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United States Department of Agriculture

Agricultural Research Service

Title: Ethanol Immersion Method for the Collection of Separate External and Internal Microbiological Spleen Samples of Baby Broiler Chicks

Authors
item Richardson, Larry
item Cox, Nelson
item Buhr, Richard
item Bailey, Joseph
item Harrison, M - UGA

Submitted to: Southern Poultry Science Society Meeting Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: November 3, 2005
Publication Date: January 23, 2006
Citation: Richardson, L.J., Cox Jr, N.A., Buhr, R.J., Bailey, J.S., Harrison, M.A. 2006. Ethanol immersion method for the collection of separate external and internal microbiological spleen samples of baby broiler chicks [abstract]. Southern Poultry Science Society Meeting. 64.

Technical Abstract: Campylobacter species have been recovered from the lymphoid tissues of avian species. However, whether the bacteria are located internally in these tissues has not been determined. The objectives of the present study were to 1) develop a method to sample the inside and outside of the spleen and 2) determine if Campylobacter can be recovered from the internal tissue of the spleen. Five experiments were conducted and for each experiment, day-of-age broiler chicks were obtained and inoculated by different routes with Campylobacter jejuni. Two days post-inoculation, necropsy was performed and the spleen and ceca were aseptically removed from each bird and individually placed into sterile sampling bags, packed on ice and transported back to the laboratory for evaluation. For external (ES) spleen sampling, 3 ml of Bolton’s enrichment broth was added to each spleen sample and shaken for 30s. The spleen was then aseptically removed from the bag. For internal spleen sampling, the spleen was submerged into a 70% ethanol solution for 10s, then removed and submerged into a saline solution to detect sterilization ability of the immersion technique. The spleen was then placed into a sterile sampling bag, macerated and 3 ml of Bolton’s enrichment broth was added and the sample was stomached for 30s. Standard laboratory procedures were then performed on the above samples (ES, IS, ethanol and saline solution) and ceca for the recovery of Campylobacter species. Overall, Campylobacter was recovered from 75% (82/109) of the ES samples, 71% (77/109) of the IS samples and from 100% (109/109) of the ceca samples. Furthermore, ethanol and saline samples were negative for Campylobacter suggesting that the ethanol immersion method is a good sterilization method of the external surface of the spleen. With Campylobacter residing on the external surface and in the internal tissue of the spleen, the organisms could be systemic inside the bird. Further research will determine the mechanisms required for this bacterium to colonize these different tissue types.

Last Modified: 10/24/2014
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