Hostname: page-component-8448b6f56d-t5pn6 Total loading time: 0 Render date: 2024-04-19T18:56:51.464Z Has data issue: false hasContentIssue false
  • English
  • Français

Role of the protozoan Isotricha prostoma, liquid-, and solid-associated bacteria in rumen biohydrogenation of linoleic acid

Published online by Cambridge University Press:  01 July 2009

C. Boeckaert ,
D. P. Morgavi ,
J.-P. Jouany ,
L. Maignien ,
N. Boon  and
V. Fievez
C. Boeckaert
Affiliation:
Laboratory for Animal Nutrition and Animal Product Quality (Lanupro), Faculty of Bioscience Engineering, Ghent University, Proefhoevestraat 10, 9090 Melle, Belgium Laboratory of Microbial Ecology and Technology (LabMET), Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000 Gent, Belgium
D. P. Morgavi
Affiliation:
INRA, Herbivore Research Unit, Research Centre of Clermont Ferrand-Theix, 63122 St Genes-Champanelle, France
J.-P. Jouany
Affiliation:
INRA, Herbivore Research Unit, Research Centre of Clermont Ferrand-Theix, 63122 St Genes-Champanelle, France
L. Maignien
Affiliation:
Laboratory of Microbial Ecology and Technology (LabMET), Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000 Gent, Belgium
N. Boon
Affiliation:
Laboratory of Microbial Ecology and Technology (LabMET), Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000 Gent, Belgium
V. Fievez*
Affiliation:
Laboratory for Animal Nutrition and Animal Product Quality (Lanupro), Faculty of Bioscience Engineering, Ghent University, Proefhoevestraat 10, 9090 Melle, Belgium
*
E-mail: Veerle.Fievez@UGent.be
Get access

Abstract

From the simultaneous accumulation of hydrogenation intermediates and the disappearance of Isotricha prostoma after algae supplementation, we suggested a role of this ciliate and/or its associated bacteria in rumen biohydrogenation of unsaturated fatty acids. The experiments described here evaluated the role of I. prostoma and/or its associated endogenous and exogenous bacteria in rumen biohydrogenation of C18:2n-6 and its main intermediates CLA c9t11 and C18:1t11. Fractions of I. prostoma and associated bacteria, obtained by sedimentation of rumen fluid sampled from a monofaunated sheep, were used untreated, treated with antibiotics or sonicated to discriminate between the activity of I. prostoma and its associated bacteria, the protozoan or the bacteria, respectively. Incubations were performed in triplicate during 6 h with unesterified C18:2n-6, CLA c9t11 or C18:1t11 (400 μg/ml) and 0.1 g glucose/cellobiose (1/1, w/w). I. prostoma did not hydrogenate C18:2n-6 or its intermediates whereas bacteria associated with I. prostoma converted a limited amount of C18:2n-6 and CLA c9t11 to trans monoenes. C18:1t11 was not hydrogenated by either I. prostoma or its associated bacteria but was isomerized to C18:1c9. A phylogenetic analysis of clones originating from Butyrivibrio-specific PCR product was performed. This indicated that 71% of the clones from the endogenous and exogenous community clustered in close relationship with Lachnospira pectinoschiza. Additionally, the biohydrogenation activity of solid-associated bacteria (SAB) and liquid-associated bacteria (LAB) was examined and compared with the activity of the non-fractioned I. prostoma monofaunated rumen fluid (LAB + SAB). Both SAB and LAB were involved in rumen biohydrogenation of C18:2n-6. SAB fractions performed the full hydrogenation reaction to C18:0 while C18:1 fatty acids, predominantly C18:1t10 and C18:1t11, accumulated in the LAB fractions. SAB and LAB sequence analyses were mainly related to the genera Butyrivibrio and Pseudobutyrivibrio with 12% of the SAB clones closely related to the C18:0 producing B. proteoclasticus branch. In conclusion, this work suggests that I. prostoma and its associated bacteria play no role in C18:2n-6 biohydrogenation, while LAB convert C18:2n-6 to a wide range of C18:1 fatty acids and SAB produce C18:0, the end product of rumen lipid metabolism.

Keywords

protozoaButyrivibrioLachnospiraconjugated linoleic acidstrans fatty acids
Type
Full Paper
Information
animal , Volume 3 , Issue 7 , July 2009 , pp. 961 - 971
Copyright
Copyright © The Animal Consortium 2009

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Abaza, MA, Abou Akkada, AR, El-Shazly, K 1975. Effect of rumen protozoa on dietary lipid in sheep. Journal of Agricultural Science 85, 135–143.CrossRefGoogle Scholar
Abou Akkada, A, Howard, BH 1961. The biochemistry of rumen protozoa. 4. Decomposition of pectic substances. Biochemical Journal 78, 512–517.CrossRefGoogle ScholarPubMed
Bauman, DE, Baumgard, LH, Corl, BA, Griinari, JM 2000. Biosynthesis of conjugated linoleic acid in ruminants. Journal of Animal Science 77, 1ae–15ae.CrossRefGoogle Scholar
Boeckaert, C, Fievez, V, Van Hecke, D, Verstraete, W, Boon, N 2007. Changes in rumen biohydrogenation intermediates and ciliate protozoa diversity after algae supplementation to dairy cattle. European Journal of Lipid Science and Technology 109, 767–777.CrossRefGoogle Scholar
Boeckaert, C, Vlaeminck, B, Fievez, V, Maignien, L, Dijkstra, J, Boon, N 2008. Accumulation of trans C18:1 fatty acids in the rumen after algae supplementation is associated with changes in the Butyrivibrio community. Applied and Environmental Microbiology 74, 6923–6930.CrossRefGoogle ScholarPubMed
Boon, N, Top, EM, Verstraete, W, Siciliano, SD 2003. Bioaugmentation as a tool to protect the structure and function of an activated-sludge microbial community against a 3-chloroaniline shock load. Applied and Environmental Microbiology 69, 1511–1520.CrossRefGoogle ScholarPubMed
Broudiscou, LP, Lassalas, B 2000. Effects of Lavandula officinalis and Equisetum arvense dry extracts and isoquercitrin on the fermentation of diets varying in forage contents by rumen microorganisms in batch culture. Reproduction Nutrition Development 40, 431–440.CrossRefGoogle ScholarPubMed
Broudiscou, LP, Papon, Y, Broudiscou, A 1999. Effects of inorganic nitrogen and amino acids on the degradation of ammonia-treated barley straw and proteosynthesis in a continuous culture of rumen microbes. Animal Feed Science and Technology 77, 149–162.CrossRefGoogle Scholar
Chalupa, W, Kutches, AJ 1968. Biohydrogenation of linoleic-1-14C acid by rumen protozoa. Journal of Animal Science 27, 1502–1508.CrossRefGoogle Scholar
Chow, TT, Fievez, V, Moloney, AP, Raes, K, Demeyer, D, De Smet, S 2004. Effect of fish oil on in vitro rumen lipolysis, apparent biohydrogenation of linoleic and linolenic acid and accumulation of biohydrogenation intermediates. Animal Feed Science and Technology 117, 1–12.CrossRefGoogle Scholar
Coleman, GS 1978. Rumen entodiniomorphid protozoa. In Methods of cultivating parasites in vitro (ed. JR Baker and AER Taylor), pp. 39–54. Academic Press, London.Google Scholar
Cornick, NA, Jensen, NS, Stahl, DA, Hartman, PA, Allison, MJ 1994. Lachnospira pectinoschiza sp. nov., an anaerobic pectinophile from the pig intestine. International Journal of Systematic Bacteriology 44, 87–93.CrossRefGoogle ScholarPubMed
Dawson, RMC, Kemp, P 1969. The effect of defaunation on the phospholipids and on the hydrogenation of unsaturated fatty acids in the rumen. Biochemical Journal 115, 351–352.CrossRefGoogle ScholarPubMed
DeSantis, TZ, Hugenholtz, P, Larsen, N, Rojas, M, Brodie, EL, Keller, K, Huber, T, Dalevi, D, Hu, P, Andersen, GL 2006. Greengenes, a chimera-checked 16S rRNA gene database and workbench compatible with ARB. Applied and Environmental Microbiology 72, 5069–5072.CrossRefGoogle ScholarPubMed
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. British Journal of Nutrition 96, 697–704.Google ScholarPubMed
Dušková, D, Marounek, M 2001. Fermentation of pectin and glucose, and activity of pectin-degrading enzymes in the rumen bacterium Lachnospira multiparus. Letters in Applied Microbiology 33, 159–163.CrossRefGoogle ScholarPubMed
Folch, J, Lees, M, Sloane Stanley, SGH 1957. A simple method for the isolation and purification of total lipids from animal tissues. Journal of Biological Chemistry 226, 497–509.CrossRefGoogle ScholarPubMed
Girard, V, Hawke, JC 1978. The role of holotrichs in the metabolism of dietary linoleic acid in the rumen. Biochimica et Biophysica Acta 528, 17–27.CrossRefGoogle Scholar
Goering, HK, Van Soest, PJ 1970. Forage fiber analysis (apparatus, reagents, procedures, and some applications). Agriculture handbook 379. USDA-ARS, US Government Printing Office, Washington, DC, USA.Google Scholar
Gutierrez, J, Williams, PP, Davis, RE, Warwick, EJ 1962. Lipid metabolism of rumen ciliates and bacteria. 1. Uptake of fatty acids by Isotricha prostoma and Entodinium simplex. Applied Microbiology 10, 548–551.CrossRefGoogle ScholarPubMed
Harfoot, CG, Noble, RC, Moore, JH 1973. Food particles as a site for biohydrogenation of unsaturated fatty acids in the rumen. Biochemical Journal 132, 829–832.CrossRefGoogle ScholarPubMed
Jouany, J-P, Zainab, B, Senaud, J, Groliere, CA, Grain, J, Thivend, P 1981. Role of the rumen ciliate protozoa Polyplastron multivesiculatum, Entodinium sp., and Isotricha prostoma in the digestion of a mixed diet in sheep. Reproduction Nutrition Development 21, 871–884.CrossRefGoogle ScholarPubMed
Kemp, P, Lander, DJ 1984. Hydrogenation in vitro of alpha-linolenic acid to stearic acid by mixed cultures of pure strains of rumen bacteria. Journal of General Microbiology 130, 527–533.Google Scholar
Kemp, P, White, RW, Lander, DJ 1975. The hydrogenation of unsaturated fatty acids by five bacterial isolates from the sheep rumen, including a new species. Journal of General Microbiology 90, 100–114.CrossRefGoogle ScholarPubMed
Legay-Carmier, F, Bauchart, D 1989. Distribution of bacteria in the rumen contents of dairy cows given a diet supplemented with soya-bean oil. British Journal of Nutrition 61, 725–740.CrossRefGoogle Scholar
Ludwig, W, Strunk, O, Westram, R, Richter, L, Meier, H, Yadhukumar, , Buchner, A, Lai, T, Steppi, S, Jobb, G, Förster, W, Brettske, I, Gerber, S, Ginhart, AW, Gross, O, Grumann, S, Hermann, S, Jost, R, König, A, Liss, T, Lüßmann, R, May, M, Nonhoff, B, Reichel, B, Strehlow, R, Stamatakis, A, Stuckmann, N, Vilbig, A, Lenke, M, Ludwig, T, Bode, A, Schleifer, K-H 2004. ARB: a software environment for sequence data. Nucleic Acids Research 32, 1363–1371.CrossRefGoogle ScholarPubMed
Moon, CD, Pacheco, DM, Kelly, WJ, Leahy, SC, Li, D, Kopecny, J, Attwood, GT 2008. Reclassification of Clostridium proteoclasticum as Butyrivibrio proteoclasticus comb. nov., a butyrate-producing ruminal bacterium. International Journal of Systematic and Evolutionary Microbiology 58, 2041–2045.CrossRefGoogle Scholar
Morgavi, DP, Boudra, H, Jouany, JP, Michalet-Doreau, B 2004. Effect and stability of gliotoxin, an Aspergillus fumigatus toxin, on in vitro rumen fermentation. Food Additives and Contaminants 21, 871–878.CrossRefGoogle ScholarPubMed
Mosley, EE, McGuire, MA 2008. Biohydrogenation of vaccenic-1-13C acid by ruminal microbes in vitro. Journal of Animal Science 86 (E-suppl. 2), 88.Google Scholar
Onodera, R, Yamaguchi, H, Eguchi, C, Kandatsu, M 1977. Limits of survival of mingled rumen bacteria in washed cell suspension of rumen ciliate protozoa. Agricultural and Biological Chemistry 41, 2465–2466.Google Scholar
Or-Rashid, MM, AlZahal, O, McBride, BW 2008. Studies on the production of conjugated linoleid acid from linoleic and vaccenic acids by mixed rumen protozoa. Applied Microbiology and Biotechnology 81, 533–541.CrossRefGoogle Scholar
Paillard, D, McKain, N, Rincon, MT, Shingfield, KJ, Givens, DI, Wallace, RJ 2007a. Quantification of ruminal Clostridium proteoclasticum by real-time PCR using a molecular beacon approach. Journal of Applied Microbiology 103, 1251–1261.CrossRefGoogle ScholarPubMed
Paillard, D, McKain, N, Chaudhary, LC, Walker, ND, Pizette, F, Koppova, I, McEwan, NR, Kopecny, J, Vercoe, PE, Louis, P, Wallace, RJ 2007b. Relation between phylogenetic position, lipid metabolism and butyrate production by different Butyrivibrio-like bacteria from the rumen. Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology 91, 417–422.CrossRefGoogle ScholarPubMed
SAS Institute In c. 2004. Output delivery system: user’s guide. SAS Institute Inc., Cary, NC, USA.Google Scholar
Singh, S, Hawke, JC 1979. The in vitro lipolysis and biohydrogenation of monogalactosyldiglyceride by whole rumen contents and its fractions. Journal of the Science of Food and Agriculture 30, 603–612.CrossRefGoogle ScholarPubMed
Wallace, RJ, Chaudhary, LC, McKain, N, McEwan, NR, Richardson, AJ, Vercoe, PE, Walker, ND, Paillard, D 2006. Clostridium proteoclasticum: a ruminal bacterium that forms stearic acid from linoleic acid. FEMS Microbiology Letters 265, 195–201.CrossRefGoogle Scholar
Wang, Q, Garrity, GM, Tiedje, JM, Cole, JR 2007. Naïve Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Applied Environmental Microbiology 73, 5261–5267.CrossRefGoogle ScholarPubMed
Ward, PFV, Scott, TW, Dawson, MRC 1964. The hydrogenation of unsaturated fatty acids in the ovine digestive tract. Biochemical Journal 92, 60–68.CrossRefGoogle ScholarPubMed
Williams, AG, Coleman, GS 1992. The rumen protozoa. Springer, New York, USA.CrossRefGoogle Scholar
Williams, PP, Gutierrez, J, Davis, RE 1963. Lipid metabolism of rumen ciliates and bacteria. 2. Uptake of fatty acids and lipid analysis of Isotricha intestinalis and rumen bacteria with further information on Entodinium simplex. Applied Microbiology 11, 260–264.CrossRefGoogle Scholar
Wright, DE 1959. Hydrogenation of lipids by rumen protozoa. Nature 184, 875–876.CrossRefGoogle ScholarPubMed