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Volume 66, Issue 9


Identification and determination of relatedness of lactobacilli using different DNA amplification methods

Kristýna Turková
  • Institute of Food Science and Biotechnology, Faculty of Chemistry, Brno University of Technology, Purkyňova 118, CZ-612 00, Brno, Czech Republic
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/ Bohuslav Rittich
  • Institute of Food Science and Biotechnology, Faculty of Chemistry, Brno University of Technology, Purkyňova 118, CZ-612 00, Brno, Czech Republic
  • Department of Experimental Biology, Faculty of Science, Masaryk University, Tvrdého 14, CZ-602 00, Brno, Czech Republic
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/ Alena Španová
  • Institute of Food Science and Biotechnology, Faculty of Chemistry, Brno University of Technology, Purkyňova 118, CZ-612 00, Brno, Czech Republic
  • Department of Experimental Biology, Faculty of Science, Masaryk University, Tvrdého 14, CZ-602 00, Brno, Czech Republic
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Published Online: 2012-06-22 | DOI: https://doi.org/10.2478/s11696-012-0206-7


Several DNA amplification-based methods were used for identification and evaluation of the relation between lactobacilli isolated from breastfed full-term infant faeces (31 strains), dairy products (5 strains) and silage (1 strain). Twenty-seven strains isolated from infant faeces were identified as Lactobacillus rhamnosus (9), Lactobacillus gasseri (6), Lactobacillus paracasei (4), Lactobacillus fermentum (4), Lactobacillus salivarius (2), Lactobacillus plantarum (1), and Lactobacillus helveticus (1) using 10 species-specific polymerase chain reactions (PCRs), multiplex PCR for the Lactobacillus casei group, and sequencing of 16S rDNA. Four strains were not identified. Six strains isolated from dairy products and silage were identified as Lactobacillus rhamnosus. A repetitive extragenic palindromic polymerase chain reaction (rep-PCR) with primer (GTG)5 and a randomly amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) with primer M13 were used for confirmation of species identification. Fingerprints were used for evaluation of the relatedness of lactobacilli. Differences between strains from infant faeces, dairy products, and silage were not detected.

Keywords: Lactobacillus; breastfed infants; dairy products; identification; PCR; fingerprinting

  • [1] Ben Amor, K., Vaughan, E. E., & de Vos, W. M. (2007). Advanced molecular tools for the identification of lactic acid bacteria. The Journal of Nutrition, 137, 741S–747S. Google Scholar

  • [2] Bernardeau, M., Guguen, M., & Vernoux, J. P. (2006). Beneficial lactobacilli food and feed: long-term use, biodiversity and proposals for specific and realistic safety assessments. FEMS Microbiology Reviews, 30, 487–513. DOI: 10.1111/j.1574-6976.2006.00020.x. http://dx.doi.org/10.1111/j.1574-6976.2006.00020.xCrossrefGoogle Scholar

  • [3] Byun, R., Nadkarni, M. A., Chhour, K. L., Martin, F. E., Jacques, N. A., & Hunter, N. (2004). Quantitative analysis of diverse Lactobacillus species present in advanced dental caries. Journal of Clinical Microbiology, 42, 3128–3136. DOI: 10.1128/jcm.42.7.3128-3136.2004. http://dx.doi.org/10.1128/JCM.42.7.3128-3136.2004CrossrefGoogle Scholar

  • [4] Dellaglio, F., Dicks, L. M. T., Du Toit, M., & Torriani, S. (1991). Designation of ATCC 334 in place of ATCC 393 (NCDO 161) as the neotype strain of Lactobacillus casei subsp. casei and rejection of the name Lactobacillus paracasei (Collins et al., 1989). International Journal of Systematic and Evolutionary Microbiology, 41, 340–342. DOI: 10.1099/00207713-41-2-340. CrossrefGoogle Scholar

  • [5] Dellaglio, F., Felia, G. E., & Torriani, S. (2002). The status of the species Lactobacillus casei (Orla-Jensen 1916) Hansen and Lessel 1971 and Lactobacillus paracasei Collins et al. 1989. International Journal of Systematic and Evolutionary Microbiology, 52, 285–287. CrossrefGoogle Scholar

  • [6] Desai, A. R., Shah, N. P., & Powell, I. B. (2006). Discrimination of dairy industry isolates of the Lactobacillus casei group. Journal of Dairy Science, 89, 3345–3351. DOI: 10.3168/jds.S0022-0302(06)72371-2. http://dx.doi.org/10.3168/jds.S0022-0302(06)72371-2CrossrefGoogle Scholar

  • [7] Dicks, L. M. T., Du Plessis, E. M., Dellaglio, F., & Lauer, E. (1996). Reclassification of Lactobacillus casei subsp. casei ATCC 393 and Lactobacillus rhamnosus ATCC 15820 as Lactobacillus zeae nom. rev., designation of ATCC 334 as the neotype of L. casei subsp. casei, and rejection of the name Lactobacillus paracasei. International Journal of Systematic and Evolutionary Microbiology, 46, 337–340. DOI: 10.1099/00207713-46-1-337. CrossrefGoogle Scholar

  • [8] Dubernet, S., Desmasures, N., & Guéguen, M. (2002). A PCR-based method for identification of lactobacilli at the genus level. FEMS Microbiological Letters, 214, 271–275. DOI: 10.1016/s0378-1097(02)00895-9. http://dx.doi.org/10.1111/j.1574-6968.2002.tb11358.xCrossrefGoogle Scholar

  • [9] Ehrmann, M. A., & Vogel, R. F. (2005). Molecular taxonomy and genetics of sourdough lactic acid bacteria. Trends in Food Science & Technology, 16, 31–42. DOI: 10.1016/j.tifs.2004.06.004. http://dx.doi.org/10.1016/j.tifs.2004.06.004CrossrefGoogle Scholar

  • [10] Ferchichi, M., Valcheva, R., Prévost, H., Onno, B., & Dousset, X. (2008). A one-step reaction for the rapid identification of Lactobacillus mindensis, Lactobacillus panis, Lactobacillus paralimentarius, Lactobacillus pontis and Lactobacillus frumenti using oligonucleotide primers designed from the 16S-23S rRNA intergenic sequences. Journal of Applied Microbiology, 104, 1797–1807. DOI: 10.1111/j.1365- 2672.2007.03712.x. http://dx.doi.org/10.1111/j.1365-2672.2007.03712.xCrossrefGoogle Scholar

  • [11] Gevers, D., Huys, G., & Swings, J. (2001). Applicability of rep-PCR fingerprinting for identification of Lactobacillus species. FEMS Microbiology Letters, 205, 31–36. DOI: 10.1016/s0378-1097(01)00439-6. http://dx.doi.org/10.1111/j.1574-6968.2001.tb10921.xCrossrefGoogle Scholar

  • [12] Klein, G. (2007). International committee on systematics of prokaryotes; Subcommittee on the taxonomy of Bifidobacterium, Lactobacillus and related organisms. International Journal of Systematic and Evolutionary Microbiology, 57, 1367–1369. DOI: 10.1099/ijs.0.65143-0. http://dx.doi.org/10.1099/ijs.0.65143-0CrossrefGoogle Scholar

  • [13] Li, Y., & Nishino, N. (2011). Bacterial and fungal communities of wilted Italian ryegrass silage inoculated with and without Lactobacillus rhamnosus or Lactobacillus buchneri. Letters in Applied Microbiology, 52, 314–321. DOI: 10.1111/j.1472-765x.2010.03000.x. http://dx.doi.org/10.1111/j.1472-765X.2010.03000.xWeb of ScienceCrossrefGoogle Scholar

  • [14] Mitsou, E. K., Kirtzalidou, E., Oikonomou, I., Liosis, G., & Kyriacou, A. (2008). Fecal microflora of Greek healthy neonates. Anaerobe, 14, 94–101. DOI: 10.1016/j.anaerobe.2007.11.002. http://dx.doi.org/10.1016/j.anaerobe.2007.11.002Web of ScienceCrossrefGoogle Scholar

  • [15] Rossetti, L., & Giraffa, G. (2005). Rapid identification of dairy lactic acid bacteria by M13-generated, RAPD-PCR fingerprint databases. Journal of Microbiological Methods, 63, 135–144. DOI: 10.1016/j.mimet.2005.03.001. http://dx.doi.org/10.1016/j.mimet.2005.03.001CrossrefGoogle Scholar

  • [16] Sambrook, J., & Russel, D. W. (2001). Molecular cloning: A laboratory manual (II) (3rd ed.). New York, NY, USA: Cold Spring Laboratory Harbor Press. Google Scholar

  • [17] Sisto, A., De Bellis, P., Visconti, A., Morelli, L., & Lavermicocca, P. (2009). Development of a PCR assay for the strain-specific identification of probiotic strain Lactobacillus paracasei IMPC2.1. International Journal of Food Microbiology, 136, 59–65. DOI: 10.1016/j.ijfoodmicro.2009.09.017. http://dx.doi.org/10.1016/j.ijfoodmicro.2009.09.017CrossrefGoogle Scholar

  • [18] Štšepetova, J., Sepp, E., Kolk, H., Löivukene, K., Songisepp, E., & Mikelsaar, M. (2011). Diversity and metabolic impact of intestinal Lactobacillus species in healthy adults and the elderly. British Journal of Nutrition, 105, 1235–1244. DOI: 10.1017/s0007114510004770. http://dx.doi.org/10.1017/S0007114510004770CrossrefWeb of ScienceGoogle Scholar

  • [19] Švec, P., Kukletová, M., & Sedláček, I. (2010). Comparative evaluation of automated ribotyping and RAPD-PCR for typing of Lactobacillus spp. occurring in dental carries. Antonie van Leeuwenhoek, 98, 85–92. DOI: 10.1007/s10482-010-9432- 6. http://dx.doi.org/10.1007/s10482-010-9432-6CrossrefGoogle Scholar

  • [20] Verdenelli, M. C., Ghelfi, F., Silvi, S., Orpianes, C., Cecchini, C., & Cresci, A. (2009). Probiotic properties of Lactobacillus rhamnosus and Lactobacillus paracasei isolated from human faeces. European Journal of Nutrition, 48, 355–363. DOI: 10.1007/s00394-009-0021-2. http://dx.doi.org/10.1007/s00394-009-0021-2CrossrefWeb of ScienceGoogle Scholar

  • [21] Wall, R., Fitzgerald, G., Hussey, S., Ryan, T., Murphy, B., Ross, P., & Stanton, C. (2007). Genomic diversity of cultivable Lactobacillus populations residing in the neonatal adult gastrointestinal tract. FEMS Microbiology Ecology, 59, 127–137. DOI: 10.1111/j.1574-6941.2006.00202.x. http://dx.doi.org/10.1111/j.1574-6941.2006.00202.xWeb of ScienceCrossrefGoogle Scholar

  • [22] Walter, J., Tannock, G. W., Tilsala-Timisjarvi, A., Rodtong, S., Loach, D. M., Munro, K., & Alatossava, T. (2000). Detection and identification of gastrointestinal Lactobacillus species by using denaturing gradient gel electrophoresis and speciesspecific PCR primers. Applied and Environmental Microbiology, 66, 297–303. DOI: 10.1128/aem.66.1.297-303.2000. http://dx.doi.org/10.1128/AEM.66.1.297-303.2000CrossrefGoogle Scholar

  • [23] Ward, L. J. H., & Timmins, M. J. (1999). Differentiation of Lactobacillus casei, Lactobacillus paracasei and Lactobacillus rhamnosus by polymerase chain reaction. Letters in Applied Microbiology, 29, 90–92. DOI: 10.1046/j.1365-2672.1999.00586.x. http://dx.doi.org/10.1046/j.1365-2672.1999.00586.xCrossrefGoogle Scholar

About the article

Published Online: 2012-06-22

Published in Print: 2012-09-01

Citation Information: Chemical Papers, Volume 66, Issue 9, Pages 842–851, ISSN (Online) 1336-9075, DOI: https://doi.org/10.2478/s11696-012-0206-7.

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© 2012 Institute of Chemistry, Slovak Academy of Sciences.

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