Common mistakes using enzymes in poultry nutrition

Hi, Dr. Piotr. Hope you are doing well. If use phytase, nsps and protease enzymes in feed, how can we get matrix from these. If we go for matrix phytase not only work on phosphorus and calcium but also release some amino acids and energy. Protease enzyme not only release amino acids but this enzyme also has some impact on calcium, p and energy. What should we do when we use these 3 enzymes in combination?

Dear Luis Fernando Luna. I became curious about your comment on defining matrices for a combination of enzymes based on growth models. Could you elaborate on that? Thank you.

Dear Luis Fernando Luna. Thanks for the explanation and offering. My point is that there are two nutrient releases that have a direct link with performance: Energy (not really a nutrient) and amino acids. There's no way, just based on the performance change, to separate how much of the impact is coming from each one. Throwing everything in only one (f.i., energy), would generate either an over or under-estimation of it's impact. What I have done in the past with our model (Panorama) is to set an expected impact on amino acids and the performance and the model predicts the necessary change in energy to get that impact. If you do a mistake on the amino acid impact then your energy estimation is also wrong.

Dear Dr. Prasad. I got curious about the maximum energy release of 125-150 kcal/kg, whatever the combination of additives and diet composition. Where does this number comes from?

Dear Dave Albin. I am not sure if I understood your explanation. Could you try to put it in another words? Sorry for my lack of understanding.

I think the ME for enzymes depends birds age, enzymes composition, dietary /feedstuffs composition and energy value of the diet but it is safe to assume that it is equal to 50 kcal per kg diet but again depends on factors mentioned above.

Hello, your content is correct. But the age of the bird and the operation or physiological activity of the digestive system. They are more important with the ingredients of the food than the ingredients of the enzyme. Because the composition of the enzyme depends on the age of the bird according to the growth or genetic development and the composition of the food. You are reminded of the activity of enzymes and the amount of calories they are able to release. It is necessary to consider the strength or activity of the endogenous enzyme inside the bird's body in relation to the bird's age. On the other hand, the activity of bacteria or fungi in probiotics or the origin of enzymes must be observed in terms of the need for nutrients and energy inside the bird's body, which is itself considered a living organism. However, it is not easy to say how much energy metabolism has.

Hello
It is respectfully recalled that according to the results of the experiment using multi-enzyme with barley in the diet of laying hens, the quality of eggs, especially the shell, decreased. Please, if your scientific colleagues have observed such results and scientific articles in their farm or research experiments, please send them to the email below. Thanks
Yaqabfar@yahoo.moshatahe

The enzyme products that have been developed are of great value to the broiler producer. We know commercial enzyme products release nutrients from the bulk ingredients that make the ingredients more valuable. Typically, a successful product improves value well beyond its cost. And we know that enzyme products named for the same enzyme can have other beneficial enzymes from the fermentation process that make them different, as well.
The best way to use an enzyme product would be to know how each bulk ingredient is affected by the product. That prevents us from limiting the choices we have when formulating in changing ingredient markets. We cannot be sure that the enzyme product will provide the same value if we see reducing one energy source or protein source for another when the prices are favorable unless we know how each ingredient will be affected.
In my opinion, companies marketing enzyme products should have a set of nutrient values for each ingredient the user may feed. Without this, we lose the ability to accurately formulate feeds. As bulk ingredient costs change, the formulas change. If more energy is released from maize, we will undervalue maize in a formulation because we fail to give it credit for the additional energy our chosen enzyme product releases. Then, when costs dictate that we replace maize with a suddenly less expensive source of energy and protein, we cannot be sure that we get the same benefit from the enzyme product.
The difficulty is knowing the actual contribution from each ingredient. The fact that the percentages of each bulk ingredient in each diet we use does vary allows us to begin to interpret the results. The only piece of data that we need to determine an enzyme product impact, after feeding, is to know what nutritional values must have been fed to get the observed outcome. Accurate prediction of flock growth from nutrition is the key and can come from accurate growth programs that do that prediction.
Obviously, the amount of energy released from maize in a starter feed cannot be the same as the energy released from maize in a finisher diet. Also, the sum of energy released from the different ingredients must be equal to the energy required to match the growth. So, a series of simultaneous equations are required, finding the matching energy expression that fits the minimum nutritional change that gives the growth.
This is the challenge, and, in my opinion, the results should be expressed and percent change of the initial ingredient values. Now, the ingredient plus its share of enzyme product cost can be fairly evaluated in formulations. Obviously, the formulation cannot contain native products and enzyme enhanced products together as the decision to add the enzyme impacts all ingredients.

Hi All. As we all know that enzymes are very heat sensitive. Let's also check our feedmill parameters. If we are using Pellet Mill let's be careful in our meal temperature not to exceed 80-85°C. Or if we are using PPLA it will be an advantage. Thanks

Hi dr. Piotr Stanislaski

When incorporating enzymes into poultry nutrition, several common mistakes can undermine their effectiveness and economic benefits. Here's a structured overview of these pitfalls:

1. Incorrect Enzyme Selection.
- Substrate Mismatch:
Using enzymes not tailored to the feed's composition (e.g., neglecting xylanase for wheat-based diets rich in NSPs).
- Phytase  misapplication.

2. Dosage Errors
Ignoring Product Potency.

3. Storage and Handling Issues:
- Heat/Moisture exposure, as prof. dr. Attia said, & Feed processing damage.

4. Compatibility Oversights:
- Additive Interactions, Neglecting antagonistic effects from minerals (e.g., copper, zinc) or pH imbalances in the gut.

5. Diet Formulation Neglect:
- Nutrient Redundancy,  Failing to adjust inorganic phosphorus or amino acid levels post-enzyme inclusion, missing cost and environmental benefits.

6. Assumption of Uniform Efficacy:
- Product Variability.

7. Ignoring Animal-Specific Factors:
- As prf. Attia said, age/health status: Not tailoring enzyme use to developmental stages (e.g., chicks) or health conditions affecting gut environment.

8. Lack of Efficacy Monitoring:
- Absence of Testing.

9. Regulatory and Quality Lapses:
- Non-Compliance, Using unapproved enzymes or inconsistent batches due to poor quality control.

10. Over-Reliance on Enzymes:
- Compensating for Poor Diets.

When focusing on other enzymes commonly used in poultry nutrition beyond phytase and NSPases (e.g., xylanase, ß-glucanase), several mistakes can occur. These enzymes include proteases, amylases, lipases, mannanases, galactosidases, and multi-enzyme blends, each requiring specific considerations. Below are the key errors and oversights:

1. Proteases.
Mistakes: 
- Over-Hydrolysis, excessive protease activity can generate bitter peptides (from hydrolyzed proteins), reducing feed palatability. 
- Ignoring Endogenous Enzymes: Overlapping with the bird’s own digestive proteases (e.g., pepsin, trypsin), leading to redundancy or nutrient imbalances. 
- Substrate Mismatch: Using proteases in low-protein diets where benefits are marginal. 
- pH Neglect: Not ensuring compatibility with the gut’s acidic pH (e.g., some fungal proteases work best at neutral pH, limiting efficacy in the proventriculus). 

2. Amylases. 
Mistakes: 
- Energy Overestimation: Assuming amylase allows for drastic reductions in dietary energy without testing actual metabolizable energy (AMEn) responses. 
- Overuse in High-Starch Diets: Adding amylase to corn-based diets already rich in digestible starch, offering minimal ROI. 
- Heat Lability: Using non-thermostable amylases in pelleted feed, leading to inactivation during processing. 

3. Lipases.
Mistakes: 
- Bile Salt Deficiency: Using lipases without ensuring sufficient bile salts (critical for emulsification), especially in young chicks. 
- Ignoring Fat Quality: Applying lipases to rancid or oxidized fats, which may worsen nutrient absorption. 
- Overdosing: Excess lipase activity can release free fatty acids that irritate the gut lining. 

4. Mannanases
Mistakes: 
- Unnecessary Use: Adding mannanase to diets with low soybean meal content or alternative protein sources (e.g., sunflower meal). 
- Immune Response Misunderstanding: Failing to recognize that ß-mannans trigger innate immune responses; overdosing mannanase may suppress this immune priming. 

5. Galactosidases (a-galactosidase).
Mistakes: 
- Ignoring Substrate Levels: Using galactosidase in corn-wheat diets with negligible GOS content. 
- Microbiome Disruption: Over-hydrolyzing prebiotic GOS, which may reduce beneficial microbial growth in the hindgut. 

6. Multi-Enzyme Blends.  Mistakes: 
- Additive Assumptions: Assuming enzyme effects are purely additive without testing synergies/antagonisms (e.g., protease + phytase interactions). 
- Redundancy: Including enzymes that target substrates not present in the diet (e.g., adding ß-glucanase to a corn-based diet). 
- Dose Imbalance: Using uniform dosing for all enzymes in the blend, despite varying substrate levels and enzyme kinetics. 

7. General Mistakes Across All Enzymes: 
- Ignoring Bird Age: Using the same enzyme dose for chicks (immature gut) vs. growers/layers.   
- Diet Formulation Rigidity: Not reformulating diets to capitalize on enzyme benefits (e.g., reducing supplemental phosphorus with phytase but keeping calcium levels unchanged, disrupting Ca:P ratios). 
- Overlooking Gut Health: Enzymes like xylanase improve gut viscosity but may alter microbial populations; abrupt removal can destabilize the microbiome. 
- Stability Misjudgments: Assuming all enzymes survive feed processing (e.g., steam pelleting) or storage without verifying thermostability data.

I think that we should look after: 
1. Substrate Analysis: Verify substrate levels (e.g., phytate, NSPs) in diets before applying enzymes. 

2. Context-Specific Ranges: Define ME ranges based on enzyme type, substrate, and bird age (e.g., 80–100 kcal/kg for xylanase in wheat-based broiler diets). 

3. Dynamic Formulation: Use matrix values as a starting point, but adjust formulations based on: 
- Feed processing conditions (e.g., pelleting temperature).  - Bird health and genetics. 
- Regulatory limits. 

4. Third-Party Validation: Demand enzyme suppliers provide peer-reviewed data or conduct on-farm trials. 

5. Holistic Nutrient Balancing: Pair ME adjustments with amino acid, calcium, and phosphorus reformulation.

Thank you doctor, really i missed it, i appreciate it, I'm so sorry!

Dear Mohamed, thanks a lot, 100% agree, actually and important.

Quite correct in your opinions. Also, adding more than one enzyme at a type can muddy the water on value. People can freeze on the bulk ingredients not knowing which substrate is benefiting from the enzymes and miss beneficial changes on the feed costs.

Elaborate set of discussion above. I have experienced first hand the unwillingness of any producer/feed miller in disclosing the current set of ingredients in use. There is always much ambiguity regarding the raw materials used, unless of course you are a consultant. The bridge to value proposition and value addition is enzymes and their specific use after determining its substrate. However either ingredients are not disclosed of the analysis thereof, making the recommendation of enzymes difficult or assumption based.

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Substrate Analysis:
Verifying Substrate Levels in Diets Before Applying Enzymes

Enzyme supplementation in animal feed is a highly effective tool for improving nutrient utilization, especially when it comes to breaking down complex compounds in raw materials like grains, oilseeds, and fibrous plant materials. However, for enzymes to be truly effective, their application must be based on a solid understanding of the substrate they are targeting. Substrate analysis, which involves evaluating the levels of specific compounds like phytates and non-starch polysaccharides (NSPs) in the diet, is a crucial first step before enzyme application.

Let’s explore why substrate analysis is vital and how it informs enzyme recommendations.

1. Why Substrate Analysis Matters
Substrate analysis is essential because it helps identify the types and concentrations of the compounds that enzymes are designed to break down. Different enzymes target different substrates, and understanding the composition of the diet ensures that the correct enzymes are chosen for the job. Without proper analysis, there’s a risk of underestimating or overestimating the necessity of enzyme supplementation, which could lead to inefficient use of resources or suboptimal animal performance.

Targeting Specific Enzyme Functions:
Enzymes like phytase, xylanase, amylase, and cellulase have specific substrates they act upon, such as phytate (in plant-based proteins) or fiber (in cereals and grains). Knowing how much of these substrates are present in the diet allows for targeted enzyme supplementation that optimizes digestion and nutrient absorption.
Improving Feed Efficiency: With proper substrate analysis, you can ensure that enzymes are used where they will have the most significant impact, leading to better feed conversion ratios (FCR), improved growth rates, and overall healthier animals.

2. Common Substrates of Interest in Feed and Their Role
a. Phytate (Phytic Acid)
Phytate is the form of phosphorus found in plant-based ingredients like grains (corn, wheat, etc.), oilseeds, and legumes. However, phytate is largely indigestible to most monogastric animals (pigs, poultry, etc.) because they lack the enzyme phytase that breaks down phytate into more bioavailable forms of phosphorus.

Importance of Phytate Analysis:
If your diet contains high levels of phytate (common in grains and oilseeds), supplementation with phytase enzymes will improve phosphorus availability, reduce the need for inorganic phosphorus supplementation, and decrease environmental phosphorus waste.
Without knowing the phytate content, it’s difficult to gauge the proper phytase dose. Too little phytase could result in insufficient phosphorus utilization, while excessive phytase could be wasteful.
b. Non-Starch Polysaccharides (NSPs)

NSPs are a major component of plant cell walls, particularly in cereal grains (e.g., wheat, barley, oats) and oilseeds. These complex carbohydrates, such as hemicellulose, xylan, and beta-glucans, are poorly digested by monogastric animals.

NSPs and Their Effects:
NSPs form highly viscous solutions in the digestive tract, which can interfere with nutrient digestion and absorption.
Xylanase and beta-glucanase are enzymes that target these polysaccharides. However, their effectiveness is heavily dependent on the NSP levels present in the diet.

Importance of NSP Analysis:
By quantifying the NSP content, you can better understand the challenge posed by fiber in the diet.
Accurate analysis allows you to decide on the type and quantity of fiber-degrading enzymes (e.g., xylanase for xylans or beta-glucanase for beta-glucans), improving the digestibility of the feed and potentially increasing the energy available to the animal.
c. Cellulose

Cellulose, a major component of plant fibers, is highly resistant to digestion in non-ruminant animals. However, cellulase enzymes can help break down cellulose, releasing more fermentable sugars and improving digestibility.

Importance of Cellulose Analysis:
Knowing the cellulose content helps determine whether cellulase supplementation will be beneficial, especially in fibrous diets.
Over-supplementation with cellulase, however, can be wasteful, especially if the diet doesn’t contain significant amounts of cellulose.

3. Methods for Substrate Analysis
To effectively assess substrates like phytate, NSPs, and cellulose, several analytical techniques can be used to determine their levels in the feed. Common methods include:

a. Chemical Extraction
For phytate, specialized chemical extraction methods can isolate the phytate from other forms of phosphorus in the diet. These methods often involve acidic solutions that break down the phosphorus compounds and allow for precise quantification.
Example: The myo-inositol hexaphosphate method is commonly used for extracting and quantifying phytate in feed ingredients.

b. Enzymatic Assays
Enzymatic assays are used to quantify the NSP content in feed. These assays use specific enzymes (e.g., xylanase or cellulase) to break down the polysaccharides, and the resulting sugars can be measured to estimate the NSP content.
Example: The sequential enzyme digestion method uses various enzymes to sequentially degrade fiber fractions, and the sugars released are measured.

c. Fiber Analysis (Neutral Detergent Fiber and Acid Detergent Fiber)
Fiber analysis techniques like NDF (Neutral Detergent Fiber) and ADF (Acid Detergent Fiber) allow for the quantification of fiber content in the diet. NDF measures the cell wall content (including hemicellulose, cellulose, and lignin), while ADF gives a measure of the more indigestible parts of the cell wall, primarily cellulose and lignin.
Example: NDF and ADF can be used in conjunction with other tests to understand the specific fiber components that may benefit from enzyme supplementation.

4. Impact of Substrate Analysis on Enzyme Recommendations
Once the substrate levels are known, enzyme recommendations can be much more accurate and tailored. Here's how substrate analysis directly impacts enzyme strategies:

a. Phytase for Phytate
If high levels of phytate are detected (e.g., in corn or wheat), the inclusion of a phytase enzyme is highly recommended to improve phosphorus availability.
The dosage and type of phytase (e.g., 7-phytase or 3-phytase) will depend on the phytate levels and the specific needs of the diet.

b. Xylanase and Beta-Glucanase for NSPs
If NSPs, such as xylans or beta-glucans, are present in high quantities (common in wheat or barley), then xylanase or beta-glucanase supplementation will help reduce viscosity in the gut, improving nutrient digestibility and energy utilization.
The quantity of enzymes needed will depend on the specific levels of these NSPs.

c. Cellulase for Cellulose
If the feed contains high levels of cellulose (e.g., in forages or fibrous grains), cellulase supplementation can be considered to enhance fiber degradation and increase the availability of fermentable sugars.
By accurately matching enzyme types and dosages to the specific substrates in the diet, feed efficiency can be maximized, and the cost of enzyme supplementation can be justified.