Using plant wax markers to estimate diet composition and intakes of mixed forages in sheep by feeding a known amount of alkane-labelled supplement

The feeding of known amounts of supplements to grazing animals can be accomplished relatively easily. If the supplement and the other diet components have distinctive profiles of cuticular wax n-alkanes, then the supplement intake and the alkane profiles of the supplement, other dietary components and faeces can be used to estimate the proportions and hence intakes of several forages by the grazing animal. However, this method requires knowledge of recoveries of n-alkanes in faeces. Twenty four wethers were fed one of four diets comprising equal combinations of 1, 2, 3 or 4 forages. Forages used were subterranean clover, phalaris, annual ryegrass and wheat straw. Forages were chopped using a chaff cutter and fed with solvent-extracted cottonseed meal (CSM) labelled with beeswax and synthetic C28 alkane to provide a characteristic alkane profile. Faecal grab samples were taken from sheep from 14 to 23 days after administration of an intra-ruminal controlled-release device (CRD) containing 1 g of each of C32 and C36 alkane. Total faeces were collected from half the sheep on each treatment in order to measure alkane recoveries in individual sheep. Faecal concentrations of the n-alkanes C25 to C31 and C33 were corrected for recovery using the individual sheep value, the treatment mean or the grand mean for all four treatments. Dietary compositions were then estimated from corrected faecal concentrations of n-alkanes using a least-squares procedure and, together with the known supplement intakes, were used to estimate the intakes of all other diet components. Estimates from this ‘labelled supplement’ method were compared with the amounts fed or those estimated using the alkanes derived from the CRD. The labelled supplement method accurately and precisely estimated dietary component proportions and intake for all treatments when measured recoveries for individual sheep were used. Precision declined when recovery was based on estimated recoveries for treatment means or the grand mean. Estimates of intake based on dietary C33 and the measured release rates of C32 or C36 alkanes from the CRD did not differ from measured intakes. Estimates based on the C32 : C31 alkane pair over-estimated intake. Estimates of whole-diet digestibility based on the various ways of estimating intake were all very close to the digestibilities calculated from directly-measured intakes and faecal outputs. It is concluded that the feeding of a known amount of supplement can be successfully used to estimate dietary proportions and hence intakes of diet components, in mixed diets with up to five ingredients, but this approach requires estimates of faecal alkane recovery.