Abstract
The present study was designed to investigate the capability of mixed rumen protozoa to synthesize conjugated linoleic acid (CLA) from linoleic (LA) and vaccenic acids (VA). Rumen contents were collected from fistulated cows. The protozoal fraction was separated and washed several times with MB9 buffer and then resuspended in autoclaved rumen fluid. The suspensions were anaerobically incubated up to 18 h at 38.5 °C with substrates in the presence (P-AB) or the absence of antibacterial-agents (P-No-AB). Neither P-AB nor P-No-AB suspensions were capable of producing CLA from VA (11t-18:1). Linoleic acid was catabolized by P-No-AB to a greater extent than P-AB. Different isomers of CLA were synthesized by P-AB from LA. The 9c11t-CLA was predominant. Thirty seven percent of the maximum accumulated 9c11t-CLA was found in the P-AB suspension as early as 0.1 h into the incubation period. Accumulation of 10t12c-CLA in P-AB suspension was approximately 10.0 times lower than that of 9c11t-CLA. There were no significant productions of VA, 10t-18:1, and 18:0 in P-AB compared with the control, indicating that rumen protozoa have no ability to biohydrogenate CLA isomers. On the other hand, the concentrations of 10t-18:1, VA, and 18:0 in P-No-AB were greater (P < 0.05) compared with those in P-AB, indicating the role of symbiotic bacteria associated with P-No-AB in biohydrogenating CLA isomers. We concluded that mixed rumen protozoa are capable of synthesizing CLA from LA through isomerization reactions. However, they are incapable of metabolizing CLA further. They are also incapable of vaccenic acid biohydrogenation and/or desaturation.
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References
Chalupa W, Kutches AJ (1968) Biohydrogenation of linoleic-1–14C acid by rumen protozoa. J Anim Sci 27:1502–1508
Chin SF, Liu W, Storkson JM, Ha YL, Pariza MW (1992) Dietary sources of conjugated dienoic isomers of linoleic acid, a newly recognized class of anticarcinogens. J Food Comp Anal 5:185–197
Cruz-Hernandez C, Deng Z, Zhou J, Hill AR, Yurawecz MP, Delmonte P, Mossoba MM, Dugan MER, Kramer JKG (2004) Methods for analysis of conjugated linoleic acids and trans-18:1 isomers in dairy fats by using a combination of gas chromatography, silver-ion thin-layer chromatography/gas chromatography, and silver-ion liquid chromatography. J AOAC Int 87:545–562
Denholm AM, Ling JR (1989) In vitro metabolism of 2,2′-diaminopimelic acid from gram-positive and gram-negative bacterial cells by ruminal protozoa and bacteria. Appl Environ Microbiol 55:212–218
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 96:697–704
Devillard E, McIntosh FM, Duncan SH, Wallace RJ (2007) Metabolism of linoleic acid by human gut bacteria: different routes for biosynthesis of conjugated linoleic acid. J Bacteriol 189:2566–2570
Emmanuel B (1974) On the origin of rumen protozoan fatty acids. Biochim Biophys Acta 337:404–413
Fukuda S, Suzuki Y, Murai M, Asanuma N, Hino T (2006) Isolation of a novel strain of Butyrivibrio fibrisolvens that isomerizes linoleic acid to conjugated linoleic acid without hydrogenation, and its utilization as a probiotic for animals. J Appl Microbiol 100:787–794
Griinari JM, Bauman DE (1999) Biosynthesis of conjugated linoleic acid and its incorporation into meat and milk in ruminants. In: Yurawecz MP, Mossoba MM, Kramer JKG, Pariza MW, Nelson GJ (eds) Advances in Conjugated Linoleic Acid Research, vol 1. AOCS, Champaign, pp 180–200
Harfoot CG (1978) Lipid metabolism in the rumen. Prog Lipid Res 17:21–54
Harfoot CG, Hazlewood GP (1997) Lipid metabolism in the rumen. In: Hobson PN, Stewart CS (eds) The rumen microbial ecosystem. Blackie and Prof., London, UK, pp 382–426
Kim YJ, Liu RH, 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 92:976–982
Lee SS, Ha JK, Cheng K-J (2000) Relative contributions of bacteria, protozoa, and fungi to in vitro degradation of orchard grass cell walls and their interactions. Appl Environ Microbiol 66:3807–3813
Müller A, Ringseis R, Düsterloh K, Gahler S, Eder K, Steinhart H (2005) Detection of conjugated dienoic fatty acids in human vascular smooth muscle cells treated with conjugated linoleic acid. Biochim Biophys Acta 1737:145–151
Nagaraja TG, Newbold CJ, Van Nevel CJ, Demeyer DI (1997) Manipulation of rumen fermentation. In: Hobson PN, Stewart CS (eds) The rumen microbial ecosystem. Blackie and Prof., London, UK, pp 523–632
Nam IS, Garnsworthy PC (2007) Biohydrogenation of linoleic acid by rumen fungi compared with rumen bacteria. J Appl Microbiol 103:551–556
Odongo NE, Or-Rashid MM, Kebreab E, France J, McBride BW (2007) Effect of supplementing myristic acid in dairy cow rations on ruminal methanogenesis and fatty acid profile in milk. J Dairy Sci 90:1851–1858
Or-Rashid MM, Onodera R, Wadud S (2001) Biosynthesis of methionine from homocysteine, cystathionine and homoserine plus cysteine by mixed rumen microorganisms in vitro. Appl Microbiol Biotechnol 55:758–764
Or-Rashid MM, Odongo NE, McBride BW (2007) Fatty acid composition of ruminal bacteria and protozoa, with emphasis on conjugated linoleic acid, vaccenic acid, and odd-chain and branched-chain fatty acids. J Anim Sci 85:1228–1234
Or-Rashid MM, Kramer JKG, Wood MA, McBride BW (2008) Supplemental algal meal alters the ruminal trans-18:1 fatty acid and conjugated linoleic acid composition in cattle. J Anim Sci 86:187–196
Pariza MW (2004) Perspective on the safety and effectiveness of conjugated linoleic acid. Am J Clin Nutr 79:1132–1136
Shingfield KJ, Ahvenjrvi S, Toivonen V, Ärölä A, Nurmela KVV, Huhtanen P, Griinari JM (2003) Effect of fish oil on biohydrogenation of fatty acids and milk fatty acid content in cows. Anim Sci 77:165–179
Toomey D, Harhen B, Roche HM, Fitzgerald D, Belton O (2005) Profound resolution of early atherosclerosis with conjugated linoleic acid. Atherosclerosis 187:40–49
Wahle KW, Heys SD, Rotondo D (2004) Conjugated linoleic acids: Are they beneficial or detrimental to health? Prog Lipid Res 43:553–587
Wallace RJ, McKain N, Shingfield KJ, Devillard E (2007) Isomers of conjugated linoleic acids are synthesized via different mechanisms in ruminal digesta and bacteria. J Lipid Res 48:2247–2254
Williams AG (1986) Rumen holotrich ciliate protozoa. Microbiol Rev 50:25–49
Williams AG, Coleman GS (1997) The rumen protozoa. In: Hobson PN, Stewart CS (eds) The rumen microbial ecosystem. Blackie and Prof., London, UK, pp 73–139
Yáñez-Ruiz DR, Scollan ND, Merry RJ, Newbold CJ (2006) Contribution of rumen protozoa to duodenal flow of nitrogen, conjugated linoleic acid and vaccenic acid in steers fed silages differing in their water-soluble carbohydrate content. Br J Nutr 96:861–869
Yurawecz MP, Kramer JKG, Ku Y (1999) Methylation procedures for conjugated linoleic acid. In: Yurawecz MP, Mossoba MM, Kramer JKG, Pariza MW, Nelson GJ (eds) Advances in Conjugated Linoleic Acid Research, vol 1. AOCS, Champaign, pp 64–82
Acknowledgements
The authors would like to thank the staff at the Elora Dairy Research Centre (University of Guelph, ON, Canada) for their technical assistance. We would also like to acknowledge the continued support received from the Ontario Ministry of Agriculture, Food and Rural Affairs (OMAFRA) and the Natural Sciences and Engineering Research Council of Canada (NSERC).
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Or-Rashid, M.M., AlZahal, O. & McBride, B.W. Studies on the production of conjugated linoleic acid from linoleic and vaccenic acids by mixed rumen protozoa. Appl Microbiol Biotechnol 81, 533–541 (2008). https://doi.org/10.1007/s00253-008-1690-0
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DOI: https://doi.org/10.1007/s00253-008-1690-0