Colonization of mature laying hens with Salmonella Enteritidis using either oral or intracloacal

inoculation

P.A. Adhikari1, D.E. Cosby2, N.A. Cox2 and W.K. Kim1

Mature laying hens are more difficult to colonize than young chicks. The objective of this study

was to compare the oral (OR) versus the intracloacal (IC) routes of inoculation for colonizing 40

week old laying hens. In the first experiment, 32 laying hens were placed one per pen in

conventional wire layer cages and allowed to adapt for a week, with ad libitum access to feed

and water under a 16L:8D light regimen. All hens were challenged OR with 0.1 mL of a 1.6 x

108 CFU of Salmonella Enteritidis (SE). Birds were euthanized after 14 days and the following

were aseptically removed: ceca, ovary, spleen and liver-gallbladder (LGB). All samples were

negative for Salmonella. In experiment 2, 14 hens were challenged OR with 1.0 mL of 3.3 x 108

CFU of SE and 14 were inoculated IC with 1.0 mL of 3.3 x 108 CFU of SE. Animal

husbandry/handling was the same as in experiment 1. After 7 d, 7 of the OR hens, 7 of the IC

hens and 4 uninoculated (control) hens were euthanized and sampled as in experiment 1 and at

14 d, the remaining birds were sampled the same. After 7d, the recovery of SE with IC versus

OR inoculation was ceca (100 vs 86%), ovary (43 vs 14%), spleen (57 vs 43%) and LGB (43 vs

14%). At 14 d, the respective recovery rate was ceca (43 vs 14%), LGB (14% for both) and no

recovery in the ovary or spleen. There was a higher incidence of recovery in the IC hens. These

results suggest that mature hens over 40 weeks of age can be colonized (at least temporarily)

after either an OR or IC challenge with a larger volume (0.1 vs 1.0 mL) and a higher number of

cells. Once internalized the SE shows some translocation into other organs including the ovary,

spleen and LGB. However, the colonization did not consistently persist either in ceca or other
internal organs at 14 days. Short term persistence of colonization and translocation limits the

ability to do research with mature hens over two weeks in duration.

Keywords: Salmonella Enteritidis, laying hen, oral, intracloacal, colonization

1
University of Georgia, Department of Poultry Science, Athens, Georgia
2
United States Department of Agriculture, The U.S. National Poultry Research Center, Athens,

Georgia