Open AccessDetection 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
Author(s) -
Le Luo Guan,
Karen E. Hagen,
Gerald W. Tannock,
D. R. Korver,
G. M. Fasenko,
Gwen E. Allison
Publication year - 2003
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.69.11.6750-6757.2003
Subject(s) - lactobacillus crispatus , biology , lactobacillus acidophilus , amplified ribosomal dna restriction analysis , lactobacillus salivarius , lactobacillus , temperature gradient gel electrophoresis , lactobacillus reuteri , lactobacillus gasseri , microbiology and biotechnology , ribosomal dna , 16s ribosomal rna , polymerase chain reaction , crop , ribosomal rna , food science , probiotic , bacteria , genetics , agronomy , gene , phylogenetics
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 10(8) to 10(9) 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.