Progress 09/01/11 to 03/23/15
Outputs
Progress Report Objectives (from AD-416): 1) Produce functional protein isolates from animal by-products. 1a. Develop a pilot scale process for producing poultry blood-based flocculant. 1b. Increase the yield and molecular weight of soluble peptides extracted from rendered animal protein. 2) Utilize animal-protein isolates in biobased applications. 2a. Broaden the application range for animal protein-based polyacrylamide (PAM) substitutes. 2b. Develop practical processes for the production of fermentation feedstocks derived from animal by-product proteins. Evaluate these feedstocks in fermentation systems. Approach (from AD-416): Meat animal processing inevitably yields by-products, which include tissues and organs that are unsuitable for human food, such as feathers and bones, or for which there is low food market demand, such as blood. The current outlets for animal by-products, either through profitable use or disposal in the environment, are limited and shrinking. The proposed project is designed to create new utilization opportunities for these substances and reduce the environmental impact of meat processing. Two groups of promising applications are targeted here, use as fermentation feedstocks and use as renewable alternatives for synthetic polymeric flocculants such as polyacrylamide (PAM). Rendered protein will be processed into forms well-adapted for use in non-pharmaceutical industrial fermentations. Poultry blood and substances derived from rendered proteins will be tested in applications including algae culture flocculation, ethanol whole stillage clarification, mine tailings flocculation, cellulosic fiber flocculation, and municipal sewage sludge settling. At all stages, emphasis will be placed on employing processing techniques that are appropriate to the relatively low value of the products to be produced. Successful completion of this project will yield technologies that can be commercialized in a relatively short time period and produce new revenue streams for renderers and meat processors, while benefiting the environment. Normally if dense particles, such as sand, are introduced into a volume of water, the particles will sink. When the particles are very small, however, their tendency to sink is diminished. If muddy water is placed in a glass, the larger soil particles will sink to the bottom within a few minutes, but the finest particles may remain suspended in the water for days. There are a wide variety of industrial, agricultural, and environmental scenarios in which fine suspended particles must be removed from large volumes of water; often when dealing with large volumes, technologies such as filtration or centrifugation are impractical. Substances called flocculants are often used in such scenarios. Flocculants cause suspended particles to stick together in clumps, which greatly facilitates their removal. Development of both bio-based and synthetic polymeric flocculants is a very active area of research. Prior to the current project, our research group showed that certain proteins recovered from animal byproducts, such as blood or rendered protein, could be used to flocculate suspensions of clay particles. A large portion of the current project was devoted to building upon this initial discovery. A large portion of our efforts were directed towards solving the technical problems of transforming blood into a marketable flocculant. If blood-based flocculant goes into commercial production, blood will probably be collected from multiple slaughterhouses and transported to a central processing facility; such an arrangement will require blood to be held for 2-4 days prior to processing. Blood is highly unstable, and once outside the animal it quickly begins to degrade in a variety of ways. It coagulates within minutes, making recovery of the flocculant protein impractical. On a longer time scale, the red cells begin to break open, releasing their contents and greatly increasing the cost to recover the protein. Finally, the action of microbial contaminants in the blood produces hydrogen sulfide which is a major workplace hazard. In our research, we have measured the extent and timescale of each mode of blood degradation under industrially relevant conditions, beginning with collection of blood directly from slaughtered birds. We have identified inexpensive chemical treatments to inhibit the degradation and studied the effect of these treatments on flocculant performance. This work reduces barriers to the commercial adoption of the blood flocculant technology by allowing the blood collection and processing to be further separated in time and place. Production of flocculant from blood involves a series of processing steps which amount to a crude purification of the hemoglobin from the blood. One of these steps involves breaking open the blood cells to release their contents. The conventional method for rupturing blood cells introduces a large amount of extra water; this approach is inappropriate for flocculant production because it would significantly increase the processing cost. To break open blood cells without increasing the water content, we designed a continuous system built around an ultrasonic probe which creates intense pressure waves. We found this system to be very effective in breaking red blood cells while retaining the flocculating properties of the hemoglobin. We modelled the system so that complete hemoglobin recovery could be achieved with a minimum input of electrical energy. Exposure times as short as 75 milliseconds had a large effect on the release of hemoglobin. Such short exposure times allow a large volume of blood cells to be processed using a small piece of equipment. Our benchtop-sized system is able to process 24 liters an hour; a full-scale version of the system would handle volumes appropriate for industrial flocculant production. This work provides a reliable, inexpensive method for accomplishing a key step in flocculant production. Past research has also shown that protein extracted from meat and bone meal (MBM) can serve as an effective flocculant. The technical difficulty with using MBM in this application is the very poor solubility of MBM protein. Attempts were made to improve the solubility by cutting the protein into smaller pieces using alkali or enzymes, but the very small protein fragments produced were poor flocculants. To overcome these limitations we utilized an enzyme that makes fewer cuts in the protein, resulting conversion of 80% of the MBM protein into large protein fragments that are both soluble and act as good flocculants. This work represents progress towards the goal of MBM protein extracts that perform as well as blood proteins in flocculant applications. Another portion of our efforts focused on topics relating to the flocculant performance. The methods available for measuring and comparing the performance of various polymeric flocculants are not standardized and are weak in a number of technical respects. We took a systematic approach to designing a superior method that can be used to produce results that are comparable between research groups. Using a variety of flocculants, we crafted a method that is robust, sensitive, and reliable. It employs a relatively inexpensive instrument, and a special buffer system adopted from another area of research. This accomplishment will make flocculant research more productive, indirectly benefitting those in agriculture who are either flocculant users or producers of the raw materials used to make flocculants. Experiments were conducted on three important variables which can affect the performance of a flocculant - acidity, salt content and flocculant concentration. Blood and hemoglobin flocculated suspended particles best under relatively acidic conditions. The same substances performed increasingly well as the salt content of the suspension was increased; this surprising finding expands the range of applications that the flocculants may be useful for, since industrial suspensions often have high salt content. Finally the useful concentration range for these experimental flocculants was shown to be quite wide. In all of the work described up to this point, the substance to be flocculated has been clay. About one-third of biomass is composed of lignin. During biomass processing this lignin is often extracted into an alkaline solution. Lignin is normally recovered by treating the solution with sulfuric acid, which causes it to precipitate. We worked with an industrial collaborator who requires a method for recovering the lignin without the acid. A variety of conventional and experimental flocculants were tested for their performance in this application. The blood-based flocculant was one of only a couple flocculants found to be highly effective; none of the other biobased flocculants nor any of the synthetic organic flocculants were effective. We demonstrated that the addition of particular salts alters the flocculant protein structure, resulting in improved performance, and at least one synthetic flocculant can be used in conjuction with blood flocculant to improve overall flocculant, but only when these substances are added to the lignin solution in a particular manner. These results highlight that animal protein based flocculants are an increasingly attractive solution for the recovery of sulfur-free lignin. Accomplishments 01 Developed blood protein-based method for recovering lignin from biomass processing wastewater. The transformation of biomass into fuel often yields lignin as a major by-product. Lignin is valuable and useful, but recovery of the lignin from the biomass processing wastewater is problematic; the conventional recovery method results in highly acidic lignin and allows a significant proportion of it to escape. ARS Researchers at Wyndmoor, Pennsylvania, developed methods to recover lignin through flocculation, that can be used to either augment or replace the conventional lignin recovery method. The results of the research provide a new application for the previously developed blood- flocculant technology, and allow biomass processors to avoid lignin acidification and improve recovery. This research was conducted in cooperation with GreenValue Enterprises, LLC, a firm specializing in the production of lignin. 306 3 B 2010 02 Quantified the savings achievable by diverting blood from wastewater streams. Chicken blood is a by-product of chicken processing; it currently has no established uses and no market value. The majority of the blood is disposed of by renderers, but a portion of the blood enters into a plant�s wastewater stream. ARS Researchers at Wyndmoor, Pennsylvania, conducted a detailed examination of the contribution that blood makes to wastewater pollutant loads. They found that even small amounts of blood can have a major impact on the water quality. The data generated was used in conjunction with a nationwide sampling of municipal wastewater surcharge formulas to estimate the expenses that can be avoided by preventing blood loss. The results will help processors justify investments in upgrades to improve blood capture, and will promote the adoption of value-added uses for chicken blood. 306 3 B 2010
Impacts
(N/A)
Publications
- Garcia, R.A., Piazza, G.J. 2015. Application of the elusieve process to the classification of meat and bone meal particles. Applied Engineering in Agriculture. 31(1):165-170. DOI: 10.13031/aea.31.10410.
- Garcia, R.A., Clevenstine, S.M., Piazza, G.J. 2015. Ultrasonic processing for recovery of chicken erythrocyte hemoglobin. Journal of Food and Bioproducts Processing. 94:1-9. DOI: 10.1016/j.fbp.2014.12.002.
- Garcia, R.A., Stein, S.D., Piazza, G.J. 2014. Poultry blood preservation and the impact of preservation on flocculant activity. Applied Engineering in Agriculture. 30(3):445-453. DOI: 10.13031/aea.30.10436.
- Piazza, G.J., Lora, J., Garcia, R.A. 2014. Flocculation of kaolin and lignin by bovine blood and hemoglobin. Journal of Chemical Technology & Biotechnology. 90:1419-1425. DOI: 10.1002/jctb.4443.
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Progress 10/01/13 to 09/30/14
Outputs
Progress Report Objectives (from AD-416): 1) Produce functional protein isolates from animal by-products. 1a. Develop a pilot scale process for producing poultry blood-based flocculant. 1b. Increase the yield and molecular weight of soluble peptides extracted from rendered animal protein. 2) Utilize animal-protein isolates in biobased applications. 2a. Broaden the application range for animal protein-based polyacrylamide (PAM) substitutes. 2b. Develop practical processes for the production of fermentation feedstocks derived from animal by-product proteins. Evaluate these feedstocks in fermentation systems. Approach (from AD-416): Meat animal processing inevitably yields by-products, which include tissues and organs that are unsuitable for human food, such as feathers and bones, or for which there is low food market demand, such as blood. The current outlets for animal by-products, either through profitable use or disposal in the environment, are limited and shrinking. The proposed project is designed to create new utilization opportunities for these substances and reduce the environmental impact of meat processing. Two groups of promising applications are targeted here, use as fermentation feedstocks and use as renewable alternatives for synthetic polymeric flocculants such as polyacrylamide (PAM). Rendered protein will be processed into forms well-adapted for use in non-pharmaceutical industrial fermentations. Poultry blood and substances derived from rendered proteins will be tested in applications including algae culture flocculation, ethanol whole stillage clarification, mine tailings flocculation, cellulosic fiber flocculation, and municipal sewage sludge settling. At all stages, emphasis will be placed on employing processing techniques that are appropriate to the relatively low value of the products to be produced. Successful completion of this project will yield technologies that can be commercialized in a relatively short time period and produce new revenue streams for renderers and meat processors, while benefiting the environment. There a wide variety of industrial, agricultural and environmental scenarios in which suspended particles must be removed from large volumes of water; often when dealing with large volumes, technologies such as filtration or centrifugation are impractical. Substances called flocculants are often used in such scenarios. Flocculants cause suspended particles to stick together in clumps, which greatly facilitates the particles� removal. Development of both bio-based and synthetic polymeric flocculants is a very active area of research. In previous reporting periods we have shown that hemoglobin from poultry blood is promising as a biobased flocculant; the source material (blood) is a by-product that is burdensome to the poultry industry, and its technical performance as a flocculant is very strong. In FY14, our flocculant research advanced along a couple of fronts. Production of flocculant from blood involves a series of processing steps which amount to a crude purification of the hemoglobin from the poultry blood. One of these steps involves breaking open the blood cells to release their contents. To accomplish this task we designed a continuous processing system built around an ultrasonic probe which creates intense pressure waves. We found this system to be very effective in breaking red blood cells, while retaining the flocculating properties of the hemoglobin. Exposure times as short as 75 milliseconds had a large effect on the release of hemoglobin. Such short exposure times allow a large volume of blood cells to be processed with a small piece of equipment. Our bench top sized system is able to process 24 liters an hour; a full- scale version of the system would handle volumes appropriate for industrial flocculant production. Lignin is a biological material found in vascular plants. When using plant biomass for ethanol production, the fermentable sugar polymers are removed, leaving lignin as a byproduct. The lignin can be processed to provide bio-based substances which can replace particular petroleum-based substances. A collaborator from a lignin-producing business required a method for recovering lignin from dilute suspensions without using acid as is normally done for lignin recovery. A variety of conventional and experimental flocculants were tested for their performance in this application. The blood based flocculant was one only a couple tested flocculants found to be highly effective; none of the other biobased flocculant nor any of the synthetic organic flocculants were effective. Further research gave insight in the reasons for the blood flocculant�s good performance; this better understanding may facilitate further improvements of the flocculant�s performance. Accomplishments 01 Application of ultrasound to chicken blood processing. Hemoglobin is a substance inside of red blood cells. In the past it has been discovered that hemoglobin from waste chicken blood is valuable for producing a particular bio-based product. Isolating the hemoglobin from the blood requires a series of processing steps, one of which is breaking open the blood cells. ARS researchers at Wyndmoor, Pennsylvania adapted ultrasound technology to design a system for breaking open blood cells at a very high rate without damaging the hemoglobin. The researchers modelled the system so that complete hemoglobin recovery could be achieved with a minimum input of electrical energy. This system is designed to be compatible with practical manufacturing systems. This accomplishment is expected to facilitate the adoption of waste blood as a feedstock for bio-based products.
Impacts
(N/A)
Publications
- Piazza, G.J., Lora, J., Garcia, R.A. 2014. Flocculation of high purity wheat straw soda lignin. Bioresource Technology. 152:548-551. DOI: org/10. 1016/j.biortech.2013.11.040.
- Garcia, R.A., Riner, S.A., Piazza, G.J. 2013. Design of a laboratory method for rapid evaluation of experimental flocculants. Applied Engineering in Agriculture. 53(2):880-886.
- Piazza, G.J., Garcia, R.A. 2013. Proteolysis of meat and bone meal to increase utilisation. Animal Production Science. 54:200-206. Available:
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Progress 10/01/12 to 09/30/13
Outputs
Progress Report Objectives (from AD-416): 1) Produce functional protein isolates from animal by-products. 1a. Develop a pilot scale process for producing poultry blood-based flocculant. 1b. Increase the yield and molecular weight of soluble peptides extracted from rendered animal protein. 2) Utilize animal-protein isolates in biobased applications. 2a. Broaden the application range for animal protein-based polyacrylamide (PAM) substitutes. 2b. Develop practical processes for the production of fermentation feedstocks derived from animal by-product proteins. Evaluate these feedstocks in fermentation systems. Approach (from AD-416): Meat animal processing inevitably yields by-products, which include tissues and organs that are unsuitable for human food, such as feathers and bones, or for which there is low food market demand, such as blood. The current outlets for animal by-products, either through profitable use or disposal in the environment, are limited and shrinking. The proposed project is designed to create new utilization opportunities for these substances and reduce the environmental impact of meat processing. Two groups of promising applications are targeted here, use as fermentation feedstocks and use as renewable alternatives for synthetic polymeric flocculants such as polyacrylamide (PAM). Rendered protein will be processed into forms well-adapted for use in non-pharmaceutical industrial fermentations. Poultry blood and substances derived from rendered proteins will be tested in applications including algae culture flocculation, ethanol whole stillage clarification, mine tailings flocculation, cellulosic fiber flocculation, and municipal sewage sludge settling. At all stages, emphasis will be placed on employing processing techniques that are appropriate to the relatively low value of the products to be produced. Successful completion of this project will yield technologies that can be commercialized in a relatively short time period and produce new revenue streams for renderers and meat processors, while benefiting the environment. Flocculants are substances which help in clarifying liquids by causing suspended particles to settle more quickly; they are employed in a wide variety of industrial, agricultural, environmental and construction applications. Development of both bio-based and synthetic polymeric flocculants is a very active area of research. In previous reporting periods we have shown that hemoglobin from poultry blood is promising as a biobased flocculant; the source material (blood) is a by-product that is burdensome to the poultry industry, and its technical performance as a flocculant is very strong. In FY13, our flocculant work advanced along a couple of fronts. Commercial production of flocculant from poultry blood is likely to involve collection of blood from multiple slaughterhouses and transport to a central processing facility; such an arrangement will require blood to held for 2-4 days prior to processing. Blood is highly unstable, and once outside the animal it quickly begins to degrade in a variety of ways. It coagulates within minutes, making recovery of the hemoglobin impractical. On a longer time scale, the red cells begin to break open, releasing their contents and greatly increasing the cost to recover the hemoglobin. Finally, the action of microbial contaminants in the blood produces hazardous hydrogen sulfide which is a major workplace safety concern. In our research we have measured the extent and timescale of each mode of blood degradation under industrially relevant conditions, beginning with collection of blood directly from slaughtered birds. We have identified inexpensive chemical treatments to inhibit the breakdown and studied the effect of these treatments on flocculant performance. Completion and refinement of this work will greatly facilitate the commercial adoption of the blood flocculant technology by allowing the blood collection and processing to be further separated in time and place. In all prior flocculant work in this project, the substance to be flocculated has been clay. About one-third of biomass is composed of lignin. During biomass processing this lignin is often extracted into an alkaline solution. Lignin is normally recovered by treating the solution with sulfuric acid, which causes it to precipitate. Over the past year, we have worked with an industrial collaborator who requires a method for recovering the lignin without the acid. We have determined appropriate dosing and conditions for using hemoglobin coagulation in place of sulfuric acid precipitation. This work expands the range of applications for hemoglobin flocculant. Accomplishments 01 Conversion of meat and bone meal protein into an effective flocculant. In certain situations, agricultural soil erosion is controlled using a synthetic polymer chemical which causes the soil particles to clump together and prevents them from being carried away in the moving water. Meat and bone meal (MBM) is an inexpensive by-product of meat processing. It has been known that protein extracted from MBM can bind soil particles similarly to the synthetic chemical; MBM protein, however, has very low solubility and consequently is impractical to use in this application. Previously described methods for making MBM protein more soluble yield a product with greatly diminished soil binding properties. ARS researchers at Wyndmoor, Pennsylvania demonstrated that with a suitable enzyme and optimized reaction conditions, high solubility and good soil binding properties can be achieved simultaneously. This accomplishment provides a new utilization opportunity for MBM and an inexpensive source for biobased erosion control agents.
Impacts
(N/A)
Publications
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Progress 10/01/11 to 09/30/12
Outputs
Progress Report Objectives (from AD-416): 1) Produce functional protein isolates from animal by-products. 1a. Develop a pilot scale process for producing poultry blood-based flocculant. 1b. Increase the yield and molecular weight of soluble peptides extracted from rendered animal protein. 2) Utilize animal-protein isolates in biobased applications. 2a. Broaden the application range for animal protein-based polyacrylamide (PAM) substitutes. 2b. Develop practical processes for the production of fermentation feedstocks derived from animal by-product proteins. Evaluate these feedstocks in fermentation systems. Approach (from AD-416): Meat animal processing inevitably yields by-products, which include tissues and organs that are unsuitable for human food, such as feathers and bones, or for which there is low food market demand, such as blood. The current outlets for animal by-products, either through profitable use or disposal in the environment, are limited and shrinking. The proposed project is designed to create new utilization opportunities for these substances and reduce the environmental impact of meat processing. Two groups of promising applications are targeted here, use as fermentation feedstocks and use as renewable alternatives for synthetic polymeric flocculants such as polyacrylamide (PAM). Rendered protein will be processed into forms well-adapted for use in non-pharmaceutical industrial fermentations. Poultry blood and substances derived from rendered proteins will be tested in applications including algae culture flocculation, ethanol whole stillage clarification, mine tailings flocculation, cellulosic fiber flocculation, and municipal sewage sludge settling. At all stages, emphasis will be placed on employing processing techniques that are appropriate to the relatively low value of the products to be produced. Successful completion of this project will yield technologies that can be commercialized in a relatively short time period and produce new revenue streams for renderers and meat processors, while benefiting the environment. Flocculants are substances which help in clarifying liquids by causing suspended particles to settle more quickly; they are employed in a wide variety of industrial, agricultural, environmental and construction applications. Development of both bio-based and synthetic polymeric flocculants is a very active area of research. The tools available for measuring and comparing the performance of polymeric flocculants, though, are non-standardized and weak in a number of technical respects. In FY2012, we took a systematic approach to designing a method better suited for research into polymeric flocculants. Using a variety of flocculants, we crafted a method that is robust, sensitive and reliable. It employs a relatively inexpensive instrument, and a special buffer system adopted from another area of research. Our own flocculant research has progressed along a couple of fronts. In previous reporting periods we have shown that hemoglobin from poultry blood is promising as a biobased flocculant; the source material (blood) is a by-product that is burdensome to the poultry industry, and its technical performance as a flocculant is very strong. In FY12, we began work on design of a practical process for producing hemoglobin-flocculant from raw poultry blood at a pilot plant scale. This task is difficult because the flocculant properties of hemoglobin are diminished if it is not processed carefully, yet the processing costs must be kept very low. Our work in the past year has focused on the initial steps of the processing, where blood is collected and stored, while preventing clotting and red blood cell breakage. In investigating existing technologies for this purpose we found that while many were too expensive for our purposes, they were also overkill. We showed that significantly lower concentrations and lesser grades of anti-coagulant and preservative chemicals were adequate for non-medical blood processing. Meat and Bone Meal (MBM) is another promising source material for producing bio-based flocculants. The technical difficulty with using MBM in this application is the material�s very poor solubility. In past reporting periods we found that MBM protein could be solubilized through harsh alkaline or non-selective enzymatic treatments, but that such treatments sacrificed much of the potential flocculant activity in the material. In FY12, we found that most of the protein in MBM can be solubilized using much milder enzymatic treatments, resulting in products with better flocculant properties. These enzymatic treatments make only very selective cuts in the protein chain, resulting in bigger, more functional protein fragments. This research is an advance towards the specific goals of the CRIS project, but it could also serve as a starting point for development of an MBM-based adhesive. Accomplishments 01 A method for measuring the performance of flocculants. Flocculants are substances which assist in clarifying liquids by causing suspended particles to settle more quickly. Flocculants are used in agriculture f applications such as erosion control or manure treatment, and experiment flocculants are also being produced from agricultural by-products. Research and development into new flocculants requires methods for measuring flocculant performance, but the available methods are poorly designed. ARS researchers at Wyndmoor, Pennyslvania, have developed a flocculant performance measurement method that is robust, sensitive and reliable, using only inexpensive equipment and chemicals. This accomplishment will make flocculant research more productive, indirectly benefitting those in agriculture who are either flocculant users or producers of the raw materials to make flocculants.
Impacts
(N/A)
Publications
- Garcia, R.A., Phillips, J.G., Adeola, O. 2012. Improved prediction of meat and bone meal metabolizable energy content for ducks through in vitro methods. Poultry Science. 91:1854-1859.
- Piazza, G.J., Nunez, A., Garcia, R.A. 2012. Identification of highly active flocculant proteins in bovine blood. Applied Biochemistry and Biotechnology. 166(5):1203-1214.
- Liang, Y., Garcia, R.A., Piazza, G.J., Wen, Z. 2011. Non-feed application of rendered animal proteins for microbial production of eicosapentaenoic acid by the fungus Pythium irregulare. Journal of Agricultural and Food Chemistry. 59(22):11990-11996.
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