Apidologie

, 42:642 | Cite as

Detection and identification of Lactobacillus bacteria found in the honey stomach of the giant honeybee Apis dorsata

  • Naser Tajabadi
  • Makhdzir Mardan
  • Mohd Yazid Abdul Manap
  • Mustafa Shuhaimi
  • Amir Meimandipour
  • Leila Nateghi
Original article

Abstract

This is the first assay that describes the isolation and identification of strains and species of Lactobacillus from the honey stomach of the Asiatic giant honeybee, Apis dorsata. Samples of honeybees were collected from A. dorsata colonies in different bee trees, and Lactobacillus was isolated from honey stomachs using selective media. The isolates were Gram-stained and tested for catalase reaction. The 16S rRNA genes from extracted DNA of bacterial colonies were amplified with polymerase chain reaction using lactobacilli genus primers (27F and 1492R). All bacterial 16S rRNA genes were sequenced and deposited in GenBank. The 34 isolated strains yielded three distinct rRNA sequences of 15 different strains. Lactobacillus sequences isolated from the bees' honey stomachs were comprised of Lactobacillus kunkeei related-sequences (56%) with other abundant sequences being related to other Lactobacillus sp. (38%) and Lactobacillus vermiform (6%). These strains can be good candidates for potential application as probiotics in honeybees and also as natural food preservatives, which, in turn, may be useful in the food industry.

Keyword

Apis dorsata honey stomach Lactobacillus bacteria probiotics 

Notes

Acknowledgments

This study had funding support from the Food Biotechnology Research Laboratory (Professor Yazid Laboratory), Faculty of Food Sciences and Technology, Universiti Putra Malaysia. The authors would like to thank Mr. A. Javanmard, Mr. M.M. Saberioon (Department of Agriculture Technology), Dr. Andony Melathopoulos (Agriculture and Agri-food of Canada) and all the honey hunters in Kedah state of Malaysia for providing honeybee samples.

Détection et identification de bactéries Lactobacillus du jabot de l’abeille géante Apis dorsata

Apis dorsata / jabot / Lactobacillus / probiotiques

Nachweis und Identifizierung von Lactobacillus Bakterien im Honigmagen der Riesenhonigbiene Apis dorsata

Apis dorsata / honigmagen / Lactobacillus Bakterien / probiotische Lebensmittel

References

  1. Bae, S., Fleet, G.H., Heard, G.M. (2006) Lactic acid bacteria associated with wine grapes from several Australian vineyards. J. Appl. Microbiol. 100, 712–727PubMedCrossRefGoogle Scholar
  2. Coeuret, V., Dubernet, S., Bernardeau, M., Guegun, M., Vernouxj, P. (2003) Isolation, characterization and identification of lactobacilli focusing mainly on cheeses and other dairy products. Lait 83, 269–306CrossRefGoogle Scholar
  3. Du Plessis, H., Dicks, L., Lambrechts, M., Pretorius, I., Du Toit, M. (2004) Identification of lactic acid bacteria isolated from South African brandy base wines. Int. J. Food Microbiol. 91, 19–29PubMedCrossRefGoogle Scholar
  4. Edwards, C.G., Haag, K.M., Collins, M.D., Hutson, R.A., Huang, Y.C. (1998) Lactobacillus kunkeei sp. nov.: A spoilage organism associated with grape juice fermentations. J. Appl. Microbiol. 84, 698–702PubMedCrossRefGoogle Scholar
  5. Gilliam, M. (1997) Identification and roles of non-pathogenic microflora associated with honeybees. FEMS Microbiol. Lett. 155, 1–10CrossRefGoogle Scholar
  6. Gilliam, M., Morton, H.L., Prest, D.B., Martin, R.D., Wickerham, L.J. (1977) The mycoflora of adult worker honeybees, Apis mellifera: effects of 2, 4, 5-T and caging of bee colonies. J. Invertebr. Pathol. 30, 50–54PubMedCrossRefGoogle Scholar
  7. Huang, Y.C., Edwards, C.G., Peterson, J.C. (1996) Relationship between sluggish fermentations and the antagonism of yeast by lactic acid bacteria. Am. J. Enol. Vitic. 47, 1–10Google Scholar
  8. Jones, J.C., Myerscough, M.R., Graham, S. (2004) Honey bee nest thermoregulation: diversity promotes stability. Science 16, 402–404CrossRefGoogle Scholar
  9. Killer, J., Kopec, J., Mrazek, J., Rada, V., Dubna, S., Marounek, M. (2010) Bifidobacteria in the digestive tract of bumblebees. Anaerobe 16, 165–170PubMedCrossRefGoogle Scholar
  10. Lane, D.J. (1991) 16S/23S rRNA sequencing. In: Stackebrandt, E., Goodfellow, M. (eds.) Nucleic acid techniques in bacterial systematics, pp. 115–175. Wiley, NewYorkGoogle Scholar
  11. Lee, C.Y., Kime, R.W. (1984) The use of honey for clarifying apple juice. J. Apic. Res. 23, 45–49Google Scholar
  12. Lyapunov, Y.E., Kuzyaev, R.Z., Khismatullin, R.G., Bezgodova, O.A. (2008) Intestinal enterobacteria of the hibernating Apis mellifera mellifera L. bees. Microbiolgy 77, 373–379CrossRefGoogle Scholar
  13. Mardan, M., Kiew, R. (1985) Flowering periods of plants visited by honeybees in two areas of Malaysia, Proc. 3 Int. Conf Apic, Trop, Climates, Nairobi, 209–216.Google Scholar
  14. Mato, I., Huidobro, J.F., Simal-Lozano, J. (2006) Rapid determination of nonaromatic organic acids in honey by capillary zone electrophoresis with direct ultraviolet detection. J. Agric. Food Chem. 54, 1541–1550PubMedCrossRefGoogle Scholar
  15. Mrazek, J., Strosova, L., Fliegerova, K., Kott, T., Kopecny, J. (2008) Diversity of insect intestinal microflora. Folia Microbiol l53, 229–233CrossRefGoogle Scholar
  16. Olofsson, T.C., Vasquez, A. (2008) Detection and identification of a novel lactic acid bacterial flora within the honey stomach of the honeybee Apis mellifera. Curr. Microbiol. 57, 356–363PubMedCrossRefGoogle Scholar
  17. Olofsson, T.C., Vasquez, A. (2009) Phylogenetic comparison of bacteria isolated from the honey stomachs of honey bees Apis mellifera and bumble bees Bombus spp. J. Apic. Res. Bee World 48, 233–237CrossRefGoogle Scholar
  18. Ouwehand, A.C., Salminen, S., Isolauri, E. (2002) Probiotics: an overview of beneficial effects. Antonie Van Leeuwenhoek 82, 279–289PubMedCrossRefGoogle Scholar
  19. Rada, V., Máchová, M., Huk, J., Marounek, M., Dušková, D. (1997) Microflora in the honeybee digestive tract, counts, characteristics and sensitivity to veterinary drugs. Apidologie 28, 357–365CrossRefGoogle Scholar
  20. Reuter, G. (2001) The Lactobacillus and Bifidobacterium microflora of the human intestine: composition and succession. Curr. Issues Intest. Microbiol. 2, 43–53PubMedGoogle Scholar
  21. Shuhaimi, M. (2003) Species classification and molecular studies of bile salt hydrolase gene in Bifidobacterium spp, Doctor of philosophy, University Putra Malaysia, 1–174Google Scholar
  22. Snowdon, J.A., Cliver (1996) Microorganism in honey. Int. J. Food Microbiol. 3, 1–26CrossRefGoogle Scholar
  23. Sokal, R., Rohlf, F. (1995) Biometry. WH Freeman and Company, New YorkGoogle Scholar
  24. Spjut, R.H. (1985) Limitations of a random screen: search for new anticancer drugs in higher plants. Econ. Bot. 39, 266–288CrossRefGoogle Scholar
  25. Stackebrandt, E., Goebel, B.M. (1994) Taxonomic note: a place for DNA: DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int. J. Syst. Bacteriol. 44, 846–849CrossRefGoogle Scholar
  26. Steinkraus K.H. (1995) Handbook of indigenous fermented foods, Dekker New York NY.Google Scholar
  27. Tamura, K., Dudley, J., Nei, M., Kumar, S. (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol. Biol. Evol. 24, 1596–1599PubMedCrossRefGoogle Scholar
  28. Vasquez, A., Olofsson, T.C., Sammataro, D. (2009) A scientific note on the lactic acid bacterial flora in honeybees in the USA—a comparison with bees from Sweden. Apidologie 40, 26–28CrossRefGoogle Scholar
  29. Ward, L.J.H., Brown, J.C.S., Davey, G.P. (1994) Application of the ligase chain reaction to the detection of nisin A and nisin Z genes in Lactococcus lactis spp. FEMS Microbiol. Lett. 117, 29–34PubMedCrossRefGoogle Scholar
  30. Yoshiyama, M., Kimura, K. (2009) Bacteria in the gut of Japanese honeybee, Apis cerana japonica and their antagonistic effect against Paenibacillus larvae, the causal agent of American foulbrood. J. Invertebr. Pathol. 102, 91–96PubMedCrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Naser Tajabadi
    • 1
    • 4
  • Makhdzir Mardan
    • 1
  • Mohd Yazid Abdul Manap
    • 2
  • Mustafa Shuhaimi
    • 3
  • Amir Meimandipour
    • 5
  • Leila Nateghi
    • 6
  1. 1.Department of Agriculture Technology, Faculty of AgricultureUniversiti Putra MalaysiaSerdangMalaysia
  2. 2.Department of Food Technology, Faculty of Food Sciences and TechnologyUniversiti Putra MalaysiaSerdangMalaysia
  3. 3.Department of Microbiology, Faculty of Biotechnology and Biomolecular SciencesUniversiti Putra MalaysiaSerdangMalaysia
  4. 4.Department of Honey BeeAnimal Science Research Institute of IranKarajIran
  5. 5.Department of Animal and Marine BiotechnologyNational Institute of Genetic Eengineering and Biotechnology (NIGEB)TehranIran
  6. 6.Department of Food Science and TechnologyVaramin-Pishva Branch, Islamic Azad UniversityVaraminIran

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