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# DIGESTIBILITY

Apparent v. true digestibility
True digestibility involves correction for endogenous losses, apparent digestion does not. Endogenous losses – Include: • Sloughed off intestinal cells • Digestive juices (enzymes) • Microbial matter – Quantified by measuring fecal output of fasted animals – Can be 9.8 to 12.9 % DMI – Should they be quantified?

In vivo digestibility methods
Direct or total/complete collection Difference method Regression method Indirect method

1. Total collection .

In vivo digestibility trials in metabolism crates .

In vivo digestibility trials in pens .

Total collection calculations Digestibility (g/kg) = Nutrient in feed .DM in feces x 1000 DM in feed Organic matter digestibility (OMD.Nutrient in feces x 1000 Nutrient in feed Dry matter digestibility (DMD. g/kg) = OM in feed . % or g/kg . g/kg) = DM in feed .OM in feces x 1000 OM in feed Can be expressed as a proportion.

25) – DCP= Digestible Crude Protein – DCF= Digestible Crude Fiber – DNFE= Digestible Nitrogen-Free Extract – DEE= Digestible Ether Extract (2.Digestibility indices that estimate energy value Digestible organic matter content (DOMD) (g/kg DM) = OM in feed .OM in feces x 1000 DM in feed TDN = DCP + DCF + DNFE + DEE(2.25) .

2.Base feed DMO) Test feed DMI Cons – Assumptions may be invalid . Difference method Allows digy calculation for 2 feeds fed simultaneously Assumptions – No interaction b/w the digy of the feeds – Must know digy & fecal DM output (DMO) of base feed Test feed DMD = Test feed DMI – (Fecal DMO.

Cons – Considerable expense and labor for estimating digy of one feed. estimation for two feeds – Feed different ratios of the two feeds – Estimate digy of each of the ratios – Fit regression of test feed inclusion vs. digy – Extrapolate to estimate digy of test feed.3. . Regression method Schneider & Flatt (1975) Also allows digy.

200 20 40 60 80 100 % inclusion of test feed in ration .Regression method 800 Base feed digy. DMD (g/kg) 600 400 Test feed digy.

• Animal physiological changes • Forage physiological changes Adaptation period – Necessary to adapt the animals to • New feed (microbial population changes) • Strange equipment • Strange housing – 6 – 14 day period is the norm .g.Digy trial issues Changeover designs – necessary if period effects are an issue e.

if a feed contains 1% Cr2O3 & feces contains 2% Cr2O3.g. E. has doubled.Marker digestibility trials Particularly useful for grazing animals Procedure – Add indigestible marker to feed eg chromic oxide – Measure concentration in feed & feces – Estimate disappearance of marker from gut. 50% of DM must have been digested . diet digestibility = 50% – Since Cr3O2 conc.

must also know the % nutrient in feed & feces %Nutrient Digestibility = 100 – 100 x % indicatorfeed % indicatorfeces X % nutrientfeces % nutrientfeed Homework: If lambs are fed a bahia grass diet containing 7% protein & 1% chromic oxide. For the digy of a specific nutrient. and their feces contains 5% CP and 2% chromic oxide.Marker trials contd. Calculate CP digy. .

Marker digestibility Pros – Total feces collection not necessary – Total intake determination not necessary – Easier. less labor Cons – Representative sampling essential – Accurate estimation of nutrient or marker conc. essential – Assumes complete excretion of marker hence Recovery of marker determines accuracy of digy .

less labor .Marker types External – Chromic oxide – Dysporium – Polyamide Can contaminate forage Internal – Lignin – AIA – ADF – n-alkanes Easier.

( s handling) Marker migration – Must not affect feed digy External markers may contaminate forage .Marker issues Difficulty of mixing marker with forages – Dose cows instead.

Problems with in vivo experiments Animal trials are: – Expensive – Protracted – Laborious – Public concerns – Animal stress ??? Must estimate nutritive value with less animal dependent techniques .

of samples – Laboratory-based .Ideal in vitro methods should be: – Rapid (one step) & routinely practicable – Accurate – Cheap & not laborious – Repeatable & robust – Biologically meaningful – Broad-based (apply to all forage types) – Handle large nos.

Rumen fluid –pepsin in vitro digestibility (IVOMD) •Developed by Tilley & Terry (1967) •Measures apparent digy in rumen fluid (48 h) and acid pepsin (48 h) •Gives accurate predictions of in vivo digy for most forages .

5 40.Prediction of silage OMD in vivo from different methods (g/kg DM) Method r2 RSD KMnO4 lignin ADF NDF (M) ADF IVOMD 21. 1989) .9 45.1 54..6 50.9 33.7 55.6 (Givens et al.8 32.1 45.8 74.

Rumen fluid problems Variation in Inoculum composition & activity due to – Host animal diet – Animal species – Collection time – Processing (blending vs. filtration) .

optimal pH. temp – High viscosity hinders filtration – Offensive odors – Hygiene – (Prevent pathogen infection) .Rumen fluid problems Analytical issues – Maintenance of anaerobic media.

24 570 550 530 530 580 630 680 Rumen fluid-pepsin DOMD (Adesogan et al.Relationship between in vivo and in vitro DOMD of wheat silage (g/kg DM) 690 670 Year One Year Two In vivo DOMD 650 630 610 590 r2 =0. 1998) .

Rumen fluid technique problems Standards needed to correct for variability in rumen fluid composition & activity Disregards / inappropriately represents: – Ruminal outflow (uses a batch process) – Digests maillard product not digested in vivo – Associative effects between feeds – Endogenous secretions – Post abomasal digestion .

values – Still requires rumen fluid 2.Alternatives to Tilley & Terry 1. Enzyme. Rumen fluid – Neutral detergent (Van Soest.based assays . 1967) – More akin to true digestibility – Gives higher digy. Feces – Gives lower digestibility estimates 3.

97 (Ohmed et al. 2001) .98 0..77 – 0.Prediction of DMD in vivo from in vitro fecal liquor DMD Spp. of feces donor Ovine Bovine r2 range 0.90 0.97 Equine Caprine 0.96-0.33 – 0.

Cellulase 2.Cell-free enzyme in vitro digestibility Examples of procedures used: 1. Pepsin cellulase Amylase pre-treatment important for starch-rich feeds Gammanase for oil-rich feeds . Neutral detergent-cellulase +gammanase 4. Neutral detergent.cellulase 3.

Relationships between DMD in vivo and enzyme predicted DMD Method Cellulase R2 0.94 0. 1986) .83 (Bughara & Sleper.83 Neutral detergent cellulase Acid pepsin – cellulase 0.88 Rumen fluid 0.

3 (McLeod & Minson.9 0.94 3.90 0.6 0.Prediction of in vivo OMD of forages from different methods Method r RSD (%) AE(+) ND + cellulase Pepsin + cellulase 0.3 2. 1982) Higher analytical error with ND – cellulase technique may outweigh shorter processing time .

6 75. 1990) .2 33.Prediction of in vivo OMD of spring grass from different methods Method r2 RSD ND + cellulase Pepsin + cellulase Rumen fluid-pepsin (M) ADF 76.20) (Givens et al.9 27.8 33..3 Poorer relationships found for autumn grass (r2 = 13.1 28.9 67.0 66.

Basidiomycete Aspergillus niger Herbage 57 48 45 Cellulose paper 69 20 10 Rhizopus spp. 1975) . 35 7 (Jones & Hayward.Effect of enzyme source on cellulase activity % DM solubilized Fungi Trichoderma spp.

ruminantium 0.25 0.14C-Casein 0. bovis S.5 hydrolysis (mg/ml) Co-culture 0.0 10 Time (h) 20 Commercial enzymes don’t fully simulate microbial activity of mixed rumen microbes .0 S.

Enzyme method problems Equations are species-specific Represent effect of a few enzymes Variability in enzyme activity – Due to enzyme source & batch .

The ANKOM equipment .

87x + 4.99x + 3. DOMD 80 y = 0.93 2 70 70 tube 60 tube 60 50 50 40 40 50 bag 60 50 55 60 bag 65 70 75 80 85 70 80 .Ankom digestibility validation Prediction of tube app. rsd=2.04 2 Prediction of tube true DOMD from bag true DOMD 80 y = 0.61 r = 0. rsd = 4.83. DOMD from bag app.93.25 r = 0.

incubation and mixing – Uses a batch process (& ash-free bags) Cons – Bag pore size may allow excess outflow or restrict microbial colonization – Bag material & pore size may affect results • Monofilamentous cloth – precise aperture • Multifilamentous cloth – pore size affected by stresses e.ANKOM pros & cons Pros – Simplifies filtration.g. dacron .

In vitro digestibility summary Pros – Predicts in vivo digy more accurately than NDF or lignin – Handles several samples & are biologically meaningful Cons – May require fistulated animals – Labor intensive & protracted – Plagued by variability in composition & activity of inoculum/enzyme – Doesn’t indicate the kinetics of digestion .

E. A comparison of filter bag methods with conventional tube methods of determining the in vitro digestibility of forages. R. pp 438-441. Evaluation of a filter bag system for NDF.F. J.H. R. P. 1963. A.M. and Owen. Journal of the British Grassland Society.D. .. 113134. Finish Grassland Association.. 2000. 39: 276-279.. E. Helsinki.. pp..J.F. S. 18: 104-111.A. CABI Publishing. 1966. Pedersen. Axford and H. Animal Feed Science and Technology.J.. and IVDMD forage analysis. and Adesogan. D. P 263 Tilley. Wilman.. K. CABI Publishing. 1999. Owen. UK. Givens. Givens D. E. Crop Science. Van Soest.A.M.P.I. and Terry. Measuring chemical composition and nutritive value in forages. R. Adesogan. 84: 33-47. A. and Toy. Masterson. J. ADF. A two stage technique for the in vitro digestion of forage crops. Field and Laboratory methods for grassland and animal production research. Omed (Editors) 2000.. Vogel.T.Digestibility references Chapters 6 – 8 In: D. Wine.I. Wallingford.A. Forage Evaluation in Ruminant Nutrition. L. Proceedings of . J. The Xth International Grassland Congress. and Moore. Estimation of the true digestibility of forages by the in vitro digestion of cell walls.