A comparative analysis of the physicochemical, nutritional, and secondary metabolite profiles of commonly used species in multi-species swards, and their contributions to animal health

Limited research characterising the nutritional and secondary metabolite profiles of individual species used in multi-species swards (MSS) has constrained our understanding of species-specific functional roles, as well as evidence-based pasture design of MSS. This study aimed to comparatively analyse the physicochemical, nutritional, and secondary metabolite profiles of five commonly used MSS species – perennial ryegrass ( Lolium perenne ), white clover ( Trifolium repens ), red clover ( Trifolium pratense ), plantain ( Plantago lanceolata ), and chicory ( Cichorium intybus ) to identify species-specific traits that may contribute to enhanced forage quality and animal health outcomes. Monocultures were grown under standardised conditions in Trim, Co. Meath, Ireland, received 20 kg of N/ha as protected urea, and were wilted for 24 h post-harvest. Moisture, pH, ash, total soluble solids, protein, peptides, fat, carbohydrates, minerals, trace elements, total phenols, flavonoids, and antioxidant activity were quantified using a range of spectrophotometric and chromatographic techniques. Significant differences between species were observed ( P < 0.05). RC displayed the highest crude protein (18.27% ± 0.51), followed by WC (18.25% ± 0.23). Although lowest in crude protein (8.35% ± 0.90), PRG ranked second in peptide content (63.53g Leu-Gly equivalents/kg ±2.91) and exhibited the greatest antioxidant activity, with the lowest IC50 (15.54 g/kg ±0.39). Plantain had the highest concentrations of total phenols (15.35g chlorogenic acid equivalents/kg ±1.38) and flavonoids (6.91g catechin equivalents/kg ±0.62). Chicory demonstrated a diverse mineral profile, with elevated levels of magnesium (4875.75 mg/kg ±359.00), copper (11.17 mg/kg ±2.15), and zinc (41.14 mg/kg ±2.78). These compositional differences highlight unique functional roles individual species may play in animal health, forage quality, and sustainability.