Novel lactic acid bacteria isolated from the bumble bee gut: Convivina intestini gen. nov., sp. nov., Lactobacillus bombicola sp. nov., and Weissella bombi sp. nov.
- 874 Downloads
Twelve isolates of lactic acid bacteria (LAB) were obtained in the course of a bumble bee gut microbiota study and grouped into four matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry clusters. Comparative 16S rRNA gene sequence analysis revealed that cluster 1 isolates, represented by strain LMG 28288T, are most closely related to Lactobacillus apis (97.0 % sequence similarity to that of L. apis LMG 26964T). Cluster 2 isolates represented by strain LMG 28290T are most closely related to Weissella hellenica (99.6 % sequence similarity to that of W. hellenica LMG 15125T). The single cluster 3 and 4 isolates had identical 16S rRNA gene sequences which were 94.8 % similar to that of Leuconostoc mesenteroides subsp. mesenteroides LMG 6893T, their nearest phylogenetic neighbour. A polyphasic taxonomic study additionally including comparative pheS sequence analysis, DNA–DNA hybridization experiments, DNA G+C content analysis, (GTG)5-PCR fingerprinting and a biochemical characterization, demonstrated that cluster 1 isolates represent a novel Lactobacillus species for which we propose the name Lactobacillus bombicola sp. nov. with LMG 28288T (= DSM 28793T) as the type strain; and that cluster 2 isolates represent a novel Weissella species for which we propose the name Weissella bombi sp. nov. with LMG 28290T (= DSM 28794T) as the type strain. Cluster 3 and 4 isolates, in contrast, represented a very distinct, novel taxon that could be distinguished from members of the genera Leuconostoc and Fructobacillus, its nearest phylogenetic neighbours, by its cellular morphology, non-fructophilic metabolism and DNA G+C content. We therefore classify both isolates into a novel species representing a novel LAB genus for which the name Convivina intestini gen. nov., sp. nov. is proposed with LMG 28291T (= DSM 28795T) as the type strain.
KeywordsLactic acid bacteria Lactobacillus Weissella Leuconostocaceae Bumble bee Gut microbiota
The authors acknowledge support by Ghent University-Special Research Fund (BOF).
Compliance with Ethical Standards
The present research involved sampling of bumble bees for which no permission was required as bumble bees are not included in the “Decree of Species (het Soortenbesluit (http://codex.vlaanderen.be/Zoeken/Document.aspx?DID=1018227¶m=informatie])” of the Flemish government with inception on 01/09/2009. The authors do not have a conflict of interest.
- De Baere S, Eeckhaut V, Steppe M, De Maesschalck C, De Backer P, Van Immerseel F, Croubels S (2013) Development of a HPLC-UV method for the quantitative determination of four short-chain fatty acids and lactic acid produced by intestinal bacteria during in vitro fermentation. J Pharm Biomed Anal 80:107–115. doi: 10.1016/j.jpba.2013.02.032 CrossRefPubMedGoogle Scholar
- De Bruyne K, Schillinger U, Caroline L, Boehringer B, Cleenwerck I, Vancanneyt M, Vandamme P (2007) Leuconostoc holzapfelii sp. nov., isolated from Ethiopian coffee fermentation and assessment of sequence analysis of housekeeping genes for delineation of Leuconostoc species. Int J Syst Evol Microbiol 57(Pt 12):2952–2959. doi: 10.1099/ijs.0.65292-0 CrossRefPubMedGoogle Scholar
- De Vuyst L, Camu N, De Winter T, Vandemeulebroecke K, Van de Perre V, Vancanneyt M, 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(1):79–90. doi: 10.1016/j.ijfoodmicro.2007.02.030 CrossRefPubMedGoogle Scholar
- Endo A, Okada S (2008) Reclassification of the genus Leuconostoc and proposals of Fructobacillus fructosus gen. nov., comb. nov., Fructobacillus durionis comb. nov., Fructobacillus ficulneus comb. nov. and Fructobacillus pseudoficulneus comb. nov. Int J Syst Evol Microbiol 58(Pt 9):2195–2205. doi: 10.1099/ijs.0.65609-0 CrossRefPubMedGoogle Scholar
- MacFaddin JF (1980) Biochemical tests for identification of medical bacteria. Lippincott Williams & Wilkins, PhiladelphiaGoogle Scholar
- Nei M, Kumar S (2000) Molecular evolution and phylogenetics. Oxford University Press, New YorkGoogle Scholar
- Olofsson TC, Alsterfjord M, Nilson B, Butler E, Vásquez A (2014) Lactobacillus apinorum sp. nov., Lactobacillus mellifer sp. nov., Lactobacillus mellis sp. nov., Lactobacillus melliventris sp. nov., Lactobacillus kimbladii sp. nov., Lactobacillus helsingborgensis sp. nov. and Lactobacillus kullabergensis sp. nov., isol. Int J Syst Evol Microbiol 64(Pt 9):3109–3119. doi: 10.1099/ijs.0.059600-0 CrossRefPubMedCentralPubMedGoogle Scholar
- Rasmont P, Pauly A, Terzo M, Patiny S, Michez D, Iserbyt S, Haubruge E (2005) The survey of wild bees (Hymenoptera, Apoidea) in Belgium and France. Retrieved from www.atlashymenoptera.net. Accessed 27 Dec 2013
- Stackebrandt E, Smith D, Casaregola S, Varese GC, Verkleij G, Lima N, Bridge P (2014) Deposit of microbial strains in public service collections as part of the publication process to underpin good practice in science. SpringerPlus 3(1):208. doi: 10.1186/2193-1801-3-208 CrossRefPubMedCentralPubMedGoogle Scholar
- Vos P, Garrity G, Jones D, Krieg NR, Ludwig W, Rainey FA, Schleifer K-H, Whitman W (Eds)(2009). Bergey’s manual of dystematic bacteriology Volume 3: The firmicutes. doi: 10.1007/978-0-387-68489-5
- Williams PH (1982) The distribution and decline of British bumble bees (Bombus Latr.). J Apic Res 21(4):236–245Google Scholar