Innovative phase-wise process for the production of animal feed rich in aglycone isoflavones

Abstract

The process for the preparation of animal feed rich in aglycone isoflavones comprises: - at least one phase of supply (I) of a legume-cereal mixture comprising soybean products in a percentage by weight of between 1% and 80% of the total weight of the legume-cereal mixture; - at least one phase of milling (II) of the legume-cereal mixture to obtain a flour; - at least one phase of mixing / incubation (III) of the flour in acidic aqueous solution at a temperature of between 20°C and 70°C for a time comprised between Ih and 16h to obtain a mix containing aglycone isoflavones; at least one phase of rapid extraction (IV) of the aqueous matrix from the mix to obtain a semisolid matrix; - at least one phase of baking / drying (V) of the semisolid matrix at a temperature of between 110°C and 250°C for a time comprised between 10 min and 45 min; - at least one phase of cooling and stabilization (VI) of the resulting product to obtain an animal feed rich in aglycone isoflavones.

Description

[0001] INNOVATIVE PHASE-WISE PROCESS FOR THE PRODUCTION OF ANIMAL FEED RICH IN AGLYCONE ISOFLAVONES

[0002] Technical Field

[0003] The present invention relates to a phase-wise process for the preparation of animal feed rich in aglycone isoflavones.

[0004] Background Art

[0005] Isoflavones are plant compounds present in nature that have been classified as phytoestrogens. They possess a wide variety of beneficial biological properties for health, including anti-inflammatory, antioxidant and immunomodulatory properties. Part of their biological activity also manifests itself in the modulation of hormonal homeostasis through their high affinity for the estrogen receptor 0 (ER-0). They are mainly found in soybeans and soy-based products such as tofu, tempeh, soy milk and soy protein.

[0006] In nature, isoflavones are conjugated to carbohydrate molecules. These carbohydrate forms have no biological activity and are poorly absorbed by the digestive system. When the bond with these carbohydrate molecules is hydrolyzed, they are referred to as aglycone isoflavones. These hydrolyzed forms are linked to the beneficial biological activities widely described in scientific literature. In fact, a diet enriched with isoflavones with a high percentage of aglycones offers a number of biological and health benefits, comprising: hormonal regulation: isoflavones can mimic or modulate the effects of estrogen in the body, helping to balance hormone levels. This is particularly useful during menopause, when a drop in estrogen levels can lead to symptoms such as hot flashes, mood swings and osteoporosis. antioxidant properties: isoflavones have greater antioxidant activity than vitamins E and C, which help neutralize free radicals and reduce oxidative stress. This can protect cells from damage and reduce the risk of chronic diseases, including cancer and cardiovascular disease, in which oxidative stress plays a key role. anti-inflammatory effects: isoflavones have been shown to have antiinflammatory properties, which can reduce inflammation and potentially alleviate conditions such as arthritis and other inflammatory diseases. cholesterol reduction: studies indicate that isoflavones can improve lipid profiles by lowering LDL (bad) cholesterol levels and increasing HDL (good) cholesterol. This contributes to improved cardiovascular health and reduces the risk of heart disease. bone health: isoflavones may increase bone mineral density and decrease bone resorption, helping to prevent osteoporosis and fractures due to age- related bone loss. cancer prevention: isoflavones have been linked to a lower risk of certain cancers, particularly breast and prostate cancer. Their ability to modulate estrogen activity and operate as antioxidants plays a role in inhibiting the growth and proliferation of cancer cells. cardiovascular health: by improving cholesterol levels, reducing inflammation and operating as antioxidants, isoflavones contribute to the overall health of the cardiovascular system, thus reducing the risk of atherosclerosis, hypertension and other heart conditions. menopausal symptoms: isoflavones may alleviate menopausal symptoms such as hot flashes and night sweats by providing a natural source of estrogen-like compounds, thus improving the quality of life for postmenopausal women. weight control: isoflavones may be useful in weight control by influencing fat metabolism and reducing body fat accumulation, particularly in postmenopausal women. cognitive function: some studies suggest that isoflavones may support the cognitive function and potentially reduce the risk of neurodegenerative diseases such as Alzheimer, thanks to their neuroprotective and antiinflammatory properties.

[0007] These benefits are also seen when a diet enriched with aglycone isoflavones is administered to animals. For example, it is known that feeding laying hens with feed enriched with pasta for human consumption with a high content of aglycone isoflavones allows increasing egg production and improving nutritional quality. However, this formulation of animal feed combined with pasta for human consumption is neither practical nor feasible for large-scale animal feeding for obvious reasons of economic and industrial sustainability. Description of the Invention

[0008] The main aim of the present invention is to devise a process for the preparation of animal feed products that allows the concentration of biologically active isoflavones (aglycones) in the feed products themselves to be maximized in an effective, simple, natural and economically sustainable way without the use of additives and chemical laboratory processes.

[0009] Another object of the present invention is to devise a process for the preparation of animal feed products that allows the aforementioned drawbacks of the prior art to be overcome within the scope of a simple, rational, easy and effective to use as well as affordable solution.

[0010] The aforementioned objects are achieved by this process for the preparation of animal feed products rich in aglycone isoflavones having the characteristics of claim 1.

[0011] Brief Description of the Drawings

[0012] Other characteristics and advantages of the present invention will become more apparent from the description of a preferred, but not exclusive, embodiment of a process for the preparation of animal feed products rich in aglycone isoflavones. In the attached drawings:

[0013] Figure 1 represents a block diagram of the process according to the invention;

[0014] Figure 2 shows a graph indicating the percentage conversion of isoflavones to aglycone isoflavones as a result of the process.

[0015] Embodiments of the Invention

[0016] The process according to the invention for the preparation of animal feed products rich in aglycone isoflavones comprises: at least one phase of supply I of a legume-cereal mixture comprising soybean-based products in a percentage by weight of between 1% and 80% of the total weight of the legume-cereal mixture; at least one phase of milling II of the legume-cereal mixture to obtain a flour; at least one phase of mixing and incubation III of the flour in an acidic aqueous solution at a temperature of between 20°C and 70°C for a time of between Ih and 16h to obtain a mix containing aglycone isoflavones; at least one phase of rapid extraction IV of the aqueous matrix from the mix to obtain a semisolid matrix with a reduced moisture content; at least one phase of baking / drying V of the semisolid matrix at a temperature of between 110°C and 250°C for a time of between 10 min and 45 min; at least one phase of cooling and stabilization VI of the resulting product to obtain an animal feed product rich in aglycone isoflavones.

[0017] The selection of specific legume-cereal mixtures, together with a controlled phase of mixing / incubation III and a phase of baking / drying V, makes it possible to obtain feed products containing a high concentration of isoflavones through a high degree of hydrolysis exclusively obtained by “natural” means.

[0018] In fact, thanks to the natural presence of 0-glucosidase enzymes in the legumecereal mixtures, the particular incubation conditions of phase III allow obtaining the enzymatic hydrolysis of the carbohydrate bonds of isoflavones and allow obtaining biologically active aglycone isoflavones.

[0019] Soybean-based products are selected since they are known to be rich in isoflavones. In this regard, it should be noted that in this disclosure, the term “soybean-based products” refers to products of the soya plant and semi-finished derivatives, specifically soy seeds, soybean germ, extracts, concentrates, etc.

[0020] Cereals, on the other hand, are very rich in 0-glucosidase enzymes and allow a high degree of hydrolysis of isoflavones to be achieved. 0-glucosidase enzymes are found in smaller quantities in legumes.

[0021] According to a possible embodiment of this process, the soybean-based products are soy seeds. In this case, the soybean-based products are present in the legume-cereal mixture in a percentage by weight of between 20% and 80% of the total weight of the legume-cereal mixture, preferably equal to 50%.

[0022] In accordance with an alternative embodiment, the soybean-based products are soybean derivatives selected from the list comprising: concentrates, extracts, germ. Soybean derivatives generally have a higher isoflavone content. In this case, soybean-based products are present in the legume-cereal mixture in a percentage by weight of between 1% and 50% of the total weight of the legume-cereal mixture.

[0023] Furthermore, it cannot be ruled out that the legume-cereal mixture may comprise at least one plant product other than a legume and a cereal, or that it may comprise extracts, concentrates and derivatives of plants with a high isoflavone content (e.g., red clover and alfalfa).

[0024] Specifically, in addition to soybean-based products, the legume-cereal mixture may comprise at least one plant product selected from the list comprising: barley, com, wheat, rye, spelt, oat, millet, rice, sorghum, triticale, quinoa, sunflower, teff, fonio, kudzu, beans, lentils, peas, chickpeas, fava beans, peanuts, lupins, green beans, or a derivative thereof, such as extracts, concentrates, etc.

[0025] The legume-cereal mixture is then subjected to the phase of milling II, during which it is ground to obtain flour.

[0026] Grinding can be carried out using different types of mills, each with its own characteristics and specificities suited to certain milling objectives.

[0027] Grinding can be carried out using a mill selected from the list comprising: stone mill, roller mill, hammer mill, ball mill, ladder mill.

[0028] Preferably, grinding is carried out using a hammer mill.

[0029] The sizes of the sieves used in grinding range from 10 mesh to 40 mesh, preferably of between 20 mesh and 30 mesh.

[0030] The flour therefore has a grain size of between 0.4 mm and 2 mm.

[0031] Preferably, the flour has a grain size of between 0.55 mm and 0.85 mm.

[0032] The flour is then mixed with the acidic aqueous solution.

[0033] The acidic aqueous solution is a buffer solution obtained from a food-grade organic acid chosen from the (non-exhaustive) list comprising citric acid and ascorbic acid.

[0034] The use of an acid solution promotes the enzyme-induced hydrolysis reaction and stabilizes the product for storage purposes.

[0035] Usefully, the aqueous solution has a pH of between 4 and 6. Preferably, the aqueous solution has a pH of 5.

[0036] The phase of mixing / incubation III is carried out with a flour / aqueous solution ratio of between 1:0.2 and 1:4.

[0037] Preferably, the phase of mixing / incubation III is carried out with a flour / acidified aqueous solution ratio of 1: 1.3.

[0038] The phase of mixing / incubation III is preferably carried out at a temperature of between 40°C and 45°C.

[0039] Usefully, the phase of mixing / incubation III is carried out for a time of between 4 hours and 12 hours.

[0040] Preferably, the phase of incubation III is carried out for a period of 10 hours. As shown in Figure 2, in fact, 10 hours after the start of the phase of mixing / incubation III, the maximum percentage of conversion from isoflavones to aglycone isoflavones is substantially reached. The percentage of conversion was determined over time using HPLC (High Performance Liquid Chromatography) analysis.

[0041] The phase of mixing / incubation III is carried out using a heated mixer designed to allow gradual and homogeneous heat transfer, avoiding contact of the mix with the walls which are heated to a temperature above 45°C.

[0042] The optimal rotational speed of the mixer is of between 5 rpm and 35 rpm, preferably 24 rpm.

[0043] At the end of the phase of mixing / incubation III, the water contained in the mix is quickly extracted.

[0044] Usefully, the phase of rapid water extraction IV is carried out by means of a technique selected from the list comprising: centrifugation, mechanical pressing, percolation, filtration.

[0045] The phase of rapid water extraction IV comprises a step of recovery VII of the aqueous matrix and reuse of the same in the phase of mixing / incubation III for the subsequent preparation processes.

[0046] In accordance with a preferred embodiment, therefore, the aqueous solution of the phase of mixing / incubation III is preferably prepared from the aqueous matrix recovered from previous production processes.

[0047] The semi-solid matrix thus obtained is then subjected to the phase of baking / drying V. During the phase of baking / drying V, in addition to the reduction of residual moisture, a further heat-promoted hydrolysis process takes place, aimed at improving the nutritional properties and preservation of the finished product.

[0048] The phase of baking / drying V is preferably carried out at a temperature of 150°C.

[0049] The phase of baking / drying V is carried out using a convection oven, a fluid bed system, a hot air system, a microwave system, a negative pressure system, an extrusion system, an expansion system or a vacuum oven.

[0050] Preferably, the phase of baking / drying V is carried out using a convection oven. The phase of baking / drying V is carried out in such a way as to obtain a product with a predefined residual water content.

[0051] The residual water content is of between 10% and 14%.

[0052] At the end of the phase of baking / drying V, the solid matrix is cooled down in order to stabilize the product obtained and, if necessary, allow its use for the formulation of balanced animal feed with a high content of aglycone isoflavones.

[0053] This invention proposes a new method for the production of a new animal feed through a unique process that provides many health benefits for animals, comprising laying hens. It has, in fact, been proven that the benefits resulting from a diet rich in aglycone isoflavones improve both the production and the nutritional quality of the eggs laid. This includes, but is not limited to, improved eggshell strength, making the eggshell stronger and reducing the likelihood of breakage. In addition, the use of this new feed improves the quality of the yolk and its nutritional profile, as proven by the reduction in the cholesterol content of the egg, with obvious repercussions on human nutrition as well.

[0054] These biological effects can then have repercussions on the general well-being of hens, such as:

[0055] Bone health: Improved bone density: Isoflavones can increase bone mineral density, which is essential for laying hens to prevent osteoporosis and fractures, which are common problems due to the high demand for calcium for eggshell production.

[0056] Immune system enhancement: Antioxidant properties: Isoflavones have antioxidant effects that can reduce oxidative stress, thereby increasing overall immune response and reducing the incidence of disease.

[0057] Anti-inflammatory effects: Reducing inflammation helps maintain overall health and prevent chronic conditions.

[0058] Digestive health: Isoflavones can positively affect gut health by promoting beneficial bacteria and modifying the gut microbiome to improve nutrient absorption and overall digestive health.

[0059] Stress reduction: Isoflavones may potentially reduce stress in hens, leading to improved behavior and reduced pecking and cannibalism.

[0060] In conclusion, this new animal feed enriched with aglycone isoflavones improves egg production and quality and the overall well-being of hens by improving reproductive function, bone health, immune response and overall vitality.

[0061] An additional application of this new animal feed enriched with aglycone isoflavones is the potential to reduce greenhouse gas emissions from animals, particularly from herbivorous ruminants in intensive farming. By improving digestive efficiency, directly inhibiting methane production and improving the overall health and productivity of animals, this new isoflavone-enriched diet can contribute to more sustainable farming. It is proposed that this new animal feed enriched with aglycone isoflavones will be effective in reducing greenhouse gases:

[0062] Improved digestive efficiency with an enhanced fermentation process: Isoflavones can modify the microbial population in the gastrointestinal tract, promoting beneficial bacteria that improve the intestinal fermentation process. This can lead to more efficient digestion and potentially reduce the production of methane, one of the main greenhouse gases emitted by ruminants.

[0063] Direct inhibition of methane: Some studies suggest that isoflavones can modify the intestinal bacterial flora and directly inhibit methanogenic archaebacteria, the microorganisms responsible for methane production in the rumen.

[0064] Furthermore, 3-nitroxypropanol, a molecule similar in chemical structure to isoflavones, stands out as the most effective molecule currently available for reducing greenhouse gas emissions from ruminants, particularly methane production. The aglycone isoflavones in this new animal diet may target a similar mechanism, making it a promising solution for sustainable livestock farming. Our preliminary investigations using virtual models that employ artificial intelligence in the field of chemistry allow us to hypothesize a possible direct role for aglycone isoflavones in reducing methane production.

[0065] By improving the overall health and productivity of animals, isoflavones can lead to a reduction in the overall environmental footprint. Healthier animals with better growth rates and feed conversion ratios produce less greenhouse gases per unit of meat or milk.

[0066] All of the above suggests the importance of improving animal feed with natural products to have an impact on the environment and on animal health that can affect greenhouse gas production with a possible reduction in animal gas emissions.

[0067] The feed product obtainable by means of this process allows improving animal health, promoting greater growth, better bone density, better immune response and an overall improvement in animal well-being.

[0068] Furthermore, this process makes it possible to obtain a feed product that can be administered to laying hens, improving egg production and quality, characterized by stronger eggshells and an improved nutritional profile as demonstrated by the reduced cholesterol concentration in the components of eggs produced by animals fed with products with a high concentration of aglycone isoflavones.

[0069] At the same time, the feed product obtainable by this process also improves the intestinal microbiota.

[0070] Finally, it should be noted that this process and the related feed product contribute to reducing greenhouse gas emissions from ruminants through improved digestive efficiency and direct inhibition of methane production.

Claims

CLAIMS1) Process for the preparation of animal feed rich in aglycone isoflavones, characterized by the fact that it comprises: at least one phase of supply (I) of a legume-cereal mixture comprising soybean products in a percentage by weight of between 1% and 80% of the total weight of said legume-cereal mixture; at least one phase of milling (II) of said legume-cereal mixture to obtain a flour; at least one phase of mixing / incubation (III) of said flour in acidic aqueous solution at a temperature of between 20°C and 70°C for a time comprised between Ih and 16h to obtain a mix containing aglycone isoflavones; at least one phase of rapid extraction (IV) of the aqueous matrix from said mix to obtain a semisolid matrix; at least one phase of baking / drying (V) of said semisolid matrix at a temperature of between 110°C and 250°C for a time comprised between 10 min and 45 min; at least one phase of cooling and stabilization (VI) of the resulting product to obtain an animal feed rich in aglycone isoflavones.2) Process according to claim 1, characterized by the fact that said soybean products are soy seeds and by the fact that said soybean products are present in said legume-cereal mixture in a percentage by weight of between 20% and 80% of the total weight of said legume-cereal mixture.3) Process according to claim 1, characterized by the fact that said soybean products are soybean derivatives selected from the list comprising: concentrates, extracts, germ and by the fact that said soybean products are present in said legume-cereal mixture in a percentage by weight of between 1% and 50% of the total weight of said legume-cereal mixture.4) Process according to one or more of the preceding claims, characterized by the fact that said legume-cereal mixture comprises at least one plant product selected from the list comprising: barley, com, wheat, rye, spelt, oat, millet, rice, sorghum, triticale, quinoa, sunflower, teff, fonio, kudzu, beans, lentils, peas, chickpeas, fava beans, peanuts, lupins, green beans, a derivative thereof.5) Process according to one or more of the preceding claims, characterized by the fact that said legume-cereal mixture comprises at least one plant product with high isoflavone content other than legumes and cereals, such as red clover and alfalfa and derivatives thereof6) Process according to one or more of the preceding claims, characterized by the fact that said flour has a grain size of between 0.4 mm and 2 mm.7) Process according to one or more of the preceding claims, characterized by the fact that said aqueous solution has a pH of between 4 and 6.8) Process according to one or more of the preceding claims, characterized by the fact that said phase of mixing / incubation (III) is carried out with a flour / aqueous solution ratio of between 1:0.2 and 1:4.9) Process according to one or more of the preceding claims, characterized by the fact that said phase of rapid extraction (IV) is carried out by means of a technique selected from the list comprising: centrifugation, mechanical pressing, percolation, filtration.10) Process according to one or more of the preceding claims, characterized by the fact that said phase of rapid extraction (IV) comprises a step of recovery (VII) of said aqueous matrix and reuse of the same in said phase of mixing / incubation (III) for the subsequent preparation processes.11) Use of a food product obtainable by the process according to one or more of the preceding claims, for the preparation of a balanced animal feed.

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