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Applied and Environmental Microbiology, November 2009, p. 6764-6776, Vol. 75, No. 21
0099-2240/09/$08.00+0     doi:10.1128/AEM.01150-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Comparison and Utilization of Repetitive-Element PCR Techniques for Typing Lactobacillus Isolates from the Chicken Gastrointestinal Tract{triangledown} ,{dagger}

David P. Stephenson,1 Robert J. Moore,2 and Gwen E. Allison1,3*

Research School of Biology, The Australian National University, Canberra, ACT 0200, Australia,1 CSIRO Livestock Industries, Australian Animal Health Laboratory, Geelong, Victoria 3220, Australia,2 ANU Medical School, The Australian National University, Canberra, ACT 0200, Australia3

Received 19 May 2009/ Accepted 7 September 2009

Three repetitive-element PCR techniques were evaluated for the ability to type strains of Lactobacillus species commonly identified in the chicken gastrointestinal tract. Enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) produced species- and strain-specific profiles for Lactobacillus crispatus, Lactobacillus gallinarum, Lactobacillus johnsonii, and Lactobacillus reuteri isolates. The technique typed strains within these species equally as well as pulsed-field gel electrophoresis. DNA concentration and quality did not affect the ERIC-PCR profiles, indicating that this method, unlike other high-resolution methods, can be adapted to high-throughput analysis of isolates. Subsequently, ERIC-PCR was used to type Lactobacillus species diversity of a large collection of isolates derived from chickens grown under commercial and necrotic enteritis disease induction conditions. This study has illustrated, for the first time, that there is great strain diversity within each Lactobacillus species present and has revealed that chickens raised under commercial conditions harbor greater species and strain diversity than chickens raised under necrotic enteritis disease induction conditions.

* Corresponding author. Mailing address: Research School of Biology, Bldg. 41 Linnaeus Way, Australian National University, Canberra, ACT 0200, Australia. Phone: 61-2-6125-4795. Fax: 61-2-6125-0313. E-mail: gwen.allison{at}anu.edu.au

{triangledown} Published ahead of print on 11 September 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, November 2009, p. 6764-6776, Vol. 75, No. 21
0099-2240/09/$08.00+0     doi:10.1128/AEM.01150-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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