Effects of including different energy sources in the diet supplemented with small peptides of cottonseed on in vitro rumen fermentation, digestibility and microbial enzymes activity

Document Type : Original Research Articles (Regular Papers)


1 Department of Animal Science, Faculty of Agriculture, Lorestan University, Khorramabad, Iran

2 Departmen of Animal Science, Lorestan University, Iran

3 Lorestan University



The aim of the present study was to investigate the effects of different sources of non-fibrous carbohydrates (NFC) in the dairy cow diet supplemented with small peptides of cottonseed (Fortide C) on in vitro ruminal gas production (GP), fermentation parameters, substrate disappearance and microbial enzyme activity. Four experimental diets were fed, which were iso-caloric and iso-nitrogenous, containing 1) maize, 2) barley, 3) wheat or 4) a maize+barley mixture as the main sources of NFC. Each experimental diet was supplemented with 7.05 g Fortide C/kg dry matter (DM) and incubated with media containing rumen liquor for 96 h in vitro. Dietary supplementation of the Fortice C-contained diet with wheat grain yielded greater gas production (GP) at 16 h of incubation, total GP and potential GP (b) than those containing maize (P<0.05), but similar to barley-containing diet (P>0.05). Other GP parameters including GP at 24, 48 and 72 h of incubation and constant rate of GP (c) were similar among the experimental diets. The highest and lowest DM disappearance, apparently degraded substrates, organic matter disappearance, estimated metabolizable energy, short chain fatty acids and microbial protein synthesis (MPS) were observed with the using wheat and maize in the diets supplemented with Fortide C, respectively (P<0.05). Using wheat in the diet decreased NH3-N compared to the maize diet (P<0.05). The inclusion of the wheat in the diet supplemented with Fortide C increased activity of carboxymethyl cellulase and α-amylase compared to the maize diet (P<0.05), while it was similar to the barley diet (P>0.05). However, microcrystalline cellulase and filter paper-degrading activities were unchanged among the dietary treatments. Overall, using wheat as the main source of NFC in the dairy cow diet supplemented with Fortide C improved in vitro ruminal fermentation profile, substrate disappearance, MPS and microbial enzyme activity compared to maize or maize+barley-based diets.


Main Subjects

Agarwal, N., 2000. Estimation of fiber degrading enzyme. In: Chaudhary, L.C., Agarwal, N., Kamra, D.N., Agarwal, D.K., (Eds), Feed Microbiology. CAS Animal Nutrition, IVRI, Izatnagar, India, pp. 278–291.
AOAC, 1995. Official Methods of Analysis. 16th Ed. Association of Official Analytical Chemists, Washington, DC, USA.
Araba, A., Byers, F.M., Guessous, F., 2002. Patterns of rumen fermentation in bulls fed barley/molasses diets. Animal Feed Science and Technology 97, 53-64.
Argyle, J.L., Baldwin, R.L., 1989. Effects of amino acids and peptides on rumen microbial growth yields. Journal of Dairy Science 72, 2017-2027.
Azizi-Shotorkhoft, A., Rouzbehan, Y., Fazaeli, H., 2013. The influence of the different carbohydrate sources on utilization efficiency of processed broiler litter in sheep. Livestock Science 148, 249-254.  
Azizi-Shotorkhoft, A., Sharifi, A., Azarfar, A., Kiani, A., 2018. Effects of different carbohydrate sources on activity of rumen microbial enzymes and nitrogen retention in sheep fed diet containing recycled poultry bedding. Journal of Applied Animal Research 46, 50-54.
Blümmel, M., Steingss, H., Becker, K., 1997. The relationship between in vitro gas production, in vitro microbial biomass yield and 15N incorporation and its implications for the prediction of voluntary feed intake of roughages. British Journal of Nutrition 77, 911-921.
Broderick, G., Kang, J.H., 1980. Automated simultaneous determination of ammonia and total amino acids in ruminal fluid and in vitro media. Journal of Dairy Science 54, 1176-1183.
Broderick, G.A., Luchini, N.D., Reynal, S.M., Varga, G.A., Ishler, V.A., 2008. Effect on production of replacing dietary starch with sucrose in lactating dairy cows. Journal of Dairy Science 91, 4801-4810.  
Carro, M.D.,  Miller, E.L., 1999. Effect of supplementing a fiber basal diet with different nitrogen forms on ruminal fermentation and microbial growth in an in vitro semi-continuous culture system (RUSITEC). British Journal of Nutrition 82, 149-157.
Chai, W.Z., Van Gelder, A.H., Cone, J.W., 2004. Relationship between gas production and starch degradation in feed samples. Animal Feed Science and Technology 114, 195-204.
Chamberlain, D.G., Robertson, S., Choung, J.J., 1993. Sugars versus starch as supplements to grass silage: effects on ruminal fermentation and the supply of microbial protein to the small intestine, estimated from the urinary excretion of purine derivatives in sheep. Journal of Science of Food andAgriculture 63,189-194. 
Chikunya, S., Newbold, C.J., Rode, L., Chen, X.B., Wallace, R.J., 1996. Influence of dietary rumen degradable protein on bacterial growth in the rumen of sheep receiving different energy sources. Animal Feed Science and Technology 63, 333-340.
Cruz Soto, R.,  Muhammed, S.A., Newbold, C.J., Stewart, C.S., Wallace, R.J., 1994. Influence of peptides, amino acids and urea on microbial activity in the rumen of sheep receiving grass hay and on the growth of rumen bacteria in vitro. Animal Feed Science and Technology 49, 151-161.
Dabrowski, K., Lee, K., Rinchard, J., 2003. The smallest vertebrate, teleost fish, can utilize synthetic dipeptide based diets. Journal of Nutrition 133, 4225-4229.
Engvall, A., 1980. α-amylase activity in rumen fluid of cows producing milk of low and normal fat content. Journal of Dairy Science 63, 2012-2019.
FASS, 2010. Guide for the Care and Use of Agricultural Animals in Research and Teaching 3rd edn. Federation of Animal Science Societies, Champaign, IL. 
Getachew, G., Makkar, H.P.S., Becker, K., 2002. Tropical browses: contents of phenolic compounds, in vitro gas production and stoichiometric relationship between short chain fatty acid and in vitro gas production. Journal of Agricultural Science 139, 341-352.
Gozho, G.N., Mutsvangwa, T., 2008. Influence of carbohydrate source on ruminal fermentation characteristics, performance, and microbial protein synthesis in dairy cows. Journal of Dairy Science 91, 2726-2735.
Huntington, G.B., 1997. Starch utilization by ruminants: From basics to the bunk. Journal of Animal Science 75, 852-867.
Ives, S.E., Titgemeyer, E.C., Nagaraja, T.G., 2002. Technical note: effect of removal of microbial cells by centrifugation on peptide and alpha-amino nitrogen concentrations in ruminal fluid. Journal of Dairy Science 85, 3059-3061.
Jones, D.F., Hoover, W.H., Miller, W.T.K., 1998. Effects of concentrations of peptides on microbial metabolism in continuous culture. Journal of Animal Science 76, 611-616.
Karimi. E., Azarfar, A., Azizi, A., 2018. Investigating the effects of supplementing fattening lamb diet with small peptides of cottonseed meal on the digestibility and fermentation of nutrients in vitro. Iranian Journal of Animal Science 48, 549-558 (in Persian).
Khezri, A., Rezayazdi, K., Danesh Mesgaran, M., Moradi-Sharbabk, M., 2009. Effect of different rumen-degradable carbohydrates on rumen fermentation, nitrogen metabolism and lactation performance of Holstein dairy cows. Asian-Australian Journal of Animal Science 22, 651-658.
Kotzamanis, Y.P., Gisbert, E., Gatesoupe, F.J., Zambonino Infante, J., Cahu, C., 2007. Effects of different dietary levels of fish protein hydrolysates on growth, digestive enzymes, gut microbiota, and resistance to Vibrio anguillarum in European sea bass (Dicentrarchus labrax) larvae. Comparative Biochemistry and Physiology Part A: Molecular and Integrative Physiology 147, 205-214.
Lanzas, C., Fox, D.G., Pell, A.N., 2007. Digestion kinetics of dried cereal grains. Animal Feed Science and Technology 136, 265-280.
Marten, G.C., Barnes, R.F., 1980. Prediction of energy digestibility of forages with in vitro rumen fermentation and fungal enzyme systems, p. 61-71. In: Pigden, W.J., Balch, C.C., Graham, M., (Eds.). Standardization of Analytical Methodology for Feeds. Int. Devel. Res. Ctr., Ottawa, ON, Canada.
Menke, K.H., Raab, L., Salewski, A., Steingass, H., Fritz, D., Schneider, W., 1979. The estimation of the digestibility and metabolizable energy content of ruminant feeding stuffs from the gas production when they are incubated with rumen liquor in vitro. Journal of Agricultural Science 93, 217-222.
Merry, R.J., McAllan, A.B., Smith, R.H., 1990. In vitro continuous culture studies on the effect of nitrogen source on microbial growth and fiber digestion. Animal Feed Science and Technology 31, 55-64. 
Miller, J.L., 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry 31, 426-428.
Nasr, H., 1950. Amylolytic activity in the rumen of the sheep. Journal of Agricultural Science 40, 308-310.
Nogueira Filho, J.C.M., Fondevila, M., Barrios Urdaneta, A., Gonzalez Ronquillo, M., 2000. In vitro microbial fermentation of tropical grasses at an advanced maturity stage. Animal Feed Science and Technology 83, 145-157.
NRC, 2001. Nutrient requirements of dairy cattle. 7th Revised Edition, Subcommittee on Dairy Cattle Nutrition, Committee on Animal Nutrition, Board on Agriculture and Natural Resources, National Academy Press, Washington, D.C.
Obara, Y., Dellow, D.W., 1993. Effect of intraruminal infusions of urea, sucrose or urea plus sucrose kinetics in sheep fed chopped lucerne hay. Journal of Agricultural Science 121, 125-130.
Ørskov, E.R., McDonald, I., 1979. The estimation of protein degradability in the rumen from incubation measurements weighed according to rate of passage. Journal of Agricultural Science 92, 499-503.
Pan, Y.L., Webb, J.K.E., 1998. Peptide-bound methionine sources for protein accretion and cell proliferation in primary cultures of ovine skeletal muscle. Journal of Nutrition 128, 251-256.
Russell, J.B., O’Connor, J.D., Fox, D.G., Van Soest, P.J., Sniffen, C.J., 1992. A net carbohydrate and protein system for evaluating cattle diets. I. Ruminal fermentation. Journal of Animal Science 70, 3551-3561.
Russi, J.P., Wallace, R.J., Newbold, C.J., 2002. Influence of the pattern of peptide supply on microbial activity in the rumen simulating fermenter (RUSITEC). British Journal of Nutrition 88, 73-80.
Sahoo, A., Agarwal, N., Kamra, D.N., Chaudhary, L.C., Pathrak, N.N., 1999. Influence of level of molasses in de-oiled rice bran-based concen- trate mixture on rumen fermentation patterns in crossbred cattle calves. Animal Feed Science and Technology 80, 83-90.
Sannes, R.A., Messman, M.A., Vagnoni, D.B., 2002. Form of rumen-degradable carbohydrate and nitrogen on microbial protein synthesis and protein efficiency of dairy cows. Journal of Dairy Science 85, 900-908. 
Satter, L.D., Slyter, L.L., 1974. Effect of ammonia concentration on rumen microbial protein production in vitro. British Journal of Nutrition 32, 199-208.
Seo, J.K., Yang, J., Kim, H.J., Upadhaya, S.D., Cho, W.M., Ha, J.K., 2010. Effect of synchronization of carbohydrate and protein supply on ruminal fermentation, nitrogen metabolism and microbial protein synthesis in Holstein steers. Asian-Australasian Journal of Animal Sciences 23, 1455-1461.
Srinivas, B., Karim, S.A., 2009. Bioactive peptides as synergistic additive in ruminant diets – an epigrammatic synthesis. Agricultural Reviews 30, 307-310.
Theodorou, M.K., Williams, B.A., Dhanoa, M.S., McAllana, A.B., France, J., 1994. A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feeds. Animal Feed Science and Technology 48, 185-197.
Tomankova, O., Homolka, P., 2004. In vitro ruminal degradability of cereal grain starch. Czech Journal of Animal Science 49, 151-155.
Van Soest, P.J., Robertson, J.B., Lewis, B.A., 1991. Methods for dietary fiber, neutral detergent fiber and non-starch polysaccharides in relation to animal nutrition. Journal of Dairy Science 74, 3583-3597.
Van Soest, P.J., 1994. Nutritional Ecology of the Ruminant. Comstock Publication, Ithaca, NY.
Vercoe, P.E., Makkar, H.P.S., Schlink, A.C., 2010. In vitro Screening of Plant Resources for Extranutritional Attributes in Ruminants: Nuclear and Related Methodologies. Springer Verlag Gmbh.
Zhang, B., Xue, L.Q., Li, L.L., Chena, Y.G., Wen, G.H., Hou, D.X., 2007. Effects of soybean small peptides on nitrogen balance, nutrient digestibility and several indices in the portal venous plasma of goats. Small Ruminant Research 72, 1-10.