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In vitro studies of the metabolism of [14C]-n-alkanes using ruminal fluid of sheep as substrate

Published online by Cambridge University Press:  01 December 2008

A. Keli
Affiliation:
The Macaulay Institute, Craigiebuckler, Aberdeen AB15 8QH, UK Departamento de Producción Animal y Ciencia de los Alimentos, Universidad de Zaragoza, Miguel Servet 177, 50013 Zaragoza, Spain
R. W. Mayes
Affiliation:
The Macaulay Institute, Craigiebuckler, Aberdeen AB15 8QH, UK
A. de Vega*
Affiliation:
Departamento de Producción Animal y Ciencia de los Alimentos, Universidad de Zaragoza, Miguel Servet 177, 50013 Zaragoza, Spain
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Abstract

Whether the rumen microbes are able to synthesize and/or degrade long-chain alkanes in anaerobic conditions remains a question to be answered before these hydrocarbons can be confidently used as duodenal flow or rumen transit markers. In this context, an experiment in vitro was carried out to establish whether within a rumen liquor fermentation system, n-alkanes can be derived from de-waxed structures of the plant or from non-alkane wax components (long-chain fatty alcohols, long-chain fatty acids and esters), or may be metabolized by bacteria to other components or to shorter-chain hydrocarbons. Ryegrass was labelled with 14C in growth chambers under controlled conditions in order to use it as a substrate. The labelled material obtained was separated in three fractions: labelled alkanes, labelled de-waxed plant and labelled wax components without the alkanes. These fractions were used for three different incubations in vitro, which objectives were as follows: 1. To check whether rumen bacteria can synthesize alkanes from carbon structures other than waxes (e.g. sugars). 2. To verify whether rumen bacteria can metabolize the n-alkanes to other compounds. 3. To check whether rumen bacteria can synthesize n-alkanes from other carbon compounds from waxes. The results showed that there was neither bacterial synthesis nor metabolism of the n-alkanes in in vitro conditions.

Type
Full Paper
Copyright
Copyright © The Animal Consortium 2008

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References

Askar, AR, Guada, JA, Balcells, J, de Vega, A, Castrillo, C 2005. Validation of use of purine bases as microbial marker by 15N labelling in growing lambs given high-concentrate diets: effects of grain processing, animal age and digesta sampling site. Animal Science 81, 5765.CrossRefGoogle Scholar
Bartley, EE, Helmer, LG, Meyer, RM 1971. Metabolism of 14C labelled octadecane by cattle. Journal of Animal Science 33, 13511355.CrossRefGoogle 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, 12151225.CrossRefGoogle Scholar
Di Muccio, A, Lintas, C, Filos, M, Bernardini, MP 1984. Occurrence and distribution of n-alkanes in sucking-calf tissues. Ecotoxicology and Environmental Safety 8, 248253.CrossRefGoogle ScholarPubMed
Dove, H, Mayes, RW 1991. The use of plant wax alkanes as marker substances in studies of the nutrition of herbivores: a review. Australian Journal of Agricultural Research 42, 913952.CrossRefGoogle Scholar
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, 123139.CrossRefGoogle 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, 16801697 (Online Journal).CrossRefGoogle ScholarPubMed
Dove H, Mayes RW, Freer M, Coombe JB and Foot JZ 1989. Faecal recoveries of the alkanes of plant cuticular waxes in penned and in grazing sheep. Proceedings of the XVI International Grassland Congress, Nice, France, pp. 1093–1094.Google Scholar
Ferreira, LMM, Oliván, M, Rodrigues, MAM, García, U, 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, 16361646.CrossRefGoogle Scholar
Giráldez, FJ, Lamb, S, López, S, Mayes, RW 2004. Effects of carrier matrix and dosing frequency on digestive kinetics of even-chain alkanes and implications on herbage intake and rate of passage studies. Journal of the Science of Food and Agriculture 84, 15621570.CrossRefGoogle Scholar
Hawke, JC 1973. Lipids. In Chemistry and biochemistry of herbage (ed. GW Butler and RW Bailey), pp. 212263. Academic Press, London, UK.Google Scholar
Hungate, RE 1966. The rumen and its microbes. Academic Press, London, UK.Google Scholar
INRA 1978. Alimentation des ruminants. INRA Publications, Versailles, France.Google Scholar
Kafilzadeh, F, Parker, DS 1990. The use of n-alkanes as indigestible markers in studies on intestinal digestion in sheep. Animal Production 50, 578579.Google Scholar
Keli A 2006. Use of alkanes in sheep: their behaviour in the digestive tract and possible alternatives. PhD, Zaragoza University, Zaragoza, Spain.Google Scholar
Keli, A, Andueza, D, de Vega, AGuada, JA 2008. Validation of the n-alkane and NIRS techniques to estimate intake, digestibility and diet composition in sheep fed mixed lucerne: ryegrass diets. Livestock Science doi: 10.1016/j.livsci.2008.02.011.CrossRefGoogle Scholar
Kolattukudy, PE 1970. Plant waxes. Lipids 5, 259275.CrossRefGoogle Scholar
Kolattukudy, PE, Hankin, L 1966. Metabolism of a plant wax paraffin (n-nonacosane) in the rat. The Journal of Nutrition 90, 167174.CrossRefGoogle ScholarPubMed
Kunst, L, Samuels, AL 2003. Biosynthesis and secretion of plant cuticular wax. Progress in Lipid Research 42, 5180.CrossRefGoogle ScholarPubMed
Lester, DE 1979. Normal paraffins in living matter-occurrence metabolism and pathology. Progress in Food and Nutrition Science 3, 166.Google ScholarPubMed
Mayes, RW, Lamb, CS, Colgrove, PM 1986. The use of dosed and herbage n-alkanes as markers for the determination of herbage intake. The Journal of Agricultural Science 107, 161170.CrossRefGoogle Scholar
Mayes RW, Lamb CS and Colgrove PM 1988. Digestion and metabolism of dosed even-chain and herbage odd-chain n-alkanes in sheep. Proceedings of the 12th General Meeting of the European Grassland Federation, Dublin, Ireland, pp. 159–163.Google Scholar
McCarthy, RD 1964. Mammalian metabolism of straight chain saturated hydrocarbons. Biochimica et Biophysica Acta 84, 7479.Google ScholarPubMed
Ohajuruka, OA, Palmquist, DL 1991. Evaluation of n-alkanes as digesta markers in dairy cows. Journal of Animal Science 69, 17261732.CrossRefGoogle ScholarPubMed
Oliván, M, Ferreira, LMM, Celaya, R, Osoro, K 2007. Accuracy of the n-alkane technique for intake estimates in beef cattle using different sampling procedures and feeding levels. Livestock Science 106, 2840.CrossRefGoogle Scholar
Petrón, MJ, Antequera, T, Muriel, E, Tejeda, JF, Ventanas, J 2004. Linear hydrocarbons content of intramuscular lipids of dry-cured Iberian ham. Meat Science 66, 295300.CrossRefGoogle ScholarPubMed
Radwan, SS, Sorkhoh, NA 1993. Lipids of n-alkane-utilizing microorganisms and their application potential. Advances in Applied Microbiology 39, 2990.CrossRefGoogle Scholar
Robaina, A, Grainger, C, Moate, P, Davis, L 1993. The use of n-alkanes for estimating diet digestibility and silage intake of stall-fed dairy cows. In The alkane workshop. Harnessing technology for the beef industry. University of New England, Armidale, 7pp.Google Scholar
Sporman, AM, Widdel, F 2000. Metabolism of alkylbenzenes, alkanes, and others hydrocarbons in anaerobic bacteria. Biodegradation 11, 85105.CrossRefGoogle Scholar
Stakelum G and Dillon P 1990. Dosed and herbage alkanes as feed intake predictors with dairy cows: the effect of feeding level and frequency of perennial ryegrass. Proceedings of the VII European Grazing Workshop, Wageningen, Netherlands.Google Scholar
Tejeda, JF, García, C, Petrón, MJ, Andrés, AI, Antequera, T 2001. n-Alkane content of intramuscular lipids of Iberian fresh ham from different feeding systems and crossbreeding. Meat Science 57, 371377.CrossRefGoogle ScholarPubMed
Tulliez, JE, Bories, GF 1975a. Métabolisme des hydrocarbures paraffiniques et naphténiques chez les animaux superieurs. I. Retention des paraffines (normal, cyclo et ramifiées) chez le rat. Annales de la Nutrition et de l’Alimentation 29, 201211.Google Scholar
Tulliez, JE, Bories, GF 1975b. Métabolisme des hydrocarbures paraffiniques et naphténiques chez les animaux supérieurs. II. Accumulation et mobilisation chez le rat. Annales de la Nutrition et de l’Alimentation 29, 213221.Google Scholar