Postbiotic piglet milk powder as well as preparation method and application thereof

Abstract

The invention discloses postbiotic piglet milk powder as well as a preparation method and application thereof, and belongs to the technical field of animal feed. The milk powder contains 30-40% of solid-state fermented feed, and the release amount of active ingredients is increased after the traditional Chinese medicines in the fermented raw materials are fermented, so that the milk powder can play a better role; fish meal and soybean concentrated protein in the fermentation raw materials are converted into small peptides and amino acids after being fermented, so that the nutritional value of the product is improved; generated butyric acid and lactic acid have pleasant acid fragrance and have a food calling effect; the generated proteinase and lipase can solve the problem of weak digestive function of piglets; generated butyric acid and antibacterial peptide have an antibacterial effect; the contained beneficial bacteria can be colonized in intestinal tracts, and the effects of regulating intestinal microecological balance, resisting bacteria, aiding digestion and the like can be continuously achieved. The milk powder is rich in nutrition, contains a large amount of proteins and immune factors similar to breast milk, and can effectively buffer weaning stress of piglets and improve body resistance of the piglets; and the dissolvability is good, no precipitate is generated, and the product can be fully utilized.

Description

Technical Field

[0001] The invention belongs to the technical field of animal feed, and in particular relates to a postbiotic piglet milk powder and a preparation method and application thereof. Background Art

[0002] In large-scale breeding farms, some sows are not adapted to the environment and suffer from heat stress and immune stress, and their feed intake is reduced, resulting in insufficient lactation. This is a common problem in the breeding industry. Piglets are very fragile from birth to weaning. Insufficient breast milk will cause diarrhea, weak vitality, decreased resistance, slow weight gain, and eventually produce weak and stiff pigs. Under general management conditions in winter, the mortality rate of weak and stiff pigs can be as high as 50% to 80%. The health level of survivors is also significantly lower than that of normal pigs, and they are easily attacked by diseases. Once they are infected with diseases, they will threaten other pigs, seriously affecting the economic benefits of the pig farm. Pig formula milk powder is an effective way to solve this problem.

[0003] At present, there are some patents on pig formula milk powder, the formula of which involves dairy products (milk powder, whey powder, whey protein powder, etc.), protein (plasma protein powder, egg powder, etc.), sugars (glucose, brown sugar, lactose, sucrose, corn dextrin and plant polysaccharides), vegetable oils, organic acids, amino acids, vitamins, inorganic salts, flavoring agents and sweeteners (CN201610827118.0, CN 201410539605.8, CN 201710421183.8, CN 201510437184.2). These products are rich in nutrition and can effectively relieve the stress response of piglets before and after weaning, improve their immunity, reduce the incidence of diarrhea, and thus improve their growth performance. There are also some pig formula milk powders with better effects on the market, but these products have poor solubility and precipitate after preparation. Piglets only drink the supernatant, and the precipitate is easy to spoil after long-term storage, causing waste and pollution.

[0004] Therefore, a new pig health milk powder product with good solubility and strong antibacterial and health-care ability is urgently needed. Summary of the invention

[0005] In view of this, the present invention provides a formula milk powder that is rich in nutrition, contains a large amount of protein and immune factors similar to breast milk, can effectively buffer the weaning stress of piglets and improve their body resistance, in order to solve the above-mentioned many common problems existing in such products.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] One of the purposes of the present invention is to provide a solid-state fermentation feed, which is prepared by fermenting Clostridium butyricum-Bacillus coagulans fermentation supernatant in a solid-state fermentation medium; the Clostridium butyricum-Bacillus coagulans fermentation supernatant is prepared by inoculating and fermenting Clostridium butyricum and Bacillus coagulans in a ratio of 1.5-2.5:1; the solid-state fermentation medium comprises, by weight percentage, 40-60% of Chinese medicinal raw materials, 10-30% of soybean protein concentrate, 3-8% of fish meal, 10-20% of corn flour, 3-8% of molasses, 1-2% of dipotassium hydrogen phosphate, 2-3% of magnesium sulfate and 0.5-2% of sodium citrate; the inoculation volume weight percentage of the Clostridium butyricum-Bacillus coagulans fermentation supernatant in the solid-state fermentation medium is 10-30%.

[0008] Furthermore, the Clostridium butyricum-Bacillus coagulans fermentation supernatant is obtained by fermentation using a two-stage flow-addition feeding method, and the pH value is adjusted to 6-7 before the two feedings. The first feeding is carried out during static culture until the beginning of the logarithmic phase, the feeding liquid component is glucose, and the feeding duration is 6-10 hours. The second feeding liquid components include glucose, peptone and dipotassium hydrogen phosphate, and the feeding is carried out until the end of the logarithmic phase.

[0009] Furthermore, the Clostridium butyricum preservation number is CCTCC No: M2022151, and the Bacillus coagulans preservation number is CGMCC No.22951.

[0010] Furthermore, the Chinese medicinal raw materials include astragalus, acanthopanax senticosus and licorice.

[0011] The second object of the present invention is to provide a method for preparing the solid-state fermented feed, comprising the following steps:

[0012] Step 1: Solid-state fermentation strain preparation

[0013] The Clostridium butyricum and the Bacillus coagulans are inoculated into the culture medium according to the inoculation ratio, the liquid filling coefficient is 60-80%, and the combined fermentation is carried out at 36-38° C. using the fermentation method, and the centrifuged supernatant after the fermentation is the solid-state fermentation strain;

[0014] Step 2: Solid-state fermentation feed preparation

[0015] First, astragalus, acanthopanax and licorice are crushed and mixed in a mass ratio of 1-3:1-3:0.5-2 to obtain a Chinese medicine raw material; then, the raw material is mixed and sterilized with other raw material components according to the formula to obtain the solid fermentation medium; then, the solid fermentation medium is loaded into a fermentation bag, the strain prepared in step 1 is inoculated according to the volume weight ratio, water is added to a material-water ratio of 1:0.5-1.0, the bag is sealed after being stirred evenly, and the bag is cultured at 36-38°C for 5-8 days, during which the mixed material is shaken every day, and after the fermentation is completed, the product is dried, crushed and sieved to obtain the solid fermentation feed.

[0016] Furthermore, in step one, the culture medium components include 8-12 g / L bran, 13-17 g / L corn steep liquor powder, 13-17 g / L peptone, 0.5-1.5 g / L dipotassium hydrogen phosphate, 0.1-1.0 g / L manganese sulfate, and a pH value of 6.5-7.5; in the fermentation method, the first feed liquid component is 4-8 g / L glucose, and the second feed liquid component includes 1-3 g / L glucose, 0.5-1.5 g / L peptone and 0.1-0.5 g / L dipotassium hydrogen phosphate.

[0017] Furthermore, in step 2, the Chinese medicine raw material is crushed to a mesh size of 50 to 80; the solid-state fermentation medium is sterilized at 115 to 125° C. for 15 to 25 minutes; the fermentation bag is provided with a one-way air release valve; and the fermentation product is sieved to a mesh size of 80 to 100.

[0018] The third object of the present invention is to provide a postbiotic milk powder based on the solid-state fermented feed, wherein the postbiotic milk powder comprises the following components by weight percentage: 30-40% of the solid-state fermented feed, 30-40% of sugars, 13-33% of dairy products, 5-7% of inorganic salts, 0.4-0.6% of vitamins and 0.4-0.6% of suspending agents, thickeners and emulsifiers.

[0019] Furthermore, the sugars include lactose, sucrose and glucose; the dairy products include whole milk powder and whey protein powder; the inorganic salts include ferrous sulfate, zinc sulfate, manganese sulfate, sodium selenite, potassium dihydrogen phosphate, calcium carbonate and sodium chloride; the vitamins include vitamin C, vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, pantothenic acid, folic acid and biotin; the suspending agents, thickeners and emulsifiers include xanthan gum, sucrose fatty acid esters, polyglycerol fatty acid esters and microcrystalline cellulose.

[0020] A fourth object of the present invention is to provide an application of the solid-state fermented feed, the preparation method or the postbiotic milk powder in piglet breeding.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] (1) The formula milk powder provided by the present invention is rich in nutrition and contains a large amount of protein and immune factors similar to breast milk, which can effectively buffer the weaning stress of piglets and improve their body resistance.

[0023] (2) The product of the present invention uses a Chinese medicine fermented by Clostridium butyricum and Bacillus coagulans. The Chinese medicine ingredients, such as Astragalus, Licorice and Acanthopanax senticosus, which have antibacterial, antiviral and immunity-enhancing effects, have their active ingredients released more efficiently after being fermented by Clostridium butyricum and Bacillus coagulans, and thus can better exert their effects. The fish meal and soybean protein concentrate in the fermentation raw materials are converted into small peptides and amino acids after fermentation, thereby improving the nutritional value of the product. The butyric acid produced can be used as a nutrient for the intestine, and the lactic acid produced has an antibacterial effect. Butyric acid and lactic acid have a pleasant sour aroma and an appetite-inducing effect. The protein enzyme and lipase produced can solve the problem of weak digestive function in piglets. The antimicrobial peptides such as Clostridium butyricum and coagulin produced have an antibacterial effect. The Clostridium butyricum and Bacillus coagulans contained in the product can be colonized in the intestine of animals, and continuously produce beneficial effects such as regulating the balance of intestinal microecology, antibacterial effects and aiding digestion.

[0024] (3) The present invention uses the Clostridium butyricum-Bacillus coagulans centrifuge liquid as a seed for solid-state fermentation to achieve waste utilization and solve the environmental problems caused by waste liquid.

[0025] (4) The formula milk powder prepared by the present invention has very good solubility and does not generate precipitation, so it can be fully utilized.

[0026] Description of the deposit certificate:

[0027] Collection certificate 1:

[0028] Depository: China Center for Type Culture Collection;

[0029] Deposit number: CCTCC No: M 2022151;

[0030] Deposit date: February 25, 2022;

[0031] Deposit address: Wuhan University, Wuhan, China;

[0032] Taxonomic name: Clostridium butyricum.

[0033] Preservation certificate 2:

[0034] Depository: General Microbiology Center, China Microbiological Culture Collection Administration;

[0035] Deposit number: CGMCC No.22951;

[0036] Deposit date: July 26, 2021;

[0037] Storage address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing;

[0038] Taxonomic name: Bacillus coagulans). DETAILED DESCRIPTION

[0039] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Without departing from the spirit and essence of the present invention, modifications or substitutions made to the methods, steps or conditions of the present invention are within the scope of the present invention. Unless otherwise specified, the products, reagents, equipment, etc. used in the following examples are all commercially available, and the methods used are consistent with conventional methods unless otherwise specified.

[0040] The Clostridium butyricum and Bacillus coagulans strains used in the joint fermentation of the present invention were preserved by the research group in the early stage, and the fermentation process was described in the previously applied patent ZL 202010482530.X.

[0041] The technical solution of the present invention is further described in detail below in conjunction with embodiments.

[0042] Example 1 Preparation of Clostridium butyricum-Bacillus coagulans solid-state fermentation feed

[0043] 1. Preparation of solid-state fermentation strains

[0044] Preparation of Clostridium butyricum seeds: The Clostridium butyricum activation medium has the same components as the seed medium, including: peptone 10g / L, beef extract 10g / L, yeast powder 3g / L, glucose 5g / L, soluble starch 1g / L, sodium chloride 5g / L, sodium acetate 3g / L, L-cysteine ​​hydrochloride 0.5g / L, pH 6.8. Culture in an anaerobic incubator at 37°C for 12h, shake flask filling coefficient 50%, inoculation amount 4%.

[0045] Preparation of Bacillus coagulans seeds: The activation medium of Bacillus coagulans has the same components as the seed medium, including: 10 g / L glucose, 5 g / L yeast extract, 10 g / L tryptone, 1000 mL distilled water, pH 6.8-7.0. Cultivate at 37°C, 180 rpm for 24 h, with a liquid volume of 20% and an inoculum volume of 4%.

[0046] Clostridium butyricum-Bacillus coagulans 10L tank joint fermentation: the culture medium components include bran 10g / L, corn steep liquor dry powder 15g / L, peptone 15g / L, potassium dihydrogen phosphate 1g / L, manganese sulfate 0.5g / L, pH 7. The joint fermentation is carried out at 37°C, the inoculation amount of Clostridium butyricum and Bacillus coagulans is 8% and 4% respectively, and the liquid filling coefficient is 70%. A two-stage flow feeding is adopted, first static culture is carried out to the logarithmic phase, the speed is adjusted to 50rpm, and 10% NaOH solution is added to adjust the pH to 6.5, and then the first feeding is carried out. The feeding liquid component is glucose 6g / L, and the feeding lasts for 8h. After the feeding is completed, 10% NaOH solution is added again to adjust the pH to 6.5, and the second feeding is carried out. The components are glucose 2g / L, peptone 1g / L, and potassium dihydrogen phosphate 0.2g / L composite feeding liquid is fed until the end of the logarithmic phase, and the fermentation time is 40h.

[0047] After the fermentation, the fermentation liquid was centrifuged at 4000 rpm for 10 min, the obtained precipitate was used to prepare bacterial powder, and the discarded supernatant was used as the strain for solid-state fermentation feed.

[0048] 2. Solid-state fermentation feed preparation

[0049] Astragalus, Acanthopanax and Licorice (mass ratio 2:2:1) were crushed to 80 meshes respectively and mixed evenly to obtain Chinese medicine raw materials. 50% Chinese medicine raw materials, 20% soybean protein concentrate, 5% fish meal, 15% corn flour, and 5% molasses were mixed evenly, and then mixed evenly with 1.5% potassium dihydrogen phosphate, 2.5% magnesium sulfate and 1% sodium citrate, and sterilized at 121°C for 20 minutes to obtain solid fermentation medium. The solid fermentation medium was loaded into a fermentation bag with a one-way air release valve, and Clostridium butyricum-Bacillus coagulans strains were added to the bag at a ratio of 20% (v / w), water was added to the bag to a material-water ratio of 1:0.8, and the bag was sealed after stirring evenly. It was placed at 37°C for 7 days, and the materials were mixed evenly by shaking every day. After the fermentation, the product was dried, crushed, and sieved through a 100-mesh sieve.

[0050] Example 2 Preparation of Clostridium butyricum-Bacillus coagulans postbiotic milk powder

[0051] 1. The raw material formula is shown in Table 1.

[0052] Table 1 Main raw material formula of post-biotic milk powder (%)

[0053]

[0054] Note: In addition to the substances in Table 1, each formula also contains 1% potassium dihydrogen phosphate, 0.5% calcium carbonate, 0.5% sodium chloride, 0.5% complex vitamins and 4.0% complex minerals, 0.15% xanthan gum, 0.1% sucrose fatty acid esters, 0.1% polyglycerol fatty acid esters and 0.15% microcrystalline cellulose. The complex minerals are composed of the following substances (g / kg): iron 20, zinc 19, manganese 0.5, selenium 0.5, the components containing iron, zinc and manganese are sulfates, and the components containing selenium are sodium selenite; vitamins include the following substances (g / kg): vitamin C 3.2, vitamin B10.3, vitamin B20.5, vitamin B60.12, vitamin B120.01, niacin 0.2, pantothenic acid 0.13, folic acid 0.5, biotin 0.04.

[0055] 2. Preparation of Formula Milk Powder

[0056] The above raw materials are crushed until they can pass through an 80-mesh sieve, and then poured into a blender in descending order of weight. After mixing evenly, they are dried at 90°C until the moisture content is less than 4%, and then inspected, measured and packaged.

[0057] The formula milk powder prepared according to the above method was dissolved in warm water in different proportions and all were in a uniform state, and no precipitation was formed after being left for 12 hours.

[0058] Example 3 Application of Clostridium butyricum-Bacillus coagulans postbiotic milk powder

[0059] 180 healthy suckling piglets with similar body shape were selected from the delivery room of the breeding base of Henan Guang'an Feed Co., Ltd. and randomly divided into 9 groups, each with 20 piglets. The experimental groups were fed with pig milk powder prepared according to Formula 1 to Formula 6 in Example 2 (experimental groups 1 to 6), "newborn suckling piglet special milk powder" containing probiotics purchased from Pinyou Animal Milk Powder Store (experimental group 7), fermented Chinese medicine with the content in Formula 1 (experimental group 8) and the same amount of Clostridium butyricum and Bacillus coagulans mixed bacterial powder in Formula 1 (experimental group 9), and the control group was fed normally.

[0060] The piglets were fed and managed according to the routine, including ventilation, heat preservation, deworming, and free access to food and water. They were fed 3-4 times a day. They were fed milk powder (milk powder for piglets: warm water = 1:3) 3 days after delivery. The experiment lasted for 30 days.

[0061] The feed intake during the experiment was recorded, and the piglets were weighed at the beginning and end of the experiment. The number of piglets with diarrhea was counted at 9 am every day, and the average daily weight gain, diarrhea rate and feed-to-meat ratio (F / G) were calculated.

[0062] Diarrhea rate = number of pigs with diarrhea / (number of pigs tested × total number of days) × 100%

[0063] SAS (Statistical Analysis) version 7.2 was used for statistical analysis and variance analysis, and the results were considered significant when P < 0.05. Significant differences were expressed by letter marking: first, all the averages were arranged in order from large to small, and then the letter a was marked on the largest average; and the average was compared with the following averages, and all those with insignificant differences were marked with the letter a, until a certain average with significant difference was marked with the letter b; then the average marked with b was used as the standard, and compared with the averages above that were larger than it, and all those with insignificant differences were also marked with the letter b; then the largest average marked with b was used as the standard, and compared with the following unmarked averages, all those with insignificant differences continued to be marked with the letter b, until a certain average with significant difference was marked with c. The results are shown in Table 2, and the average value of each group of data was taken.

[0064] Table 2 Comparison of animal feeding experimental effects

[0065]

[0066]

[0067] From the statistical analysis results of Table 2, it can be seen that compared with the control group, all test groups have significantly increased average daily weight gain, and feed-to-meat ratio and diarrhea rate are significantly reduced. The daily weight gain and feed-to-meat ratio results of test groups 5 and 6 are slightly better than those of other test groups, but the difference is not significant. Test groups 5 and 6 are more effective than other test groups in reducing the diarrhea rate of piglets, which is comparable to the results of commercial formula milk powder, but the production cost is much lower than that of commercial milk powder. The formula milk powder of the present invention has the best effect, because in addition to the dairy products that can strengthen nutrition and improve immunity, the product of the present invention also contains Clostridium butyricum-Bacillus coagulans fermented Chinese medicine, in which Astragalus, Licorice and Acanthopanax have antibacterial, antiviral and immunity-enhancing effects; the small peptides and amino acids produced by fermentation improve the nutritional value of the product; the butyric acid and lactic acid with a pleasant sour aroma have an appetizing effect; the digestive enzymes produced can improve the digestive ability of piglets; the butyric acid and antimicrobial peptides produced have antibacterial effects. Only adding fermented Chinese medicine or only adding probiotics lacks the effect of dairy products, and the effect is obviously not as good as the formula milk powder of the present invention.

[0068] In addition, this product has also achieved good results in rescuing weak, sick piglets and early weaning stress piglets.

[0069] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A solid-state fermented feed, characterized in that: The invention discloses a method for preparing the fermentation of Clostridium butyricum-Bacillus coagulans by fermenting the fermentation supernatant of Clostridium butyricum-Bacillus coagulans in a solid fermentation medium; the fermentation supernatant of Clostridium butyricum-Bacillus coagulans is prepared by inoculating and fermenting Clostridium butyricum and Bacillus coagulans in a ratio of 1.5-2.5:1; the solid fermentation medium comprises, by weight percentage, 40-60% of Chinese medicinal materials, 10-30% of soybean protein concentrate, 3-8% of fish meal, 10-20% of corn flour, 3-8% of molasses, 1-2% of dipotassium hydrogen phosphate, 2-3% of magnesium sulfate and 0.5-2% of sodium citrate; the inoculation volume weight percentage of the fermentation supernatant of Clostridium butyricum-Bacillus coagulans in the solid fermentation medium is 10-30%.

2. The solid-state fermented feed according to claim 1, characterized in that: The Clostridium butyricum-Bacillus coagulans fermentation supernatant is obtained by fermentation in a two-stage flow-addition feeding mode, the pH value is adjusted to 6-7 before two feedings, the first feeding is carried out at the beginning of the logarithmic phase after static culture, the feeding liquid component is glucose, and the feeding duration is 6-10 hours, and the second feeding liquid component includes glucose, peptone and dipotassium hydrogen phosphate, and the feeding is carried out until the end of the logarithmic phase.

3. The solid-state fermented feed according to claim 2, characterized in that: The Clostridium butyricum preservation number is CCTCC No: M2022151, and the Bacillus coagulans preservation number is CGMCC No.22951.

4. The solid-state fermented feed according to claim 3, characterized in that: The Chinese medicinal raw materials include astragalus, acanthopanax senticosus and liquorice.

5. The method for preparing the solid-state fermented feed according to any one of claims 1 to 4, characterized in that: The following steps are involved: Step 1: Solid-state fermentation strain preparation The Clostridium butyricum and the Bacillus coagulans are inoculated into the culture medium according to the inoculation ratio, the liquid filling coefficient is 60-80%, and the combined fermentation is carried out at 36-38° C. using the fermentation method, and the centrifuged supernatant after the fermentation is the solid-state fermentation strain; Step 2: Solid-state fermentation feed preparation First, astragalus, acanthopanax and licorice are crushed and mixed in a mass ratio of 1-3:1-3:0.5-2 to obtain a Chinese medicine raw material; then, the raw material is mixed and sterilized with other raw material components according to the formula to obtain the solid fermentation medium; then, the solid fermentation medium is loaded into a fermentation bag, the strain prepared in step 1 is inoculated according to the volume weight ratio, water is added to a material-water ratio of 1:0.5-1.0, the bag is sealed after being stirred evenly, and the bag is cultured at 36-38°C for 5-8 days, during which the mixed material is shaken every day, and after the fermentation is completed, the product is dried, crushed and sieved to obtain the solid fermentation feed.

6. The preparation method according to claim 5, characterized in that: In step one, the culture medium components include 8-12 g / L bran, 13-17 g / L corn steep liquor powder, 13-17 g / L peptone, 0.5-1.5 g / L dipotassium hydrogen phosphate, 0.1-1.0 g / L manganese sulfate, and a pH value of 6.5-7.5; in the fermentation method, the first feed liquid component is 4-8 g / L glucose, and the second feed liquid component includes 1-3 g / L glucose, 0.5-1.5 g / L peptone and 0.1-0.5 g / L dipotassium hydrogen phosphate.

7. The preparation method according to claim 6, characterized in that: In step 2, the Chinese medicine raw material is crushed to a mesh size of 50 to 80; the solid-state fermentation medium is sterilized at 115 to 125° C. for 15 to 25 minutes; the fermentation bag is provided with a one-way air release valve; and the fermentation product is sieved to a mesh size of 80 to 100.

8. A postbiotic milk powder based on the solid-state fermented feed according to any one of claims 1 to 4, characterized in that: The postbiotic milk powder comprises the following components by weight percentage: 30-40% of the solid fermentation feed, 30-40% of sugars, 13-33% of dairy products, 5-7% of inorganic salts, 0.4-0.6-3% of vitamins and 0.4-0.6% of suspending agents, thickeners and emulsifiers.

9. The postbiotic milk powder according to claim 8, characterized in that: The sugars include lactose, sucrose and glucose; the dairy products include whole milk powder and whey protein powder; the inorganic salts include ferrous sulfate, zinc sulfate, manganese sulfate, sodium selenite, potassium dihydrogen phosphate, calcium carbonate and sodium chloride; the vitamins include vitamin C, vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, pantothenic acid, folic acid and biotin; the suspending agents, thickeners and emulsifiers include xanthan gum, sucrose fatty acid esters, polyglycerol fatty acid esters and microcrystalline cellulose.

10. Use of the solid-state fermented feed according to any one of claims 1 to 4, the preparation method according to any one of claims 5 to 7, or the postbiotic milk powder according to any one of claims 8 to 9 in piglet breeding.

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