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Applied and Environmental Microbiology, November 2003, p. 6750-6757, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6750-6757.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Detection and Identification of Lactobacillus Species in Crops of Broilers of Different Ages by Using PCR-Denaturing Gradient Gel Electrophoresis and Amplified Ribosomal DNA Restriction Analysis

Le Luo Guan,1 Karen E. Hagen,1 Gerald W. Tannock,1,2 Doug R. Korver,1 Gaylene M. Fasenko,1 and Gwen E. Allison1*

Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta, Canada T6G 2P5,1 Department of Microbiology, University of Otago, Dunedin, New Zealand2

Received 2 June 2003/ Accepted 19 August 2003

The microflora of the crop was investigated throughout the broiler production period (0 to 42 days) using PCR combined with denaturing gradient gel electrophoresis (PCR-DGGE) and selective bacteriological culture of lactobacilli followed by amplified ribosomal DNA restriction analysis (ARDRA). The birds were raised under conditions similar to those used in commercial broiler production. Lactobacilli predominated and attained populations of 108 to 109 CFU per gram of crop contents. Many of the lactobacilli present in the crop (61.9% of isolates) belonged to species of the Lactobacillus acidophilus group and could not be differentiated by PCR-DGGE. A rapid and simple ARDRA method was developed to distinguish between the members of the L. acidophilus group. HaeIII-ARDRA was used for preliminary identification of isolates in the L. acidophilus group and to identify Lactobacillus reuteri and Lactobacillus salivarius. MseI-ARDRA generated unique patterns for all species of the L. acidophilus group, identifying Lactobacillus crispatus, Lactobacillus johnsonii, and Lactobacillus gallinarum among crop isolates. The results of our study provide comprehensive knowledge of the Lactobacillus microflora in the crops of birds of different ages using nucleic acid-based methods of detection and identification based on current taxonomic criteria.

* Corresponding author. Mailing address: Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta, Canada T6G 2P5. Phone: (780) 492-9841. Fax: (780) 492-4265. E-mail: Gwen.Allison{at}ualberta.ca.

Applied and Environmental Microbiology, November 2003, p. 6750-6757, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6750-6757.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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