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Background  Porcine proliferative enteropathy in pigs is caused by the obligate, intracellular bacterium  Lawsonia intracellularis .  In vitro  studies have shown close bacterium-cell interaction followed by cellular uptake of the bacterium within 3 h post inoculation (PI). However, knowledge of the initial  in vivo  interaction between porcine intestinal epithelium and the bacterium is limited. The aims of the present study were to evaluate the usefulness of a ligated small intestinal loop model to study  L. intracellularis  infections and to obtain information on the very early  L. intracellularis -enterocyte interactions.    Methods  A ligated small intestinal loop model using three different  L. intracellularis  inocula was applied to 10-11-week-old pigs. The inocula were 1) wild type bacteria derived from overnight incubation of  L. intracellularis  bacteria from spontaneous disease, 2) crude vaccine bacteria (Enterisol ®  Ileitis Vet), and 3) vaccine bacteria propagated in cell culture. The bacteria-enterocyte interaction was visualised using immunohistochemistry on specimens derived 1, 3 and 6 h PI respectively.    Results  Although at a low level, close contact between bacteria and the enterocyte brush border including intracellular uptake of bacteria in mature enterocytes was seen at 3 and 6 h PI for the vaccine and the propagated vaccine inocula. Interaction between the wild-type bacteria and villus enterocytes was scarce and only seen at 6 h PI, where a few bacteria were found in close contact with the brush border.    Conclusions  The ligated intestinal loop model was useful with respect to maintaining an intact intestinal morphology for up to 6 h. Furthermore, the study demonstrated that  L. intracellularis  interacts with villus enterocytes within 3 to 6 h after inoculation into intestinal loops and that the bacterium, as shown for the vaccine bacteria, propagated as well as non-propagated, was able to invade mature enterocytes. Thus, the study demonstrates the early intestinal invasion of  L. intracellularis in vivo .

Application of a pig ligated intestinal loop model for early Lawsonia intracellularis infection