their production and storage of honey, and as having clear relevance for future honeybee and human probiotics.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3


  1. 1.

    Babendreier D, Joller D, Romeis J et al (2007) Bacterial community structures in honeybee intestines and their response to two insecticidal proteins. FEMS Microbiol Ecol 59:600–610

    PubMed  Article  CAS  Google Scholar 

  2. 2.

    Bae S, Fleet GH, Heard GM (2006) Lactic acid bacteria associated with wine grapes from several Australian vineyards. J Appl Microbiol 100:712–727

    PubMed  Article  CAS  Google Scholar 

  3. 3.

    Coenye T, Vandamme P (2003) Extracting phylogenetic information from whole-genome sequencing projects: the lactic bacteria as a test case. Microbiology 149:3507–3517

    PubMed  Article  CAS  Google Scholar 

  4. 4.

    Cox-Foster DL, Conlan S, Holmes EC et al (2007) A metagenomic survey of microbes in honey bee colony collapse disorder. Science 12:283–287

    Article  Google Scholar 

  5. 5.

    Edwards CG, Haag KM, Collins MD et al (1998) Lactobacillus kunkeei sp. nov.: a spoilage organism associated with grape juice fermentations. J Appl Microbiol 84:698–702

    PubMed  Article  CAS  Google Scholar 

  6. 6.

    Evans JD, Lopez DL (2004) Bacterial probiotics induce an immune response in the honey bee (Hymenoptera: Apidae). J Econ Entomol 97:752–756

    PubMed  Article  CAS  Google Scholar 

  7. 7.

    Evans JD, Armstrong TN (2006) Antagonistic interactions between honey bee bacterial symbionts and implications for disease. BMC Ecol 21:6:4

    Google Scholar 

  8. 8.

    Food and Agriculture Organisation/World Health Organisation. Joint FAO/WHO (2002) Expert consultation on evaluation of health and nutritional properties of probiotics in food. Available at: Accessed 15 July 2008

  9. 9.

    Genersch E, Ashiralieva A, Fries I (2005) Strain- and 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

    PubMed  Article  CAS  Google Scholar 

  10. 10.

    Huang YC, Edwards CG, Peterson JC et al (1996) Relationship between sluggish fermentations and the antagonism of yeast by lactic acid bacteria. Am J Enol Vitic 47:1–10

    CAS  Google Scholar 

  11. 11.

    Jeyaprakash A, Hoy MA, Allsopp MH (2003) Bacterial diversity in worker adults of Apis mellifera capensis and Apis mellifera scutellata (Insecta: Hymenoptera) assessed using 16S rRNA sequences. J Invertebr Pathol 84:96–103

    PubMed  Article  CAS  Google Scholar 

  12. 12.

    Jones JC, Myerscough MR, Graham S et al (2004) Honey bee nest thermoregulation: Diversity promotes stability. Science 16:402–404

    Article  Google Scholar 

  13. 13.

    Ludwig W, Strunk O, Klugbauer S et al (1998) Bacterial phylogeny based on comparative sequence analysis. Electrophoresis 19:554–568

    PubMed  Article  CAS  Google Scholar 

  14. 14.

    Lusby PE, Coombes AL, Wilkinson JM (2005) Bactericidal activity of different honeys against pathogenic bacteria. Arch Med Res 36:464–467

    PubMed  Article  CAS  Google Scholar 

  15. 15.

    Mato I, Huidobro JF, Simal-Lozano J et al (2006) Rapid determination of nonaromatic organic acids in honey by capillary zone electrophoresis with direct ultraviolet detection. J Agric Food Chem 54:1541–1550

    PubMed  Article  CAS  Google Scholar 

  16. 16.

    Van der Meulen R, Adriany T, Verbrugghe K et al (2006) Kinetic analysis of bifidobacterial metabolism reveals a minor role for succinic acid in the regeneration of NAD + through its growth-associated production. Appl Environ Microbiol 72:5204–5210

    PubMed  Article  Google Scholar 

  17. 17.

    Molan PC (2001) Honey as a topical antibacterial agent for treatment of infected wounds. World Wide Wounds. Available at: Accessed 15 July 2008

  18. 18.

    Olofsson TC, Ahrné S, Molin G (2007) The bacterial flora of vacuum-packed cold-smoked salmon stored at 7 degrees C, identified by direct 16S rRNA gene analysis and pure culture technique. J Appl Microbiol 103:109–119

    PubMed  Article  CAS  Google Scholar 

  19. 19.

    Ouwehand AC, Salminen S, Isolauri E (2002) Probiotics: an overview of beneficial effects. Antonie van Leeuwenhoek 82:279–289

    PubMed  Article  CAS  Google Scholar 

  20. 20.

    Reynaldi FJ, De Giusti MR, Alippi AM (2004) Inhibition of the growth of Ascosphaera apis by Bacillus and Paenibacillus strains isolated from honey. Rev Argent Microbiol 36:52–55

    PubMed  CAS  Google Scholar 

  21. 21.

    Scardovi V, Trovatelli LD (1969) New species of bifid bacteria from Apis mellifica L. and Apis indica F. A contribution to the taxonomy and biochemistry of the genus Bifidobacterium. Zentralbl Bakteriol Parasitenkd Infektionskr Hyg 123:64–88

    PubMed  CAS  Google Scholar 

  22. 22.

    Snowdon JA, Cliver (1996) Microorganism in honey. Int J Food Microbiol 3:1–26

  23. 23.

    Steinkraus KH (1995) Handbook of indigenous fermented foods. Dekker, New York, NY

    Google Scholar 

  24. 24.

    Taormina PJ, Niemira BA, Beuchat LR (2001) Inhibitory activity of honey against foodborne pathogens as influenced by the presence of hydrogen peroxide and level of antioxidant power. Int J Food Microbiol 69:217–225

    PubMed  Article  CAS  Google Scholar 

  25. 25.

    de Vuyst L, Vandamme E J (1994) Bacteriocins of lactic acid bacteria. Oxford, UK: Blackie Academic and Professional, Chapman & Hall

  26. 26.

    White JW, Subers MH, Schepartz A (1963) The identification of inhibine, the antibacterial factor in honey, as hydrogen peroxide and its origin in a honey glucose-oxidase system. Biochem Biophys Acta 73:57–70

    PubMed  Article  CAS  Google Scholar 

  27. 27.

    White JW, Doner LW (2006) Honey composition and properties. Available at: Accessed 15 July 2008

  28. 28.

    White R, Cooper R, Molan P (2005) Honey: a modern wound management product. Wounds UK, Aberdeen, UK

    Google Scholar 

  29. 29.

    Winston ML (1987) The biology of the honey bee. Harvard University Press, Cambridge, MA

    Google Scholar 

Download references


This study was financed by Gyllenstiernska Krapperupstiftelsen, Ekhagastiftelsen and Sparbankstiftelsen Skåne. We are grateful for the help of the beekeeper Tage Kimblad, late assistant Prof. Sten Ståhl and others for their knowledge and experience, and for their comments and reflections on our work.

Author information



Corresponding author

Correspondence to Tobias C. Olofsson.

Additional information

The laboratory work was partly performed at the Department of Food Technology, Engineering and Nutrition, Lund University, Lund, Sweden.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Olofsson, T.C., Vásquez, A. Detection and Identification of a Novel Lactic Acid Bacterial Flora Within the Honey Stomach of the Honeybee Apis mellifera . Curr Microbiol 57, 356–363 (2008).

Download citation


  • Lactobacillus
  • Lactic Acid Bacterium
  • Lactic Acid Bacterium Isolate
  • Nectar Sugar
  • Fermented Food Product