An evaluation of greenhouse gas emissions reduction potential of plantain (Plantago lanceolata L.) in pastoral dairy production systems : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Agriculture Systems Management at Massey University, Manawatu, New Zealand

Abstract

There is increasing interest in the ability of plantain (PL) to reduce nitrogen (N) leaching losses and mitigate nitrous oxide (N₂O) emissions, while maintaining milk and pasture production. While PL’s role in lowering urinary N concentration is well established, the results regarding the effect of PL on N₂O emissions have been inconsistent. Furthermore, evidence has shown that cows fed pure PL produce less methane (CH₄) emissions compared to those fed ryegrass. However, whether this CH₄ reduction can be achieved with PL in mixed pasture, along with a clear understanding of the mechanism(s) behind those reductions, are still to be determined. This thesis evaluates PL’s potential to mitigate CH₄ and N₂O emissions through a series of in vitro and a field experiment, focusing on mixed pastures with moderate PL levels. When pastures, either a conventional ryegrass-white clover (RWC) or an RWC mix containing ~40% of PL (PLM), were collected during different seasons and tested in an in vitro rumen batch culture system, differences in their chemical composition led to significant differences in CH₄ and rumen ammonia (NH₃) production. Compared to RWC, PLM had lower fibre (neutral detergent fibre and acid detergent fibre), higher lignin, more fermentable carbohydrates (non-structural carbohydrates), and plant secondary metabolites (PSM, acteoside and aucubin) detected only in PLM, while maintaining similar digestibility and crude protein (CP) levels. Consequently, PLM produced up to 27% less net NH₃ in spring, up to 19% less CH₄ in summer, and 17% less net NH₃ in autumn compared to RWC (p<0.05) in vitro. Plant secondary metabolites found in PL, have been associated with reducing N losses from grazed pastures. However, their influence on enteric CH₄ emissions remains unexplored. Additionally, the dose-response relationship between CH₄ and NH₃ production at different concentrations of PSM needs to be established. To address this, purified compounds (>99% purity) of acteoside and aucubin were incubated with perennial ryegrass (RG) as a basal substrate, and gas and CH₄ production were measured in vitro. The addition of acteoside to RG increased gas production (GP) by up to 12%, with a similar quantity of CH₄ production, but a 5–15% lower proportion of CH₄ in gas (%CH₄), compared to the control. Aucubin addition resulted in a longer lag phase for GP and CH₄ production. On addition of aucubin, it took up to 15% more time to reach the halftime (T1/2) GP and up to 20% longer to reach the T1/2 CH₄ production. The combined treatments of acteoside and aucubin produced up to 13% greater GP with similar CH₄ production and reduced %CH₄ by around 9%. These reductions are attributed to the modification of the hydrogen utilisation pathway (less hydrogen to produce CH₄) affected by acteoside. Aucubin reduced rumen net NH₃ production by up to 46%, with a similar reduction observed when acteoside was combined with aucubin. These reductions are attributed to the possible antimicrobial activity of aucubin. These results suggest that PL influences rumen fermentation in vitro, resulting in lower CH₄ and NH₃ production. Since higher rumen NH₃ correlates with greater urinary N excretion into the environment, reducing NH₃ levels in the rumen is advantageous. Previous studies have shown that N₂O emissions from PL pastures may be reduced due to smaller N concentrations in urine and/or biological nitrification inhibition (BNI) activity. In this study, urine collected from cows fed diets containing 0% PL, ~20% PL, and diluted urine from PL-fed cows, was applied to pastures containing 0% PL, 30% PL, and 40% PL during spring. The N₂O emissions were measured over 55 days. Results indicated a trend toward lower N₂O emissions as assessed using the emission factor (EF₃) metric, with increasing PL content (p<0.09), with an average reduction of around 28% for pastures containing 30–40% PL compared to RWC pastures (p=0.03). This reduction in N₂O emissions from PL pastures was attributed to BNI activity rather than differences in urine-N concentrations per se. These results enhance our understanding of PL’s role in mitigating environmental impacts from grazing ruminants in temperate systems. This thesis concludes that medium PL pastures (30–40% PL) exhibit significant environmental benefits compared to RWC pastures in vitro, with reductions in CH₄ and rumen NH₃ influenced by PSM in PL and the seasonal variability in chemical composition. Moreover, under conditions conducive to higher N₂O emissions (in spring), maintaining 30–40% PL in the pasture could reduce N₂O emissions more effectively than excluding PL.