Abstract

Proanthocyanidins are plant specialized metabolites which are beneficial to animal nutrition and health. This study determined how proanthocyanidin profiles of sainfoin (Onobrychis viciifolia) and birdsfoot trefoil (Lotus corniculatus) change during the forage conservation process and along the digestive tract of lamb. We determined soluble, protein- and fiber-bound proanthocyanidins by spectrophotometric methods and soluble proanthocyanidin profiles by UPLC-MS/MS. During the conservation process, the total proanthocyanidin contents reduced in both forages and the relative proportion of insoluble proanthocyanidins increased, especially in sainfoin. The soluble proanthocyanidins, their mean degree of polymerization and the relative prodelphinidin share declined in both feed species. In the abomasum of lambs fed sainfoin silage, most of the proanthocyanidins were in insoluble form bound to proteins and fibers, but in the small and large intestines, the proportion of soluble proanthocyanidins increased again. For lambs fed birdsfoot trefoil, the trend was not so clear as proanthocyanidins were already mainly soluble in the abomasum. Nevertheless, a large part of soluble proanthocyanidins was recovered in the digestive tract but could not be detected by the UPLC-MS/MS method used. This study suggests that proanthocyanidins have probably been metabolized in the digestive tract by the resident microbiota.

Highlights

  • The plant tannins, i.e., proanthocyanidins (PAs, syn. condensed tannins), hydrolysable tannins and phlorotannins, are a unique group of plant specialized metabolites

  • PAs are present in soluble and insoluble forms, for example over 90% of PAs in watermelon pilea leaves (Pilea cadierei) and in Turkey oak heartwood (Quercus cerris), and over 75% of watermelon pilea leaves (Pilea cadierei) and in Turkey oak heartwood (Quercus cerris), and over 75%

  • Sainfoin had on average six-fold greater PA contents in fresh, wilted and silage materials than birdsfoot trefoil, which is in agreement with previous experiments [36,37]

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Summary

Introduction

The plant tannins, i.e., proanthocyanidins (PAs, syn. condensed tannins), hydrolysable tannins and phlorotannins, are a unique group of plant specialized metabolites. The PAs with a high number of hydroxyl groups (hydrogen bonds) or aryl groups interactions) or a high mean degree of polymerization (mDP, more binding sites) bind stronger to (hydrophobic interactions) or a high mean degree of polymerization (mDP, more binding sites) bind proteins [12,13]. BuOH/HCl method by using acetone as cosolvent with BuOH/HCl Another common method) and thereby increasing the anthocyanidin yields, especially when used directly on the plant method for the analysis of insoluble PAs is their acid catalyzed depolymerization in the presence of material. Reaction products can be analyzed by LC-MS These methods were initially developed for soluble PAs, but have been later utilized for insoluble ones [24,25,26,27]. We determined the soluble and insoluble PAs by the well-established Terrill’s method [35] and by the more recent BuOH/HCl/Ace assay [21], and UPLC-MS/MS method [7]

Results and Discussion
Unidentifiable
Unidentifiable Role of Insoluble PAs
Preparation of the Experimental Forages
Experimental Lambs and Treatments
Nutrient Analysis of the Feed
Methods
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