Slide 1 Hello, my name is Kurtis Goebel, Master's student with Dr. Stein here at the University of Illinois. I will be presenting phosphorus and energy digestibility in enzyme-treated soybean meal. Slide 2 So as a review, for the production of soybean meal, we'll first start with raw soybeans. Then the next steps will be, they will be cleaned, cracked, dehulled, and flaked, followed by hexane treatment to remove soybean oil, then we'll get our dehulled and defatted soybean meal. Then our next process will be heat treatment. And lastly we will produce our end product, which is our heat treated, dehulled and defatted soybean meal. Slide 3 For the development of Hamlet Protein products, we'll take this same heat treated, dehulled, and defatted soybean meal that we get from our convetional soybean meal process, and we'll treat this with a proprietary blend of enzymes. These enzymes will then be inactivated by enzyme inactivation process. Then these products will be dried, milled, and lastly we will get our Hamlet Protein products. So, when we measured our phosphorus and energy digestibility in these two experiments I will present later, we used several Hamlet Protein products. These products were sourced from the same soybeans that our conventaional soybean meal that we used in these experiments as well. Slide 4 So the first Hamlet Protein product that we used is HP-200, which is the standard enzyme-treated soybean meal. This is a product we used in our energy digestibility experiment. This product was processed at a faster rate, therefore we left more sugars and antinutritional factors as well as an added fraction of fibers was added back to the product. So we'll see an increase in crude fiber percentage in this product. Slide 5 The next three products that we used: First, it was HP-310, which is an enzyme-treated soybean meal. And we used this product both in the phosphorus and energy experiments. And this was sourced from a non-GMO soybean, and the reason why this is a non-GMO soybean is that Hamlet Protein Company is from Denmark, therefore the UK has restrictions in using genetically modified organisms. So to be able to satisfy their regulations, they have to use soybeans that were from a non-GMO source, so as you remember from the HP-200, this was actually a GMO sourced soybean to make that product. However, these three products were sourced from a non-GMO soybean. The second enzyme-treated soybean is HP-340, which is the same enzyme-treated soybean meal as in HP-310, however, this had added phytase. And as you can expect, we used this in our phosphorus experiment to determine if the phytase improves phosphorus digestibility. And lastly, as I mentioned before, we used a conventioanl soybean meal. And again, this is the same soybean meal source as our HP product that was sourced from the same soybeans. Slide 6 So the overall benefits of using enzyme treatment is, first we inactivate our trypsin inhibitors. Secondly, we removed our antinutritional factors, first being antigens, as I'll express as beta-conglycinin or glycinin. Secondly, we removed our sugars, such as our sucrose. And lastly, we removed oligosaccharides, as I'll express as stachyose or raffinose. Third, we have increased our crude protein percent, and lastly you'll see a tendency for fat percentage to also increase. Slide 7 So looking into our analysis of our enzyme-treated soybean meal, as you can see, over the top we have HP-200, HP-310, HP-340, and conventional soybean meal. When you look at dry matter percentages, you see a tendency for the enzyme-treated soybean meal to have higher dry percentages. When you have more water taken out of these products as you see, you'll have a tendency for the energy concentration to increase as well as your crude protein percent. Our GE, expressed as kcal/kg, we have 4400 kcal/kg in our enzyme-treated soybean meal, roughly, compared to our 4100 for soybean meal. When you look at crude protein percentage, you see a much greater crude protein percentage for enzyme-treated soybean meal, and this is again because we have removed more water so we should expect to see higher crude protein percentage when compared to our conventional soybean meal. When you look at total lysine percent in these products, we see an increase, actually, in our lysine percent when compared to our conventional soybean meal, and this again is because we have an increase in total crude protein percentage. For fat percentage in these products, we see a tendency for an increase, but not a substantial increase when compared to our conventional soybean meal. Slide 8 When we continue with our other analysis that we performed on these enzyme-treated soybean meals, we'll first look at total phosphorus. And as you can see, we see a similar total phosphorus percentage when compared to conventional soybean meal, and maybe even an increase in total phosphorus. As I mentioned before, we were able to remove the trypsin inhibitors by using enzyme-treated soybean treatment. So when you compare to conventional soybean meal at 5.7 trypsin inhibitor units per mg, after enzyme treatment with our HP-200, we see a reduction almost in 2 trypsin inhibitor units, as well as when we look at HP-340, we see a very substantial, even, decrease when compared to our conventional soybean meal. The two antigens that we expressed and analyzed for were beta-conglycinin and glycinin. These were expressed in parts per million. When you first look at conventional soybean meal, we have 130,000 parts per million, whereas when we enzyme-treat the soybean meal, we see significant reduction in 3, 4, and 5 parts per million for HP-200, 310, and 340. Similarly, in our glycinin concentration, for conventional soybean meal, we have 420,000 parts per million, whereas when we enzyme-treat these products, we see 17,000, 3300, and 90 parts per million for HP-200, 310, and 340. So we can conclude that the enzyme treatment was very successful in removing these antigen concentrations. Slide 9 So the additional benefit of using enzyme treatment was to not only remove the antigens, but like I said, we were able to remove the sucrose, stachyose, and raffinose. I first want to direct your attention to the bottom of the graph, where we analyze the conventional soybean meal that hasn't been processed with the enzyme treatment. So for the sucrose, we see 5.78% sucrose in conventional soybean meal. And as you look up across the HP-200, 310, and 340, we see a substantial decrease in the amount of sucrose. Similarly, for oligosaccharides of stachyose and raffinose, we see soybean meal at the bottom of the graph at 3.8%, roughly, for stachyose and 1% for raffinose. And as you look across your HP-200, 310, and 340, we see a substantial decrease in our concentrations for these oligosaccharides when we have enzyme-treated. Slide 10 So some of the implications: Again we can decrease sugars and oligosaccharide concentration in these enzyme-treated soybean meals. However, by decreasing the sugars and oligosaccharides we saw an increase in crude protein and fat. Slide 11 So when we compare how the conventional soybean meal and compare against how the Hamlet Protein products can be fed, as we know the conventional soybean meal is restricted in weanling pig diets, and this is because of the presence of antigens. However, when we look at the Hamlet Protein products, we can suggest that they could replace the high-priced animal proteins, and this is because we have removed the antigens. At the current time, amino acid digestibility has been measured in these products, however, phosphorus and energy digestibility have not. Slide 12 Therefore, it was our thought to measure phosphorus digestibility. Slide 13 For our objective of this experiment, we wanted to measure phosphorus digestibility in enzyme treated soybean meal. Next, we wanted to determine if enzyme treamtent of soybean meal compromises phosphorus digestibility compared to conventional soybean meal. Slide 14 I'll first discuss the Hamlet soybean products that we used. We used two of the Hamlet Protein products, one being Hamlet Protein 310 as I'll express as HP-310, and this is our standard enzyme-treated soybean meal. The second is HP-340, which is the same HP-310 but we've added phytase. And last, we'll test this against our conventional soybean meal, which again is sourced from the same soybeans. However, this product hasn't been put to the enzyme treatment like HP-310 and HP-340. Slide 15 So for our materials and methods, we placed 36 barrows in metabolism cages. We used six diets, with six pigs per diet. They were fed at three times their maintenance energy requirement. Slide 16 Feces were collected underneath a screen that was placed underneath the cage that they were housed in. Slide 17 So getting into our results, when you look at the total phosphorus in the feces, we first will set up the slide. At the bottom of the graph, you'll see HP-310, HP-340, and conventional soybean meal. On the left side is the percentage of phosphorus in the feces. The yellow bar is the diet without phytase. The blue bar is the diet with phytase. So as you can see, the diet that contained HP-310, when we have added phytase, we see significant reduction from 3.5% phosphorus to 2% phosphorus in the feces. However, when you look at HP-340, we do not see a statistical difference between the diets that had phytase added. Similarly to the HP-310, our conventional soybean meal had the same results as we have 3.5% phosphorus in the feces taken down to 2% phosphorus in the feces when we added phytase to the diet. Slide 18 So when we look at the apparent total tract digestibility of phosphorus, we see again similar results with HP-310 and conventional soybean meal as when we added phytase: we have increased our apparent total tract digestibility of phosphorus. However, with our HP-340, again is the enzyme-treated soybean meal with phytase, we do not see an additional benefit when we add 500 units of phytase to the diet. So you can conclude already that the phytase added to the HP-340 during the enzyme treatment is successful in maintaining the apparent total tract digestibility of phosphorus. Slide 19 So as we discuss some of the same points that I just went over in the last two graphs, again, our phosphorus in our feces was reduced when phytase was added. Pigs fed the Hamlet Protein 340 had a lower concentration of phosphorus in the feces than pigs fed HP-310 and conventional soybean meal regardless of phytase inclusion. Slide 20 Without dietary phytase, the apparent total tract digestibility of phosphorus was greater for HP-340 than for HP-310 and conventional soybean meal. With dietary phytase, the apparent total tract digestibility of phosphorus in HP-340 was greater than in HP-310. Slide 21 So our second experiment we looked at energy digestibility in enzyme treated soybean meal. Slide 22 So our objective again was to test whether enzyme treatment of soybean meal compromises the digestibility of energy compared to conventional soybean meal. Slide 23 We used two Hamlet soybean products in this experiment. One being HP-200, which is enzyme-treated soybean meal that was put through the enzyme treatment at a faster rate, leaving more crude fiber but decreasing our crude protein percentage to 52% compared to 56% as in our HP-310 and 340. The second Hamlet Protein product that we used was HP-310 and again, this is a standard enzyme treated soybean meal. And lastly, we compared this to our conventional soybean meal that we used as a standard that, again, was sourced from the same soybeans that was taken out from the process prior to enzyme treatment. Slide 24 For our materials and methods, we placed 28 barrows in metabolism cages. We used four diets with seven pigs per diet. We had total collection of urine and fecal materials. These pigs were fed at three times their maintenance energy requirement. Slide 25 So as in our phosphorus digestibility experiment, our collection of fecals was collected underneath a screen that was placed under their cages. And then for urine, another screen that had a funnel built into it collected our urine. Slide 26 So when we look at our apparent total tract digestibility of energy, our yellow bar symbolized our corn diet, our red bar is our HP-200 diet, our white bar is our HP-310 diet, and our blue bar is our conventional soybean meal diet. And you see, there are not statistical differences regardless of the diet we fed. Slide 27 However, on a dry matter basis, when you look at digestible energy for the individual ingredients, we see a reduction in digestible energy in corn when compared to our soy products. Then when you examine the soy products, we do not compromise our digestible energy in HP-200 and HP-310 that have been enzyme treated compared to our conventional soybean meal that has not been enzyme treated. Slide 28 However, on a dry matter basis, our metabolizable energy in these same ingredients, we do not see a statistical difference in all these ingredients. However, there is a tendency, maybe, for corn to be slightly less than our soy products. Slide 29 So for a few points to wrap up, our HP-200 and HP-310 and conventional soybean meal had similar digestible energy but all were greater than corn. And this is surprising because even with decreases in sugars, oligosaccharides, and an increase in crude fiber, we were still able to maintain the same level of digestibility than we did with our conventional soybean meal when we used an enzyme-treated soybean meal. Secondly, our metabolizable energy was not different among all treatments. Slide 30 So my take home message for both experiments is: Hamlet Protein soy products can be fed to young pigs -- and this is because we have removed the sugars (as I express with sucrose), oligosaccharides, antigens, and trypsin inhibitors -- and still be able to have similar phosphorus and energy digestibility as conventional soybean meal. Therefore, these soybean products can be used successfully in diets fed to young pigs. Slide 31 Thank you for your attention.