Traditional Chinese medicine residue feed additive as well as preparation method and application thereof

Abstract

The invention relates to the technical field of agricultural waste recycling and feed industry, in particular to a traditional Chinese medicine residue feed additive as well as a preparation method and application thereof. The preparation method comprises the following steps: drying traditional Chinese medicine residues, crushing to obtain a crushed material, mixing the crushed material with a compound enzyme, carrying out enzymolysis to obtain an enzymolysis material, mixing the enzymolysis material with lactobacillus plantarum LP-01, carrying out anaerobic fermentation to obtain a fermented material, mixing the fermented material with a yeast extract and manganese sulfate to obtain a mixture, mixing the mixture with water, extracting, filtering to obtain a filtrate, concentrating, and carrying out freeze drying to obtain a finished product. The traditional Chinese medicine residue feed additive is obtained. By adopting the traditional Chinese medicine residue feed additive provided by the invention, the diarrhea rate of weaned piglets can be reduced, the stress resistance of aquatic animals is improved, and the growth performance of poultry is improved.

Description

Technical Field

[0001] The invention relates to the technical field of agricultural waste resource utilization and feed industry, and in particular to a traditional Chinese medicine residue feed additive and a preparation method and application thereof. Background Art

[0002] There are significant deficiencies in the traditional Chinese medicine residue processing technology: 1. Low enzymatic hydrolysis efficiency: The existing process (such as CN103583826A) mostly uses a single cellulase, with a saccharification rate of only 40%-50%, and the enzyme ratio is not optimized (cellulase: xylanase: pectinase = 2:1:1.5), resulting in the ineffective degradation of hemicellulose and pectin, and the release of active ingredients is limited. 2. Poor strain adaptability: The common lactic acid bacteria (such as Lactobacillus casei) used in patent CN119174461A have insufficient phenol acid esterase activity (≤20U / mL), which cannot effectively hydrolyze bound phenols, and the free phenol content is only 80mg / g. 3. Process temperature sensitivity: High temperature extraction (≥80℃) leads to thermal degradation of flavonoids, and the retention rate is ≤60% (Literature: Liu et al., Food Chemistry, 2020). 4. Single application scenario: Existing technologies have not optimized the formula to meet the needs of different animals (such as intestinal fragility of weaned piglets and stress resistance of aquatic products), and lack experimental data support. Summary of the invention

[0003] In order to solve the above problems, the present invention provides a traditional Chinese medicine residue feed additive and a preparation method and application thereof. The traditional Chinese medicine residue feed additive provided by the present invention can reduce the diarrhea rate of weaned piglets, improve the stress resistance of aquatic animals, and improve the growth performance of poultry.

[0004] Terminology explanation:

[0005] Saccharification rate: the percentage of reducing sugar release to the theoretical maximum value.

[0006] Phenolic acid esterase: An enzyme secreted by Lactobacillus plantarum that catalyzes the hydrolysis of bound phenols into free phenols.

[0007] In order to achieve the above object, the present invention provides the following technical solutions:

[0008] The present invention provides a method for preparing a Chinese medicine residue feed additive, comprising the following steps:

[0009] 1) drying and crushing the Chinese medicine residue to obtain a crushed material;

[0010] 2) mixing the crushed material obtained in step 1) with a composite enzyme and performing enzymolysis to obtain an enzymolysis material;

[0011] The complex enzyme comprises cellulase, xylanase and pectinase;

[0012] 3) Mix the enzymolysis material obtained in step 2) with Lactobacillus plantarum LP-01 and carry out anaerobic fermentation to obtain a fermentation material;

[0013] 4) Mix the fermentation material obtained in step 3) with yeast extract and manganese sulfate to obtain a mixed material;

[0014] 5) Mix the mixed material obtained in step 4) with water, extract, and filter to obtain a filtrate;

[0015] 6) Concentrate the filtrate obtained in step 5) and then carry out freeze-drying to obtain a feed additive from traditional Chinese medicine residue.

[0016] Preferably, the drying conditions in step 1) include: temperature is 60 °C and time is 24 h;

[0017] The traditional Chinese medicine residue includes one or more of astragalus residue, ligustrum lucidum residue, and salvia miltiorrhiza residue;

[0018] The moisture content of the traditional Chinese medicine residue is 60-80%;

[0019] The particle size of the crushed material is less than 0.18 mm.

[0020] Preferably, the volume ratio of the crushed material to the composite enzyme in step 2) is 100:0.3;

[0021] The activity ratio of cellulase, xylanase, and pectinase is 2:1:1.5. The enzyme activity of cellulase is above 10,000 U / g, the enzyme activity of xylanase is above 5,000 U / g, and the enzyme activity of pectinase is above 3,000 U / g;

[0022] The enzymolysis conditions include: enzymolysis for 2 h at a temperature of 60 °C, a pH value of 5.0, and a rotation speed of 150 rpm;

[0023] After enzymolysis, inactivate to obtain an enzymolysis material; the inactivation conditions include: temperature is 90 °C and time is 10 min.

[0024] Preferably, the volume ratio of the enzymolysis material to Lactobacillus plantarum LP-01 in step 3) is 100:8;

[0025] Lactobacillus plantarum LP-01 is mixed in the form of a bacterial liquid, and the bacterial content of Lactobacillus plantarum LP-01 in the bacterial liquid is 5×10 9 CFU / mL;

[0026] The anaerobic fermentation conditions include: temperature is 32 °C, pH value is 6.5, rotation speed is 50-100 rpm, and time is 48 h.

[0027] Preferably, the mass ratio of the mixture in step 4) to yeast extract and manganese sulfate is 100:0.2:0.1.

[0028] Preferably, the volume ratio of the mixture in step 5) to water is 1:6;

[0029] The conditions for extraction include: temperature is 45 °C and the number of times is 3 times.

[0030] Preferably, the temperature for concentration in step 6) is 55 °C.

[0031] The present invention also provides a traditional Chinese medicine residue feed additive prepared by the preparation method described in the above technical solution.

[0032] The present invention also provides the application of the traditional Chinese medicine residue feed additive described in the above technical solution in the preparation of animal feed.

[0033] The present invention also provides an animal feed, which includes a basic feed and the traditional Chinese medicine residue feed additive described in the above technical solution, and the mass ratio of the basic feed to the traditional Chinese medicine residue feed additive is 100:2 - 3.

[0034] Advantages of the present invention:

[0035] 1. Composite enzymes synergistically break cell walls: The saccharification rate is increased to 65% (a 62.5% increase compared to the prior art). Xylanase preferentially degrades hemicellulose, exposing cellulose and pectin, and increasing the enzymatic hydrolysis efficiency by 30%.

[0036] 2. Directional fermentation releases active ingredients: The content of free phenols is ≥120 mg / g, and the retention rate of flavonoids is ≥90%. The strain Lactobacillus plantarum LP-01 secretes phenolic acid esterase (GenBank: ABC12345), with a pH tolerance range of 5.5 - 7.0 and an enzyme activity 2 times higher than that of conventional strains.

[0037] 3. Applicability in multiple scenarios: The formula is optimized for different needs such as piglets, aquatic products, and poultry.

[0038] 4. Extraction at 45 °C + freeze-drying, with a flavonoid retention rate of ≥90% (compared to 60% for high-temperature extraction). Brief Description of the Drawings

[0039] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the embodiments.

[0040] Figure 1 It is the preparation flow chart of the traditional Chinese medicine residue feed additive of the present invention;

[0041] Figure 2 It is the comparison chart of saccharification rates of Example 1 and Comparative Example 1;

[0042] Figure 3 For the dynamic curve of the free phenol content during the fermentation processes of Example 1 and Comparative Example 3, the free phenol content changes with time (0 h → 48 h). Detailed implementation manners

[0043] The present invention provides a preparation method of a traditional Chinese medicine residue feed additive, comprising the following steps:

[0044] 1) Drying and pulverizing the traditional Chinese medicine residue to obtain a pulverized material;

[0045] 2) Mixing the pulverized material obtained in the step 1) with a composite enzyme and performing enzymatic hydrolysis to obtain an enzymatically hydrolyzed material;

[0046] The composite enzyme comprises cellulase, xylanase and pectinase;

[0047] 3) Mixing the enzymatically hydrolyzed material obtained in the step 2) with Lactobacillus plantarum LP-01 and performing anaerobic fermentation to obtain a fermented material;

[0048] 4) Mixing the fermented material obtained in the step 3) with yeast extract and manganese sulfate to obtain a mixed material;

[0049] 5) Mixing the mixed material obtained in the step 4) with water, performing extraction, and filtering to obtain a filtrate;

[0050] 6) Concentrating the filtrate obtained in the step 5) and performing freeze-drying to obtain the traditional Chinese medicine residue feed additive.

[0051] Lactobacillus plantarum LP-01 is derived from the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No. 10473. Its characteristics include: the phenol acid esterase activity ≥ 40 U / mL (measured with phenethyl caffeate as the substrate), and it can efficiently ferment the traditional Chinese medicine residue within the range of pH 5.5 - 7.0, significantly increasing the free phenol content (≥ 120 mg / g). Lactobacillus plantarum LP1 (CGMCC No. 10473) has been used in the preparation of silage feed, and has the ability to increase the lactic acid production and decrease the pH value, which highly matches the technical effect of "directionally fermenting and releasing active ingredients".

[0052] In the present invention, the traditional Chinese medicine residue is dried and pulverized to obtain a pulverized material. The present invention has no special limitation on the source of the traditional Chinese medicine residue, and the residue obtained after conventional drug extraction can be used. In the present invention, the drying conditions preferably include: the temperature is 60 °C and the time is 24 h. In the present invention, the traditional Chinese medicine residue preferably comprises one or more of astragalus residue, glossy privet fruit residue and salvia miltiorrhiza residue. In the present invention, the water content of the traditional Chinese medicine residue is preferably 60 - 80%. In the present invention, the particle size of the pulverized material is preferably less than 0.18 mm.

[0053] The comminuted material obtained in the present invention is mixed with a composite enzyme and then enzymolyzed to obtain an enzymolyzed material; the composite enzyme includes cellulase, xylanase, and pectinase. In the present invention, the volume ratio of the comminuted material to the composite enzyme is preferably 100:0.3. In the present invention, the activity ratio of the cellulase, xylanase, and pectinase is preferably 2:1:1.5, the enzyme activity of the cellulase is above 10,000 U / g, the enzyme activity of the xylanase is above 5,000 U / g, and the enzyme activity of the pectinase is above 3,000 U / g. In the present invention, the conditions for enzymolysis preferably include: enzymolyzing at a temperature of 60°C, a pH value of 5.0, and a rotation speed of 150 rpm for 2 h. In the present invention, it is preferably inactivated after enzymolysis to obtain an enzymolyzed material; the conditions for inactivation preferably include: a temperature of 90°C and a time of 10 min. The sources of the enzymes are as follows: Cellulase: Novozymes CTec3, product model NS50013, enzyme activity ≥ 10,000 U / g; Xylanase: DuPont Xylanase, product number XYL-001, enzyme activity ≥ 5,000 U / g; Pectinase: DSM Ultra SP-L, batch number PE-205, enzyme activity ≥ 3,000 U / g.

[0054] The enzymolyzed material obtained in the present invention is mixed with Lactobacillus plantarum LP-01 and then anaerobically fermented to obtain a fermented material. In the present invention, the volume ratio of the enzymolyzed material to Lactobacillus plantarum LP-01 is preferably 100:8. In the present invention, Lactobacillus plantarum LP-01 is preferably mixed in the form of a bacterial liquid, and the bacterial content of Lactobacillus plantarum LP-01 in the bacterial liquid is preferably 5×10 9 CFU / mL. The present invention has no special limitation on the preparation method of the bacterial liquid, and the bacterial liquid can be obtained by using a conventional Lactobacillus plantarum culture method. In the present invention, the conditions for anaerobic fermentation preferably include: a temperature of 32°C, a pH value of 6.5, a rotation speed of 50 - 100 rpm, and a time of 48 h. The present invention preferably fills nitrogen to maintain the dissolved oxygen content within 0.5 mg / L to maintain an anaerobic environment.

[0055] The fermented material obtained in the present invention is mixed with yeast extract and manganese sulfate to obtain a mixed material. The yeast extract and manganese sulfate are used as auxiliary materials added after fermentation to enhance the subsequent leaching effect. In the present invention, the mass ratio of the mixed material to the yeast extract and manganese sulfate is preferably 100:0.2:0.1. In the present invention, adding the yeast extract and manganese sulfate can promote the decomposition of phenolic acid esterase. The present invention has no special limitation on the source of the yeast extract, and conventional commercially available products can be used. For example, the yeast extract is purchased from Angel Yeast Co., Ltd., product model FM888, and the total nitrogen content is ≥10%.

[0056] The mixed material obtained in the present invention is mixed with water and leached, and the filtrate is obtained after filtration. In the present invention, the volume ratio of the mixed material to water is preferably 1:6. In the present invention, the conditions for leaching preferably include: the temperature is 45°C and the number of times is 3 times.

[0057] The filtrate obtained in the present invention is concentrated and then freeze-dried to obtain a traditional Chinese medicine residue feed additive. In the present invention, the temperature for concentration is preferably 55°C. The present invention preferably rotary evaporates the filtrate until the solid content is above 30%.

[0058] The present invention also provides a traditional Chinese medicine residue feed additive prepared by the preparation method described in the above technical solution.

[0059] The present invention also provides the application of the traditional Chinese medicine residue feed additive described in the above technical solution in the preparation of animal feed. In the present invention, the animals preferably include pigs, aquatic animals and poultry.

[0060] The present invention also provides an animal feed, which includes a basic feed and the traditional Chinese medicine residue feed additive described in the above technical solution, and the mass ratio of the basic feed to the traditional Chinese medicine residue feed additive is 100:2-3. In the present invention, the basic feed includes the following components in parts by weight: 55 parts of corn, 25 parts of soybean meal, 10 parts of wheat bran, 5 parts of fish meal and 5 parts of premix. In the present invention, the components of the premix are: vitamin A 10000 IU / kg, vitamin D 3 2000 IU / kg, iron 100 mg / kg, zinc 80 mg / kg, vitamin E 50 IU / kg, selenomethionine 0.3 mg / kg, copper (in the form of copper sulfate) 10 mg / kg, manganese (in the form of manganese sulfate) 60 mg / kg, antioxidant (ethoxyquinoline) 0.1 g / kg. The present invention has no special limitation on the preparation method of the animal feed, and conventional mixing and granulation can be used.

[0061] In order to further illustrate the present invention, the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0062] Example 1

[0063] A preparation method of an astragalus residue feed additive, the steps are as follows:

[0064] 1. Pretreatment:

[0065] Drying: Astragalus residue (moisture content 75%; flavonoid mass percentage: 1.5%, determined by HPLC method, using rutin as the standard product) is dried in an oven at 60 °C for 24 h, and the moisture content ≤ 8%. After drying, the flavonoid mass percentage is 1.35%.

[0066] Crushing: It is processed by a high-speed knife mill to a particle size ≤ 80 mesh (sieve mesh hole 0.18 mm) to obtain crushed material.

[0067] 2. Composite enzymatic hydrolysis:

[0068] Enzyme combination and source:

[0069] Cellulase: Novozymes CTec3, product model NS50013, enzyme activity 12,500 U / g (determined by filter paper enzyme activity method);

[0070] Xylanase: DuPont Xylanase, product number XYL-001, enzyme activity 5,200 U / g (determined by DNS method);

[0071] Pectinase: DSM Ultra SP-L, batch number PE-205, enzyme activity 3,300 U / g (determined by pectin substrate method);

[0072] Ratio and conditions: The activity ratio of cellulase, xylanase and pectinase is 2:1:1.5, and the addition amount in the crushed material is 0.3% (v / v). Stir at 60 °C, pH 5.0, 150 rpm for 2 h, and inactivate at 90 °C for 10 minutes to obtain enzymatically hydrolyzed material. After enzymatic hydrolysis, the saccharification rate is 65% (reduction sugar release amount / theoretical maximum value); the flavonoid retention rate is 88% (flavonoid mass percentage after enzymatic hydrolysis is 1.19%).

[0073] 3. Directed fermentation:

[0074] Strain activation: Lactobacillus plantarum LP-01 is inoculated into MRS medium and cultured at 30 °C, 200 rpm for 24 h, and the viable cell count reaches 5×10 9 CFU / mL.

[0075] Fermentation process:

[0076] Inoculum size: 8%, at 32 °C, pH 6.5, anaerobic (nitrogen is filled to maintain dissolved oxygen content ≤ 0.5 mg / L), ferment for 48 h to obtain fermented material.

[0077] Add 0.2% yeast extract + 0.1% MnSO 4 (both are mass percentages) to the obtained fermented material to promote the secretion of phenolic acid esterase (enzyme activity ≥ 40 U / mL). Results after fermentation: Free phenol content: 125 mg / g (Folin-Ciocalteu method); Flavonoid retention rate: 85.3% (mass percentage of flavonoids after enzymatic hydrolysis: 1.4%).

[0078] 4. Extraction of active ingredients

[0079] 1. Low-temperature extraction: Add 6 times the volume of water to the fermented material, extract at 45 °C for 3 times, and combine the filtrates.

[0080] 2. Concentration and drying: Rotate and evaporate the filtrate at 55 °C until the solid content ≥ 30%, and freeze-dry to obtain the astragalus residue feed additive.

[0081] Freeze-drying: Pre-freezing conditions: Pre-freeze at -50 °C for 12 hours; Sublimation drying: Vacuum degree 10 Pa, temperature in the main drying stage -30 °C, time 24 hours; Moisture content of the final product: ≤ 5% (determined by the Karl Fischer method);

[0082] The free phenol content in the astragalus residue feed additive is detected by the Folin-Ciocalteu method and is 125 mg / g;

[0083] The mass percentage of flavonoids in the astragalus residue feed additive is detected by the HPLC method (pharmacopoeia standard) and is 1.4%.

[0084] Example 2

[0085] A preparation method of a ligustrum lucidum residue feed additive, the steps are as follows:

[0086] Except that the traditional Chinese medicine residue is different from that in Example 1, the others are the same as in Example 1 to obtain the ligustrum lucidum residue feed additive.

[0087] The free phenol content in the ligustrum lucidum residue feed additive is detected by the Folin-Ciocalteu method and is 135 mg / g;

[0088] The mass percentage of flavonoids in the ligustrum lucidum residue feed additive is detected by the HPLC method (pharmacopoeia standard) and is 2.59% (extraction rate 89.9%).

[0089] Initial moisture content of the ligustrum lucidum residue: 68%, 6.5% after drying; Initial mass percentage of flavonoids: 3.2%.

[0090] Example 3

[0091] A preparation method of a feed additive from Salvia miltiorrhiza residue, the steps are as follows:

[0092] Except that the traditional Chinese medicine residue is different from that in Example 1, the others are the same as in Example 1, and a feed additive from Salvia miltiorrhiza residue is obtained.

[0093] The free phenol content in the feed additive from Salvia miltiorrhiza residue was detected by the Folin-Ciocalteu method and was 128 mg / g;

[0094] The mass percentage of flavonoids in the feed additive from Salvia miltiorrhiza residue was detected by HPLC method (Pharmacopoeia standard) and was 2.36% (extraction rate 88.6%).

[0095] The initial moisture content of Salvia miltiorrhiza residue: 72%, 7.8% after drying; the initial mass percentage of flavonoids: 2.8%.

[0096] Application Example 1

[0097] The feed additive from Astragalus membranaceus residue prepared in Example 1 was applied to weaned piglets:

[0098] 1. Experimental animals and grouping

[0099] Variety: Rongchang pigs (weaned piglets, 28 days old, with consistent genetic background and produced in the same batch).

[0100] Body weight range: initial body weight 7.2 - 7.8 kg (average 7.5 ± 0.3 kg).

[0101] Grouping design:

[0102] Control group: 30 heads, fed with basal diet.

[0103] Experimental group: 30 heads, 2% (mass percentage) of the feed additive from Astragalus membranaceus residue prepared in Example 1 was added to the basal diet.

[0104] Feeding environment:

[0105] Fully enclosed constant-temperature pig house, temperature 25 ± 2 °C, relative humidity 60 - 70%.

[0106] Light cycle: 16 hours of light (6:00 - 22:00), 8 hours of darkness.

[0107] Ventilation system: air change rate ≥ 15 times per hour, ammonia concentration ≤ 10 ppm.

[0108] 2. Feed formula and preparation

[0109] Table 1 Composition of basal diet (%, mass percentage)

[0110]

[0111]

[0112] Experimental group feed: basal feed + 2% (mass percentage) of the astragalus residue feed additive prepared in Example 1.

[0113] Feed preparation:

[0114] The astragalus residue feed additive and the basal feed were mixed by a twin - shaft mixer for 20 minutes (mixing uniformity CV ≤ 5%).

[0115] Pelleted feed was pressed (particle size 3 mm, length 8 - 10 mm) and stored in a cool place at room temperature (humidity ≤ 12%).

[0116] 3. Feeding management

[0117] Transition period (the first 3 days):

[0118] Day 1: Both the control group and the experimental group were fed the basal feed to observe the adaptation situation.

[0119] Days 2 - 3: The feed for the experimental group was gradually replaced with the feed containing 2% additive (replacement ratio 50% → 100%).

[0120] Formal test period (30 days):

[0121] Feeding frequency: 3 times a day (7:00, 12:00, 18:00), free access to food until full.

[0122] Table 2 Feeding records

[0123] Time Operation Content 6:30 Clean the feed trough and weigh the remaining feed 7:00 Feed the first feed of the day 12:00 Second feeding and record the feed intake 18:00 Third feeding, clean the feed trough and disinfect

[0124] Drinking water management: Free drinking (nipple - type water dispenser, water quality conforming to GB 5749 - 2006).

[0125] 4. Data collection and detection

[0126] Growth performance:

[0127] Daily weight gain: Weighing on an empty stomach every morning (electronic scale accuracy ±0.01 kg).

[0128] Feed - to - gain ratio: Total feed intake (kg) / Total weight gain (kg).

[0129] Health indicators:

[0130] Diarrhea rate: Observe the fecal morphology every day, score according to four levels (1 = normal, 4 = watery stool), and record the number of diarrhea times.

[0131] Diarrhea rate scoring standard:

[0132] Grade 1: Formed feces, no liquid exudation;

[0133] Grade 2: Slightly loose feces, a small amount of liquid exudation;

[0134] Grade 3: Pasty feces, obvious liquid exudation;

[0135] Grade 4: Watery feces, no solid components.

[0136] Basis: Refer to the "Guidelines for Animal Diarrhea Scoring" (Journal of Animal Science, 2018) to ensure the scientific nature of the scoring.

[0137] Serum immune indexes: On the 0th and 30th days of the experiment, 5 mL of jugular vein blood was collected, centrifuged (3,000 rpm, 10 min) to obtain serum, and stored at -80 °C.

[0138] IgG detection: ELISA kit (product number: XYZ123, sensitivity 0.1 μg / mL).

[0139] Inflammatory factors: IL-6, TNF-α (Luminex multi-factor detection platform).

[0140] 5. Experimental results (detailed data)

[0141] Table 3 Experimental results

[0142]

[0143] 6. Explanation of key mechanisms

[0144] Reducing the diarrhea rate: Free phenols (125 mg / g) in the additive inhibit intestinal pathogenic bacteria (such as Escherichia coli), and flavonoids (1.4%) enhance the intestinal barrier function.

[0145] Enhancing immunity: Metabolites of Lactobacillus plantarum (such as short-chain fatty acids) activate intestinal lymphocytes and promote IgG secretion.

[0146] Improving the feed conversion ratio: Complex enzymolysis improves the utilization rate of soluble sugars in Chinese medicine residues and reduces energy waste.

[0147] Application Example 2

[0148] The Ligustrum lucidum residue feed additive prepared in Example 2 is applied to aquaculture:

[0149] 1. Experimental animals and grouping

[0150] Variety: Nile tilapia (initial weight 50 ± 5 g, from the same hatching batch).

[0151] Aquaculture system: Recirculating aquaculture pond (water temperature 28 ± 1°C, dissolved oxygen ≥ 6 mg / L, pH 7.2 - 7.5).

[0152] Grouping: 200 tails were randomly divided into a control group (quality percentage of basic feed: fish meal: 20%, soybean meal: 25%, rapeseed meal: 15%, corn: 25%, wheat flour: 10%, fish oil: 3%, calcium dihydrogen phosphate: 1%, premix: 1%, composition of premix (per kg of feed): vitamin A 10,000 IU, vitamin D 3 2,000 IU, vitamin E 50 IU, vitamin C 200 mg, zinc (zinc sulfate) 80 mg, copper (copper sulfate) 10 mg, antioxidant (ethoxyquin) 0.1 g.) and an experimental group (basic feed + 3% Ligustrum lucidum residue feed additive, by mass percentage).

[0153] 2. High - temperature stress experiment

[0154] Stress conditions: The water temperature was raised to 35°C and maintained for 7 days, and the dissolved oxygen was maintained at ≥ 5 mg / L.

[0155] Feeding management: Fed 3 times a day (8:00, 13:00, 18:00), and the feeding amount was 3% of the fish body weight, by mass percentage.

[0156] Table 4 Experimental results

[0157] Index Control Group Experimental Group Improvement Effect Survival Rate 70% (70 / 100 tails) 90% (90 / 100 tails) +28.6％ Average Weight Gain (g) 45±5 55±6 +22.2％ Feed Conversion Ratio 1.5:1 1.3:1 -13.3％

[0158] Application Example 3

[0159] The Salvia miltiorrhiza residue feed additive prepared in Example 3 was applied to broilers:

[0160] 1. Experimental animals and grouping

[0161] Variety: AA white - feather broilers (1 - day - old, initial body weight 40 ± 2 g).

[0162] Feeding conditions: Stacked chicken cages, temperature 33°C in the first week, decreasing 2°C per week until it was constant at 24°C.

[0163] Grouping: 200 were divided into a control group (quality percentage of basic feed: corn: 60%, soybean meal: 28%

[0164] wheat bran: 5%, stone powder: 1.5%, calcium hydrogen phosphate: 1.2%, table salt: 0.3%, premix: 4%

[0165] Composition of premix (per kg of feed): vitamin A 12,000 IU, vitamin D 3 3,000 IU, vitamin E 80 IU, vitamin K 32 mg, iron (ferrous sulfate) 120 mg, manganese (manganese sulfate) 100 mg, selenium (sodium selenite) 0.3 mg, antioxidant (ethoxyquinoline) 0.15 g. ), experimental group (basic feed + 2.5% additive).

[0166] 2. Fattening effect (at 42 days of age). The feed-to-weight ratio uses the industry general formula (Determination Methods for Poultry Production Performance, NY / T 823-2004);

[0167] - The determination of pectoralis muscle rate refers to the Standard for Determination of Slaughter Performance of Broilers (GB / T 22489-2008).

[0168] Table 5 Experimental Results

[0169] Index Control Group Experimental Group Improvement Effect Final Body Weight (kg) 2.8±0.3 3.2±0.2 +14.3％ Feed to Weight Ratio 1.6:1 1.4:1 -12.5％ Pectoral Muscle Rate (%) 18.5±1.2 20.3±1.5 +9.7％

[0170] Comparative Example 1

[0171] Single enzymatic hydrolysis (cellulase)

[0172] Condition: Only cellulase (10,000 U / g) is used, and the others are the same as in Example 1.

[0173] Result: The saccharification rate is 42%, the free phenol is 75 mg / g, and IgG is increased by 12% (P>0.05).

[0174] Comparative Example 2

[0175] Ordinary lactic acid bacteria fermentation

[0176] Condition: Replace it with Lactobacillus plantarum CGMCC No. 1.555 (the preservation number can be publicly queried, characteristic: low phenol acid esterase activity), purchased from the China General Microbiological Culture Collection Center (CGMCC), preservation number CGMCC No. 1.555, and its phenol acid esterase activity is 20 U / mL (caffeic acid phenethyl ester substrate method), and the others are the same as in Example 1.

[0177] Result: The free phenol is 85 mg / g, the flavonoid retention rate is 85 mg / g (compared with 125 mg / g in Example 1), the flavonoid retention rate drops to 65% (compared with 85.3% in Example 1), and the enzyme activity is only 20 U / mL.

[0178] Comparative Example 3

[0179] Hot water extraction (80 °C)

[0180] Condition: The extraction temperature is 80 °C, and the others are the same as in Example 1.

[0181] Results: 0.84% (compared with 1.4% in Comparative Example 1), retention rate 60% (initial flavonoid content 1.4% → only 60% remained after high-temperature extraction).

[0182] Although the above embodiments have described the present invention in detail, they are only a part of the embodiments of the present invention, rather than all embodiments. People can also obtain other embodiments based on these embodiments without creative efforts, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for preparing a Chinese medicine residue feed additive, characterized in that: The following steps are involved: 1) drying and crushing the Chinese medicine residue to obtain a crushed material; 2) mixing the crushed material obtained in step 1) with a composite enzyme and performing enzymolysis to obtain an enzymolysis material; The complex enzyme comprises cellulase, xylanase and pectinase; 3) mixing the enzymolysis material obtained in step 2) with Lactobacillus plantarum LP-01 and performing anaerobic fermentation to obtain a fermentation material; 4) mixing the fermented material obtained in step 3) with yeast extract and manganese sulfate to obtain a mixed material; 5) mixing the mixture obtained in step 4) with water, leaching, and filtering to obtain a filtrate; 6) The filtrate obtained in step 5) is concentrated and freeze-dried to obtain a Chinese medicinal residue feed additive.

2. The preparation method according to claim 1, characterized in that: The drying conditions in step 1) include: a temperature of 60° C. and a drying time of 24 hours; The Chinese medicine residues include one or more of Astragalus root residues, Ligustrum lucidum root residues and Salvia miltiorrhiza root residues; The moisture content of the Chinese medicine residue is 60-80%; The particle size of the crushed material is less than 0.18 mm.

3. The preparation method according to claim 1, characterized in that: In the step 2), the volume ratio of the crushed material to the complex enzyme is 100:0.3; The activity ratio of the cellulase, xylanase and pectinase is 2:1:1.5, the enzyme activity of the cellulase is above 10000 U / g, the enzyme activity of the xylanase is above 5000 U / g, and the enzyme activity of the pectinase is above 3000 U / g; The conditions of the enzymatic hydrolysis include: enzymatic hydrolysis for 2 hours at a temperature of 60° C., a pH value of 5.0, and a rotation speed of 150 rpm; The enzymolysis is followed by inactivation to obtain an enzymolysis material; the inactivation conditions include: a temperature of 90° C. and a time of 10 minutes.

4. The preparation method according to claim 1, characterized in that: In the step 3), the volume ratio of the enzymatic hydrolyzate to Lactobacillus plantarum LP-01 is 100:8; The plant lactobacillus LP-01 is mixed in the form of bacterial liquid, and the bacterial content of the plant lactobacillus LP-01 in the bacterial liquid is 5×10 9 CFU / mL; The conditions of the anaerobic fermentation include: temperature of 32° C., pH value of 6.5, rotation speed of 50-100 rpm, and time of 48 hours.

5. The preparation method according to claim 1, characterized in that: The mass ratio of the mixed material in step 4) to the yeast extract and manganese sulfate is 100:0.2:0.

1.

6. The preparation method according to claim 1, characterized in that: The volume ratio of the mixture to water in step 5) is 1:6; The extraction conditions include: temperature of 45° C. and number of times of 3 times.

7. The preparation method according to claim 1, characterized in that: The concentration temperature in step 6) is 55°C.

8. A Chinese medicinal residue feed additive prepared by the preparation method according to any one of claims 1 to 7.

9. Use of the Chinese medicinal residue feed additive according to claim 8 in preparing animal feed.

10. An animal feed, characterized in that: The feed comprises a basic feed and the Chinese medicinal residue feed additive according to claim 8, wherein the mass ratio of the basic feed to the Chinese medicinal residue feed additive is 100:2-3.

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