Skip to main content Accessibility help

Effect of fibrolytic enzymes added to a Andropogon gayanus grass silage-concentrate diet on rumen fermentation in batch cultures and the artificial rumen (Rusitec)

  • G. O. Ribeiro (a1) (a2), L. C. Gonçalves (a1), L. G. R. Pereira (a3), A. V. Chaves (a4), Y. Wang (a2), K. A. Beauchemin (a2) and T. A. McAllister (a2)...


In vitro batch cultures were used to screen four fibrolytic enzyme mixtures at two dosages added to a 60 : 40 silage : concentrate diet containing the C4 tropical grass Andropogon gayanus grass ensiled at two maturities – vegetative stage (VS) and flowering stage (FS). Based on these studies, one enzyme mixture was selected to treat the same diets and evaluate its impact on fermentation using an artificial rumen (Rusitec). In vitro batch cultures were conducted as a completely randomized design with two runs, four replicates per run and 12 treatments in a factorial arrangement (four enzyme mixtures×three doses). Enzyme additives (E1, E2, E3 and E4) were commercial products and contained a range of endoglucanase, exoglucanase and xylanase activities. Enzymes were added to the complete diet 2 h before incubation at 0, 2 and 4 μl/g of dry matter (DM). Gas production (GP) was measured after 3, 6, 12, 24 and 48 h of incubation. Disappearance of DM (DMD), NDF (NDFD) and ADF (ADFD) were determined after 24 and 48 h. For all four enzyme mixtures, a dosage effect (P<0.05) was observed for NDFD and ADFD after 24 h and for DMD, NDFD and ADFD after 48 h of incubation of the VS diet. For the FS diet, a dosage effect was observed for GP and NDFD after 24 h and for GP, DMD, NDFD and ADFD after 48 h of incubation. There was no difference among enzyme mixtures nor was there an enzyme×dose interaction for the studied parameters. Because of the greatest numerical effect on NDF disappearance and the least cost price, enzyme mixture E2 at 4 µl/g of diet DM was selected for the Rusitec experiment. The enzyme did not impact (P>0.05) DM, N, NDF or ADF disappearance after 48 h of incubation nor daily ammonia-N, volatile fatty acids or CH4 production. However, enzyme application increased (P<0.05) microbial N production in feed particle-associated (loosely-associated) and silage feed particle-bound (firmly associated) fractions. With A. gayanus silage diets, degradation may not be limited by microbial colonization, but rather by the ability of fibrolytic enzymes to degrade plant cell walls within this recalcitrant forage.


Corresponding author


Hide All
Association of Official Analytical Chemists (AOAC) 2006. Official methods of analysis, 18th edition. AOAC, Arlington, VA, USA.
Avellaneda, JH, Pinos-Rodríguez, JM, González, SS, Bárcena, R, Hernández, A, Cobos, M, Hernández, D and Montañez, O 2009. Effects of exogenous fibrolytic enzymes on ruminal fermentation and digestion of Guinea grass hay. Animal Feed Science and Technology 149, 7077.
Bailey, RW 1967. Quantitative studies of ruminant digestion of ingested plant carbohydrates from the reticulo-rumen. New Zealand Journal of Agricultural Research 10, 1532.
Beauchemin, KA, Colombatto, D, Morgavi, DP and Yang, W 2003. Use of exogenous fibrolytic enzymes to improve feed utilization by ruminants. Journal of Animal Science 81, E37E47.
Canadian Council on Animal Care 1993. Guide to the care and use of experimental animals. Canadian Council on Animal Care, Ottawa, Ontario, Canada.
Centro Internacional de Agricultura Tropical 1990. Andropogon gayanus Kunth: a grass for tropical acid soils. Centro Internacional de Agricultura Tropical, Cali, Colombia.
Cheng, K-J and McAllister, TA 1997. Compartmentation in the rumen. In The rumen microbial ecosystem (ed. PN Hobson and CS Stewart), pp. 492552. Blackie Academic & Press, London, UK.
Colombatto, D and Beauchemin, KA 2003. A proposed methodology to standardize the determination of enzymic activities present in enzyme additives used in ruminant diets. Canadian Journal of Animal Science 83, 559568.
Colombatto, D, Mould, FL, Bhat, MK and Owen, E 2003a. Use of fibrolytic enzymes to improve the nutritive value of ruminant diets: a biochemical and in vitro rumen degradation assessment. Animal Feed Science and Technology 107, 201209.
Colombatto, D, Mould, FL, Bhat, MK, Morgavi, DP, Beauchemin, KA and Owen, E 2003b. Influence of fibrolytic enzymes on the hydrolysis and fermentation of pure cellulose and xylan by mixed ruminal microorganisms in vitro . Journal of Animal Science 81, 10401050.
Czerkawski, JW and Breckenridge, G 1977. Design and development of a long-term rumen simulation technique (Rusitec). British Journal of Nutrition 38, 371384.
Dean, DB, Adesogan, AT, Krueger, NA and Littell, RC 2008. Effects of treatment with ammonia or fibrolytic enzymes on chemical composition and ruminal degradability of hays produced from tropical grasses. Animal Feed Science and Technology 145, 6883.
Eun, JS and Beauchemin, KA 2007. Assessment of the efficacy of varying experimental exogenous fibrolytic enzymes using in vitro fermentation characteristics. Animal Feed Science and Technology 132, 298315.
Facchini, FDA, Reis, VRA, Roth, AP, Magalhães, KA, Peixoto-Nogueira, SC, Casagrande, DR, Reis, RA and MDLTM, Polizeli 2012. Effects of Aspergillus spp. exogenous fibrolytic enzymes on in vitro fermentation of tropical forages. Journal of Science Food and Agriculture 92, 25692573.
Fedorak, PM and Hrudey, SE 1983. A simple apparatus for measuring gas-production by methanogenic cultures in serum bottles. Environmental Technology Letters 4, 425432.
Giraldo, LA, Ranilla, MJ, Tejido, ML and Carro, MD 2007a. Influence of exogenous fibrolytic enzymes and fumarate on methane production, microbial growth and fermentation in Rusitec fermenters. British Journal of Nutrition 98, 753761.
Giraldo, LA, Tejido, ML, Ranilla, MJ and Carro, MD 2007b. Effects of exogenous cellulase supplementation on microbial growth and ruminal fermentation of a high-forage diet in Rusitec fermenters. Journal of Animal Science 85, 19621970.
Jung, HG and Allen, MS 1995. Characteristics of plant cell walls affecting intake and digestibility of forages by ruminants. Journal of Animal Science 73, 27742790.
Karkalas, J 1985. An improved enzymatic method for determination of native or modified starch. Journal of Science Food and Agriculture 36, 10191027.
Mandebvu, P, West, JW, Froetschel, MA, Hatfield, RD, Gates, RN and Hill, GM 1999. Effect of enzyme or microbial treatment of bermudagrass forages before ensiling on cell wall composition, end products of silage fermentation and in situ digestion kinetics. Animal Feed Science and Technology 77, 317329.
McDougall, EI 1948. Studies on ruminant saliva. 1. The composition and output of sheep’s saliva. Biochemical Journal 43, 99109.
Meale, SJ, Beauchemin, KA, Hristov, AN, Chaves, AV and McAllister, TA 2014. BOARD-INVITED REVIEW: opportunities and challenges in using exogenous enzymes to improve ruminant production. Journal of Animal Science 92, 427442.
Menke, KH, Raab, L, Salewski, A, Steingass, H, Fritz, D and Schneider, W 1979. Estimation of the digestibility and metabolizable energy content of ruminant feedingstuffs from the gas-production when they are incubated with rumen liquor in vitro . Journal of Agricultural Science 93, 217222.
Morgavi, DP, Beauchemin, KA, Nsereko, VL, Rode, LM, Iwaasa, AD, Yang, WZ, McAllister, TA and Wang, Y 2000. Synergy between ruminal fibrolytic enzymes and enzymes from Trichoderma longibrachiatum . Journal of Dairy Science 83, 13101321.
Nsereko, VL, Morgavi, DP, Rode, LM, Beauchemin, KA and McAllister, TA 2000. Effects of fungal enzyme preparations on hydrolysis and subsequent degradation of alfalfa hay fiber by mixed rumen microorganisms in vitro . Animal Feed Science and Technology 88, 153170.
Ouédraogo-Koné, S, Kaboré-Zoungrana, CY and Ledin, I 2008. Intake and digestibility in sheep and chemical composition during different seasons of some West African browse species. Tropical Animal Health and Production 40, 155164.
Phakachoed, N, Suksombat, W, Colombatto, D and Beauchemin, KA 2013. Use of fibrolytic enzymes additives to enhance in vitro ruminal fermentation of corn silage. Livestock Science 157, 100112.
Phengvichith, V and Ledin, I 2007. Effect of feeding different levels of wilted cassava foliage (Manihot esculenta, Crantz) on the performance of growing goats. Small Ruminant Research 71, 109116.
Playne, MJ 1985. Determination of ethanol, volatile fatty acids, lactic and succinic acids in fermentation liquids by gas chromatography. Journal of Science Food and Agriculture 36, 638644.
Rhine, ED, Mulvaney, RL, Pratt, EJ and Sims, GK 1998. Improving the berthelot reaction for determining ammonium in soil extracts and water. Soil Science Society of America Journal 62, 473480.
Ribeiro, GO Jr, Teixeira, AM, Velasco, FO, Faria, WG Jr, Pereira, LGR, Chaves, AV, Gonçalves, LC and McAllister, TA 2014. Production, nutritional quality and in vitro methane production from Andropogon gayanus grass harvested at different maturities and preserved as hay or silage. Asian Australasian Journal of Animal Science 27, 330341.
Ribeiro, GO Jr, Teixeira, AM, Velasco, FO, Faria, WG Jr, Jayme, DJ, Maurício, RM, Gonçalves, LC and McAllister, TA 2015. Methane production and energy partitioning in sheep fed Andropogon gayanus grass ensiled at three regrowth stages. Canadian Journal of Animal Science 95, 103110.
Robertson, JB and Van Soest, PJ 1981. The detergent system of analysis and its application to human foods. In The analysis of dietary fiber in food (ed. WPT James and O Theander), pp. 123158. Marcel Dekker, New York, NY, USA.
Van Soest, PJ, Robertson, JB and Lewis, BA 1991. Methods for dietary fiber, neutral detergent fiber and nonstarch polysachharides in relation to animal nutrition. Journal of Dairy Science 74, 35833597.
Wang, Y and McAllister, TA 2002. Rumen microbes, enzymes and feed digestion – a review. Asian Australasian Journal of Animal Science 15, 16591676.
Wang, YX, McAllister, TA, Yanke, LJ, Xu, ZJ, Cheeke, PR and Cheng, K-J 2000. In vitro effects of steroidal saponins from Yucca Schidigera extract on rumen microbial protein synthesis and ruminal fermentation. Journal of Science Food and Agriculture 80, 21142122.
Wang, Y, McAllister, TA, Rode, LM, Beauchemin, KA, Morgavi, DP, Nsereko, VL, Iwaasa, AD and Yang, W 2001. Effects of an exogenous enzyme preparation on microbial protein synthesis, enzyme activity and attachment to feed in the Rumen Simulation Technique (Rusitec). British Journal of Nutrition 85, 325332.
Wilson, JR 1993. Organization of forage plant tissues. In Forage cell wall structure and digestibility (ed. HG Jung, DR Buxton, RD Hatfield and J Ralph), pp. 132. American Society of Agronomy, Crop Science Society of America, Soil Science Society of America, Madison, WI, USA.
Wilson, JR and Mertens, DR 1995. Cell wall accessibility and cell structure limitations to microbial digestion of forage. Crop Science 35, 251259.
Wylie, MJ, Ellis, WC, Matis, JH, Bailey, EM, James, WD and Beever, DE 2000. The flow of forage particles and solutes through segments of the digestive tracts of cattle. British Journal of Nutrition 83, 295306.



Altmetric attention score

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