Two Gram-positive bacterial strains, LMG 24284 and LMG 24285, were isolated from different spontaneous cocoa bean heap fermentations in Ghana. Analysis of their 16S rRNA gene sequences indicated that they were members of the and species groups, respectively. DNA–DNA hybridization experiments with their nearest phylogenetic neighbours demonstrated that both strains represented novel species that could be differentiated from their nearest neighbours by sequence analysis, whole-cell protein electrophoresis, fluorescent amplified fragment length polymorphism analysis and biochemical characterization. Therefore, two novel species are proposed, sp. nov. (type strain LMG 24284 =DSM 21115) and sp. nov. (type strain LMG 24285 =DSM 21116).


Article metrics loading...

Loading full text...

Full text loading...



  1. Bringel, F., Castioni, A., Olukoya, D. K., Felis, G. E., Torriani, S. & Dellaglio, F.(2005).Lactobacillus plantarum subsp. argentoratensis subsp. nov., isolated from vegetable matrices. Int J Syst Evol Microbiol 55, 1629–1634.[CrossRef] [Google Scholar]
  2. Camu, N., De Winter, T., Verbrugghe, K., Cleenwerck, I., Vandamme, P., Takrama, J. S., Vancanneyt, M. & De Vuyst, L.(2007). Dynamics and biodiversity of populations of lactic acid bacteria and acetic acid bacteria involved in spontaneous heap fermentation of cocoa beans in Ghana. Appl Environ Microbiol 73, 1809–1824.[CrossRef] [Google Scholar]
  3. Camu, N., González, Á., De Winter, T., Van Schoor, A., De Bruyne, K., Vandamme, P., Takrama, J. S., Addo, S. K. & De Vuyst, L.(2008). Influence of turning and environmental contamination on the dynamics of populations of lactic acid and acetic acid bacteria involved in spontaneous cocoa bean heap fermentation in Ghana. Appl Environ Microbiol 74, 86–98.[CrossRef] [Google Scholar]
  4. Carr, J. G. & Davies, P. A.(1970). Homofermentative lactobacilli of ciders including Lactobacillus mali nov. spec. J Appl Bacteriol 33, 768–774.[CrossRef] [Google Scholar]
  5. Carr, J. G., Davies, P. A. & Dougan, J.(1979).Cocoa fermentation in Ghana and Malaysia I. Chatham, UK: Natural Resources Institute.
  6. Curk, M. C., Hubert, J. C. & Bringel, F.(1996).Lactobacillus paraplantarum sp. nov., a new species related to Lactobacillus plantarum. Int J Syst Bacteriol 46, 595–598.[CrossRef] [Google Scholar]
  7. De Bruyne, K., Camu, N., Lefebvre, K., De Vuyst, L. & Vandamme, P.(2008).Weissella ghanensis sp. nov., isolated from a Ghanaian cocoa fermentation. Int J Syst Evol Microbiol 58, 2721–2725.[CrossRef] [Google Scholar]
  8. De Vuyst, L., Camu, N., De Winter, T., Vandemeulebroecke, K., Van de Perre, V., Vancanneyt, M., De Vos, P. & Cleenwerck, I.(2008). Validation of the (GTG)5-rep-PCR fingerprinting technique for rapid classification and identification of acetic acid bacteria, with a focus on isolates from Ghanaian fermented cocoa beans. Int J Food Microbiol 125, 79–90.[CrossRef] [Google Scholar]
  9. Ezaki, T., Hashimoto, Y. & Yabuuchi, E.(1989). Fluorometric deoxyribonucleic acid-deoxyribonucleic acid hybridization in microdilution wells as an alternative to membrane filter hybridization in which radioisotopes are used to determine genetic relatedness among bacterial strains. Int J Syst Bacteriol 39, 224–229.[CrossRef] [Google Scholar]
  10. Franz, C. M. A. P., Vancanneyt, M., Vandemeulebroecke, K., De Wachter, M., Cleenwerck, I., Hoste, B., Schillinger, U., Holzapfel, W. H. & Swings, J.(2006).Pediococcus stilesii sp. nov., isolated from maize grains. Int J Syst Evol Microbiol 56, 329–333.[CrossRef] [Google Scholar]
  11. Gevers, D., Huys, G. & Swings, J.(2001). Applicability of rep-PCR fingerprinting for identification of Lactobacillus species. FEMS Microbiol Lett 205, 31–36.[CrossRef] [Google Scholar]
  12. Goris, J., Suzuki, K., De Vos, P., Nakase, T. & Kersters, K.(1998). Evaluation of a microplate DNA-DNA hybridization method compared with the initial renaturation method. Can J Microbiol 44, 1148–1153.[CrossRef] [Google Scholar]
  13. Hansen, C. E., del Olmo, M. & Burri, C.(1998). Enzyme activities in cocoa beans during fermentation. J Sci Food Agric 77, 273–281.[CrossRef] [Google Scholar]
  14. Kato, Y., Sakala, R. M., Hayashidani, H., Kiuchi, A., Kaneuchi, C. & Ogawa, M.(2000).Lactobacillus algidus sp. nov., a psychrophilic lactic acid bacterium isolated from vacuum-packaged refrigerated beef. Int J Syst Evol Microbiol 50, 1143–1149.[CrossRef] [Google Scholar]
  15. Lagunes Gálvez, S., Loiseau, G., Paredes, J. L., Barel, M. & Guiraud, J.-P.(2007). Study on the microflora and biochemistry of cocoa fermentation in the Dominican Republic. Int J Food Microbiol 114, 124–130.[CrossRef] [Google Scholar]
  16. Mesbah, M., Premachandran, U. & Whitman, W. B.(1989). Precise measurement of the G+C content of deoxyribonucleic acid by high-performance liquid chromatography. Int J Syst Bacteriol 39, 159–167.[CrossRef] [Google Scholar]
  17. Meyer, C. P. & Paulay, G.(2005). DNA barcoding: error rates based on comprehensive sampling. PLoS Biol 3, e422[CrossRef] [Google Scholar]
  18. Naser, S. M., Dawyndt, P., Hoste, B., Gevers, D., Vandemeulebroecke, K., Cleenwerck, I., Vancanneyt, M. & Swings, J.(2007). Identification of lactobacilli by pheS and rpoA gene sequence analyses. Int J Syst Evol Microbiol 57, 2777–2789.[CrossRef] [Google Scholar]
  19. Nielsen, D. S., Teniola, O. D., Ban-Koffi, L., Owusu, M., Andersson, T. S. & Holzapfel, W. H.(2007). The microbiology of Ghanaian cocoa fermentations analysed using culture-dependent and culture-independent methods. Int J Food Microbiol 114, 168–186.[CrossRef] [Google Scholar]
  20. Passos, F. M. L., Lopez, A. S. & Silva, D. O.(1984a). Aeration and its influence on the microbial sequence in cacao fermentations in Bahia, with emphasis on lactic acid bacteria. J Food Sci 49, 1470–1474.[CrossRef] [Google Scholar]
  21. Passos, F. M. L., Silva, D. O., Lopez, A., Ferreira, C. L. L. F. & Guimarães, W. V.(1984b). Characterization and distribution of lactic acid bacteria from traditional cocoa bean fermentations in Bahia. J Food Sci 49, 205–208.[CrossRef] [Google Scholar]
  22. Pot, B., Vandamme, P. & Kersters, K.(1994). Analysis of electrophoretic whole-organism protein fingerprints. In Chemical Methods in Prokaryotic Systematics, pp. 493–521. Edited by M. Goodfellow & A. G. O'Donnell. Chichester: Wiley.
  23. Schwan, R. F. & Wheals, A. E.(2004). The microbiology of cocoa fermentation and its role in chocolate quality. Crit Rev Food Sci Nutr 44, 205–221.[CrossRef] [Google Scholar]
  24. Stackebrandt, E. & Kandler, O.(1979). Taxonomy of the genus Cellulomonas, based on phenotypic characters and deoxyribonucleic acid-deoxyribonucleic acid homology, and proposal of seven neotype strains. Int J Syst Bacteriol 29, 273–282.[CrossRef] [Google Scholar]
  25. Thompson, S. S., Miller, K. B. & Lopez, A. S.(2001). Cocoa and coffee. In Food Microbiology: Fundamentals and Frontiers, 2nd edn, pp. 721–733. Edited by M. P. Doyle, L. R. Beuchat & T. J. Montville. Washington, DC: American Society for Microbiology.
  26. Zanoni, P., Farrow, J. A. E., Philips, B. A. & Collins, M. D.(1987).Lactobacillus pentosus (Fred, Peterson, and Anderson) sp. nov., nom. rev. Int J Syst Bacteriol 37, 339–341.[CrossRef] [Google Scholar]

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error