SOD (superoxide dismutase) compound bacteria fermented feed as well as preparation method and application thereof

Abstract

The invention provides an SOD (superoxide dismutase) compound bacteria fermented feed as well as a preparation method and application thereof, and belongs to the technical field of feed products. Through a specific raw material combination and a compound bacteria fermentation process, the nutritional value and functionality of the feed can be remarkably improved. The prepared fermented feed is rich in high-activity beneficial components, can effectively improve the sensory quality of the feed and the ingestion willingness of animals, can also promote the absorption and utilization of nutrient substances, has positive effects on enhancing the health state of animal bodies and improving the breeding benefits, and provides a feasible scheme for developing novel functional feed products.

Description

Technical Field

[0001] This invention relates to the field of feed product technology, and in particular to a fermented feed containing SOD complex bacteria, its preparation method, and its application. Background Technology

[0002] As animal husbandry and aquaculture become more intensive and large-scale, animal production performance continues to improve, but the problem of oxidative stress caused by metabolism is also becoming increasingly prominent. Oxidative stress can damage animal cells, affect immune function and overall health, and consequently lead to a decline in production performance. Therefore, adding exogenous antioxidants to feed to help animals maintain redox balance has become an important direction in modern animal nutrition research and feed industry practice.

[0003] Superoxide dismutase (SOD), a key metalloenzyme, is the first line of defense in organisms against superoxide anion free radicals, possessing highly efficient antioxidant capabilities. Theoretically, its application in animal feed could help animals directly resist oxidative damage. Traditional methods of addition primarily involve directly using SOD preparations extracted from animal blood or plants, or adding natural raw materials rich in SOD. However, directly added SOD may face problems such as unstable activity and easy inactivation during feed processing, storage, and in the animal's digestive tract, and its high cost limits its large-scale application.

[0004] On the other hand, microbial fermentation technology, as a green and efficient feed processing method, has been widely used in the development of new feed resources. Fermenting feed ingredients with specific probiotics (such as lactic acid bacteria, yeast, and Bacillus) can produce various beneficial metabolites such as organic acids, digestive enzymes, and vitamins, while simultaneously degrading anti-nutritional factors and improving palatability and digestibility. Fermented feed itself also possesses the potential to regulate the intestinal microecology due to its high content of live bacteria. How to organically combine antioxidant functions with the multiple benefits of fermented feed, and generate or enrich stable active substances in situ through the fermentation process, is a potential pathway currently being explored. Summary of the Invention

[0005] The purpose of this invention is to provide a SOD compound microbial fermented feed, its preparation method and application, which solves the technical problems of existing feed additives having single function and insufficient synergistic effect in effectively improving feed superoxide dismutase activity, palatability and digestibility.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0007] This invention provides a SOD compound microbial fermented feed, which is made from the following raw materials in parts by weight: 70-90 parts Zhonghua Liangmu forage grass, 5-15 parts corn, 5-15 parts soybean meal, and 0.05-1.5 parts compound microbial fermentation agent.

[0008] Preferably, in the above-mentioned SOD compound microbial fermented feed, the compound microbial fermenting agent includes lactic acid bacteria, Lactobacillus plantarum, Bacillus subtilis, Rhodopseudomonas palustris, Propionibacterium propionate, Lactobacillus brunelli, Lactobacillus paracasei, Lactococcus lactis, Bacillus licheniformis, yeast, Saccharomyces cerevisiae, and prions.

[0009] Preferably, in the above-mentioned SOD compound microbial fermented feed, the nominal total viable count of the compound microbial fermenting agent is 20 billion to 40 billion CFU / g.

[0010] This invention also provides a method for preparing the above-mentioned SOD compound bacteria fermented feed, comprising the following steps: providing Zhonghua Liangmu forage and pressing it to separate forage juice and forage residue; crushing the forage residue; mixing the crushed forage residue, corn and soybean meal to obtain a mixed main material; mixing the compound bacteria fermentation agent with a liquid carrier to obtain an inoculum; mixing the inoculum with the mixed main material evenly to obtain a material to be fermented; and subjecting the material to be fermented to sealed anaerobic fermentation.

[0011] Preferably, in the above method, the pasture juice accounts for 25%-35% of the total mass of Zhonghua Liangmu pasture; the pulverizing step makes the particle size of the pasture residue less than 3mm.

[0012] Preferably, in the above method, the liquid carrier includes water or pasture juice; and when the liquid carrier includes pasture juice, the mixing ratio of the compound microbial fermentation agent and the pasture juice is 25-35L of pasture juice per 1kg of compound microbial fermentation agent.

[0013] Preferably, in the above method, after the inoculum is mixed with the main mixed material, the overall moisture content of the material is controlled between 50% and 60%; the sealed anaerobic fermentation is carried out in a ton bag.

[0014] Preferably, in the above method, the temperature of the sealed anaerobic fermentation is 22℃-35℃; and the time of the sealed anaerobic fermentation is 60 hours-144 hours.

[0015] The present invention also provides a feed composition comprising a basic feed and the above-mentioned SOD complex fermented feed, wherein the amount of SOD complex fermented feed added accounts for 10%-80% of the total mass of the feed composition.

[0016] The present invention also provides the application of the above-mentioned SOD compound bacteria fermented feed or the above-mentioned feed composition in the preparation of feed for raising animals, including poultry, mammals or aquatic animals.

[0017] The beneficial effects of this invention are: This invention, through a specific combination of raw materials and a compound microbial fermentation process, can significantly enhance the nutritional value and functionality of feed. The prepared fermented feed is not only rich in highly active beneficial components, effectively improving the sensory quality of the feed and animals' willingness to eat, but also promotes the absorption and utilization of nutrients. It plays a positive role in enhancing the health of animals and improving breeding efficiency, providing a practical solution for developing new functional feed products. Attached Figure Description

[0018] Figure 1 Results of the test report for laying hen feed: 1; Figure 2 Result 2 of the test report for laying hen feed; Figure 3 Result 3 of the test report for laying hen feed; Figure 4 The result of the fish feed test report is 4; Figure 5 The result of the fish feed test report is 5; Figure 6 The results of the fish feed test report are as follows: 6; Figure 7 Result 1 of the mutton test report; Figure 8 Result 2 of the mutton test report; Figure 9 Result 3 of the mutton test report; Figure 10 The results of SOD activity detection in the compound bacterial nutrient solution; Figure 11 Nutritional component test report for compound probiotic health liquid 1; Figure 12 Nutritional component test report for compound probiotic health liquid 2; Figure 13 3. Nutritional component test report for compound bacterial health liquid; Figure 14 Egg test report result 1; Figure 15 Egg test report result 2; Figure 16 Egg test report result 3; Figure 17 The result of the beef test report is 1; Figure 18 Result 2 of the beef test report; Figure 19 The results of the pig feed test report; Figure 20 The first result of the pork test report; Figure 21 The second result of the pork test report; Figure 22 For pork test report result 3, note: Figures 1-22 The test report images shown are all directly from the original report documents issued by the testing institutions, therefore the institution logos, watermarks, and other information contained in the images cannot be removed. Detailed Implementation

[0019] This invention provides a SOD compound microbial fermented feed, comprising the following raw materials in parts by weight: 70-90 parts Zhonghua Liangmu forage grass, 5-15 parts corn, 5-15 parts soybean meal, and 0.05-1.5 parts compound microbial fermentation agent.

[0020] In this invention, Zhonghua Liangmu forage refers to a specific forage variety suitable for animal feed. After conventional processing such as harvesting and drying, it can be used in feed processing and is a commercially available product that can be purchased through conventional channels. Zhonghua Liangmu forage is rich in nutrients such as crude fiber, vitamins, and minerals. Its use in fermented feed helps provide structural substances and promotes intestinal health. Its dosage range is, for example, 75-85 parts, more preferably 78-82 parts, and most preferably 80 parts.

[0021] The corn mentioned is a common energy feed ingredient, usually used after being crushed, and can be yellow corn, white corn, etc. It mainly provides carbohydrates such as starch in the feed. The soybean meal mentioned is a common plant protein feed ingredient, a byproduct of soybean oil extraction, rich in protein and amino acids. The combined use of corn and soybean meal helps to balance the energy-to-protein ratio of the feed. The amount of corn used can range, for example, 8-12 parts, more specifically, 9-11 parts, or even 10 parts. The amount of soybean meal used can also range, for example, 8-12 parts, more specifically, 9-11 parts, or even 10 parts.

[0022] The compound microbial fermentation agent refers to a preparation containing multiple active microorganisms with synergistic effects. It can be in the form of powder, granules, or liquid, and its purpose is to modify feed ingredients through the metabolic activities of microorganisms. The dosage range of the compound microbial fermentation agent is, for example, 0.8-1.2 parts, further, for example, 0.9-1.1 parts, and even, for example, 1.0 part.

[0023] In some preferred embodiments of the present invention, the compound microbial fermentation agent comprises at least three strains selected from lactic acid bacteria, *Lactobacillus plantarum*, *Bacillus subtilis*, *Rhodopseudomonas palustris*, *Propionibacterium propionate*, *Lactobacillus brunelli*, *Lactobacillus paracasei*, *Lactococcus lactis*, *Bacillus licheniformis*, yeast, *Saccharomyces cerevisiae*, and prions. Lactic acid bacteria are a collective term for bacteria that produce large amounts of lactic acid, commonly found in fermented foods and feeds, and help lower pH and inhibit harmful bacteria. Yeasts are single-celled fungi, such as *Saccharomyces cerevisiae*, that can produce various enzymes and vitamins. Bacillus (such as *Bacillus subtilis* and *Bacillus licheniformis*) are characterized by heat resistance and enzyme production. The use of a compound of at least three strains aims to utilize the complementary functions of different microorganisms to achieve a more comprehensive fermentation effect, such as simultaneously achieving acid production, enzyme production, and vitamin production—a common technical approach in the field of compound microbial preparations. More preferably, the compound microbial fermentation agent comprises four, five, or more strains selected from the above list.

[0024] In some preferred embodiments of the present invention, the nominal total viable count of the compound microbial fermentation agent is 20 billion to 40 billion CFU / g. The viable count (Colony-Forming Units per gram) is a key indicator for measuring the activity of microbial preparations; a sufficient viable count is fundamental to ensuring rapid initiation of the fermentation process and achieving the desired effect. The nominal total viable count is further preferably 25 billion to 35 billion CFU / g, and for example, 28 billion to 32 billion CFU / g, with 30 billion CFU / g being the most preferred.

[0025] This invention also provides a method for preparing the above-mentioned SOD compound bacteria fermented feed, the method comprising the following core steps: providing Zhonghua Liangmu forage and pressing it to separate forage juice and forage residue; crushing the forage residue; mixing the crushed forage residue, corn and soybean meal to obtain a mixed main material; mixing the compound bacteria fermentation agent with a liquid carrier to obtain an inoculum; mixing the inoculum with the mixed main material evenly to obtain the material to be fermented; and performing sealed anaerobic fermentation on the material to be fermented.

[0026] The pressing process refers to the step of separating the juice from Zhonghua Liangmu forage through mechanical pressure. Conventional equipment such as screw presses and roller presses can be used. The purpose is to obtain forage juice rich in soluble nutrients and reduce the moisture content of the hay residue, facilitating subsequent crushing and fermentation. The forage juice separated by the pressing process accounts for 25%-35% of the total mass of the Zhonghua Liangmu forage. This proportion range can better balance the juice yield and the physical properties of the hay residue. More preferably, the forage juice accounts for 28%-32% of the total mass of the Zhonghua Liangmu forage, and for example, 29%-31%, with the most preferred being 30%.

[0027] The pulverizing step refers to the process of reducing the particle size of the pressed forage residue through mechanical action, using equipment such as hammer mills or blade mills. Pulverizing the forage residue increases its specific surface area, promotes contact between microorganisms and materials during fermentation, thereby accelerating the fermentation process and improving fermentation uniformity. Preferably, the pulverizing step reduces the particle size of the forage residue to below 3 mm. More preferably, the particle size is below 2.5 mm, and even more preferably below 2.2 mm, and most preferably below 2 mm.

[0028] The main mixed material is composed of crushed forage residue, corn and soybean meal mixed in the aforementioned proportions. The mixing operation can be carried out in a horizontal mixer, a vertical mixer or a twin-shaft paddle mixer to ensure that the components are evenly distributed.

[0029] The liquid carrier is used to dissolve or suspend the compound microbial fermentation agent to facilitate its uniform distribution in the mixed main ingredients. The liquid carrier includes water or pasture juice. Using water as a carrier is a conventional and economical choice; while using pasture juice as a carrier allows for the direct introduction of nutrients into the fermentation system, which may be beneficial for the initial colonization of microorganisms. When the liquid carrier includes pasture juice, the mixing ratio of the compound microbial fermentation agent to the pasture juice is 25-35 L of pasture juice per 1 kg of compound microbial fermentation agent. This ratio helps to form a suitable concentration of bacterial solution. More preferably, the mixing ratio is 28-32 L of pasture juice per 1 kg of compound microbial fermentation agent, and for example, 29-31 L, with 30 L being the most preferred.

[0030] After the inoculant is mixed with the main raw material, the overall moisture content of the material is controlled between 50% and 60%. This moisture content range is suitable for most beneficial microorganisms to carry out solid-state fermentation, ensuring the water required for microbial growth while avoiding material sticking and poor aeration due to excessive moisture. More preferably, the moisture content is controlled between 52% and 58%, and for example, between 54% and 56%, with 55% being the most preferred (in this invention, no external water is required; the suitable moisture content can be achieved using the raw material formula described in this invention).

[0031] The sealed anaerobic fermentation refers to the process of fermenting materials in an environment that is essentially isolated from air. This sealed anaerobic fermentation is carried out in ton bags. Ton bags (also known as bulk bags) are a common type of large-capacity flexible transport packaging container. The fermentation space they create facilitates the handling and stacking of materials and effectively creates an anaerobic environment. Before fermentation, the bag opening must be tightly tied or heat-sealed.

[0032] The temperature for the sealed anaerobic fermentation is 22℃-35℃. This temperature range covers the activity temperature range of most mesophilic beneficial microorganisms. More preferably, the fermentation temperature is 22℃-30℃, and for example, 22℃-28℃, more preferably 22℃-25℃. The time for the sealed anaerobic fermentation is 60 hours-144 hours. The fermentation time needs to be matched with factors such as temperature to ensure sufficient fermentation. More preferably, the fermentation time is 72 hours-120 hours, and for example, 84 hours-108 hours, most preferably 72 hours, 96 hours, or 120 hours. The endpoint of the sealed anaerobic fermentation can be determined by at least one of the following methods: the material produces a distinct sour aroma, and / or the pH value of the material decreases by 40%-55% compared to before fermentation. The sour aroma is a typical sensory characteristic of the metabolic activity of acid-producing bacteria such as lactic acid bacteria. The pH decrease is a direct quantitative indicator of acid production during fermentation. More preferably, the pH decrease is 45%-50%.

[0033] The present invention also provides a feed composition comprising a basic feed and the above-mentioned SOD complex fermented feed, wherein the amount of SOD complex fermented feed added accounts for 10%-80% of the total mass of the feed composition.

[0034] The basal feed refers to a general term for a class of feeds that can provide animals with the necessary nutrients. It typically includes energy feeds (such as corn and wheat), protein feeds (such as soybean meal and fishmeal), mineral feeds, vitamin feeds, and various feed additives. Mixing the SOD-containing fermented feed with the basal feed aims to introduce the functional components of the fermented feed into the regular feeding system. The proportion of the SOD-containing fermented feed can be adjusted according to the species, growth stage, and feeding goals of the target animal. For example, the initial replacement ratio can be 10%-30%, which can then be gradually increased to a higher proportion, such as 40%-60%, and up to a maximum of 80%. Using a gradual transition feeding method helps animals adapt.

[0035] The present invention also provides the application of the above-mentioned SOD complex fermented feed or the above-mentioned feed composition in the preparation of feed for raising animals.

[0036] The animals include poultry, mammals, or aquatic animals. Poultry may include, for example, chickens (e.g., laying hens, broilers), ducks, geese, etc. Mammals may include, for example, pigs (e.g., fattening pigs, sows), cattle, sheep, etc. Aquatic animals may include, for example, fish (e.g., carp, crucian carp, tilapia, etc.), shrimp, crabs, etc. Using the SOD compound fermented feed or feed compositions containing it as described in this invention to feed the animals is expected to leverage the potential benefits of the fermented feed, such as improving palatability, promoting nutrient digestion and absorption, regulating intestinal microecological balance, and enhancing the animal's antioxidant capacity, thereby positively impacting animal health and production performance.

[0037] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0038] Example 1 1. Raw material composition The SOD compound microbial fermented general feed of this embodiment is prepared by means of basic feed ingredients and compound microbial fermentation agent.

[0039] The basic feed ingredients consist of Zhonghua Liangmu forage, corn, and soybean meal. Zhonghua Liangmu forage accounts for 80% of the total weight, corn accounts for 10%, and soybean meal accounts for 10%. Before use, the Zhonghua Liangmu forage needs to undergo a pressing pretreatment to extract 30% of the forage juice by weight. The pressed forage residue is then pulverized, with the particle size controlled to be below 2 mm.

[0040] The compound microbial fermentation agent has a nominal total viable count of 30 billion CFU / g. This compound microbial fermentation agent comprises lactic acid bacteria, *Lactobacillus plantarum*, *Bacillus subtilis*, *Rhodopseudomonas palustris*, *Propionibacterium propionate*, *Lactobacillus brunelli*, *Lactobacillus paracasei*, *Lactococcus lactis*, *Bacillus licheniformis*, yeast, *Saccharomyces cerevisiae*, and prions. This compound microbial fermentation agent achieves the combination of enzymes and bacteria, with lactic acid bacteria as the main component, and works synergistically with compound probiotics and enzyme preparations.

[0041] 2. Preparation method The preparation method of this embodiment includes the following steps: The first step is raw material pretreatment: Based on the requirement of 800 kg of forage residue in the main fermentation material, and considering that pressing will separate 30% of the original juice, the total amount of Zhonghua Liangmu forage to be fed is calculated to be 1143 kg. The 1143 kg of Zhonghua Liangmu forage is pressed, yielding 343 kg of forage juice for later use; the remaining 800 kg of forage residue is then pulverized, ensuring the particle size of the pulverized material is below 2 mm.

[0042] The second step, ingredient preparation and inoculation: Mix 800 kg of crushed forage residue, 100 kg of corn, and 100 kg of soybean meal from the first step evenly to form a total fermentation base of 1000 kg. Weigh 1.0 kg of the compound microbial fermentation agent and mix it evenly with 343 kg of forage juice obtained in the first step to prepare the inoculation solution.

[0043] The third step is mixing and bagging: The inoculum solution prepared in the second step is evenly sprayed onto 1000 kg of fermentation material, and thoroughly mixed mechanically or manually until the material is uniformly mixed. The uniformly mixed material is then packed into special ton bags with sealing strips, and the air inside the bags is expelled before the bag openings are completely sealed to create a strict anaerobic fermentation environment.

[0044] Step 4, Sealed Fermentation: Transfer the sealed ton bags to the fermentation site and stack or lay them flat on a clean surface. Control the ambient temperature at 22℃ for anaerobic solid-state fermentation for 72 hours. During the fermentation initiation stage, i.e., within 24 hours after fermentation begins, the odor emanating from the bag opening can be detected by smell. When a distinct aroma is present, it indicates that fermentation has started normally.

[0045] Step 5, determining the fermentation endpoint: After fermentation, open the ton bag. The material emits a strong sour aroma. Measurement shows that the pH value of the material has decreased by 45% compared to before fermentation. The material has a uniform color, a loose feel, and no mold spots or rotten odors.

[0046] Step 6, Product Acquisition: After the fermentation is determined to be successful in Step 5, the SOD compound bacteria fermented universal feed product is obtained.

[0047] Example 2 1. Raw material composition The SOD compound microbial fermented general feed of this embodiment is prepared by means of basic feed ingredients and compound microbial fermentation agent.

[0048] The basic feed ingredients consist of Zhonghua Liangmu forage, corn, and soybean meal. Zhonghua Liangmu forage accounts for 80% of the total weight, corn accounts for 12%, and soybean meal accounts for 8%. Before use, the Zhonghua Liangmu forage needs to undergo a pressing pretreatment to extract 30% of the forage juice by weight. The pressed forage residue is then pulverized, with the particle size controlled to be below 2 mm.

[0049] The compound microbial fermentation agent has a nominal total viable count of 30 billion CFU / g. This compound microbial fermentation agent comprises lactic acid bacteria, *Lactobacillus plantarum*, *Bacillus subtilis*, *Rhodopseudomonas palustris*, *Propionibacterium propionate*, *Lactobacillus brunelli*, *Lactobacillus paracasei*, *Lactococcus lactis*, *Bacillus licheniformis*, yeast, *Saccharomyces cerevisiae*, and prions. This compound microbial fermentation agent achieves the combination of enzymes and bacteria, with lactic acid bacteria as the main component, and works synergistically with compound probiotics and enzyme preparations.

[0050] 2. Preparation method The preparation method of this embodiment includes the following steps: The first step is raw material pretreatment: Based on the requirement of 800 kg of forage residue in the main fermentation material, and considering that pressing will separate 30% of the original juice, the total amount of Zhonghua Liangmu forage to be fed is calculated to be 1143 kg. The 1143 kg of Zhonghua Liangmu forage is pressed, yielding 343 kg of forage juice for later use; the remaining 800 kg of forage residue is then pulverized, ensuring the particle size of the pulverized material is below 2 mm.

[0051] The second step, ingredient preparation and inoculation: Mix 800 kg of crushed forage residue, 120 kg of corn, and 80 kg of soybean meal from the first step evenly to form a total fermentation base of 1000 kg. Weigh 1.0 kg of the compound microbial fermentation agent and mix it evenly with 343 kg of forage juice obtained in the first step to prepare the inoculation solution.

[0052] The third step is mixing and bagging: The inoculum solution prepared in the second step is evenly sprayed onto 1000 kg of fermentation material, and thoroughly mixed mechanically or manually until the material is uniformly mixed. The uniformly mixed material is then packed into special ton bags with sealing strips, and the air inside the bags is expelled before the bag openings are completely sealed to create a strict anaerobic fermentation environment.

[0053] Step 4, Sealed Fermentation: Transfer the sealed ton bags to the fermentation site and stack or lay them flat on a clean surface. Control the ambient temperature at 26℃ for anaerobic solid-state fermentation for 96 hours. During the fermentation initiation stage, i.e., within 24 hours after fermentation begins, the odor emanating from the bag opening can be detected by smell. When a distinct aroma is present, it indicates that fermentation has started normally.

[0054] Step 5, determination of fermentation endpoint: After fermentation, open the ton bag. The material emits a strong sour aroma. After measurement, the pH value of the material has decreased by 48% compared with that before fermentation. The material has a uniform color, a loose feel, and no mold spots or rotten smells.

[0055] Step 6, Product Acquisition: After the fermentation is determined to be successful in Step 5, the SOD compound bacteria fermented universal feed product is obtained.

[0056] Example 3 1. Raw material composition The SOD compound microbial fermented general feed of this embodiment is prepared by means of basic feed ingredients and compound microbial fermentation agent.

[0057] The basic feed ingredients consist of Zhonghua Liangmu forage, corn, and soybean meal. Zhonghua Liangmu forage accounts for 80% of the total weight, corn accounts for 8%, and soybean meal accounts for 12%. Before use, the Zhonghua Liangmu forage needs to undergo a pressing pretreatment to extract 30% of the forage juice by weight. The pressed forage residue is then pulverized, with the particle size controlled to be below 2 mm.

[0058] The compound microbial fermentation agent has a nominal total viable count of 30 billion CFU / g. This compound microbial fermentation agent comprises lactic acid bacteria, *Lactobacillus plantarum*, *Bacillus subtilis*, *Rhodopseudomonas palustris*, *Propionibacterium propionate*, *Lactobacillus brunelli*, *Lactobacillus paracasei*, *Lactococcus lactis*, *Bacillus licheniformis*, yeast, *Saccharomyces cerevisiae*, and prions. This compound microbial fermentation agent achieves the combination of enzymes and bacteria, with lactic acid bacteria as the main component, and works synergistically with compound probiotics and enzyme preparations.

[0059] 2. Preparation method The preparation method of this embodiment includes the following steps: The first step is raw material pretreatment: Based on the requirement of 800 kg of forage residue in the main fermentation material, and considering that pressing will separate 30% of the original juice, the total amount of Zhonghua Liangmu forage to be fed is calculated to be 1143 kg. The 1143 kg of Zhonghua Liangmu forage is pressed, yielding 343 kg of forage juice for later use; the remaining 800 kg of forage residue is then pulverized, ensuring the particle size of the pulverized material is below 2 mm.

[0060] The second step, ingredient preparation and inoculation: Mix 800 kg of crushed forage residue, 80 kg of corn, and 120 kg of soybean meal from the first step evenly to form a total fermentation base of 1000 kg. Weigh 1.0 kg of the compound microbial fermentation agent and mix it evenly with 343 kg of forage juice obtained in the first step to prepare the inoculation solution.

[0061] The third step is mixing and bagging: The inoculum solution prepared in the second step is evenly sprayed onto 1000 kg of fermentation material, and thoroughly mixed mechanically or manually until the material is uniformly mixed. The uniformly mixed material is then packed into special ton bags with sealing strips, and the air inside the bags is expelled before the bag openings are completely sealed to create a strict anaerobic fermentation environment.

[0062] Step 4, Sealed Fermentation: Transfer the sealed ton bags to the fermentation site and stack or lay them flat on a clean surface. Control the ambient temperature at 30℃ and carry out anaerobic solid-state fermentation for 120 hours. During the fermentation start-up phase, that is, within 24 hours after fermentation begins, the odor emanating from the bag opening can be judged by smell. When a distinct aroma appears, it indicates that fermentation has started normally.

[0063] Step 5, determining the fermentation endpoint: After fermentation, open the ton bag. The material emits a strong sour aroma. Measurement shows that the pH value of the material has decreased by 50% compared to before fermentation. The material has a uniform color, a loose feel, and no mold spots or rotten odors.

[0064] Step 6, Product Acquisition: After the fermentation is determined to be successful in Step 5, the SOD compound bacteria fermented universal feed product is obtained.

[0065] Experimental Example To verify the technical effectiveness of this invention, following the core process described in Example 1, universal SOD compound microbial fermented feeds suitable for different feeding objectives were prepared (the formula was consistent, only the appropriate substitution amount was adjusted according to the feeding conditions for different feeding objectives), and parallel tests were conducted by an authoritative testing institution. Key data showed that the prepared fermented feed for laying hens had an SOD activity of 64.5 U / g, a crude protein content of 15.95%, and a total of 18 amino acids of 15.38 g / 100g; the prepared fermented feed for fish had an SOD activity of 68.5 U / g, a crude protein content of 28.27%, and a total of 18 amino acids of 26.2 g / 100g. Both test results confirmed that the method described in this invention can successfully produce fermented feeds rich in SOD and various nutrients.

[0066] Furthermore, the samples related to the compound bacterial fermentation agent used in this invention were tested, and the results showed that the SOD activity in the compound bacterial nutrient solution was 23.2 U / mL; and the viable lactic acid bacteria count in the compound bacterial health care solution reached 6.6 × 10⁻⁶. 8 The sample contained CFU / g and various functional components such as amino acids, minerals, and vitamins (e.g., vitamin E 0.789 mg α-TE / 100g, vitamin K 115.6 μg / 100g, pantothenic acid 0.237 mg / 100g, etc.), verifying the functional activity and nutritional composition of the starter culture.

[0067] Furthermore, animal products fed with the SOD compound microbial fermented feed described in this invention also exhibit excellent quality. Test data shows that eggs fed this feed contain 13.2 g / 100g of protein, 0.343 mg / kg of selenium, and are rich in various amino acids; Zhonghua Liangmu ecological pork has an SOD activity as high as 576 U / g, a protein content of 21.9 g / 100g, and a total amino acid content of 20.20 g / 100g; Zhonghua Shengshi Jinmu forage-fed ecological beef contains 15.6 g / 100g of protein, 25.3 g / 100g of fat, and a total amino acid content of 19.10%; Tianmu Liangcao lamb rolls contain 18.8 g / 100g of protein and only 1.4 g / 100g of fat, and are rich in various minerals such as iron, zinc, and selenium. These results collectively demonstrate that the SOD compound microbial fermented feed provided by this invention not only effectively enhances the nutritional and functional activity of the feed itself but also further improves the nutritional value and health properties of animal products.

[0068] For detailed test data, please refer to... Figures 1-22 The official test report shown ( Figure 1-3 This is a test report for laying hen feed. Figure 4-6 For fish feed testing reports, Figure 7-9 For mutton testing reports, Figure 10-13 For test reports related to microbial agents, Figure 14-16 For egg testing reports, Figure 17-18 For beef testing reports, Figure 19 This is a report on the SOD activity test in pig feed. Figure 20-22 (Pork test report).

[0069] Other technical effects The compound microbial fermented general-purpose feed prepared in Example 1 of this invention, after fermentation by the method described above, has the following comprehensive technical effects: Fermentation at temperatures above 22°C can produce aroma within 24 hours, which can be used to accelerate the start of fermentation.

[0070] This fermented feed is characterized by rapid fermentation, inhibition of unwanted bacteria, and improved aroma and digestibility.

[0071] This product combines enzymes and bacteria, primarily lactic acid bacteria, with compound probiotics and enzyme preparations to achieve rapid degradation of crude fiber, improve palatability and digestibility.

[0072] This product can be used as a substitute in the complete feed of laying hens, fish and other animals, with a gradual transition from 10% to 80% replacement rate in the complete feed.

[0073] The functional ingredients in this compound probiotic animal health liquid mainly include: beneficial bacteria such as lactic acid bacteria, organic acidifiers, enzymes, a small amount of alcohol, small peptides, amino acids, multivitamins, and unknown growth factors. Clearly, this is a liquid probiotic additive product, a liquid organic acidifier product, a liquid appetite stimulant, a metabolism improver, and an immune enhancer.

[0074] Microscopic examination and live bacteria testing revealed that the health supplement made from fermented broken rice (within one week of fermentation) contained over 1 billion probiotic lactic acid bacteria and approximately 800 million yeast cells per milliliter, for a total bacterial count exceeding 1.6 billion per milliliter. This dosage, according to the recommended usage, far exceeds the dosage for commercially available probiotic products. Furthermore, the supplement primarily consists of live bacteria, with a diverse range of strains including Bifidobacterium, Enterococcus faecalis, Lactococcus faecium, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus casei, Bacillus natto, Bacillus licheniformis, and yeast.

[0075] The acid content, calculated as lactic acid, is 5%, with a pH of around 3. It is primarily lactic acid, but also contains various microbial metabolic acids such as malic acid, fumaric acid, and citric acid. The dosage specified for this health supplement solution already meets or exceeds the level required for adding commercially available feed acidifiers.

[0076] The test data for the health supplement made with the fermentation agent are as follows: 42 units / mL of protease activity (pH 3.6, 40°C, Folin method); 102 units / mL of amylase activity (pH 5.8, 40°C, colorimetric method); and a very rich microbial enzyme system. The dosage specified for use in the health supplement meets and exceeds the levels required for adding commercially available professional enzyme preparations to animal feed. These are levels that are generally not found or cannot be achieved by probiotic products.

[0077] It also contains approximately 4% alcohol, which not only promotes blood circulation, relaxes the skin, promotes growth, accelerates metabolism, increases appetite, reduces animal activity, and significantly increases sleep time in animals, but also relaxes the skin, thus improving growth rate. As long as it is not fed in excess, there will be no side effects, and it can be used on pregnant sows (provided it is not overdosed).

[0078] It is rich in vitamins, especially B vitamins, and special vitamins such as biotin and B12. It is very effective for fur-bearing animals, thus significantly improving the appearance of their fur.

[0079] The health supplement also contains 1%–3% free amino acids, dipeptides, low-peptides, and other bioactive substances, 1% monosaccharides, and is rich in B vitamins and VB12, which can significantly enhance animals' disease resistance and growth promotion. It contains a large amount of aromatic esters, giving it a fragrant aroma that promotes saliva secretion, increases appetite, and improves digestion. These are benefits that are generally absent or unattainable in probiotics and EM products.

[0080] As can be seen from the above embodiments, the present invention provides a compound microbial fermented feed that can stably impart high SOD activity to feed and simultaneously achieve multiple functions such as improving feed palatability, increasing nutrient digestibility and regulating animal intestinal health, as well as its efficient preparation method.

[0081] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A fermented feed containing SOD (superoxide dismutase) and other microorganisms, characterized in that, It is made from the following raw materials in parts by weight: 70-90 parts Zhonghua Liangmu forage grass, 5-15 parts corn, 5-15 parts soybean meal, and 0.05-1.5 parts compound microbial fermentation agent.

2. The SOD compound microbial fermented feed according to claim 1, characterized in that, The compound microbial fermentation agent contains lactic acid bacteria, Lactobacillus plantarum, Bacillus subtilis, Rhodopseudomonas palustris, Propionibacterium propionate, Lactobacillus brycetes, Lactobacillus paracasei, Lactococcus lactis, Bacillus licheniformis, yeast, Saccharomyces cerevisiae, and prions.

3. The SOD compound microbial fermented feed according to claim 1 or 2, characterized in that, The nominal total viable count of the compound microbial fermentation agent is 20 billion to 40 billion CFU / g.

4. A method for preparing SOD complex fermented feed as described in any one of claims 1-3, characterized in that, Includes the following steps: Zhonghua Liangmu forage is provided and pressed to separate forage juice and forage residue; the forage residue is crushed; the crushed forage residue, corn and soybean meal are mixed to obtain mixed main material; the compound bacterial fermentation agent is mixed with a liquid carrier to obtain an inoculum; the inoculum is mixed evenly with the mixed main material to obtain the material to be fermented; the material to be fermented is sealed and anaerobic fermented.

5. The method according to claim 4, characterized in that, The sap of the forage grass accounts for 25%-35% of the total mass of the Zhonghua Liangmu forage grass; the crushing step makes the particle size of the forage grass residue less than 3mm.

6. The method according to claim 4, characterized in that, The liquid carrier includes water or pasture juice; and when the liquid carrier includes pasture juice, the mixing ratio of the compound microbial fermentation agent to the pasture juice is 25-35L of pasture juice per 1kg of compound microbial fermentation agent.

7. The method according to claim 4, characterized in that, After the inoculant is mixed with the main mixed material, the overall moisture content of the material is controlled between 50% and 60%; the sealed anaerobic fermentation is carried out in a ton bag.

8. The method according to claim 4, characterized in that, The temperature for the sealed anaerobic fermentation is 22℃-35℃; the time for the sealed anaerobic fermentation is 60 hours-144 hours.

9. A feed composition, characterized in that, The feed composition comprises a basic feed and a fermented feed containing SOD complex bacteria as described in any one of claims 1-3, wherein the amount of SOD complex bacteria fermented feed added accounts for 10%-80% of the total mass of the feed composition.

10. The use of the SOD compound fermented feed as described in any one of claims 1-3 or the feed composition as described in claim 9 in the preparation of feed for raising animals, said animals including poultry, mammals or aquatic animals.

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