Skip to main content

The effect of absence of protozoa on rumen biohydrogenation and the fatty acid composition of lamb muscle

  • David R. Yáñez-Ruiz (a1), Selwyn Williams (a1) and Charles J. Newbold (a1)

The effects of the absence of protozoa in the rumen of lambs on animal growth, rumen fermentation, microbial diversity and fatty acid profiles in abomasal fluid and intramuscular fat were measured in ten control and ten protozoa-free (PF) lambs. PF lambs were prepared by isolating them from ewes within the first 24 h after birth. The PF and control lambs were kept for 4 months in two different fields and received a daily supplement of 250 g concentrate. The bacterial populations visualised by denaturing gradient gel electrophoresis differed between the two groups of animals and showed a higher bacterial diversity in control lambs than in PF lambs. Abomasal contents from control lambs contained higher concentrations of 22 : 5n-3 and 22 : 6n-3 and lower concentrations of vaccenic acid (trans-11–18 : 1) and 20 : 3n-6 than PF lambs. The rest of the fatty acids, including the conjugated linoleic acid (CLA) isomers, were present at the same concentrations in abomasal contents from both experimental groups. Fatty acid composition in intramuscular fat showed differences between the groups. PF lambs had higher proportions of 18 : 0, 18 : 3, trans-10, cis-12-CLA and total SFA than control lambs. Control lambs had higher proportions of cis-9–18 : 1, cis-9, cis-12–18 : 2, 20 : 3n-6, 22 : 6n-3 (DHA) and MUFA. In conclusion, rumen defaunation led to higher tissue levels of the trans-10, cis-12-CLA isomer and SFA and lower PUFA:SFA ratio and n-3 PUFA in lamb muscle.

  • View HTML
    • Send article to Kindle

      To send this article to your Kindle, first ensure is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about sending to your Kindle. Find out more about sending to your Kindle.

      Note you can select to send to either the or variations. ‘’ emails are free but can only be sent to your device when it is connected to wi-fi. ‘’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

      Find out more about the Kindle Personal Document Service.

      The effect of absence of protozoa on rumen biohydrogenation and the fatty acid composition of lamb muscle
      Available formats
      Send article to Dropbox

      To send this article to your Dropbox account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Dropbox.

      The effect of absence of protozoa on rumen biohydrogenation and the fatty acid composition of lamb muscle
      Available formats
      Send article to Google Drive

      To send this article to your Google Drive account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Google Drive.

      The effect of absence of protozoa on rumen biohydrogenation and the fatty acid composition of lamb muscle
      Available formats
Corresponding author
*Corresponding author: Professor C. J. Newbold, fax +44 1970 611264, email
Hide All
Abaza, MA, Abou Akkada, AR & El Shazly, K (1975) Effect of rumen protozoa on dietary lipid in sheep. J Agric Sci 85, 135143.
Abou Akkada, AR & El Shazly, K (1964) Effect of absence or presence of rumen ciliate protozoa on some blood components, nitrogen retention and digestibility of food constituents in lambs. J Agric Sci 64, 251255.
Ankom (2000) Procedures for fibre and in vitro analysis. Accessed 14 December 2000.
Aurousseau, B, Bauchart, D, Calichon, E, Micol, D & Priolo, A (2004) Effect of grass or concentrate feeding systems and rate of growth on triglyceride and phospholipids and their fatty acids in the M. longissimus thoracis of lambs. Meat Sci 66, 531541.
Bauman, DE & Griinari, JM (2003) Nutritional regulation of milk fat synthesis. Annu Rev Nutr 23, 203227.
Bauman, DEL, Baumgard, BA, Corl, BA & Griinari, JM (1999) Biosynthesis of conjugated linoleic acids in ruminants. Proc Am Soc Anim Sci, Accessed 1 September 2002.
Bird, SH & Leng, RA (1984) Further studies on the effects of the presence or absence of protozoa in the rumen of live-weight gain and wool growth of sheep. Br J Nutr 52, 607611.
Blankson, H, Stakkestad, JA, Fagertun, H, Thom, E, Wadstein, J & Gudmundsen, O (2000) Conjugated linoleic acid reduces body fat mass in overweight and obese humans. J Nutr 130, 29432948.
Boles, JA, Kott, RW, Hatfield, PG, Bergman, JW & Flynn, CR (2005) Supplemental safflower oil affects the fatty acid profile, including conjugated linoleic acid, of lamb. J Anim Sci 83, 21752181.
Bolte, MR, Hess, BW, Means, WJ, Moss, GE & Rule, DC (2002) Feeding lambs high-oleate or high-linoleate safflower seeds differentially influences carcass fatty acid composition. J Anim Sci 80, 609616.
Bonhomme, A (1990) Rumen ciliates: their metabolism and relationship with bacteria and their hosts. Anim Feed Sci Technol 30, 203266.
Chalupa, A & Kutches, AJ (1968) Biohydrogenation of linoleic 1-C14 acid by rumen protozoa. J Anim Sci 27, 15021508.
Chin, SF, Liu, W, Storkson, JM, Ha, YL & Pariza, WM (1992) Dietary sources of conjugated dienoic isomers of linoleic acid, a newly recognized class of anticarcinogens. J Food Compos Anal 5, 185197.
Coleman, GS (1964) The metabolism of Escherichia coli and other bacteria by Entodinium caudatum. J Gen Microbiol 47, 449464.
Coleman, GS (1986) The metabolism of rumen ciliate protozoa. FEMS Microbiol Rev 39, 321344.
Coleman, GS (1988) The importance of rumen ciliate protozoa in the growth and metabolism of the host ruminant. Int J Anim Sci 3, 7595.
Collombier, J (1981) Contribution à l'étude du role des protozoaires cilies du rumen dans l'apport d'azote microbien entrant dans le duodenum du ruminant (Contribution to the study of the role of rumen ciliate protozoa in the microbial nitrogen contribution entering the duodenum of ruminants). PhD Thesis, Université Clermont-Ferrand II.
Cooper, SL, Sinclair, LA, Wilkinson, RG, Hallet, KG, Enser, M & Wood, JD (2004) Manipulation of the n-3 polyunsaturated fatty acid content of muscle and adipose tissue in lambs. J Anim Sci 82, 14611470.
Dawson, RMC & Kemp, P (1969) The effect of defaunation on the phospholipids and on the hydrogenation of unsaturated fatty acids in the rumen. Biochem J 115, 351352.
De la Torre, A, Debiton, E, Durand, D, Chardigny, JM, Berdeaux, O, Loreau, O, Barthomeuf, C, Bauchart, D & Gruffat, D (2005) Conjugated linoleic acid isomers and their conjugated derivatives inhibit growth of human cancer cell lines. Anticancer Res 25, 39433949.
Demeyer, DI, Van Nevel, CJ & Van de Voorde, G (1982) The effect of defaunation on the growth of lambs fed three urea-containing diets. Arch Tierernähr 32, 595604.
Department of Health (1994) Nutritional Aspects of Cardiovascular Disease. Report on Health and Social Subjects no. 46. London: H.M. Stationery Office.
Devillard, E, McIntosh, FM, Newbold, CJ & Wallace, RJ (2006) Rumen ciliate protozoa contain high concentrations of conjugated linoleic acids and vaccenic acid, yet do not hydrogenate linoleic acid or desaturate stearic acid. Br J Nutr 96, 697704.
Díaz, MT, Álvarez, I, De la Fuente, J, et al. (2005) Fatty acid composition of meat from typical lamb production systems of Spain. United Kingdom, Germany and Uruguay. Meat Sci 71, 256263.
Duckett, SK, Andrae, JG & Owens, FN (2002) Effect of high-oil corn or added corn oil on ruminal biohydrogenation of fatty acids and conjugated linoleic acid formation in beef steers fed finishing diets. J Anim Sci 80, 33533360.
Dunbar, J, Barns, SM, Ticknor, LO & Kuske, CR (2002) Empirical and theoretical bacterial diversity in four Arizona soils. Appl Environ Microbiol 68, 30353045.
Eadie, JM & Gill, JC (1971) The effect of the absence of rumen ciliate protozoa on growing lambs fed on a roughage-concentrate diet. Br J Nutr 26, 155167.
Enser, M, Hallet, K, Hewitt, B, Fursey, GAJ, Wood, JD & Harrington, G (1998) Fatty acid composition of UK beef anf lamb muscle in relation to production system and implications for human nutrition. Meat Sci 49, 329341.
Felsenstein, F (2002) PHYLIP (Phylogeny Inference Package) version 3·6a. Seattle: University of Washington.
Folch, J, Lees, M & Sloane Stanley, GH (1957) A simple method for the isolation and purification of lipides from animal tissues. J Biol Chem 226, 497509.
Gardan, D, Gondret, F & Louveau, I (2006) Lipid metabolism and secretory function of porcine intramuscular adipocytes compared with subcutaneous and perineal adipocytes. Am J Physiol 291, E372E380.
Girard, V & Hawke, JC (1978) The role of holotrichs in the metabolism of dietary linoleic acid in the rumen. Biochim Biophys Acta 528, 404413.
Griinari, JM, Corl, BA, Lacy, SH, Chouinard, PY, Nurmela, KV & Bauman, DE (2000) Conjugated linoleic acid is synthesized endogenously in lactating dairy cows by Δ9-desaturase. J Nutr 130, 22852291.
Hall, FJ, West, J & Coleman, GS (1974) Fine structural studies on the digestion of chloroplasts in the rumen ciliate Entodinium caudatum. Tissue Cell 6, 243253.
Harfoot, GC & Hazlewood, GP (1997) Lipid metabolism in the rumen. In The Rumen Microbial Ecosystem, pp. 383425 [Hobson, PN and Stewart, CS, editors]. London: Blackie Academic and Professional.
Hsu, JT, Fahey, GC, Berger, LL, Mackie, RI & Menden, NR (1991) Manipulation of nitrogen digestion by sheep using defaunation and various nitrogen supplementation regimes. J Anim Sci 69, 12901297.
Ivan, M, Koening, KM, Morgavi, DP, Rode, LM, Newbold, CJ & Entz, T (2006) Duodenal flow and digestibility in fauna-free sheep and in sheep monofauntaed with Entodinium caudatum or Polypastron multivesiculatum. Br J Nutr 95, 469476.
Ivan, M, Neill, L, Foster, R, Alimon, R, Rode, LM & Entz, T (2000) Effects of Isotricha, Dasytricha, Entodinium, and total fauna on ruminal fermentation and duodenal flow in wethers fed different diets. J Dairy Sci 83, 776787.
Jouany, JP (1995) Effect of rumen protozoa on nitrogen utilization by ruminants. J Nutr 1335S1346S.
Jouany, JP & Lassalas, B (2003) Effect of rumen protozoa on biohydrogenation of polyunsaturated fatty acids. In Proceedings of the International Conference on the Molecular Biology and Biotechnology of Ciliates and Anaerobic Protozoa, p. 94. Nijmegen, The Netherlands: University of Nijmegen.
Jouany, JP, Demeyer, DI & Grain, J (1988) Effect of defaunating the rumen. Anim Feed Sci Technol 21, 229265.
Kemp, P & Lander, DJ (1984) Hydrogenation in vitro of α-linolenic acid to stearic acid by mixed rumen bacteria. J Gen Microbiol 130, 527533.
Kim, YJ, Liu, RM, Rychlik, JL & Russell, JB (2002) The enrichment of a ruminal bacterium (Megasphaera elsdenii YJ-4) that produces the trans-10, cis-12 isomer of conjugated linoleic acid. J Appl Microbiol 92, 976982.
Klopfenstein, TJ, Purser, DB & Tyznik, WJ (1966) Effects of defaunation on feed digestibility, rumen metabolism and blood metabolites. J Anim Sci 25, 765–773.
Koenig, KM, Newbold, CJ, McIntosh, FM & Rode, LM (2000) Effects of protozoa on bacterial nitrogen recycling in the rumen. J Anim Sci 78, 24312445.
Kramer, JKG & Zhou, JQ (2001) Conjugated linoleic acid and octadecenoic acids: extraction and isolation of lipids. Eur J Lipid Sci Technol 103, 594–600.
Kurihara, Y, Eadie, JM, Hobson, PN & Mann, SO (1968) Relationship between bacteria and ciliate protozoa in the sheep rumen. J Gen Microbiol 51, 267–288.
Kurihara, Y, Takechi, T & Shibata, F (1978) Relationship between bacteria and ciliate protozoa in the rumen of sheep fed on a purified diet. J Agric Sci 90, 373381.
Labarca, C & Paigen, K (1980) A simple, rapid, and sensitive DNA assay procedure. Anal Biochem 102, 344–352.
Lough, AK (1968) Component of fatty acids of plasma lipids of lambs with and without rumen ciliate protozoa. Proc Nutr Soc 27, 30A–31A.
McNabb, WC, Waghorn, GC, Barry, TN & Shelton, ID (1993) The effect of condensed tannins in Lotus pedunculatus on the digestion and metabolism of methionine, cystine and inorganic sulphur in sheep. Br J Nutr 70, 647661.
Maeda, H, Fujimoto, C, Haruki, Y, Maeda, T, Kokeguchi, S, Petelin, M, Arai, H, Tanimoto, I, Nishimura, F & Takashiba, S (2003) Quantitative real-time PCR using TaqMan and SYBR Green for Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, tetQ gene and total bacteria. FEMS Immunol Med Microbiol 39, 81–86.
Marrugan, A (1988) Ecological Diversity and its Measurement. London: Chapman and Hall.
Medina, EA, Horn, WF, Keim, NL, Havel, PJ, Benito, P, Kelley, DS, Nelson, GJ & Erickson, KL (2000) Conjugated linoleic acid supplementation in humans: effects on circulating leptin concentrations and appetite. Lipids 35, 783788.
Muyzer, G, de Waal, EC & Uitterlinden, GA (1993) Profiling of complex populations by denaturing gradient gel electrophoresis of polymerase chain reaction-amplified genes coding for 16S rRNA. App Environ Microbiol 59, 695–700.
Nubel, U, Engelen, B, Felske, A, Snaidr, J, Wieshuber, A, Amann, RI, Ludwing, W & Backhaus, H (1996) Sequence heterogeneities of genes encoding 16S rRNAs in Paenibacillus polymyxa detected by temperature gradient gel electrophoresis. J Bacteriol 178, 56365643.
Ogawa, J, Kishino, S, Ando, A, Sugimoto, S, Mihara, K & Shimizu, S (2005) Production of conjugated fatty acids by lactic acid bacteria. J Biosci Bioeng 100, 355–364.
Ozutsumi, Y, Tajima, K, Takenaka, A & Itabashi, H (2005) The effect of protozoa on the composition of rumen bacteria in cattle using 16S rRNA gene clone libraries. Biosci Biotechnol Biochem 69, 499506.
Raes, K, De Smet, S & Demeyer, D (2004) Effect of dietary fatty acids on incorporation of long chain polyunsaturated fatty acids and conjugated linoleic acid in lamb, beef and pork meat: a review. Anim Feed Sci Technol 113, 199–221.
Ramprasad, J & Raghacan, GV (1981) Note of the growth rate and body composition of faunated and defaunated lambs. Indian J Anim Sci 51, 570572.
Regensbogenova, MP, Pristas, P, Javorsky Hackstein, JHP, Newbold, CJ & McEwan, N (2004) Assessment of ciliates in the sheep rumen by DGGE. Lett Appl Microbiol 39, 144–147.
Riserus, U, Berglund, L & Vessby, B (2001) Conjugated linoleic acid (CLA) reduced abdominal adipose tissue in obese middle-aged men with signs of the metabolic syndrome: a randomised controlled trial. Int J Obes 25, 11291135.
Roche, HM, Noone, E, Sewter, C, Mc Bennett, S, Savage, D, Gibney, MJ, O'Rahilly, S & Vidal-Puig, AJ (2002) Isomer-dependent metabolic effects of conjugated linoleic acid: insights from molecular markers sterol regulatory element-binding protein-1c and LXRα. Diabetes 51, 20372044.
Rooke, JA, Alison, J & Borman Armstrong, DG (1990) The effect of inoculation with Lactobacillus plantarum on fermentation in laboratory silos of herbage low in water-soluble carbohydrate. Grass Forage Sci 45, 143–152.
Rowe, A, Macedo, FA, Visentainer, JV, Souza, NE & Matsushita, M (1999) Muscle composition and fatty acid profile in lambs fattened in drylot or pasture. Meat Sci 51, 283288.
Sackmann, JR, Duckett, SK, Gillis, MH, Realini, CE, Parks, AH & Eggelston, RB (2003) Effects of forage and sunflower oil levels on ruminal biohydrogenation of fatty acids and conjugated linoleic acid formation in beef steers fed finishing diets. J Anim Sci 81, 31743181.
Santra, A & Karim, SA (2002) Influence of ciliate protozoa on biochemical changes and hydrolytic enzyme profile in the rumen ecosystem. J Appl Microbiol 92, 801–811.
Stewart, CS, Flint, HJ & Bryant, MP (1997) The rumen bacteria. In The Rumen Microbial Ecosystem, pp. 10–72 [Hobson, PN and Stewart, CS, editors]. London: Blackie Academic and Professional.
Stockdale, CR & King, KR (1983) A comparison of two techniques used to estimate the herbage intake of lactating dairy cows in a grazing environment. J Agric Sci 100, 227230.
Sukhija, PS & Palmquist, DL (1988) Rapid method for determination of total fatty acid content and composition of feedstuffs and feces. J Agric Food Chem 36, 12021206.
Sutton, JD, Knight, R, McAllan, AB & Smith, RH (1983) Digestion and synthesis in the rumen of sheep given diets supplemented with free and protected oils. Br J Nutr 49, 419–432.
Sylvester, JT, Karnati, SK, Yu, Z, Morrrison, M & Firkins, JL (2004) Development of an assay to quantify ciliate protozoal biomass in cows using real-time PCR. J Nutr 134, 33783384.
Sylvester, JT, Karnati, SK, Yu, Z, Newbold, CJ & Firkins, JL (2005) Evaluation of a real time PCR assay quantifying the ruminal pool size and duodenal flow of protozoal nitrogen. J Dairy Sci 88, 2083–2095.
Toomey, S, Harhen, B, Roche, HM, Fitzgerald, D & Belton, O (2005) Profound resolution of early atherosclerosis with conjugated linoleic acid. Atherosclerosis 20, 232–238.
Turpeinen, AM, Mutanen, M, Aro, A, Salminen, I, Basu, S, Palmquist, DL & Griinari, JM (2002) Bioconversion of vaccenic acid to conjugated linoleic acid by humans. Am J Clin Nutr 76, 504510.
Ushida, K, Jouany, JP & Thivend, P (1986) Role of protozoa in nitrogen digestion in sheep given two isonitrogenous diets. Br J Nutr 56, 407–419.
Van de Vossenberg, JLCM & Joblin, KN (2003) Biohydrogenation of C18 unsaturated fatty acids to stearic acid by a strain of Butyrivibrio hungatei from the bovine rumen. Lett App Microbiol 37, 424428.
Van Nevel, CJ, Demeyer, DI & Van de Voorde, G (1985) Effect of defaunating the rumen on growth and carcass composition of lambs. Arch Anim Nutr 35, 331–337.
van Soest, PJ & Mason, VC (1991) The influence of the Maillard reaction upon the nutritive value of fibrous feeds. Animal Feed Sci Technol 32, 45–53.
Veira, DM (1986) The role of ciliate protozoa in nutrition of the ruminant. J Anim Sci 63, 15471560.
Vogels, GD, Hoppe, WF & Stumm, CK (1980) Association of methahanogenic bacteria with rumen ciliates. Appl Environ Microbiol 40, 608–612.
Wachira, AM, Sinclair, LA, Wilkinson, RG, Enser, M, Wood, JD & Fisher, V (2002) Effects of dietary fat source and breed on the carcass composition n-3 polyunsaturaed fatty acid and conjugated linoleic acid content of sheep meat and adipose tissue. Br J Nutr 88, 697709.
Wahle, WJK, Heys, SD & Rotondo, D (2004) Conjugated linoleic acids: are they beneficial or detrimental to health? Prog Lipid Res 43, 553–587.
Weatherburn, MW (1967) Phenol hypochlorite reaction for determination of ammonia. Anal Chem 89, 971974.
Whitelaw, FG, Eadie, JM, Mann, SO & Reid, RS (1972) Some effects of rumen ciliate protozoa in cattle given restricted amounts of barley diet. Br J Nutr 27, 425–437.
Williams, AG & Coleman, GS (1992) The Rumen Protozoa. New York: Springer-Verlag.
Williams, AG & Withers, SE (1990) Effect of ciliate protozoa on the activity of polysaccharide-degrading enzymes and fibre breakdown in the rumen ecosystem. J App Bacteriol 70, 144–155.
Wolin, MJ, Miller, TL & Stewart, CS (1997) Microbe-microbe interactions. In The Rumen Microbial Ecosystem, pp. 467491 [Hobson, PN and Stewart, CS, editors]. London: Blackie Academic and Professional.
Yáñez-Ruiz, DR, Scollan, ND, Merry, RJ & Newbold, CJ (2006) Contribution of rumen protozoa to duodenal flow of nitrogen, conjugated linoleic acid and trans-vaccenic acid in steers fed silages differing in their water-soluble carbohydrate content. Br J Nutr 96, 861869.
Zhang, H, Guo, Y & Yuan, J (2005) Conjugated linoleic acid enhanced the immune function in broiler chicks. Br J Nutr 94, 746–752.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

British Journal of Nutrition
  • ISSN: 0007-1145
  • EISSN: 1475-2662
  • URL: /core/journals/british-journal-of-nutrition
Please enter your name
Please enter a valid email address
Who would you like to send this to? *



Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed