Method of producing spray dried blood products from poultry blood

IDENTIFICATION OF RELEVANT DOMAIN OF SCIENCE

The invention relates to a method for the production of spray dried poultry blood products, i.e., the domain of Biotechnology

DESCRIPTION OF CURRENT STATE OF RELEVANT TECHNOLOGY

To the best knowledge of the applicant there is no existing entity in the world that is producing spray dried poultry blood products. This is mainly due to problems with obtaining and securing adequate quality poultry blood raw material in a manner that guarantees the preservation of the full value of protein fractions of poultry blood plasma, problems with removing debris falling into the raw material blood from the carcasses of poultry, and most importantly problems with securing and maintaining hygienic-quality of poultry blood in a slaughterhouse environment in a way that allows for further transport to the processing plant without the risk of propagation of pathogens. After transporting the blood to the processing plant, the fractionation process of blood and following spray-drying process may follow standards used commonly in the preparation of blood products from other animal groups like bovine and porcine blood.

DESCRIPTION OF THE MAIN PREMISE OF THE INVENTION

In the patent application contained herein the method for producing spray dried poultry blood products requires the following steps that begin with the way that blood is collected in slaughterhouses, and end in the processing facility. In slaughterhouses where birds are stunned with electrocution prior to slaughter, before the bird is slaughtered and blood released, we must remove excess water from the head, neck, and shoulders of the slaughtered birds. Per our own proprietary method, excess water will be blown off birds via nozzles deploying compressed air installed before the slaughter area. In slaughter facilities that stun birds with CO2, compressed air nozzles will not be necessary. Blood flowing from slaughtered birds flows into a blood collection trough where it is exposed to a sprayed mixture of a 10% solution of anti-coagulant in an amount equal to 4% of the total blood collected (by mass). Additionally, to the blood flowing into the collection trough we also add a solution of Volatile Fatty Acid Monoglycerides of carbon chain length C3 to C12 (propionic, butyric, decanoic, caprylic and lauric acid ) and glycerol in an amount equal to 0.1% - 10% of the total blood collected (by mass) to inhibit the growth of microflora. The anti-coagulant solution is intended to raise the pH levels significantly above pH 7. Short-chain fatty acids that are traditionally used to conserve feed components are only effective in environments with pH levels below 4.5. When pH levels are above 7, the degree of dissociation of organic acids increases dramatically and their antibacterial properties drop several hundred- to several thousand fold. Volatile Fatty Acid Monoglycerides (carbon chain length C3 - CI 2) do not hydrolyze and therefore maintain the same antibacterial qualities in environments with pH levels of 4.5 as well as above pH 7. Blood from the collection trough is purified on a set of sieves to remove mechanical impurities, chilled via a set of heat exchangers to a temperature between 3 ° C and + 10 ° C and transported to the processing plant while maintaining this temperature. Blood is then separated into poultry plasma and red blood cells (hemoglobin) by means of a high speed centrifuge. Poultry red blood cells are subjected to the spray drying process without additional processing. Poultry blood plasma subjected to a process of concentration and later purified using ultrafiltration / nanofiltration / dialysis. Afterwards it is also spray dried.

DESCRIPTION OF POTENTIAL BENEFITS OF THE INVENTION

The production and processing of poultry meat is among the most dynamically developing industries in Poland. This extremely dynamic development of the poultry industry has led Poland to become a leader in poultry production in the European Union. According to estimates by the Polish Institute of Agricultural Economics and Food Economy - National Research Institute, in 2015 Poland produced 2920 thousand tons of poultry for slaughter which is an increase of 9.4% versus 2014 and an increase of 23.1% versus 2013 performance (Poultry Market 2015). According to a report published by the European Commission in December 2014 ("Prospects for agricultural markets and income in the EU 2014-2024") in the next ten years poultry production and consumption is expected to further increase in the European Union.

Given the above, and in particular forecasts for the poultry industry in Poland and in the EU, it is relevant and particularly valuable to apply poultry blood - a byproduct of the process of poultry meat production - as a source of feed for the poultry industry. Blood accounts for up to about 10% of the bird's body weight. Thus, following the bleeding, with the current number of broilers chickens in Poland, about 70,000 - 80,000 tons of chicken blood only are produced annually. This quantity is significantly higher if we consider blood from other poultry animals. In total, poultry blood can be considered a substantial raw material source that is currently not utilized to its full potential. Poultry blood in the form of spray-dried plasma ("SDP") can serve as a source of high quality proteins, but most importantly also as a source of all biologically active substances (gamma globulin, albumin, and others) that are critical to animal production. At this time however poultry blood is not used in animal nutrition. Despite the fact that the plasma of poultry blood has a similar nutritional value to blood products from porcine and bovine animal species, its nutritional potential is wasted. Thus the invention contributes to the creation of a new market for new products that reduce the waste of valuable biological material contained in poultry blood.

Spray-dried plasma is a component of animal blood drawn and processed in such a way as to maintain the functional properties of the protein. SDP is a heterogeneous mixture of functional ingredients, enzymes and other compounds biologically active in the intestine of animals fed with the feed additive.

Following the 2006 ban on the use of antibiotic growth promoters in animal feed, steps have been taken to find new solutions that could be used to prevent gastrointestinal disorders and improve animal production (10). One good solution has been the use of feed additives in the form of spray- dried blood plasma. Spray-dried plasma is a protein rich diet supplement used in a variety of animal species (2, 7, 28). The main components of SDP are albumin, immunoglobulin, and fibrinogen. In addition, plasmids contain many biologically active proteins, such as insulin-like growth factors I, II, or transforming growth factor β (TGF-β), responsible for stimulating intestinal development, protein synthesis and intestinal regeneration ( 7, 14, 28). The amino acid composition of proteins in spray dried blood plasma is well balanced and does not differ significantly between plasma derived from pigs (SPDP - spray dried porcine plasma), plasma of bovine origin (SDBP - spray dried bovine plasma) or from poultry blood plasma (SDPP - spray dried poultry plasma).

A summary of available studies indicates that SDP modulates immune system activity, thereby protecting the body from excessive or too weak immune responses (2, 14, 25). SDP biological activity against enteric pathogens is partly due contained in the plasma of the immunoglobulin class IgG. The plasma for SDP production is derived from a very large number of animals with differentiated immune status. As a result, SDP includes a wide range of antibodies against pathogens present in the environment of livestock farming, such as E. coli, rotavirus, and other circoviruses (1, 27).

There is also specific research on the impact of porcine and bovine SDP on poultry production. It has been shown that broiler feed containing 1% SDP fed during the first 14 days of rearing, followed by feed containing a 0.5% and 0.25% addition of the plasma during the rearing period between 15 to 28 days and day 29 to day 42 respectively, improves weight gain and shape of the carcass, increases the efficiency of the pectoralis growth up to 5% (3). Campbell et al. (6) also showed that turkeys infected with Pasteurella multocida that were fed a diet containing 1 % SDBP in the first week, 0.5% from 2 to 4 weeks and 0.25% in 5 to 8 weeks experiences a milder form of the disease and showed greater survival rates. This is undoubtedly due to the immunomodulating properties of SDP (7). It has been shown that SDP fed turkeys achieve a 6.3% higher final body weight with a 3.7% lower feed consumption per kilogram of growth and a 15% reduction in the number of slaughter rejections (5).

Taking this information into account, it can be stated that adding porcine or bovine SDP to the feed of broiler chickens or turkeys improves the daily mass gains of the birds, the intake of feed, reduces the severity of diseases and in turn allows for a reduced reliance on antibiotics. It is also worth noting that porcine and bovine SDP significantly improves the body mass growth of broiler chickens, even if only used as a 1-2% feed additive during the first 10 days post hatching.

While general benefits of SDP are well documented, it should be strongly emphasized that the production and use of spray dried poultry plasma in animal feeds is innovative, and the review of existing literature confirms the lack of prior industrial production of poultry SDP. Producing such plasma and using it in animal feed will have a positive impact on the environment and will bring economic benefits to the poultry industry and other animal production industries.

Poultry blood plasma and porcine blood plasma are characterized by a similar ability to inhibit the development of pathogens, similar protein regulatory activity, and similar digestibility. As it was noted in the introduction, Poland is a leader in the production of poultry in Europe, and the growth in poultry production is increasing as opposed to the swine industry in Poland which is shrinking. Given the specificity of the way that animal immune systems develop and function as well as the unique properties of poultry blood plasma, it can be concluded that Poultry Spray Dried Plasma will bring greater economic benefits for the animal production industry, especially in the poultry sector, than the use of porcine and bovine plasma. In light of the above, the market potential of spray dried poultry blood products is even greater than that of porcine blood products.

Additionally, the Volatile Fatty Acid Monoglycerides (C3 - CI 2) added to the blood to stabilize the quality of the collected raw material, are present in the finished Spray Dried Poultry Plasma product at a concentration sufficient to exert a synergistic effect on the feed of animals. The Volatile Fatty Acid Monoglycerides not only inhibit the development of pathogens but also, like blood plasma, reduce the amount of proinflammatory cytokines secreted in response to pathogens, their toxins or other stress vectors. The Volatile Fatty Acid Monoglycerides improve energy balance and antioxidant potential in the small intestine and the body (4, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 , 24, 26, 29, 30).

BIBLIOGRAPHY

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2. Bosi P., Casini L., Finamore A., Cremokolini C, Merialdi G., Trevisi P., Nobili F., Mengheri E.: Spray - dried plasma improves growth performance and reduces inflammatory status of weaned pigs challenged with enterotoxigenic Escherichia coli K88. J. Anim. Sci. 2004, 82, 1764 -1772.

3. Bregendahl, K., Ahn D. U., Trampel D. W., Campbell J. M.: Effects of dietary spray- dried bovine plasma protein on broiler growth performance and breast-meat yield. J. Appl. Poult. Res. 2005, 14:560-568.

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6. Campbell, J.M., Quigley J.D. Ill, Russell L.E., Koehnk L.D.: Efficacy of spray-dried bovine serum on health and performance of Turkey challenged with Pasteurella multocida. J. Appl. Poult. Res. 2004, 13:388-393.

7. Campbell J.: Influence of dietary plasma proteins on supporting animal immunity systems. 19th Annual meeting. Florida Ruminant Nutrition Symposium. Florida 2008. 8. Cresci, G., Nagy, L. E., Ganapathy, V., 2013. Lactobacillus GG and tributirin supplementation reduce antibiotic-induced intestinal injury. JPEN J Parenter Enteral Nutr. 37 (6) doi: 10.1177/01486071134868

9. Czerwinski, J., O. ttojberg, S. Smulikowska, R. M. Engberg, and A. Mieczkowska. 2012. Effects of sodium butyrate and salinomycin upon intestinal microbiota, mucosal morphology and performance of broiler chickens. Arch. Anim. Nutr.

10. Davis, R. E. 1930. The metabolism of tributyrin. J. Biol. Chem. 88:67-75.

11. Fusunyan, R. D., J. J. Quinn, M. Fujimoto, R. P. MacDermott, and I. R. Sanderson. 1999. Butyrate switches the pattern of chemokine secretion by intestinal epithelial cells through histone acetylation. Mol. Med. 5:631-640.

12. Jerzsele, A., K. Szeker, R. Csizinsky, E. Gere, C. Jakab, J. J. Mallo, and P. Galfi. 2012. Efficacy of protected sodium butyrate, a protected blend of essential oils, their combination, and Bacillus amyloliquefaciens spore suspension against artificially induced necrotic enteritis in broilers. Poult. Sci. 91 :837-843.

13. Jiaolong Li, Yongqing Hou, Dan Yi, Jun Zhang, Lei Wang, Hongyi Qiu, Binying Ding, and Joshua Gong: Effect of tributirin on intestinal energy status, antioxidative capacity and immune response to liposaccharide response in chicken. Asian Austr. J. Animal Sci., 2015, vol. 28, 12: 1784 - 93.

14. Jiang R., Chang X., Stoll B., Fan M. Z., Arthington J., Weaver E., Campbell J., Burrin D. G.: Dietary plasma protein reduces small intestinal growth and lamina propria cell density in early weaned pigs. 2000, 130, 21-26.

15. Kotunia, A., J. Wolinski, D. Laubitz, M. Jurkowska, V. Rome, P. Guilloteau, and R. Zabielski. 2004. Effect of sodium butyrate on the small intestine development in neonatal piglets fed by artificial sow. J. Physiol. Pharmacol. 55(Suppl 2):59-68.

16. Leeson, S., H. Namkung, M. Antongiovanni, and E. H. Lee. 2005. Effect of butyric acid on the performanceand carcass yield of broiler chickens. Poult. Sci. 84: 1418-1422.

17. Lu, J. J., X. T. Zou, and Y. M. Wang. 2008. Effects of sodium butyrate on the growth performance, intestinal microflora and morphology of weanling pigs. J. Anim. Feed Sci. 17:568- 578.

18. Mahdavi, R. and M.Torki. 2009. Study on usage period of dietary protected butyric acid on performance, carass characteristics, serum metabolite levels and humoral immune response of broiler chickens. J. Anim. Vet. Adv. 8: 1702-1709.

19. Mallo, J. J., A. Balfagon, M. I. Gracia, P. Honrubia, and M. Puyalto. 2012. Evaluation of different protections of butyric acid aiming for release in the last part of the gastrointestinal tract of piglets. J. Anim. Sci. 90(Suppl4):227-229.

20. Namkung, H., H. Yu, J. Gong, and S. Leeson. 2011. Antimicrobial activity of butyrate glycerides toward Salmonella Typhimurium and Clostridium perfringens. Poult. Sci. 90:2217- 2222.

21. Nancey, S., J. Bienvenu, B. Coffin, F. Andre, L. Descos, and B. Flourie. 2002. Butyrate strongly inhibits in vitro stimulated release of cytokines in blood. Dig. Dis. Sci. 47:921-928.

22. Ogawa, H., P. Rafiee, P. J. Fisher, N. A. Johnson, M. F. Otterson, and D. G. Binion. 2003. Butyrate modulates gene and protein expression in human intestinal endothelial cells. Biochem. Biophys. Res. Commun. 309:512-519.

23. Panda, A. K., S. V. Rama Rao, M. V. L. N. Raju, and G. Sunder Sunder. 2009. Effect of butyric acid on performance, gastrointestinal tract health and carcass characteristics in broiler chickens. Asian Australas. J. Anim. Sci. 22: 1026-1031.

24. Parka., J. S., M. S. Woo, S. Y. Kim, W. K. Kim, and H. S. Kim. 2005. Repression of interferon-y-induced inducible nitric oxide synthase (iNOS) gene expression in microglia by sodium butyrate is mediated through specific inhibition of ERK signaling pathways. J. Neuroimmunol. 168:56-64.

25. Perez-Bosque A., Miro L., Polo J., Russell L., Campbell J., Weaver E., Crenshaw J., Moreto M. : Dietary plasma proteins modulate the immune response of diffuse gut - associated lymphoid tissue in rats challenged with Staphylococcus aureus enterotoxin. B. J. Nutr. 2008, 138, 533-537.

26. Sauer, J., K. K. Richter, and B. L. Pool-Zobel. 2007. Physiological concentrations of butyrate favorably modulate genes of oxidative and metabolic stress in primary human colon cells. J. Nutr. Biochem. 18:736-745.

27. Shen H. G, Schalk S., Halbur P. G., Campbell J. M., Russell L. E., Opriessnig T.: Commercially produced spray dried porcine plasma contains high levels of porcine circovirus type 2 (PCV2) DNA but not transmit PCV2 when fed to naive pigs. J. Anim. Sci. 2011, 89, 1930-1938.

28. Torrallardona D.: Spray dried plasma as an alternative to antibiotics in weaning pigs. Asian- Aust. J. Anim. Sci. 2010, 23, 131-148. 29. Yi, D., Y. Hou, L. Wang, B. Ding, Z. Yang, J. Li, M. Long, Y. Liu, and G. Wu. 2014. Dietary N-acetylcysteine supplementation alleviates liver injury in lipopolysaccharide-challenged piglets. Br. J. Nutr. 111:46-54.

30. Zhang, W. H., Y. Jiang, Q. F. Zhu, F. Gao, S. F. Dai, J. Chen, and G. H. Zhou. 2011. Sodium butyrate maintains growth performance by regulating the immune response in broiler chickens. Br. Poult. Sci. 52:292-301.

In summary, spray dried poultry plasma produced with the application of proprietary technology as contained in this application is expected to yield the following benefits:

Reduction of the need to rely on antibiotic interventions in animal production.

Reduction in the rate and severity of the animals' immune response to environmental stress and a resulting reduction in production losses caused by deaths, reduced weight gain, and poor feed conversion due to excessive activation of the cytokine cascade in response to stress

Improving the yield of meat from production animals as a result of lower energy losses on immune tissue activation and adverse effects of proinflammatory cytokines on muscle tissue growth

EXPLANATION OF THE FIGURES (if relevant)

No figures included

EXAMPLE OF IMPLEMENTATION OF THE INVENTION

Example 1

In slaughterhouses where birds are stunned with electrocution prior to slaughter, before the bird is slaughtered and blood released, we must remove excess water from the head, neck, and shoulders of the slaughtered birds. Per our own proprietary method, excess water will be blown off birds via nozzles deploying compressed air installed before the slaughter area. In the process of collecting poultry blood poultry a pre-mixed 10% sodium citrate solution will be added as an anticoagulant. To the anticoagulant solution a mixture of monoglycerides C3 - C12 and glycerol will be added at rate of 10%. The resulting solution of anticoagulant and monoglyceride is sprayed onto the blood flowing in the collection trough in the amount of 4% of the total blood collected (by mass). The resulting blood with an added anticoagulant (0.4%) and monoglycerides (0.4%) is preliminarily filtered on the collection trough, and then is cooled down in the heat exchanger to a temperature of 5 ° C. This process results in the collection of blood deemed to be hygienic. The hygienic blood is stored and transported in isothermal tanks to a processing plant where the centrifuge separates the fluid into red blood cells and blood plasma. The red blood cells are directed to spray drying and poultry blood plasma containing the anticoagulant and monoglycerides undergoes concentration and is processed to remove the excess salt via specially configured ultrafiltration, nanofiltration, and dialysis membranes. Concentrated plasma (concentrated above a level of 25% dry mass) is directed to spray drying which yields a spray-dried poultry blood plasma containing about 72% protein and 9% fat.

Example 2

In slaughterhouses where birds are stunned with electrocution prior to slaughter, before the bird is slaughtered and blood released, we must remove excess water from the head, neck, and shoulders of the slaughtered birds. Per our own proprietary method, excess water will be blown off birds via nozzles deploying compressed air installed before the slaughter area. In the process of collecting poultry blood poultry a pre-mixed 10% sodium citrate solution will be added as an anticoagulant. To the anticoagulant solution a mixture of monoglycerides C3 - C12 and glycerol will be added at rate of 2.5%. The resulting solution of anticoagulant and monoglyceride is sprayed onto the blood flowing in the collection trough in the amount of 4% of total blood collected (by mass). The resulting blood with an added anticoagulant (0.4%) and monoglycerides (0.1%) is preliminarily filtered on the collection trough, and then is cooled down in the heat exchanger to a temperature of 5 ° C. This process results in the collection of blood deemed to be hygienic. The hygienic blood is stored and transported in isothermal tanks to a processing plant where the centrifuge separates the fluid into red blood cells and blood plasma. The red blood cells are directed to spray drying and poultry blood plasma containing the anticoagulant and monoglycerides undergoes concentration and is processed to remove the excess salt via specially configured ultrafiltration, nanofiltration, and dialysis membranes. Concentrated plasma (concentrated above a level of 25% dry mass) is directed to spray drying which yields a spray-dried poultry blood plasma containing about 77% protein and 4% fat.

Example 3

In slaughterhouses where birds are stunned with electrocution prior to slaughter, before the bird is slaughtered and blood released, we must remove excess water from the head, neck, and shoulders of the slaughtered birds. Per our own proprietary method, excess water will be blown off birds via nozzles deploying compressed air installed before the slaughter area. In the process of collecting poultry blood poultry a pre-mixed 10% sodium citrate solution will be added as an anticoagulant and sprayed onto the blood flowing in the collection trough in the amount of 4% of total blood collected (by mass). A mixture of monoglycerides C3 - C12 and glycerol will added to the blood in the collection trough via an independent spray applicator in the amount of 10% of the total blood collected (by mass). The resulting solution of anticoagulant and monoglyceride is sprayed onto the blood flowing in the collection trough in the amount of 4% of total blood collected (by mass). The resulting blood with an added anticoagulant (0.4%) and monoglycerides (0.1%) is preliminarily filtered on the collection trough, and then is cooled down in the heat exchanger to a temperature of 5 ° C. This process results in the collection of blood deemed to be hygienic. The hygienic blood is stored and transported in isothermal tanks to a processing plant where the centrifuge separates the fluid into red blood cells and blood plasma. The red blood cells are directed to spray drying and poultry blood plasma containing the anticoagulant and monoglycerides undergoes concentration and is processed to remove the excess salt via specially configured ultrafiltration, nanofiltration, and dialysis membranes. Concentrated plasma (concentrated above a level of 25% dry mass) is directed to spray drying which yields a spray-dried poultry blood plasma containing about 27% protein and 62% fat.