Evaluation of n-alkanes, long-chain alcohols, and carbon stable isotope enrichments of n-alkanes as diet composition markers in free-grazing animals
Teklu Wegi A B , Abubeker Hassen
A * , Melkamu Bezabih C , Ajebu Nurfeta D , Sintayehu Yigrem D and Adugna Tolera D
A Department of Animal Sciences, University of Pretoria, Pretoria 0002, South Africa.
B Present address: Oromia Agricultural Research Institute, Sinana Agricultural Research Center, PO Box 208, Bale Robe, Ethiopia.
C International Livestock Research Institute, PO Box 5689, Addis Ababa, Ethiopia.
D School of Animal and Range Sciences, Hawassa University, PO Box 5, Hawassa, Ethiopia.
Animal Production Science 62(13) 1280-1289 https://doi.org/10.1071/AN21497
Submitted: 1 October 2021 Accepted: 24 March 2022 Published: 20 May 2022
© 2022 The Author(s) (or their employer(s)). Published by CSIRO Publishing
Abstract
Context: Plant species exhibit different patterns of plant cuticular wax profiles, which can potentially be used as diet composition markers in free-grazing herbivores.
Aims: Evaluate the suitability of the plant cuticular n-alkanes, long-chain alcohol (LCOH) profiles and carbon stable isotope enrichment (δ13C) of n-alkanes as markers in the estimation of diet composition of grazing animals.
Methods: Forage samples were collected from 100 representative quadrats of 0.5 m × 0.5 m at 10 m transects and sorted by species and pooled from different quadrats to obtain enough quantities of representative individual species. In total, 10 dominant forage species were identified and analysed for n-alkanes and LCOH by gas chromatography, and the isotopic ratio by using gas chromatography–combustion–isotope ratio mass spectrometry. Principal component analysis (PCA) was used to identify inter-species differences in the concentration patterns of plant wax components.
Key results: Odd-chain n-alkanes comprised the highest proportion of the total n-alkane concentration, ranging from 79% in Ischaemum afrum to 95% in Haplocarpha hastata. n-Alkanes C31, C29 and C33 were the most abundant with an average of 167, 80 and 61 mg/kg DM, in that order, in all species. Even-chain LCOH comprised the highest proportion of the total LCOH concentration, accounting for 92% in Brachiaria scalaris to 97% in Ischaemum afrum. The dominant even-chain LCOH were C30OH, C32OH, C28OH and C26OH, with an average concentration of 362, 348, 266 and 237 mg/kg DM respectively, across species. The δ13C of n-alkanes showed large variations among forage species, ranging from −19.7‰ in Andropogon amethystinus to −40.6‰ in Trifolium mattirolianum. The result of the PCA showed that 81% of the variance in the pattern of concentrations of n-alkanes was explained by the first two principal components compared with 69.3% and 82.9% in the case of LCOH and δ13C of n-alkanes, respectively.
Conclusions: Noticeable variations were observed for forage species studied in the patterns of plant wax components.
Implications: The differences in the patterns of concentrations of n-alkanes, LCOH and δ13C of n-alkanes could be suitable as markers for diet composition estimation of grazing animals.
Keywords: assessment, diet estimation, forage species profile, location specific, marker concentrations, pasture, plant cuticular wax, principal component analysis.
References
Ali HAM, Mayes RW, Hector BL, Orskov ER (2005) Assessment of n-alkanes, long-chain fatty alcohols and long-chain fatty acids as diet composition markers: the concentrations of these compounds in rangeland species from Sudan. Animal Feed Science and Technology 121, 257–271.| Assessment of n-alkanes, long-chain fatty alcohols and long-chain fatty acids as diet composition markers: the concentrations of these compounds in rangeland species from Sudan.Crossref | GoogleScholarGoogle Scholar |
Bezabih M, Pellikaan WF, Tolera A, Hendriks WH (2011) Evaluation of n-alkanes and their carbon isotope enrichments (δ13C) as diet composition markers. Animal 5, 57–66.
| Evaluation of n-alkanes and their carbon isotope enrichments (δ13C) as diet composition markers.Crossref | GoogleScholarGoogle Scholar | 22440702PubMed |
Body DR (1974) Neutral lipids of leaves and stems of Trifolium repens. Phytochemistry 13, 1527–1530.
| Neutral lipids of leaves and stems of Trifolium repens.Crossref | GoogleScholarGoogle Scholar |
Brosh A, Henkin Z, Rothman SJ, Aharoni Y, Orlov A, Arieli A (2003) Effects of faecal n-alkane recovery in estimates of diet composition. Journal of Agricultural Science 140, 93–100.
| Effects of faecal n-alkane recovery in estimates of diet composition.Crossref | GoogleScholarGoogle Scholar |
Bugalho MN, Dove H, Kelman W, Wood JT, Mayes RW (2004) Plant wax alkanes and alcohols as herbivore diet composition markers. Journal of Range Management 57, 259–268.
| Plant wax alkanes and alcohols as herbivore diet composition markers.Crossref | GoogleScholarGoogle Scholar |
Charmley E, Dove H (2007) Using plant wax markers to estimate diet composition and intakes of mixed forages in sheep by feeding a known amount of alkane labelled supplement. Australian Journal of Agricultural Research 58, 1215–1225.
| Using plant wax markers to estimate diet composition and intakes of mixed forages in sheep by feeding a known amount of alkane labelled supplement.Crossref | GoogleScholarGoogle Scholar |
Dove H, Charmley E (2008) Using the alkanes and long-chain alcohols of plant cuticular wax to estimate diet composition and the intakes of mixed forages in sheep consuming a known amount of alkane-labelled supplement. Animal 2, 1474–1485.
| Using the alkanes and long-chain alcohols of plant cuticular wax to estimate diet composition and the intakes of mixed forages in sheep consuming a known amount of alkane-labelled supplement.Crossref | GoogleScholarGoogle Scholar | 22443905PubMed |
Dove H, Mayes RW (1996) Plant wax components: a new approach to estimating intake and diet composition in herbivores. Journal of Nutrition 126, 13–26.
| Plant wax components: a new approach to estimating intake and diet composition in herbivores.Crossref | GoogleScholarGoogle Scholar | 8558293PubMed |
Dove H, Mayes RW (2005) Using n-alkanes and other plant wax components to estimate intake, digestibility and diet composition of grazing/browsing sheep and goats. Small Ruminant Research 59, 123–139.
| Using n-alkanes and other plant wax components to estimate intake, digestibility and diet composition of grazing/browsing sheep and goats.Crossref | GoogleScholarGoogle Scholar |
Dove H, Mayes RW (2006) Protocol for the analysis of n-alkanes and other plant–wax compounds and for their use as markers for quantifying the nutrient supply of large mammalian herbivores. Nature Protocols 1, 1680–1697.
| Protocol for the analysis of n-alkanes and other plant–wax compounds and for their use as markers for quantifying the nutrient supply of large mammalian herbivores.Crossref | GoogleScholarGoogle Scholar | 17487151PubMed |
Dove H, Mayes RW, Freer M (1996) Effects of species, plant part, and plant age on the n-alkane concentrations in the cuticular wax of pasture plants. Australian Journal of Agricultural Research 47, 1333–1347.
| Effects of species, plant part, and plant age on the n-alkane concentrations in the cuticular wax of pasture plants.Crossref | GoogleScholarGoogle Scholar |
Ferreira LMM, Oliván M, Garcia U, Rodrigues MAM, Osoro K (2005) Validation of the alkane technique to estimate diet selection of goats grazing heather–gorse vegetation communities. Journal of the Science of Food and Agriculture 85, 1636–1646.
| Validation of the alkane technique to estimate diet selection of goats grazing heather–gorse vegetation communities.Crossref | GoogleScholarGoogle Scholar |
Ferreira LMM, Garcia U, Rodrigues MAM, Celaya R, Dias-da-Silva A, Osoro K (2007) The application of the n-alkane technique for estimating the composition of diets consumed by equines and cattle feeding on upland vegetation communities. Animal Feed Science and Technology 138, 47–60.
| The application of the n-alkane technique for estimating the composition of diets consumed by equines and cattle feeding on upland vegetation communities.Crossref | GoogleScholarGoogle Scholar |
Ferreira LMM, Celaya R, Santos AS, Mayes RW, Rodrigues MAM, Osoro K (2013) Application of long-chain alcohols as diet composition markers in sheep fed on grass–white clover and heather–gorse plant species. Grass and Forage Science 70, 30–43.
| Application of long-chain alcohols as diet composition markers in sheep fed on grass–white clover and heather–gorse plant species.Crossref | GoogleScholarGoogle Scholar |
Ferreira LMM, Daniela JB, Celaya R, Santos AS, Osoro K, Rodrigues MAM, Pellikaan WF (2014) Utilization of carbon isotope enrichments (δ13C) of alkanes as faecal markers to estimate diet composition of goats fed with heath land vegetation. Animal Feed Science and Technology 191, 26–38.
| Utilization of carbon isotope enrichments (δ13C) of alkanes as faecal markers to estimate diet composition of goats fed with heath land vegetation.Crossref | GoogleScholarGoogle Scholar |
Genstat (2008) ‘Genstat for windows.’ 11th edn. (VSN International: Oxford)
Heublein C, Südekum K-H, Gill FL, Dohme-Meier F, Schori F (2017) Using plant wax markers to estimate the diet composition of grazing Holstein dairy cows. Journal of Dairy Science 100, 1019–1036.
| Using plant wax markers to estimate the diet composition of grazing Holstein dairy cows.Crossref | GoogleScholarGoogle Scholar | 27939548PubMed |
Institute of Biodiversity Conservation (IBC) (2005) ‘National biodiversity strategy and action plan, Government of the Federal Democratic Republic of Ethiopia. Vol. 115.’ (Institute of Biodiversity Conservation (IBC)).
| Crossref |
Kelman W, Bugalho M, Dove H (2003) Cuticular wax alkanes and alcohols used as markers to estimate diet composition of sheep (Ovis aries). Biochemical Systematics and Ecology 31, 919–927.
| Cuticular wax alkanes and alcohols used as markers to estimate diet composition of sheep (Ovis aries).Crossref | GoogleScholarGoogle Scholar |
Laredo MA, Simpson GD, Minson DJ, Orpin CG (1991) The potential for using n-alkanes in tropical forages as a marker for determination of dry matter intake by grazing ruminants. Journal of Agricultural Science 117, 355–361.
| The potential for using n-alkanes in tropical forages as a marker for determination of dry matter intake by grazing ruminants.Crossref | GoogleScholarGoogle Scholar |
López López C, Celaya R, Santos AS, Rodrigues MAM, Osoro K, Ferreira LMM (2015) Application of long-chain alcohols as faecal markers to estimate diet composition of horses and cattle fed with herbaceous and woody species. Animal 9, 1786–1794.
| Application of long-chain alcohols as faecal markers to estimate diet composition of horses and cattle fed with herbaceous and woody species.Crossref | GoogleScholarGoogle Scholar | 26160068PubMed |
Mayes RW (1998) New potential markers for determining diet composition. In ‘Proceeding of the IXth European intake workshop’. pp. 63–66. (Institute of Grassland and Environmental Research North Wyke, UK)
Mayes RW, Dove H (2000) Measurement of dietary nutrient intake in free-ranging mammalian herbivores. Nutrition Research Reviews 13, 107–138.
| Measurement of dietary nutrient intake in free-ranging mammalian herbivores.Crossref | GoogleScholarGoogle Scholar | 19087435PubMed |
Mengistu A, Kebede G, Feyissa F, Assefa G (2017) Review on major feed resources in Ethiopia: conditions, challenges and opportunities. Academic Research Journal of Agricultural Science and Research 5, 176–185.
| Review on major feed resources in Ethiopia: conditions, challenges and opportunities.Crossref | GoogleScholarGoogle Scholar |
NDA (2011) ‘Natural database for Africa, version 2.0.’ (NDA: Addis Ababa, Ethiopia)
Oliván M, Dove H, Mayes RW, Hoebee SE (1999) Recent developments in the use of alkanes and other plant wax components to estimate intake and diet composition in herbivores. Revista Portuguesa de Zootecnia VI, 1–25.
Oliván M, Ferreira LMM, García U, Celaya R, Osoro K (2007) Application of n-alkanes as diet composition markers in grazing/browsing goats and sheep: effect of using different faecal recovery corrections and plant species grouping approaches. Australian Journal of Agricultural Research 58, 1013–1022.
| Application of n-alkanes as diet composition markers in grazing/browsing goats and sheep: effect of using different faecal recovery corrections and plant species grouping approaches.Crossref | GoogleScholarGoogle Scholar |
Phillips DL, Gregg JW (2003) Source partitioning using stable isotopes: coping with too many sources. Oecologia 136, 261–269.
| Source partitioning using stable isotopes: coping with too many sources.Crossref | GoogleScholarGoogle Scholar | 12759813PubMed |
Piasentier E, Sacca E, Bovolenta S (2007) Dietary selection and ingestive behaviour of fallow deer and sheep grazing on adjacent monocultures of white clover and tall fescue. Small Ruminant Research 71, 222–233.
| Dietary selection and ingestive behaviour of fallow deer and sheep grazing on adjacent monocultures of white clover and tall fescue.Crossref | GoogleScholarGoogle Scholar |
Samuels L, Kunst L, Jetter R (2008) Sealing plant surfaces: cuticular wax formation by epidermal cells. Annual Review of Plant Biology 59, 683–707.
| Sealing plant surfaces: cuticular wax formation by epidermal cells.Crossref | GoogleScholarGoogle Scholar | 18251711PubMed |
Schweizer M, Fear J, Cadisch G (1999) Isotopic (13C) fractionation during plant residue decomposition and its implications for soil organic matter studies. Rapid Communications in Mass Spectrometry 13, 1284–1290.
| Isotopic (13C) fractionation during plant residue decomposition and its implications for soil organic matter studies.Crossref | GoogleScholarGoogle Scholar | 10407311PubMed |
