Comparative analysis of Campylobacter populations within individual market-age broilers using Fla gene typing method

Campylobacter species is the most common human pathogen causing gastrointestinal infections in humans, and poultry is considered a major source of this pathogen. In this project, we aim to study the genetic diversity of Campylobacter populations within individual chickens using cecal samples to understand the nature of intestinal colonization in chickens by Campylobacter species. Genotyping was conducted based on the DNA sequence of short variable regions (SVR) in the flaA gene. Cecal samples were collected from 9 market-age broiler chickens and used for isolation of Campylobacter genomic DNA. The SVR fragments of 400 bp were amplified using SVR-specific primers, cloned, and sequenced. Sequencing results obtained from 86 clones (~10 clones/bird) showed that on an average 23.25% of clones had mutations within individuals. The mutations did not show any consistent pattern, suggesting a random nature of the mutations. When translated SVR sequences were analyzed, on average 20.57% of strains carried altered amino acid sequences in SVR within individuals. Four translated sequences had nonsense mutations to produce truncated proteins. These results suggest that there are multiple genotypes colonizing in a cecum and the occurrence of truncated FlaA protein may represent a novel mechanism for evasion of adaptive immune responses.