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A modified rinsing method for the determination of the S, W–S and D + U fraction of protein and starch in feedstuff within the in situ technique

  • L. H. de Jonge (a1), H. van Laar (a1) (a2), W. H. Hendriks (a1) (a3) and J. Dijkstra (a1)

Abstract

A modified rinsing method for the in situ technique was developed to separate, isolate and characterise the soluble (S), the insoluble washout (W–S) and the non-washout fractions (D + U) within one procedure. For non-incubated bags (t = 0 h), this method was compared with the conventional, Combined Fractionation (CF) method that measures the D + U and S fractions in separate steps and subsequently calculates the W–S fraction. The modified method was based on rinsing of nylon bags in a closed vessel containing a buffer solution (pH 6.2) during 1 h, where shaking speeds of 40, 100, and 160 strokes per minutes (spm) were evaluated, and tested for six feed ingredients (faba beans, maize, oats, peas, soya beans and wheat) and four forages (two ryegrass silages and two maize silages). The average recoveries as the sum of all fractions were 0.972 ± 0.041 for N and 0.990 ± 0.050 for starch (mean ± s.d.). The mean W–S fraction increased with increasing shaking speed and varied between 0.017 (N) and 0.083 (starch) at 40 spm and 0.078 (N) and 0.303 (starch) at 160 spm, respectively. For ryegrass silages, the W–S fraction was absent at all shaking speeds, but was present in the CF method. The modified method, in particular at 40 and 100 spm, reduced the loss of small particles during rinsing, resulting in lower W–S and higher D + U fractions for N and starch compared with the CF method. For soya beans and ryegrass silage, the modified method reduced the S fraction of N compared with the CF method. The results obtained at 160 spm showed the best comparison with those from the CF method. The W–S fraction of the feedstuff obtained at 160 spm contained mainly particles smaller than 40 μm (0.908 ± 0.086). In most feedstuff, starch was the most abundant chemical component in the W–S fraction and its content (726 ± 75 g/kg DM) was higher than in the D + U fraction (405 ± 177 g/kg DM). Alkaline-soluble proteins were the dominant N-containing components in the W–S fraction of dry feed ingredients and its relative content (0.79 ± 0.18 of total N in W–S) was higher than in the D + U fraction (0.59 ± 0.07 of total N in D + U) for all feedstuff except maize. The molecular weight distribution of the alkaline-soluble proteins differed between the W–S and the D + U fractions of all dry feed ingredients, except soya beans and wheat.

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Aufrère, J, Garces, C, Graviou, D, Hernando, I, Demarquilly, C 1999. Degradation in the rumen of treated and untreated soya bean meal proteins. Annales de Zootechnie 48, 263273.
Chang, YW, Alli, I, Konishi, Y, Ziomek, E 2011. Characterization of protein fractions from chickpea (Cicer arientinum L.) and oat (Avena sativa L.) seeds using proteomics techniques. Food Research International 44, 30943104.
Chaudhry, AS, Webster, AJF 2001. Electrophoresis to determine the molecular weight distribution in soluble proteins from various foods and their rumen-resistant residue in cattle. Journal of the Science of Food and Agricultural 81, 10871093.
Cherney, DJR, Patterson, JA, Lemenager, RP 1990. Influence of in situ bag rinsing technique on determination of dry matter disappearance. Journal of Dairy Science 73, 391397.
Chiou, PWS, Yu, B, Wu, SS 1999. Protein sub-fractions and amino acid profiles of rumen-undegradable protein in dairy cows from soybean, cottonseed and fish meals. Animal Feed Science and Technology 78, 6580.
Cockburn, JE, Dhanoa, MS, France, J, López, S 1993. Overestimation of solubility by dacron bag methodology. Animal Production 56, 466467.
CVB 2003. Protocol for in situ rumen incubations: determination of degradation rate and washable fractions of protein, starch, cell walls and organic residual fraction. In Dutch [Protocol voor in situ pensincubatie: bepaling van afbraaksnelheid en uitwasbare fracties van eiwit, zetmeel, celwanden en organische restfractie]. Centraal Veevoeder Bureau, Lelystad, pp. 56.
de Jonge, LH, Spek, JW, van Laar, H, Dijkstra, J 2009. Effects of pH, temperature and osmolality on the level and composition of soluble N in feedstuffs for ruminants. Animal Feed Science and Technology 153, 249262.
Dewhurst, RJ, Hepper, D, Webster, AJF 1995. Comparison of in sacco and in vitro techniques for estimating the rate and extent of rumen fermentation of a range of dietary ingredients. Animal Feed Science and Technology 51, 211229.
Dhanoa, MS, France, J, Lopez, S, Dijkstra, J, Lister, SJ, Davies, DR, Bannink, A 1999. Correcting the calculation of extent of degradation to account for particulate matter loss at zero time when applying the polyester bag method. Journal of Animal Science 77, 33853391.
ISO 6496 1999. Animal feeding stuffs – determination of moisture and other volatile matter content. International Organization for Standardization, Geneva, Switzerland.
ISO 5983-2 2005. Animal feeding stuffs – determination of nitrogen content and calculation of crude protein content – Part 2: block digestion/steam distillation method. International Organization for Standardization, Geneva, Switzerland.
ISO 15914 2004. Animal feeding stuffs – enzymatic determination of total starch content. International Organization for Standardization, Genève, Switzerland.
Kandylis, K, Nikokyris, PN 1997. Relationship between nitrogen solubility and in situ protein degradability in ruminant feedstuffs. Journal of the Science of Food and Agriculture 75, 205211.
Licitra, G, Hernandez, TM, van Soest, PJ 1996. Standardization of procedures for nitrogen fractionation of ruminant feed. Animal feed Science and Technology 57, 347358.
López, S 2005. In vitro and in situ techniques for estimating digestibility. In Quantitative aspects of ruminant digestion and Metabolism, 2nd edition (ed. Dijkstra J, Forbes JM and France J), pp. 87122. CABI Publishing, Wallingford, UK.
Madsen, J, Hvelplund, T 1994. Prediction of in situ protein degradability in the rumen. Results of a European ringtest. Livestock Production Science 39, 201212.
Michalet-Doreau, B, Ould-Bah, MY 1992. In vitro and in sacco methods for the estimation of dietary nitrogen degradability in the rumen: a review. Animal Feed Science and Technology 40, 5786.
NRC 2001. Nutrient Requirements of Dairy Cattle, 7th revised edition. National Academy Press, Washington, DC, USA.
Ørskov, ER 2000. The in situ technique for the estimation of forage degradability in ruminants. In Forage Evaluation in Ruminant Nutrition (ed. DI Givens, E Owen, RFE Axford and HM Omed), pp. 175188. CABI Publishing, Wallingford, UK.
Ørskov, ER, McDonald, I 1979. The estimation of protein degradability in the rumen from incubation measurements weighted according to rate of passage. Journal of Agricultural Science 92, 499503.
Romagnolo, D, Polan, CE, Barbeau, WE 1994. Electrophoretic analysis of ruminal degradability of corn proteins. Journal of Dairy Science 77, 10931099.
SAS Institute 2002. SAS/STAT User's Guide 2002. Version 9. SAS Institute Inc., Cary, NC, USA.
Spencer, D, Higgins, TJV, Freer, M, Dove, H, Coombe, JB 1988. Monitoring the fate of dietary proteins in rumen fluid using gel electrophoresis. British Journal of Nutrition 60, 241247.
Tas, BM, Taweel, HZ, Smit, HJ, Elgersma, A, Dijkstra, J, Tamminga, S 2006. Rumen degradation characteristics of perennial ryegrass cultivars during the growing season. Animal Feed Science and Technology 131, 102119.
Thomas, C (ed.) 2004. Feed into Milk. Nottingham University Press Nottingham, UK.
van Duinkerken, G, Blok, MC, Bannink, A, Cone, JW, Dijkstra, J, van Vuuren, AM, Tamminga, S 2011. Update of the Dutch protein evaluation system for ruminants: the DVE/OEB2010 system. Journal of Agricultural Science 149, 351367.
van Soest, PJ, Robertson, JB 1985. Analysis of forage and fibrous foods. In a laboratory manual for animal science 613. Cornell University, Ithaca, New York, USA.
Verité, R, Journet, M, Jarrige, R 1979. A new system for the protein feeding of ruminants: the PDI system. Livestock Production Science 6, 349367.
Volden, H (ed.) 2011. Norfor the Nordic feed evaluation system. EAAP publication No 130. Wageningen Academic Publishers, Wageningen, The Netherlands.
Weisbjerg, MR, Bhargava, PK, Hvelplund, T, Madsen, J 1990. Use of degradation curves in feed evaluation. 679. Report from the National Institute of Animal Science, Denmark, 33pp.
Yang, H-J, Tamminga, S, Williams, BA, Dijkstra, J, Boer, H 2005. In vitro gas and volatile fatty acids production profiles of barley and maize and their soluble and washout fractions after feed processing. Animal Feed Science and Technology 120, 125140.

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