Using Proteases Today: A Commercial Nutritionists Perspective Rick Kleyn SPESFEED (Pty) Ltd.
Introduction Commercial nutrition all about balance: science vs. practicality performance vs. profitability Enzymes encompasses both.
Introduction Protein (80%) less digestible than starch (90%) in corn/soy diets. (Zanella, 1999). For each 10% improvement in the digestibility of a feed, there is a 50% decrease in the effectiveness of any enzyme that is offered (Cowieson and Bedford, 2009).
Introduction Proteases represent opportunity. Enhance digestibility - reduce feed cost. Difficult to measure true impact: Direct impact on protein digestion. Reduction in endogenous losses.
Introduction Protease improves gut health: Litter quality Improvement in welfare Improves on farm performance Adequate formulation relatively simple. Optimal formulation more difficult.
Introduction Require a basic understanding: Amino acid digestibility (and requirements). Enzyme functionality. Consider other enzymes/additives. Use a formulation system to apply our knowledge in practice.
Ingredients and Amino Acids 7
Amino Acid Digestibility Must know AA digestibility to use any enzyme effectively. Need to know what to measure: Ileal digestibility true indicator. Available, Digestible, True Ileal Digestible and Standard Ileal Digestible (SID). Used synonymously and interchangeably. 8
Amino Acid Digestibility Two main types of digestibility assays precision feeding of caecectomised roosters - measurement of amino acid disappearance; contents of the last third of the ileum compared to the amino acid intake to estimate the disappearance. Roosters value widely used. Assume digestibility unchanged with age? 9
Amino Acid Digestibility Digestibility estimates don't differentiate: undigested and unabsorbed AA s ileal endogenous amino acids. Correct for ileal endogenous content. Standardised ileal digestibility (SID). more likely to be additive in mixed diets. cornerstones of feed formulation.(adeola, 2013) DSM values calculated using SID
Amino Acid Digestibility Tahir and Pesti (2012) compared data Ajinomoto Heartland (AH) roosters. Evonik Degussa (ED) - full fed chicks (SID) AH values 6% higher than ED. Differences common ingredients < 3% Animal protein > 8% Other sources are available. 11
Other Sources Roosters SID 12
Different Values Ingredient Lysine SID Roosters Australian Evonik INRA Heartland Maize 81 92 85 83 Soya 87 90 91 91 Full Fat Soya 90 87 88 88 Sunflower 79 87 83 80 Fish Meal 77 86 89 90 Feather Meal 54 57 63 66 Cotton 57 65 63 63 Bran 73 73 74 75
Different Values Ingredient Methionine SID Roosters Australian Evonik INRA Heartland Maize 91 94 94 92 Soya 94 91 91 92 Full Fat Soya 88 88 86 86 Sunflower 88 92 93 91 Fish Meal 78 86 92 93 Feather Meal 61 61 72 74 Cotton 76 72 75 72 Bran 81 80 74 78
Amino Acid Digestibility Clear differences between laboratories. Databases are not interchangeable? Better to use intuition. Use different values for broilers & layers? 15
AA Digestibility and Broilers 16
AA Digestibility and Broilers AA digestibility further complicated by: Age plays a role. Birds utilize AA s differentially. Endogenous losses (mucin) - high in certain AA s (Threonine).
AA Digestibility and Broilers (Yegani & Korver, 2013) CP Lys Meth Cys Thre Starter 63.3 75.4 88.0 36.2 49.3 Grower 82.3 90.3 94.4 73.4 77.3 Finisher 80.6 88.2 94.2 70.0 75.2 * Contents of ileum collected 18
The Bird and it s Feed Formulation is made up of 2 components: LHS or the Matrix RHS or Feed Specification Mostly all make same mistake.
Matrix and Spec Miss match
Protease Protease appears to be more efficacious in young birds: Levels of protein in diet higher. Endogenous system immature. Acts differentially on Amino Acids
Protease and Age (Bertechini, 2009). RONOZYME ProAct Matrix Value Pre Starter Feed Starter Feed Grower Feed Withdraw Feed Mean CP (%) 4663* 4639 4178 3747 4307 Lys Dig. (%) 104 98 82 69 88 Met Dig. (%) 62 60 53 44 54 M+C Dig. (%) 196 187 175 158 179 Thr Dig. (%) 361 340 336 319 339 Trp Dig. (%) 42 40 41 38 40 Arg Dig. (%) 317 299 264 234 279 Val Dig. (%) 219 208 200 186 203 Leu Dig. (%) 121 118 124 113 119 Ile Dig. (%) 148 141 137 130 139 * Suggested matrix values for enzymes 22
Digestibility Improvement Factor (%) (Smith, 2014) Crude Protein Lysine TSAA Threonine Maize 3.4 3.6 4.7 7.2 Wheat 3.5 6.4 2.5 4.8 Soybean 2.2 2.0 3.7 4.2 Rapeseed 1.6 2.0 1.6 1.9 Sunflower 2.0 3.3 0.7 3.4
Protease and Energy Protease no energy recommendation? But: We use AME n. The little "n" represents the correction for nitrogen balance. Basic energetics - a 2% release of protein must result in a 2% increase in energy contribution. (Leeson, 2014). Our understanding far from complete.
Other Benefits of Protease
Protease and Gut Health Reduces total protein in diet. (Decous, 2013) Improves inflammatory status of GIT Reduces damage to villi Removes indigestible (fermentable) protein & anti-nutrients entering caecum: Deprives undesirable bacteria of nutrients. Prevents shift to more undesirable strains. Reduce trypsin inhibitor in soy (Mayorga, 2012) All lead to improved gut health 26
Effect of Urease and Protease (after Mayorga et al., 2012) Urease Weight (Kg) FCR 0.13 1.535 a 1.46 a 0.23 1.509 a 1.49 a 0.72 1.433 b 1.49 a 1.52 1.370 b 1.60 b Protease 0 g/ton 1.431 b 1.54 a 200 g/ton 1.491 a 1.47 b * Broilers to 28 days of age
Protein, Protease and Welfare (Leleu et al., 2012) 60 59.5 NS 1.61 1.6 59 1.59 58.5 1.58 58 NS 1.57 1.56 57.5 1.55 57 Standard Negative Nve+Protease Gain (g/d) FCR 1.54
Protein, Protease and Welfare (Leleu et al., 2012) 80 70 60 50 40 30 20 10 0 Standard Negative Nve+Protease Clean Bird% No Foot lesion%
Practical Use of Protease Select the correct enzyme. Are they single-component enzymes? Little evidence that cocktails are better. Side activities may not be thermostable. Use scientific methodology: Were experiments properly conducted? Can you identify response from components? Are the trial conditions/results relevant? 30
Practical Use of Protease Is it practical to work with/apply? Heat stable & granulation (particle size). What other parameters are important? gut health/ litter quality/ uniformity Does it pay (spend $1, expect $ 2 back)?
Formulating with Protease Must be model within formulation system. Approach must be simple and robust. If you can t do it with a pencil and paper, a formulation system can t solve the problem. Focus on undigested protein fraction Will rarely use a single enzyme.
Multiple Enzymes Need to try and understand additivity. Phytase - used in nearly all diets. Use reasonable lift values to enzymes: Too high - animal performance suffers: Too low lost opportunity (financial) Information is scant.
Phytase Alone Energy Protein/AA Minerals Phytase NSP Amylase Protease
Phytase & Carbohydrases Energy Protein/AA Minerals Phytase NSP Amylase Protease
Phytase, Carbohydrase and Protease Energy Protein/AA Minerals Phytase NSP?? Amylase Protease?
Phytase and Protease Energy Protein/AA Minerals Phytase? NSP Amylase Protease?
Formulating with Protease Not straight forward: Decide on which nutrients to consider. Decide which enzyme/s to use. Response not linear. Substrate/additive dependant. Birds do not use enzyme consistently.
Formulating with Protease Methods that can be used: Add on top mostly does not work. Increase nutrient profile of ingredient. Use additional nutrient - DIF value. Create a matrix for an enzyme/s (model). Reduce feed specifications when added.
Adding on Top Existing Specification Weight Gain Reduce 3% Add 3% Increasing Protein
Use of DIF Values Maize SBM Sun Limit Spec Cost 220 550 350 MIN AME 14.1 10.4 8.2 >= 12.8 Avl Lys 2.1 26.5 15 >= 11.5 Factor 103.6% 102%103.3% Lys DIF 2.17 27.03 15.5 >=
Use of DIF Values Maize SBM Sun Limit Spec Cost 220 550 350 MIN AME 14.1 10.4 8.2 >= 12.8 Avl Lys 2.1 26.5 15 >= Factor 103.6% 102%103.3% Lys DIF 2.17 27.03 15.5 >= 11.5
DIF Values Accurate and effective for single enzyme. Problematic with multiple enzymes? Age differences not considered. High/complex data requirement.
Models to Calculate Lift Models to calculate AA lift are required: Take expected formulation into account. Take bird age into account. Ideally, different matrix for each class. Can model multiple enzymes Model generated matrix figures added to a premix robust and effective.
Examples of Matrix Values (Nell, 2014) HiPhos HiPhos HiPhos Proact Proact NSP ase CP% 0.290 0.946 0.946 ME (MJ/kg) 0.36 0.36 0.84 Avl Lys (g) 0.13 0.48 0.48 Dig Phos (g) 1.34 1.34 1.34
Examples of Matrix Values (Nell, 2014) Pre- Starter Starter Grower Finisher Proact Proact Proact Proact 0.946 0.906 0.877 0.829 CP% Avl Lys (g) 0.48 0.45 0.43 0.4 Avl TSAA (g) 0.36 0.33 0.32 0.3
Formulating with Protease All methods discussed are a compromise. Assigning a 3 or 4% lift across protein and amino acids works adequately. DIF data hungry complicated. Model generated matrix a good option.
Ending Off Ingredient AA digestibility - starting point. Bird requirements - just as important. Protease effective in feed and on farm. Effective formulation a challenge. Use conservative values. Still highly cost effective.