Czech J. Anim. Sci., 2009, 54(7):307-314 | DOI: 10.17221/1722-CJAS

Differentiation of porcine wild-type lactobacilli strains, with ERIC-PCR and PFGE band patterns included in polyphasic taxonomy

E. Bolado-Martínez, E. Acedo-Félix
Centro de Investigación en Alimentación y Desarrollo, A.C. Carretera a la Victoria, Hermosillo, Sonora, México

Probiotic administration is an alternative to antibiotic supplementation in pig management. However, potential probiotic microorganisms included in foods must be well defined and correctly named, on the basis of a valid taxonomic system. The aim of the present study was to analyze band patterns generated by the polymerase chain reaction (PCR) amplification of enterobacterial repetitive intergenic consensus sequences (ERIC-PCR), obtained from porcine wild-type Lactobacillus strains, as a rapid alternative for genotypic characterization in polyphasic taxonomy. In the present study 36 porcine wild-type Lactobacillus strains were analyzed by polyphasic taxonomy which included API 50 CHL system, ERIC-PCR, and pulsed field gel electrophoresis (PFGE) band analysis, after digestion with XbaI or SpeI restriction enzymes. Polyphasic taxonomy discriminated among 23 strains of Lactobacillus reuteri, 12 strains of Lactobacillus salivarius and one strain of Lactobacillus mucosae. None of the tested methods was able to reliably resolve the three selected species of lactobacilli at a strain level. However, results improved considerably when ERIC-PCR results were combined with phenotypic characterization, and those results were comparable to the taxonomy that included PFGE.

Keywords: Lactobacillus; porcine lactobacilli phenotypic characterization; genotypic characterization

Published: July 31, 2009  Show citation

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Bolado-Martínez E, Acedo-Félix E. Differentiation of porcine wild-type lactobacilli strains, with ERIC-PCR and PFGE band patterns included in polyphasic taxonomy. Czech J. Anim. Sci. 2009;54(7):307-314. doi: 10.17221/1722-CJAS.
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    References

    1. Abs El-Osta Y., Hillier A.J., Davidson B.E., Dobos M. (2002): Pulsed-field gel electrophoretic analysis of the genome of Lactobacillus gasseri ATCC33323, and construction of a physical map. Electrophoresis, 23, 3321-3331. Go to original source... Go to PubMed...
    2. Bernardeau M., Vernoux J.P., Henri-Dubernet S., Guéguen M. (2007): The Lactobacillus genus. International Journal of Food Microbiology, doi:10.1016/j.ijfoodmicro.2007.08.015. Go to original source... Go to PubMed...
    3. Darbre P.D. (1999): Preparation of DNA. In: Darbre P.D. (ed.): Basic Molecular Biology. Essential Techniques. John Wiley and Sons, London, UK, 15-27.
    4. De Angelis M., Siragusa S., Berloco M. (2006): Selection of potential probiotic lactobacilli from pig feces to be used as additives in pelleted feeding. Research in Microbiology, 157, 792-801. Go to original source... Go to PubMed...
    5. Delfederico L., Hollman A., Martínez M. (2006): Molecular identification and typing of lactobacilli isolated from kefir grains. Journal of Dairy Research, 73, 20-27. Go to original source... Go to PubMed...
    6. Huaynate R.A.R., Thomaz M.C., Kronka R.N. (2006): Effect of adding macro and micro minerals in pig faeces fed diets with different levels of probiotic. Brazilian Archives of Biology and Technology, 49, 385-392. Go to original source...
    7. Kandler O., Weiss N. (1986). Regular, Nonsporing, GramPositive Rods. In: Bergey's Manual of Systematic Bacteriology (eds.): Sneath P.H.A., Mair N.S., Sharpe M. E., Holt J.G.. Williams and Wilkins, Baltimore, USA, 2, 1209-1234.
    8. Maxwell F.J., Stewart C.S. (1995). The microbiology of the gut and the role of probiotics. En Varley M.A. (ed.): The Neonatal Pig. Development and Survival. Cabi Publishing, Wallingford, UK, 155-186.
    9. Mikkelsen L.L., Bendixen C., Jakobsen M., Jensen B.B. (2003): Enumeration of bifidobacteria in gastrointestinal samples from piglets. Applied and Environmental Microbiology, 69, 654-658. Go to original source... Go to PubMed...
    10. Patterson J.A., Burkholder K.M. (2003): Application of prebiotics and probiotics in poultry production. Poultry Science, 82, 627-631. Go to original source... Go to PubMed...
    11. Reuter G., Klein G., Goldberg M. (2002): Identification of probiotic cultures in food samples. Food Research International, 35, 117-124. Go to original source...
    12. Roselli M., Finamore A., Britti M.S. (2005): Alternatives to in-feed antibiotics in pigs: evaluation of probiotics, zinc, or organic acids as protective agents for the intestinal mucosa. A comparison of in vitro and in vivo results. Animal Research, 54, 203-218. Go to original source...
    13. Roy D., Ward P., Champagne G. (1996): Differentiation of bifidobacteria by use of pulse-field gel electrophoresis and polymerase chain reaction. International Journal of Food Microbiology, 29, 11-29. Go to original source... Go to PubMed...
    14. Sakata S., Ryu C.S., Kitahara M. (2006): Characterization of the genus Bifidobacterium by automated ribotyping and 16S rRNA gene sequences. Microbiology and Immunology, 50, 1-10. Go to original source... Go to PubMed...
    15. Shim S.B., Verstegen M.W.A., Kim I.H., Kwon O.S., Verdonk J.M.A.J. (2005): Effects of feeding antibiotic-free creep feed supplemented with oligofructose, probiotics or synbiotics to suckling piglets increases the preweaning weight gain and composition of intestinal microbiota. Archives in Animal Nutrition, 59, 419-427. Go to original source... Go to PubMed...
    16. Vancanneyt M., Huys G., Lefebvre K., Vankerckhoven V., Goossens H., Swings J. (2006): Intraspecific genotypic characterization of Lactobacillus rhamnosus strains intended for probiotic use and isolates of human origin. Applied and Environmental Microbiology, 72, 5376- 5382. Go to original source... Go to PubMed...
    17. Vandamme P., Pot B., Gillis M. (1996): Polyphasic taxonomy, a consensus approach to bacterial systematics. Microbiology Reviews, 60, 407-438. Go to original source... Go to PubMed...
    18. Ventura M., Zink R. (2002): Specific identification and molecular typing analysis of Lactobacillus johnsonii by using PCR-based methods and pulsed-field gel electrophoresis. FEMS Microbiology Letters, 217, 141-154. Go to original source... Go to PubMed...
    19. Versalovic J., Koeuth T., Lupski J.R. (1991): Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes. Nucleic Acids Research, 19, 6823-6831. Go to original source... Go to PubMed...

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