Effect of different feed ingredients and additives on IPEC-J2 cells challenged with an enterotoxigenic Escherichia coli strain

Abstract

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.

Lythrum salicaria Ellagitannins Stimulate IPEC-J2 Cells Monolayer Formation and Inhibit Enteropathogenic Escherichia coli Growth and Adhesion

“…The proposed mechanism, however, requires further verification. Until now, only a few feed additives such as extracts from wheat bran, casein glycomacropeptide, mannan-oligosaccharides, locust bean extract, and Aspergillus oryzae fermentation product or autolyzed yeast product were shown to reduce EPEC attachment to IPEC-J2 cells. , However, due to a high composition heterogeneity, none of them could serve as a referenced positive control in the adhesion experiments.…”

Synergistic Effect of B-Type Lotus Seedpod Oligomeric Procyanidin and Probiotics against Adhesion of Enterotoxigenic Escherichia coli In Vitro

“…TEER values can also be used as a marker of cell permeability. Te adhesion of pathogenic bacteria to intestinal epithelial cells can lead to a decrease in the TEER value of host cells [34]. Te changes in cellular TEER values during the 6 h incubation of LSPC and LGG with IPEC-J2 cells are shown in Figure 6(c), the TEER values of LSPC, LGG, and LSPC + LGG cells increased signifcantly (P < 0.05) after 2 h of incubation, and the efect of LGG alone on cellular TEER was greater than that of the LSPC and LSPC + LGG groups.…”

Porcine and Chicken Intestinal Epithelial Cell Models for Screening Phytogenic Feed Additives—Chances and Limitations in Use as Alternatives to Feeding Trials

“…An adhesion inhibiting potential has also been demonstrated for locust bean ( Ceratonia silique ) exopolysaccharide and wheat bran [ 67 ] whereas soybean hulls, sugar-beet pulp, locust gum, fructooligosaccharides, inulin, mushroom, mannanoligosaccharides, a fermented product from Aspergillus oryzae as well as an additive containing thyme extract had no influence on adhesion of pathogenic E. coli to IPEC-J2 cells [ 67 , 68 ]. None of the additive was able to protect the IPEC-J2 cells from E. coli -induced decrease in TEER [ 68 ].…”