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Trypsin Inhibitor, the hidden enemy in Soyabean Meal

Published on: 06/28/2021
Author/s : Dr. Koushik De, Director-Technical Services, SCA Novus International

As global animal production has rapidly shifted towards reduced Antibiotic free, “Gut health” has become a popular expression and all-encompassing concept in the scientific community. The gastro-intestinal tract must provide a barrier function protecting against harmful environmental elements (e.g. toxins and pathogenic microbes), while simultaneously permitting appropriate nutrient ab...

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brendon muko brendon muko
Industrial Engineer
June 28, 2021

What is the relationship between UA and over processed meal?

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Reena Rani L C Reena Rani L C
SCA Sr. Marketing Communication Specialist
Novus International Novus International
St Charles, Missouri, United States
June 30, 2021

brendon muko, Urease values of soybean meal should not be used as the best measure of optimum processing. measurement of the urease activity do not reflect the intensity of heat treatment of soybean meal beyond the threshold of urease. Chen et.al (2019) conducted a study to determine the levels of TI and UA in 414 SBM samples from 19 different countries and to validate whether TI and UA are correlated. They found that TI were poorly correlated with UA in solvent extracted SBM samples, suggesting that UA should not be used as a surrogate indicator for TI content in soybean products.

Refer the studies in the article: Araujo et al (2019) conducted similar study to determine the correlation of TI and KOH Protein Solubility.

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brendon muko brendon muko
Industrial Engineer
June 30, 2021
Reena Rani L C thanks, l understand. Can I please have the laboratory experiment procedure for KOH Protein solubility. I have been using the UA only but the challenge have been to see if it has been over processed and that's when the UA reaches its limitations and KOH Protein solubility is supposed to come up.
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Taha Taibeh Taha Taibeh
Veterinary Doctor
June 28, 2021
Thanks for valuable information..
But what if I already add protease shall I increase the dose, if yes what is the recommended..
Reply
Reena Rani L C Reena Rani L C
SCA Sr. Marketing Communication Specialist
Novus International Novus International
St Charles, Missouri, United States
June 30, 2021
Taha Taibeh That depends on what type of protease you are using and how efficiently it can degrade the TI in Soyabean meal even when TI are present in higher amount. A good quality protease enzyme improves the amino acid digestibility and at the same time reduce the pancreas weight also indicating a potential amelioration of the negative effect of TI. The dose recommendation is very much product specific.

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June 29, 2021
Hello Doc,
Why do you think we need to increase SBM level in a diet when we consider raising birds with no antibiotic/free antibiotic! What is the relationship between increasing CP level and limiting antibiotic?
Your quote "The shift to antibiotic free production or better gut health often results in the increase of soybean meal inclusion as ..."
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Reena Rani L C Reena Rani L C
SCA Sr. Marketing Communication Specialist
Novus International Novus International
St Charles, Missouri, United States
June 30, 2021
Dr. Abdolreza Kamyab : The shift to antibiotic free production or better gut health often results in the increase of soybean meal inclusion as there are limited in the number of efficacious protein sources that successfully reduce soybean meal content. Soybean meal is the most widely used major protein source in poultry production across the world. Sometimes we don’t give much importance to CP level unless there is country regulations. We give much emphasis to amino acid balancing. Higher CP will lead to higher amount of undigested protein to hind gut which become a source of food for the harmful pathogen.
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June 30, 2021
Dear Reena,
Thanks much for the reply.
My quest is what is the relationship between SBM level in poultry feed and antibiotic free diet? I do formulate both antibiotic free diet (with or without pre or pro biotic) and duets with antibiotic based on producers request. In either case my SBM level does not change!
Speaking on formulating diet based on aa digestibility is totally different issue.
Reply
Martin Smith Martin Smith
Animal Nutritionist
Evonik Animal Nutrition Evonik Animal Nutrition
Rellinghauser, Hessen, Germany
July 1, 2021

Dr. Abdolreza Kamyab, Dear Sir, I agree with your point. The first response to removing AGPs should always be to try and reduce "crude protein" as much as costs allow. Use of pure amino acids will always improve the possibility to do this and also the cost situation. In doing so we reduce nutrient supply for potential harmful overgrowth of the likes of C. perfringens etc. However, this article also points to the benefits from evaluating and controlling soya quality. Use of tools like AMINORED from Evonik make this very rapid and easy, and are directly correlated to SID values for amino acids. MPS

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July 2, 2021
Reena Rani L C emphasis should be on lowering the CP while balancing for the limiting amino acids. Such an approach has even been shown to reduce incidences of post-weaning diarrhoea.
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July 16, 2021
Reena Rani L C very interesting topic. What is group opinion about inclusion of SPC (Soybean Protein Concentrate) in the initial diets of the birds as a strategy on removing the AGP?
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July 30, 2021
Reena Rani L C Hi Dr Reena ,higher inclusions of SBM also causes higher NSPs ,its negative effect on gut health?
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June 29, 2021
Excellent article! Everybody knows about protein advantages of soya bean meal, but not enough about disadvantages of soya bean meal.
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Ana Gavrӑu Ana Gavrӑu
Global Technical Product Manager
July 2, 2021
Dr. Fiodor S. Marchenkov,
that's true. We need to pay more attention on the quality of the CP - it's real nutritional value based on the amino acids / percent CP, meaning gram Lysin in 100g CP- as Lysin is the most impacted AA during heat treatments.
Best regards, Ana Gavrau
Reply
July 2, 2021
Dear Ana, thanks for your reply, you are absolutely right. Lysine is key AA, and all PC-feed software calculating the rest of AA, according to lysine content. The main idea about reducing of SBM is very good. AA balance make possible to reach better CP digestion, and less substrate for harmful bacteria. But probably you know that in Ukraine SBM quality is not main feed problem because we use for poultry feed mainly domestic sunflower meal (SFM). It is cheaper but need some lysine addition, but any case final feed is cheaper. SFM dosn't contain TI or lectins. SFM chlorogenic acid is less problem if to compare it to TI.
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June 30, 2021
"The shift to antibiotic free production or better gut health often results in the increase of soybean meal inclusion as there are limited in the number of efficacious protein sources that successfully reduce soybean meal content"..... Could you please elaborate a bit more on how shift to AGP-free production has led to use of more SBM?
Reply
Reena Rani L C Reena Rani L C
SCA Sr. Marketing Communication Specialist
Novus International Novus International
St Charles, Missouri, United States
June 30, 2021

Oketch Elijah Ogola: As mentioned Soybean meal is the most widely used major protein source in poultry production across the world. And there are limited in the number of efficacious protein sources that successfully reduce soybean meal content. And now in many countries meat and bone meal/animal protein usage has been either stopped or been reduced significantly because of chances of microbial contamination. So more and more people are looking for feed without animal protein sources.

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July 2, 2021
Reena Rani L C thanks for the clarification
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Dr Kibiike David Dr Kibiike David
Bachlor. Veterinary Medicine
July 14, 2021
Am looking for testing unit to test the presence of trypsin inhibitor in roasted soy.
Reply
Martin Smith Martin Smith
Animal Nutritionist
Evonik Animal Nutrition Evonik Animal Nutrition
Rellinghauser, Hessen, Germany
July 27, 2021
Dear Dr. David

Please contact myself or John Owaga from Evonik; we will be able to discuss some assistance MPS
Reply
Dr. Rama Prasad Chakraborty Dr. Rama Prasad Chakraborty
Specialist in Animal Nutrition
July 27, 2021
Trypsin inhibitor so looming large to become a threat to use SBM in bulk albeit the same ingredient is the rich source of protein and a fair source of both L Lysine and DL Methionine. Urease activity in SBM doesn't reveal much significance in poultry feeds but it restricts the rampant use of SBM along with Urea in cattle feed. The measure to reduce TI activity in SBM in poultry feeds like heat treatment or any chemical exposure requires more exploration in near future in order to ensure more growth in poultry to meet out true marketable demand. It is to reiterate that any other source as source of protein to poultry feed other than SBM is deoiled mustard cake or rape cake though they too have had their limiting factors prior using in greater scale.
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July 30, 2021
Dr. Rama Prasad Chakraborty Dear Rama,During feed production heat treatment in conditioning is not sufficient to inactivate TI ?
Reply
Dr. Rama Prasad Chakraborty Dr. Rama Prasad Chakraborty
Specialist in Animal Nutrition
August 3, 2021
Dr Hafiz Rizwan Akram
I am confident that Trypsin inhibitor is heat liable. If SBM is boiled for thirty minutes the trypsin inhibitory function gets diluted upto 90 % and boiling SBM for 15 minutes would dilute same by 80% . Therefore review your opinion first on same before commenting.
Reply
July 29, 2021

Dear Dr. Koushik De. I find interesting your discussion on trypsin inhibitors and their role as antinutritional factors in broiler nutrition. It would be nice however, if you add a list of all the literature cited and provide credit where credit is due. Havenstein et al. (2003) didn't discuss trypsin inhibitors in their comparison of the 1957 and 2001 genetics, the slide that you have in your paper comes from my discussion in 2012 at the Arkansas Nutrition Conference [Ruiz, N., 2012. New insights on the urease activity range for soybean meal: a worldwide opportunity for the poultry industry. 2012 Arkansas Nutrition Conference Proceedings]. In my opinion, and of course, it is only my opinion, trypsin inhibitors are not "the hidden enemy" in soybean meal. Since the 1940's they started to be clearly identified by Kunitz (1945), Bowman (1944), and Birk (1961). And between 1969 and 1974 Kakade et al. developed the necessary analytical chemistry for their quantification. A considerable amount of the work on trypsin inhibitors has been done in soybeans and their products such as soybean meal.

I would say that rather than hidden, trypsin inhibitors in soy have been ignored for a variety of reasons. One important reason why during decades the industry didn't refer directly to trypsin inhibitors in soybean meal was the high correlation between urease activity and the inhibitors although it was not explicitly recognized. For decades the range of adequacy measured with urease activity was 0.05 to 0.20 pH units, and millions of tons of excellent quality SBM was produced and utilized in animal nutrition worldwide including broilers. This was true until the 1990's when the rapid feed passage syndrome showed up in broiler farms, particularly out of the United States [Kouwenhoven, B., R.M. Dwars, J.F.M. Smeets. 1992. Wet litter and high feed conversion, a new problem in broilers. Pages 558-561. Vol. 1. In: Proceedings XIX World’s Poultry Congress. WPSA. Amsterdam.] The rapid feed passage syndrome was the result of the dramatic increase in feed intake of broilers which accompanied with the predominance of soybean meal worldwide as the number one source of digestible amino acids exceeded the tolerance of broilers to trypsin inhibitors in the diet.
In 2005 [Ruiz, N., and F. de Belalcázar. 2005. Field observation: Trypsin inhibitors in soybean meal are correlated with outbreaks of feed passage in broilers. Poultry Science 84(Suppl. 1): 70.] we presented field data that indicated that commercial soybean meal displaying urease activity values within the range of adequacy were clearly involved in rapid feed passage in broilers older than 21 days of age. But in 2005 we measured simultaneously trypsin inhibitors and urease activity in lots of soybean meal that resulted in excellent performance of broilers as well as in lots that generated rapid feed passage. It became clear that the old range of adequacy had changed, and by 2012 at the Arkansas Nutrition Conference we proposed a new range: 0.000-0.050 pH units. Using the ISO 14902:2001 method to determine trypsin inhibitors we correlated this new proposed range for urease with a 1.65 - 2.35 mg/g range of adequacy for trypsin inhibitors. Since 2005 we also proposed the importance of measuring trypsin inhibitors as QC parameter in soybean meal and full-fat soybeans.

Yes, trypsin inhibitors is a QC parameter for both soybean meal and full-fat soybeans. Solubility in KOH is the parameter to measure if overprocessing has occurred in SOYBEAN MEAL and it is highly correlated to amino acid digestibility in soybean meal. We have reported that KOH protein solubility is not a QC parameter for full-fat soybeans [Ruiz, N., and C.M. Parsons. 2016. The absence of correlation between protein solubility and digestible amino acids in full-fat soybeans emphasizes focusing on trypsin inhibitors reduction. International Poultry Scientific Forum, Abstract T195 (p. 58); Poult. Sci. 95(E-Suppl. 1):251.]. Soybean meal and full-fat soybeans are two different ingredients each with distinctly quality control parameters [80th Minnesota Nutrition Conference, p. 169-175, 2019].

As far as how to deal with trypsin inhibitors in the feed formulation of broilers there are several alternatives beyond proteases. Nutritionists and formulators under commercial conditions have to think in overall cost, and formulating to minimize trypsin inhibitors in a broiler finisher diet is expensive because limiting soybean meal, the economic number one provider of digestible amino acids as already said, would result in higher price per metric ton of feed. As suggested by Dr. Marchenkov there are alternative ingredients such as sunflower meal that compete against soybean meal when a maximum spec for trypsin inhibitors in the formula is in play. Canola is another one. Per our experience, 0.58-0.60 mg of TI/g of FEED in a broiler finisher feed is roughly the upper limit.

Another way to effectively control trypsin inhibitors in broiler feeds is the utilization of full-fat soybeans (FFSB) which occur in several countries out of the United States. Given the fact that FFSB in practice is a no-commodity because is processed by the end user, then it is possible to decrease trypsin inhibitors in FFSB well below the trypsin inhibitor content of commercial soybean meal (a commodity). Soybean meal cannot reach by the solvent extraction process values below 1.60-1.65 mg TI/g without being overprocessed, while FFSB, PROPERLY PROCESSED may reach values as low as 1.0 mg TI/g (measured with the ISO 14902:2001 method). Consequently, by generating FFSB of superior quality than commercial soybean meal as far as trypsin inhibitors are concerned the only limitations are cost and inventory. In our experience FFSB competes excellently with soybean meal, reduces cost per MT of feed and allows to control trypsin inhibitors below the 0.58-0.60 mg TI/g of FEED. Nelson Ruiz Nutrition, LLC, Suwanee, GA USA.

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Dr. Rama Prasad Chakraborty Dr. Rama Prasad Chakraborty
Specialist in Animal Nutrition
August 3, 2021
Nelson Ruiz
As I do appreciate your opinion. But I would like to pay heed on doing any chemical treatment to reduce TI activities in SBM rather than switching over to FFSB and to replace partially with Canola and sunflower cakes which are low in protein and rich in either fat or fibre and that truly compromise the prescribed protein values in final composition. Hence it would be prudent to find out solution in view of reducing TI acitivies in commercial available SBM only by exploring various means and ways.
Reply
August 16, 2021
Rama Prasad Chakraborty
Yes, absolutely, I also appreciate your opinion. However, unfortunately many in the industry have not explored the advantages of full-fat soybeans (FFSB) not only from the energy content standpoint of view, but also, as I wrote, from the opportunity to considerably decrease residual trypsin inhibitors to almost zero by proper processing. It is not easy to overprocess FFSB under commercial conditions while soybean meal (solvent extracted) starts undergoing overprocessing when residual trypsin inhibitors descend below 1.50-1.55 mg/g (as determined with the ISO 14902:2001 method).
Now, the frequent criticism that either canola meal or sunflower meal will add excessive fiber to the diet is not that valid, in my opinion, because you are not replacing 100% of the soybean meal. You are just offering these ingredients so that least-cost formulation would take the minimum amounts to maintain the trypsin inhibitor content of the diet for broilers after 21 days of age under control. Evidently, canola meal and sunflower meal have lower crude protein content than soybean meal, but there are plenty of synthetic amino acids in the market. The suggestion to offer alternative ingredients to soybean meal is to SOLVE or PREVENT OUTBREAKS OF RAPID FEED PASSAGE IN THE FIELD. Nutritionists and management have to make a decision under the circumstances: is it more economical to lose feed conversion, have wet litter, and lack of carcass uniformity? Or accept that in order to dilute trypsin inhibitors in the diet because a specific lot of soybean meal arrived in the feed mill with 2.5-3.0 mg TI/g (instead of 1.80-2.00 mg/g) the cost of feed is going to increase but trypsin inhibitors will be controlled? Alternatively, it may be that proteases are more economic.
In our experience, however, for those poultry and feed companies in many countries (Turkey, Colombia, Mexico, Brazil, Peru, Argentina to mention just a few) that process raw soybeans into FFSB, optimization of their processes to minimized trypsin inhibitors is the way to go and Nelson Ruiz Nutrition, LLC offers the technical advice to reach such an optimization. NRN, LLC, Suwanee, GA, USA.
Reply
Dave Albin, Ph.D. Dave Albin, Ph.D.
VP, Nutrition & Extrusion Technologies
August 16, 2021
Thank you for this article. A couple of issues must be raised, however.
First, the correlations of urease activity and TI, measured as mg/g, may not be appropriate. Urease activity is a measure enzyme activity by assessing a rise in pH, while TI, in this case, is assessing the absolute amount of TI remaining in the meal. So, it's perhaps not surprising that a measurement of enzyme activity and a measurement of absolute amount of enzyme remaining would not correlate well, especially considering that these are two different molecules (urease vs. TI). It may be more appropriate to measure TI activity, vs. mg/g, and see how these values correlate with urease activity.
Second, the use of proteases poses one major problem, namely that these enzymes could degrade other exogenous enzymes, such as carbohydrases or fiber-degrading enzymes, making them less effective.
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August 17, 2021
Dear Dr. Albin, I am glad that you bring the issue of urease activity and trypsin inhibitors because it is clearly established in the work of several authors [McNaughton and Reece (1980); Mustakas et al., (1981); Ruiz (2012); Liu and Ruiz (2021)] that a high correlation exists between urease activity (delta pH) and trypsin inhibitors (expressed either as mg/g or TUI/mg) for SOYBEAN MEAL, particularly solvent extracted soybean meal.
On the other hand, we have demonstrated that urease activity and trypsin inhibitors are not correlated in full-fat soybeans (FFSB) accross different processing methods. You are right, trypsin inhibitors is the parameter to measure in evaluating FFSB for adequacy reference trypsin inhibitors [Ruiz, N., F. de Belalcázar, and J. Castillo. 2018. Field observations: Neither KOH protein solubility nor urease activity are adequate quality predictors of commercial full-fat soybeans for poultry. International Poultry Scientific Forum, Abstract T157 (p. 46); Poult. Sci. 97(E-Suppl. 1):311]. Nelson Ruiz Nutrition, LLC, Suwanee, GA USA
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brendon muko brendon muko
Industrial Engineer
August 24, 2021
Nelson Ruiz As you said that the relationship between UA and TI holds well when it comes to solvent extracted soya meal, so is it right to use the say relationship on Full fat Soyabean meal when the relationship of UA(?pH ) doesn't greatly really correlate with TI?.
Reply
August 24, 2021
Brendon Muko. Apparently I have not been clear: there is a correlation between urease activity and trypsin inhibitors in SOYBEAN MEAL. There is no such a relationship across different processing methods for FULL-FAT SOYBEANS.
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brendon muko brendon muko
Industrial Engineer
August 24, 2021
Nelson Ruiz Thanks and l really appreciate it.
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brendon muko brendon muko
Industrial Engineer
August 24, 2021
Nelson Ruiz if we can't they is no correlation between UA and TI in full fat Soyabean then how do we determine TI in Full fat Soyabean in an industry since it's expensive to perform TI quantity analysis during production since we can't use UA.
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September 7, 2021
Really good discussion. Certainly will be great to have such discussion face-to-face once again!
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September 18, 2021

Very nice and educative topic, I learned a lot.
Thank you to all the contributors.

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dan hofer dan hofer
Poultry farmer
November 1, 2021
very good topic so would whole soybean roasted be trypsin free
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