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Immunological relevant factors in Gastro-intestinal tract
  “ An apple a day keeps the doctor away ! ”   Influence of different flavanol rich feeding regimens on the villi and crypt morphology in jejunum, ileum, and colon of young piglets.

Julia HERZIG,  Claudia DUMMER  &  Michael W. PFAFFL

Introduction:  The intestinal tract of piglets change during growth and especially in the time around weaning. The change from sow milk to food causes diversification in gut crypts and gut villi morphology. In this time the piglets have often diarrhea which produce high costs in breeding. One facility is to give antibiotic with the feed ingredients, but from 2006 on the antibiotics in feed are forbidden in the EU. Therefore farmers have to look after another possibilities to improve gut health. In this approach two different flavanol rich feeding regimens were applied in young growing piglets (freeze dried apple and red wine pomace). This compounds have high amounts of polyphenols which are known as healthy drugs. The average of the total polyphenol content in apple is 3 g per kg fresh weight (0.5 – 11 g/kg) and 2 mg/liter red wine.

Methods:  In this study 78 piglets were fed over 4 weeks with three different feeding regimens. 26 animals were fed with normal piglet starter, 26 got additionally 3.5% dry matter (dm) apple pomace and further 26 got 3.5% dm red wine pomace. Pigs were killed by electric gripper and bleeding. Samples of jejunum, ileum, and colon were collected for the histology investigation. Tissues were fixed two days in 3.7% formalin, dewatered and embedded automatically in paraffin wax. Paraffin wax embedded tissues were sectioned (9 µm in thickness, Microm, Walldorf, Germany), mounted on 2% silane (Sigma, Deisenhofen, Germany) coated slides, dewaxed in Xylol and rehydrated in descending alcohols. Slices were stained with Mayer`s Hemalaun (C.I. N°: 75290; Merck, Darmstadt, Germany), dehydrated in descending alcohols, rinsed in Xylol and embedded in Eukitt (Kindler, Freiburg, Germany). All sections were viewed under the axioskop (Zeiss microscope, Munich, Germany). Gut villi length, crypt depth and breath, and peyer patches area were imaged and measured in triplicates (n = 234 each) by using a digital camera and axiovision software (Carl Zeiss Axio Cam MR, axiovision software, 3.0.6 SP3).

Results:  The villi area in jejunum increase strongest in the control animals and apple feed animals. No significant growth could be shown in the red wine pomace group. No significant changes could be shown in ileum. Peyer patch were highly enlarged in control group after 28 days. In colon the villi and crypt area growth was significantly increased in apple as well in red wine pomace feeding groups.
Tables:   Villi, crypt, and peyer patch morphology in jejunum, ileum, and colon of piglets. Linear regression of the growth (y) in days (x) in mm or mm². The percentage value shows the relative growth in 28 days, compared to the starting day 0. Significant growth over 28 days is indicated by p value.

Discussion:   Results show that in jejunum the villi growth is inhibited by red wine pomace, and slightly reduced by apple pomace, compared to control animals. In Ileum no effect on the gut morphology were evident, but the incorporated peyer patch were enlarged in control group. This indicates a peyer patch activation in the flavanoid lacking feeding regimen and suggest an immuno-preventive effect of flavanols in the ileum. In colon both flavanol feeding regimen can enhance the villi and crypt growth.
In a further experiment the size of the epithelial cells was measured, to test if the villi or the cells itself are growing under the influence of flavanols. Cells sizes stay constant under all three feeding regimes as well over 28 days. This indicates that the villi itself is growing and the epithelial cells sizes stay constant.

Conclusions:   Apple and red wine pomace can reduce the immune system activation via the peyer patches in the ileum. Different fruits have diverse effects in the gastrointestinal tract. Red wine pomace has inhibitory effects in the jejunum villi and apple pomace has a stimulating effect in colon villi and crypt. Flavanoid rich feeding regimen showed positive effects on gut health and is a good alternative to improve pig health in agriculture after the ban on antibiotics.