Effect of different feed ingredients and additives on IPEC-J2 cells challenged with an enterotoxigenic Escherichia coli strain.
The intestinal porcine epithelial cell line IPEC-J2 was used as an in vitro model to assess effects of additives on the adhesion and cell toxic effects of a F4-positive (ETEC) and a F4-negative Escherichia coli (DSM 2840) strain. Bacterial adhesion was examined using flow cytometry in IPEC-J2 cells infected with bacteria stained with 5,6-carboxymethyl fluorescein diacetate succinimidyl ester. Measurement of transepithelial electrical resistance (TEER) was performed to characterize the impact on IPEC-J2 monolayer integrity. The feed additives were prepared as aqueous extract and tested in different dilutions and incubation times. The F4-positive ETEC strain had a high adhesion to IPEC-J2 cells and reduced TEER shortly after the in vitro infection. The nonpathogenic E. coli strain DSM 2840 showed only low adhesion capacity and no TEER impairment. Infection with ETEC with added test extracts showed a reduction of bacterial adhesion to IPEC-J2 cells by an autolyzed yeast product (p < 0.05). Bovine colostrum, an additive containing thyme extract and an organic acid mix did not interfere with the ETEC adherence. The TEER decrease of the IPEC-J2 monolayer after ETEC infection was not affected by the added substances. In conclusion, interference with epithelial adhesion might be a protective mechanism of the tested yeast extract, indicating that the cell culture model might be suitable as screening tool to complement in vivo challenge trials with piglets.
Department of Veterinary Medicine, Institute of Animal Nutrition, Freie Universität Berlin, Königin-Luise-Str. 49, 14195, Berlin, Germany.
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Spitzer
Department of Veterinary Medicine, Institute of Animal Nutrition, Freie Universität Berlin, Königin-Luise-Str. 49, 14195, Berlin, Germany.
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Speiser
Department of Veterinary Medicine, Institute of Animal Nutrition, Freie Universität Berlin, Königin-Luise-Str. 49, 14195, Berlin, Germany. wilfried.vahjen@fu-berlin.de.
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Vahjen
Department of Veterinary Medicine, Institute of Animal Nutrition, Freie Universität Berlin, Königin-Luise-Str. 49, 14195, Berlin, Germany.
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Zentek
Diarrhoea; Feed additives; IPEC-J2 cells; Piglets; Weaning