Western
Meeting of Poultry
Clinicians and Pathologists
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Clinicians and Pathologists
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Development of a Campylobacter jejuni Colonization Model for Pathogenomics Studies in Poultry 1P. Willson, 1K. Mirakhur, 1B. Sampathkumar, 1E. Mace, 1C. Reiman, 2S. Gomis, 1L. Babiuk,1A. Potter and 1B. Allan 1Vaccine & Infectious Disease Organization, University of Saskatchewan, Saskatoon, SK, S7N 5E3; Veterinary Pathology, 2Western College of Veterinary medicine, University of Saskatchewan, Saskatoon, SK, S7N 5B4. Colonization of poultry by C. jejuni preferentially occurs in the ceca, which are two blind segments at the lower part of the intestine. This presents a unique opportunity to study colonization and gene expression in both pathogen and host during colonization in vivo since one cecum can act as a control for the other. In order to evaluate the impact of antibiotic treatment and determine the optimum length post surgery that the bacteria can remain in the cecum, birds were euthanized and the ceca removed for quantification of C. jejuni and RNA extraction at 6 or 24 hours after surgery. The normal saline that was used to suspend the bacteria was absorbed and by 24 hours no fluid remained. To recover the bacteria we introduced 1 mL of saline and then clipped off one end of the cecum to recover the contents. The material recovered was viscous and contained particulate matter. This made accurate viable cell counts difficult. Between 108 and 109 CFU’s of C. jejuni (mixed with other bacterial species) were recovered at all time points. In all cases the cecum that received only normal saline was heavily colonized by C. jejuni. Treatment of birds with trimethoprim-sulphadoxine (to which C. jejuni is resistant) did not affect the concentration of C. jejuni in the ceca. TriZol® treatment of the ceca resulted in recovery of at least 10 μg of (predominantly eukaryotic) RNA per cecum. This surgical method can be applied to other species of bacteria that colonize poultry, such as Salmonella spp., and is a suitable way to investigate colonization and gene expression in the natural context of host and microorganism. |
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