Enhancement of immunohistochemical detection of Salmonella in tissues of experimentally infected pigs

  • J. Rieger | Juliane.Rieger@fu-berlin.de Freie Universität Berliny, Germany.
  • P. Janczyk Federal Institute for Risk Assessment, Germany.
  • H. Hünigen Freie Universität Berlin, Germany.
  • J. Plendl Freie Universität Berlin, Germany.

Abstract

Salmonella Typhimurium is one of the main pathogens compromising porcine and human health as well as food safety, because it is a prevailing source of foodborne infections due to contaminated pork. A prominent problem in the management of this bacteriosis is the number of subclinically infected carrier pigs. As very little is known concerning the mechanisms allowing Salmonella to persist in pigs, the objective of this study was to develop an immunohistochemical approach for the detection of salmonellae in tissue of pigs experimentally infected with Salmonella Typhimurium. Samples were obtained from a challenge trial in which piglets of the German Landrace were intragastrically infected with Salmonella enterica serovar Typhimurium DT104 (1.4-2.1x1010 CFU). Piglets were sacrificed on days 2 and 28 post infection. Tissue samples of jejunum, ileum, colon, ileocecal mesenteric lymph nodes (Lnn. ileocolici), and tonsils (Tonsilla veli palatini) were fixed in Zamboni’s fixative and paraffin-embedded. Different immunohistochemical staining protocols were evaluated. Salmonella was detected in varying amounts in the tissues. Brown iron-containing pigments in the lymph nodes interfered with the identification of Salmonella if DAB was used as a staining reagent. Detergents like Triton X-100 or Saponin enhanced the sensitivity. It seems advisable not to use a detection system with brown staining for bacteria in an experimental setup involving intestinal damage including haemorrhage. The use of detergents appears to result in a higher sensitivity in the immunohistochemical detection of salmonellae.

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Author Biographies

J. Rieger, Freie Universität Berliny
Department of Veterinary Medicine, Institute of Veterinary Anatom
P. Janczyk, Federal Institute for Risk Assessment
Department of Biological Safety, Unit Molecular Diagnostics and Genetic
H. Hünigen, Freie Universität Berlin
epartment of Veterinary Medicine, Institute of Veterinary Anatomy
J. Plendl, Freie Universität Berlin
Department of Veterinary Medicine, Institute of Veterinary Anatomy
Published
2015-07-09
Info
Issue
Section
Technical Notes
Supporting Agencies
German Research Foundation (DFG) through the Collaborative Research Centre 852 (Grant no. SFB852/1)
Keywords:
Swine, immunohistochemistry, histochemistry, bacteria, detection limit.
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How to Cite
Rieger, J., Janczyk, P., HünigenH., & Plendl, J. (2015). Enhancement of immunohistochemical detection of Salmonella in tissues of experimentally infected pigs. European Journal of Histochemistry, 59(3). https://doi.org/10.4081/ejh.2015.2516