Fermented feed for promoting intestinal health of beef cattle and preparation method of fermented feed

Abstract

The invention belongs to the technical field of fermented feed, and relates to fermented feed for promoting intestinal health of beef cattle and a preparation method of the fermented feed. Step 2, pre-enzymolysis of soybean meal; step 3, extracting active ingredients of the fresh jerusalem artichoke; step 4, performing double-layer embedding on the tea polyphenol; step five, carrying out mixed fermentation on the pre-enzymolysis basal feed, the pre-enzymolysis soybean meal, the jerusalem artichoke powder extract, the tea polyphenol microcapsules, the molasses and the compound bacterium powder; the key problems of incomplete enzymolysis, single strain function, inactivation of active ingredients, extensive parameter control and the like in the existing fermented feed are systematically solved through core technologies such as multi-stage enzymolysis, complex flora compatibility, active ingredient microencapsulation, dynamic fermentation regulation and the like.

Description

Technical Field

[0001] The present invention belongs to the technical field of fermented feed, and relates to a fermented feed for promoting the intestinal health of beef cattle and a preparation method thereof. Background Art

[0002] The existing fermented feed technology mainly ferments raw materials (such as corn, soybean meal) by microorganisms (such as lactic acid bacteria, yeast), decomposes anti-nutritional factors (such as phytic acid, cellulose), and improves the feed digestibility and nutritional value. However, the existing fermented feed has the following key defects:

[0003] 1) The enzymatic hydrolysis efficiency is low. Most processes use a single enzyme or a simple enzyme combination, which is difficult to fully decompose complex polysaccharides (such as cellulose, xylan), resulting in insufficient release of substrates and limited fermentation efficiency. For example, if β-mannan and phytic acid in soybean meal are not completely degraded, the protein utilization rate will be reduced.

[0004] 2) The microbial compatibility is single. The existing technology mostly relies on a single strain or an unoptimized strain ratio, resulting in a single function of the fermentation product and difficulty in synergistically promoting intestinal health.

[0005] 3) The active ingredients are easily inactivated. Direct addition of functional ingredients (such as tea polyphenols) is easily degraded by the fermentation environment (high temperature, high humidity, microbial metabolism), resulting in the loss of efficacy.

[0006] 4) The fermentation parameters are crudely controlled. The fermentation process mostly uses fixed temperature, humidity or oxygen concentration, which cannot dynamically adapt to the metabolic requirements of different strains, resulting in fluctuations in the activity of the microbial community and poor product consistency.

[0007] 5) The raw material pretreatment is insufficient. Raw materials such as crop straws and soybean meal are not deeply pretreated, resulting in low fiber utilization rate and significant resource waste.

[0008] 6) The by-products are not recycled. The enzymatic hydrolysis filtrate is often treated as waste, and the soluble sugars and peptides in it are not effectively recovered, resulting in nutritional loss. Summary of the Invention

[0009] In order to solve the above problems, the present invention provides a fermented feed for promoting the intestinal health of beef cattle and a preparation method thereof, which specifically includes the following steps:

[0010] Step 1: Mix and crush each component of the basic feed, pass through a 40-50 mesh sieve, then add water to a water content of 25-30%, stand for 1-2 h, sterilize with steam at 110-120 °C and 1.8-2.0 Mpa for 10-20 min, complete the pressure relief within 0.5-1 s, then mix with Enzymatic Hydrolysis Solution 1 in a mass ratio of 1:3, carry out enzymatic hydrolysis at 50-60 °C for 2-3 h, filter after the enzymatic hydrolysis is completed, collect Filtrate 1 for standby, and the filter residue is the pre-enzymatically hydrolyzed basic feed.

[0011] Preferably, the basic feed includes corn, forage grass, wheat, oats, wheat bran, crop straws, rice bran, fish meal and bone meal in a mass ratio of 10:7:5:3:2:1:2:1:1. Most preferably, the crop straws are one or more of corn straws, rice straws and wheat straws.

[0012] Preferably, the first enzyme hydrolyzate includes 400 - 500 U / g of α-amylase, 300 - 400 U / g of cellulase, 200 - 300 U / g of pectinase and 200 - 250 U / g of xylanase based on water.

[0013] Step 2: Crush the soybean meal to a particle size ≤ 0.15 mm, then mix it with the second enzyme hydrolyzate in a mass ratio of 1:4, and perform enzyme hydrolysis at 40 - 55 °C for 3 - 4 h. After the enzyme hydrolysis is completed, filter it, collect the filtrate II for standby, and the filter residue is the pre-enzyme-hydrolyzed soybean meal.

[0014] Preferably, the second enzyme hydrolyzate includes 300 - 400 U / g of β-mannanase, 100 - 200 U / g of phytase and 5800 - 6200 U / g of protease based on water.

[0015] Preferably, the enzyme hydrolysis process is carried out in two stages.

[0016] The first stage: Only β-mannanase and phytase are contained in the second enzyme hydrolyzate, and enzyme hydrolysis is carried out at 40 - 45 °C for 1 - 1.5 h.

[0017] The second stage: Add protease to the second enzyme hydrolyzate, and perform enzyme hydrolysis at 50 - 55 °C for 2 - 2.5 h.

[0018] Step 3: Wash the fresh Jerusalem artichokes, dry them at 60 - 70 °C until the moisture content ≤ 8%, crush them through a 90 - 100 mesh sieve, then mix them with the extractant, perform ultrasonic extraction at 40 - 50 kHz and 63 - 67 °C for 25 - 30 min, then centrifuge at 4000 - 5000 rpm for 10 - 20 min, take the supernatant, concentrate the supernatant until the soluble solids ≥ 45 °Brix, and then perform spray drying at an inlet air temperature of 180 - 185 °C and an outlet air temperature of 80 - 85 °C to obtain the Jerusalem artichoke extract.

[0019] Preferably, the ratio of the fresh Jerusalem artichokes to the extractant is 1 g:12 mL.

[0020] Preferably, the extractant is a citric acid solution with a mass fraction of 0.5% - 0.6%.

[0021] Step 4: Mix tea polyphenols with the inner wall material, stir at 40 - 50 °C and 150 - 200 rpm for 30 - 40 min, then inject the mixed solution into a high-voltage electrostatic microcapsule preparation device, with a voltage of 15 - 20 kV and a propulsion speed of 0.8 - 1.0 mL / min to prepare primary microspheres. Then mix the primary microspheres with the outer wall material, stir at 40 - 50 °C and 120 - 150 rpn for 1 - 1.5 h to complete coating. Filter to remove the filtrate, mix the filter residue with the curing agent, soak for 30 - 40 min, and obtain tea polyphenol microcapsules after vacuum drying.

[0022] Preferably, the mass ratio of the tea polyphenols, the inner wall material, the outer wall material, and the curing agent is 1:4:5:3.

[0023] Preferably, the inner wall material includes sodium alginate, chitosan, acetic acid, and water, with a ratio of 3 g:1 g:1 mL:100 mL.

[0024] Preferably, the outer wall material includes β-cyclodextrin, whey protein, and water, with a mass ratio of 1:2:50.

[0025] Preferably, the curing agent is a calcium chloride solution with a mass fraction of 2% - 3%.

[0026] Step 5: Mix the pre-hydrolyzed basic feed, pre-hydrolyzed soybean meal, Jerusalem artichoke powder extract, tea polyphenol microcapsules, and molasses, and add compound bacterial powder, and carry out fermentation in three stages:

[0027] The first stage: Ferment at a temperature of 35 - 40 °C, a humidity of 60 - 65%, and an oxygen supply of 0.3 - 0.4 m 3 / h for 23 - 25 h;

[0028] The second stage: Add all the filtrate 1 and filtrate 2 to the fermented material, ferment at a temperature of 30 - 35 °C, a humidity of 70 - 75%, and an oxygen concentration of 4 - 5% for 3 - 4 h, and then ferment at an oxygen concentration of 8 - 10% for 1 - 2 h, with a total fermentation time of 23 - 25 h;

[0029] The third stage: Ferment at a temperature of 25 - 28 °C, a humidity of 45 - 50%, and an oxygen concentration of <5% for 23 - 25 h. After fermentation is completed, dry at 45 - 50 °C until the water content ≤ 8%, and then add salt and stir evenly to obtain the fermented feed.

[0030] Preferably, the mass ratio of the pre-hydrolyzed basic feed, pre-hydrolyzed soybean meal, Jerusalem artichoke powder extract, tea polyphenol microcapsules, molasses, compound bacterial powder, and salt is 100:60:40:20:10:2:1.

[0031] Preferably, the compound bacterial powder includes lactic acid bacteria powder, bacillus subtilis powder, and yeast powder, with a mass ratio of 4:3:2.

[0032] Preferably, the salts include sodium chloride, sodium bicarbonate and calcium carbonate, and the mass ratio is 4:2:1.

[0033] The present invention has the following advantages:

[0034] (1) The present invention uses a composite enzyme of α-amylase, cellulase, pectinase and xylanase to enzymatically hydrolyze the basic feed, specifically decomposing the starch and fiber in corn and straw to release fermentable sugars. When enzymatically hydrolyzing soybean meal, β-mannanase and phytase are used in the first stage to remove anti-nutritional factors, and protease is added in the second stage to release small peptides, improving the protein digestibility. The enzymolysis filtrate is recycled for fermentation, realizing the full utilization of resources, significantly improving the utilization rate of soybean meal protein, and avoiding the problems of low existing enzymolysis efficiency and insufficient nutrient release.

[0035] (2) The present invention uses Lactobacillus to produce acid and inhibit bacteria, Bacillus subtilis to produce enzymes to promote digestion, and yeast to provide metabolites to form a synergistic effect, and optimizes the fermentation process. Three-stage fermentation regulation is adopted. In the first stage, lactic acid bacteria are activated to produce acid rapidly to inhibit miscellaneous bacteria; in the second stage, low oxygen is first used to promote the metabolism of yeast, and then high oxygen is used to activate Bacillus subtilis to produce enzymes; in the third stage, the products are stabilized and the shelf life is extended, significantly increasing the lactic acid content and antibacterial rate, and solving the problem of single microbial compatibility and limited function.

[0036] (3) The present invention adopts a double-layer embedding process. The inner layer of sodium alginate-chitosan forms a pH-sensitive slow-release layer, and the outer layer of β-cyclodextrin-whey protein resists mechanical damage. The calcium chloride solution enhances the mechanical strength of the microcapsule, ensuring the slow release of tea polyphenols during fermentation and avoiding early degradation, significantly increasing the retention rate of tea polyphenols after fermentation, and avoiding the problem of easy degradation of active ingredients.

[0037] (4) The present invention uses a citric acid solution and ultrasound to efficiently extract the active ingredients in Jerusalem artichoke, and the extraction rate is significantly increased. The active ingredients in Jerusalem artichoke promote the proliferation of Lactobacillus, significantly increasing the content of short-chain fatty acids in the feed, thereby improving the intestine.

[0038] (5) The present invention optimizes the basic feed formula, taking into account the balance of energy and fiber, and the enzymolysis filtrate is rich in reducing sugars and peptides, which are added as fermentation substrates in the second stage, significantly increasing the total protein content of the fermentation products, and solving the problems of insufficient raw material pretreatment and by-product waste. Detailed implementation manners

[0039] The technical solutions in the embodiments of the invention are clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0040] The sources of the microbial agents in the following embodiments are as follows

[0041] The Lactobacillus powder was purchased from Gansu Yishengxiang Biotechnology Co., Ltd., the Bacillus subtilis powder was purchased from Jinan Haoyunlai Chemical Technology Co., Ltd., and the yeast powder was purchased from Shanghai Zhongfeng Biotechnology Co., Ltd.

[0042] Example 1

[0043] Step 1: Mix corn, ryegrass, wheat, oats, wheat bran, corn straw, rice bran, fish meal and bone meal according to a mass ratio of 10:7:5:3:2:1:2:1:1, crush them, pass through a 45-mesh sieve, then add water to a water content of 30%, let stand for 1 h, sterilize with steam at 120 °C and 1.8 Mpa for 15 min, complete pressure relief within 0.5 s, and then mix with Enzyme Hydrolysis Solution 1 according to a mass ratio of 1:3, and perform enzyme hydrolysis at 55 °C for 2.5 h. After the enzyme hydrolysis is completed, filter, collect Filtrate 1 for standby, and the filter residue is the pre-enzyme-hydrolyzed basic feed. The Enzyme Hydrolysis Solution 1 includes 450 U / g of α-amylase, 350 U / g of cellulase, 250 U / g of pectinase and 200 U / g of xylanase based on water.

[0044] Step 2: Crush soybean meal to a particle size of ≤0.15 mm, then mix it with Enzyme Hydrolysis Solution 2 according to a mass ratio of 1:4, and perform enzyme hydrolysis at 50 °C for 3 h. After the enzyme hydrolysis is completed, filter, collect Filtrate 2 for standby, and the filter residue is the pre-enzyme-hydrolyzed soybean meal. The enzyme hydrolysis process is carried out in two stages.

[0045] The first stage: Enzyme Hydrolysis Solution 2 only contains β-mannanase and phytase, and perform enzyme hydrolysis at 43 °C for 1 h;

[0046] The second stage: Add protease to Enzyme Hydrolysis Solution 2 and perform enzyme hydrolysis at 53 °C for 2 h.

[0047] The Enzyme Hydrolysis Solution 2 includes 350 U / g of β-mannanase, 150 U / g of phytase and 6000 U / g of protease based on water.

[0048] Step 3: Wash fresh Jerusalem artichoke, dry it at 65 °C until the water content is ≤8%, crush it and pass through a 100-mesh sieve, then mix it with a 0.5% citric acid solution by mass ratio of 1 g:12 mL, perform ultrasonic extraction at 40 kHz and 65 °C for 30 min, then centrifuge at 5000 rpm for 15 min, take the supernatant, concentrate the supernatant until the soluble solids content is ≥45 °Brix, and then perform spray drying at an inlet air temperature of 180 °C and an outlet air temperature of 80 °C to obtain Jerusalem artichoke extract.

[0049] Step 4: Mix tea polyphenols with the inner wall material, stir at 45 °C and 150 rpm for 30 min, then inject the mixture into a high-voltage electrostatic microcapsule preparation instrument, with a voltage of 15 kV and a propulsion speed of 0.8 mL / min to prepare primary microspheres. Then mix the primary microspheres with the outer wall material, stir at 45 °C and 120 rpn for 1 h to complete coating. Filter to remove the filtrate, mix the filter residue with a 2% calcium chloride solution by mass, soak for 30 min, and obtain tea polyphenol microcapsules after vacuum drying. The mass ratio of the tea polyphenols, inner wall material, outer wall material, and calcium chloride solution is 1:4:5:3. The inner wall material includes sodium alginate, chitosan, acetic acid, and water, with a ratio of 3 g:1 g:1 mL:100 mL. The outer wall material includes β-cyclodextrin, whey protein, and water, with a mass ratio of 1:2:50.

[0050] Step 5: Mix the pre-hydrolyzed basal feed, pre-hydrolyzed soybean meal, Jerusalem artichoke powder extract, tea polyphenol microcapsules, and molasses, and add the compound bacterial powder, and ferment in three stages:

[0051] The first stage: Ferment at a temperature of 38 °C, humidity of 63%, and oxygen ventilation of 0.3 m 3 / h for 24 h;

[0052] The second stage: Add all the filtrate 1 and filtrate 2 to the fermented materials, ferment at a temperature of 32 °C, humidity of 75%, and oxygen concentration of 4% for 3 h, and then ferment at an oxygen concentration of 8% for 1 h, with a total fermentation time of 24 h;

[0053] The third stage: Ferment at a temperature of 26 °C, humidity of 45%, and oxygen concentration <5% for 24 h. After fermentation is completed, dry at 45 °C until the water content ≤8%, and then add salt and stir evenly to obtain the fermented feed. The mass ratio of the pre-hydrolyzed basal feed, pre-hydrolyzed soybean meal, Jerusalem artichoke powder extract, tea polyphenol microcapsules, molasses, compound bacterial powder, and salt is 100:60:40:20:10:2:1. The compound bacterial powder includes lactic acid bacterium powder, Bacillus subtilis powder, and yeast powder, with a mass ratio of 4:3:2. The salt includes sodium chloride, sodium bicarbonate, and calcium carbonate, with a mass ratio of 4:2:1.

[0054] Test Example 1

[0055] 1. Experimental animals and grouping

[0056] Select 60 healthy beef cattle with similar weights (initial weight 300 ± 20 kg), and randomly divide them into two groups:

[0057] Control group (30 heads): Fed with traditional fermented cattle feed purchased from Shandong Xucheng Biological Feed Co., Ltd.

[0058] Experimental group (30 heads): Fed with the fermented feed prepared by the method of Example 1.

[0059] The feeding environments (temperature, humidity, light) of the two groups of beef cattle were the same. They had free access to water, and the daily feeding amount was 3.0% of their body weight.

[0060] 2. Experimental period

[0061] The preliminary feeding period was 7 days (for environmental and feed adaptation), and the formal experimental period was 90 days.

[0062] 3. Data collection and index determination

[0063] Daily weight gain: The cattle were weighed on an empty stomach every 15 days, and the average daily weight gain was calculated.

[0064] Nutrient utilization rate: Fecal samples were collected, and the apparent digestibility of crude protein, neutral detergent fiber (NDF), and acid detergent fiber (ADF) was determined using the acid-insoluble ash method.

[0065] Feed samples were collected, and the contents of crude protein, crude fat, NDF, and ADF were determined (referring to the national standard method).

[0066] Diarrhea rate: The fecal status of the beef cattle was observed daily, and the number of diarrhea occurrences was recorded.

[0067] Intestinal health indicators: After the experiment ended, samples were taken at slaughter, and the pH of rumen fluid and the content of short-chain fatty acids (SCFAs) were determined.

[0068] The results of the above indicators are shown in Table 1.

[0069] Table 1

[0070]

[0071]

[0072] As can be seen from Table 1, the crude protein content, and the digestibility of NDF and ADF in the experimental group's feed were significantly improved compared to the traditional feed. For the beef cattle fed with the experimental group's feed, compared with the control group, the daily weight gain was significantly increased, the feed-to-weight ratio was significantly decreased, the diarrhea rate was significantly decreased, and the content of short-chain fatty acids was significantly increased. Thus, it can be seen that the fermented feed prepared by the present invention is not only significantly superior to the traditional feed in terms of nutritional content and utilization rate, but also superior to the traditional feed in improving the intestinal tract.

[0073] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present invention. Various modifications to these embodiments will be obvious to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the broadest scope consistent with the principles and novel features disclosed herein.

Claims

1. A preparation method of a fermented feed for promoting the intestinal health of beef cattle, characterized in that, It includes the following steps: Step 1: Mix and crush each component of the basic feed, sterilize it with steam, then mix and enzymatically hydrolyze it with Enzymolysis Liquid 1, filter it, collect the filtrate 1 for standby, and the filter residue is the pre-enzymatically hydrolyzed basic feed; Step 2: Crush soybean meal, mix and enzymatically hydrolyze it with Enzymolysis Liquid 2, filter it, collect the filtrate 2 for standby, and the filter residue is the pre-enzymatically hydrolyzed soybean meal; Step 3: Dry and crush fresh Jerusalem artichoke, mix it with an extractant, perform ultrasonic extraction, centrifuge it, take the supernatant, concentrate and dry the supernatant to obtain Jerusalem artichoke extract; Step 4: Mix tea polyphenols with the inner wall material and stir, then use microcapsule manufacturing equipment to prepare primary microspheres, mix and coat the primary microspheres with the outer wall material, filter to remove the filtrate, soak the filter residue with a curing agent, and dry to obtain tea polyphenol microcapsules; Step 5: Mix the pre-enzymatically hydrolyzed basic feed, pre-enzymatically hydrolyzed soybean meal, Jerusalem artichoke powder extract, tea polyphenol microcapsules and molasses, and add compound bacterial powder, and perform fermentation in three stages: In the first stage, ferment at a temperature of 35 - 40 °C, a humidity of 60 - 65%, and an oxygen ventilation volume of 0.3 - 0.4 m 3 / h for 23 - 25 h; The second stage: Add all the filtrate 1 and filtrate 2 to the fermented material, ferment at a temperature of 30 - 35 °C, humidity of 70 - 75%, and oxygen concentration of 4 - 5% for 3 - 4 h, and then ferment at an oxygen concentration of 8 - 10% for 1 - 2 h, with a total fermentation time of 23 - 25 h; The third stage: Ferment at a temperature of 25 - 28 °C, humidity of 45 - 50%, and oxygen concentration < 5% for 23 - 25 h. After fermentation is completed, dry it at 45 - 50 °C until the water content ≤ 8%, and then add salt and stir evenly to obtain the fermented feed.

2. The preparation method of a fermented feed for promoting the intestinal health of beef cattle according to claim 1, characterized in that, The basic feed described in Step 1 includes corn, forage grass, wheat, oats, bran, crop straw, rice bran, fish meal and bone meal, and the mass ratio is: 10:7:5:3:2:1:2:1:

1.

3. The preparation method of a fermented feed for promoting the intestinal health of beef cattle according to claim 1, characterized in that, The Enzymolysis Liquid 1 described in Step 1 includes 400 - 500 U / g of α-amylase, 300 - 400 U / g of cellulase, 200 - 300 U / g of pectinase and 200 - 250 U / g of xylanase based on water.

4. The preparation method of a fermented feed for promoting the intestinal health of beef cattle according to claim 1, characterized in that, The Enzymolysis Liquid 2 described in Step 2 includes 300 - 400 U / g of β-mannanase, 100 - 200 U / g of phytase and 5800 - 6200 U / g of protease based on water.

5. The preparation method of a fermented feed for promoting the intestinal health of beef cattle according to claim 1, characterized in that, The enzymatic hydrolysis process in Step 2 is carried out in two stages. The first stage: Only β-mannanase and phytase are contained in Enzymolysis Liquid 2, and enzymatically hydrolyze at 40 - 45 °C for 1 - 1.5 h; The second stage: Add protease to Enzymolysis Liquid 2, and enzymatically hydrolyze at 50 - 55 °C for 2 - 2.5 h.

6. The preparation method of a fermented feed for promoting the intestinal health of beef cattle according to claim 1, characterized in that, The extractant described in Step 3 is a citric acid solution with a mass fraction of 0.5% - 0.6%.

7. The preparation method of a fermented feed for promoting the intestinal health of beef cattle according to claim 1, wherein, The inner wall material described in Step 4 includes sodium alginate, chitosan, acetic acid and water, and the ratio is 3 g:1 g:1 mL:100 mL. Preferably, the outer wall material includes β-cyclodextrin, whey protein and water, and the mass ratio is 1:2:

50. Preferably, the curing agent is a calcium chloride solution with a mass fraction of 2% - 3%.

8. The preparation method of a fermented feed for promoting the intestinal health of beef cattle according to claim 1, characterized in that, The mass ratio of the pre-enzymatically hydrolyzed basic feed, pre-enzymatically hydrolyzed soybean meal, Jerusalem artichoke powder extract, tea polyphenol microcapsules, molasses, compound bacterial powder and salt in Step 5 is 100:60:40:20:10:2:

1.

9. The preparation method of a fermented feed for promoting the intestinal health of beef cattle according to claim 1, characterized in that, The compound bacterial powder described in step five includes Lactobacillus bacterial powder, Bacillus subtilis bacterial powder, and yeast bacterial powder, with a mass ratio of 4:3:2; the salts include sodium chloride, sodium bicarbonate, and calcium carbonate, with a mass ratio of 4:2:

1.

10. The fermented feed prepared by the method according to any one of claims 1-9.

Images