Potato residue fermented feed and preparation method thereof

Abstract

The invention provides a potato residue fermented feed and a preparation method thereof. The feed is prepared by performing microbial fermentation on the potato residues, the cellulose content of the original potato residues is effectively reduced by adjusting microbial composition and fermentation steps and adjusting addition of trace elements and the like, and the obtained feed contains a large number of active substances which can be effectively absorbed by animals, so that the feed is beneficial to improving the immunity of the animals. The animal immune regulation capability is effectively improved, and animal growth and development are promoted; the growth of aspergillus flavus is effectively inhibited, and the storage time of the feed is prolonged.

Description

Technical Field

[0001] The invention belongs to the technical field of feed processing, and in particular relates to a potato residue fermented feed and a preparation method thereof. Background Art

[0002] The statements in this section merely provide background information related to the present invention and do not necessarily constitute prior art.

[0003] Potatoes are high-quality crops in my country. After potato processing and starch extraction, some residual substances are usually left, namely potato slag. Potato slag is rich in starch and dietary fiber, and also contains a certain amount of protein, which can be used to feed animals. However, potato slag has a high water content, is easy to rot and rancid, and produces a bad smell. Its protein content is low, and it is not enough to meet the requirements of feeding animals when used alone. It is also easily contaminated by aflatoxin and other pollutants, producing toxic substances and causing harm to animal health.

[0004] Research has found that potato residues can be treated with enzymes or bacterial enzymes to reduce the cellulose content and increase the protein content, and then used as feed, which can reduce the use of soybean meal and expand feed reserves. However, there are several problems: first, the cost of enzymes is high and the operation is complicated, and the use of enzymes to treat potato residues greatly increases production costs; second, it can regulate animal immunity to a certain extent, but it often requires the addition of a large amount of Chinese herbal medicine ingredients, which increases the cost and is not conducive to large-scale processing and production; third, the palatability and preservation of feed are often ignored. Summary of the invention

[0005] In order to overcome the deficiencies of the above-mentioned prior art, the present invention provides a fermented feed of potato residue and a preparation method thereof. One of the purposes of the present invention is to reuse the industrial production waste of potato residue and reduce its pollution to the environment; the second purpose of the present invention is to prepare a feed that can effectively improve the immune regulation ability of animals; the third purpose of the present invention is to improve the problem that the existing feed is not easy to preserve for a long time.

[0006] To achieve the above objectives, one or more embodiments of the present invention provide the following technical solutions: In a first aspect, the present invention provides a potato residue fermented feed, which comprises 150-200 parts of potato residue, 120-150 parts of auxiliary materials, 100-120 parts of purified water, 2-4 parts of Trichoderma reesei fermentation liquid, 8-15 parts of composite bacteria fermentation liquid, 2-4 parts of plant extract, 1-3 parts of vitamins and 1-3 parts of trace element additives; The auxiliary materials include 90-120 parts of bran, 40-60 parts of yam, and 30-50 parts of purslane; The composite bacteria fermentation liquid comprises 3-5 parts of yeast, 2-5 parts of Lactobacillus plantarum, 3-5 parts of Bacillus licheniformis, 1-2 parts of Bacillus velezaceus, and 1-3 parts of Pediococcus pentosaceus; The plant extract comprises 1-3 parts of curcumin and 1-3 parts of quercetin.

[0007] Yam is a plant that can be used as both medicine and food. It can be used to prepare feed. It is rich in nutrients such as starch, protein, polysaccharides, etc., which can improve animal intestinal health, enhance immunity, and promote animal growth and development.

[0008] Purslane is also a plant with both medicinal and edible properties. It can be used in feed preparation. It is rich in nutrients and can improve the intestinal health of animals and enhance immunity. Purslane extracts are used in animal breeding and production, and can also reduce and prevent livestock and poultry diseases and reduce the use of antibiotics. However, its fermented products are rarely used in animal immunity.

[0009] Plant extracts refer to substances extracted or processed from plants (whole or part of a plant) using appropriate solvents or methods, and can be used to prepare feed additives. Curcumin and quercetin, as feed additives, can improve the production performance of livestock and poultry, enhance immunity, improve meat and egg quality, and enhance antioxidant capacity. They are effective substitutes for antibiotics and help achieve antibiotic-free farming.

[0010] In one or more specific embodiments of the present invention, the vitamins include 0.3-0.8 parts of vitamin A, 0.3-0.8 parts of vitamin E, and 1.5-3 parts of vitamin C. Vitamins can help improve animal resistance and regulate their immune ability. Curcumin and quercetin work together with vitamins and active substances produced by microbial fermentation to improve animal immunity and prolong feed storage time.

[0011] In one or more specific embodiments of the present invention, the trace element additive includes 1-2 parts of ammonium sulfate, 0.5-1 parts of dipotassium hydrogen phosphate, 0.5-1 parts of sodium chloride, and 0.5-1 parts of magnesium sulfate. The addition of trace elements is not only conducive to fermentation, but also provides support for subsequent animal nutrition.

[0012] In one or more specific embodiments of the present invention, the feed comprises 180-200 parts of potato residue, 140-150 parts of auxiliary materials, 100-120 parts of purified water, 2.5-3.5 parts of Trichoderma reesei fermentation broth, 8-13 parts of composite bacteria fermentation broth, 2.5-3.5 parts of plant extracts, 1.5-2 parts of vitamins and 1.5-2.5 parts of trace element additives; The auxiliary materials include 100-120 parts of bran, 45-60 parts of yam, and 40-50 parts of purslane; The composite bacteria fermentation liquid comprises 3-4 parts of yeast, 2-4 parts of Lactobacillus plantarum, 2-4 parts of Bacillus licheniformis, 1-2 parts of Bacillus velezaceus, and 1-3 parts of Pediococcus pentosaceus; The plant extract comprises 2-3 parts of curcumin and 1.5-2 parts of quercetin.

[0013] In one or more specific embodiments of the present invention, the feed comprises 180 parts of potato residue, 140 parts of auxiliary materials, 120 parts of purified water, 3 parts of Trichoderma reesei fermentation broth, 10 parts of composite bacteria fermentation broth, 3 parts of plant extracts, 2 parts of vitamins and 2 parts of trace element additives; The auxiliary materials include 100 parts of bran, 50 parts of yam and 40 parts of purslane; The composite bacteria fermentation liquid comprises 4 parts of yeast, 3 parts of Lactobacillus plantarum, 3 parts of Bacillus licheniformis, 1 part of Bacillus velezaceus, and 2 parts of Pediococcus pentosaceus; The plant extracts are 2 parts of curcumin and 2 parts of quercetin.

[0014] A second aspect of the present invention provides a method for preparing a fermented potato residue feed, comprising the following steps: 1) Add 2-4 parts of Trichoderma reesei fermentation liquid to 150-200 parts of potato residue, 120-150 parts of auxiliary materials, 100-120 parts of purified water, and 1-3 parts of trace element additives, and ferment at 24-28℃ with ventilation for 48-72 hours until the primary fermentation is completed with white mycelium; 2) Stir 8-15 portions of the compound bacteria fermentation liquid into the product of the primary fermentation, ferment at 30-37°C with ventilation for 24-48 hours, and then ferment in a sealed environment for 24-72 hours to complete the secondary fermentation; 3) Mix the secondary fermentation product with 2-4 parts of plant extract and 1-3 parts of vitamins, dry to a water content of less than 15%, crush and package.

[0015] In the present invention, it is found through experiments that yeast, Lactobacillus plantarum, Bacillus licheniformis, Bacillus velez, Pediococcus pentosaceus and Trichoderma reesei can coexist, especially after Trichoderma reesei is fermented first, it is more conducive to the subsequent fermentation of composite bacteria, and the total bacterial count is increased to 10 8 CFU / g. The reason is that after cultivation, Trichoderma reesei produces a large amount of cellulase, hemicellulase, protease and amylase. Later, during the secondary fermentation, the fermentation conditions have a certain stress effect on it, causing some bacteria to release a variety of enzymes, which is more conducive to reducing the cellulose in potato residues and auxiliary materials, and at the same time, it can provide more favorable conditions for the fermentation of other strains.

[0016] The active substances produced by fermentation of Trichoderma reesei, yeast, Lactobacillus plantarum, Bacillus licheniformis, Bacillus velez, and Pediococcus pentosaceus can also effectively inhibit the growth of other bacteria, especially Aspergillus flavus. These mixed bacteria can promote each other's growth, effectively decompose cellulose, produce a large number of small molecule peptides and sugars, and effectively regulate animal immunity; moreover, these strains can colonize in the animal intestines, regulate the health of intestinal flora, and improve the performance of feed in immune regulation.

[0017] The use of Lactobacillus plantarum not only ensures the production of nutrients, but also can effectively adjust the pH, adjust the taste of feed, and increase the palatability of feed.

[0018] In one or more specific embodiments of the present invention, the auxiliary material is crushed to 0.5-1 cm.

[0019] In one or more specific embodiments of the present invention, in step 1), the potato residue and auxiliary materials are first steamed at 100°C for 20-30 minutes, cooled to room temperature and then set aside. Heating the potato residue and auxiliary materials can kill the bacterial strains attached to them to a certain extent, and inhibit the growth and reproduction of bacterial strains such as Aspergillus flavus that can produce harmful substances, which is more conducive to the later fermentation and produces more beneficial substances; in addition, the heating step can effectively prolong the storage time of the feed.

[0020] In one or more specific embodiments of the present invention, in step 1) and step 2), the ventilation volume is 0.1-0.3 vvm.

[0021] In one or more specific embodiments of the present invention, in the step 3), the drying temperature is 40-60°C.

[0022] In the present invention, the parts are all calculated by weight.

[0023] One or more of the above technical solutions have the following beneficial effects: The present invention aims to turn potato residue into "treasure" and realize resource recycling. The potato residue fermented feed provided by the present invention is rich in nutrition, good in palatability, and suitable for a variety of animals. Different fermented materials are combined with each other, and under the synergistic effect of mixed microorganisms, a large amount of active substances are produced, which are more easily absorbed by animals; it can improve animal resistance, promote animal growth and development, and reduce the production cost of animal feed; at the same time, it inhibits Aspergillus flavus and effectively prolongs the storage time of feed.

[0024] Advantages of additional aspects of the present invention will be given in part in the following description, and in part will become obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The accompanying drawings in the specification, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0026] Figure 1 The effect of different treatment methods on the total bacterial count (unit: ×10 6 CFU / g).

[0027] Figure 2 The effect of different treatment methods on the number of each bacterial cell (unit: ×10 6 CFU / g). DETAILED DESCRIPTION

[0028] It should be noted that the following detailed descriptions are exemplary and are intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present invention belongs.

[0029] In the present invention, fermentation liquid refers to the liquid containing the microorganism itself and its metabolites after the microorganism is inoculated into the culture medium or fermentation material and cultured for a period of time.

[0030] In the present invention, the Trichoderma fermentation broth and the composite bacteria fermentation broth can be cultured using existing technologies. Specifically, the present invention adopts the following method: Preparation method of Trichoderma reesei fermentation broth: Using distilled water slant spore suspension, pick 1 ring of spores to 5 mL of liquid seed culture medium in a 25 mL test tube and incubate at 28 °C and 180 r / min. -1 Culture on an air shaker for 2-3 days and measure the OD 600 The value is between 0.6-0.8, and it is used as liquid seed after microscopic examination. -1 ): Glucose 10, peptone 3, ammonium sulfate 2, yeast extract 0.5, KH 2 PO 4 4. Ca 2 Cl 2 ·2H 2 O 0.3, MgSO 4 7H 2 O 0.3, sterilize at 121℃ for 40min before use.

[0031] Preparation method of composite bacterial fermentation broth: Mix the fermentation broths of the following strains in parts by weight.

[0032] Preparation of yeast fermentation broth: Use distilled water slant spore suspension, pick 1 ring of spores to 5mL liquid seed culture medium in a 25mL test tube, and incubate at 28℃ 180 r·min-1 Culture on an air shaker for 2-3 days and measure the OD 600 The value was between 0.6 and 0.8, and the seeds were used as liquid seeds after microscopic examination. YPD medium (g·L -1 ): Yeast extract 10.0, peptone 20.0, D-glucose 20.0, 115℃, high temperature steam sterilization for 20 minutes.

[0033] Preparation method of Lactobacillus plantarum fermentation broth: Use distilled water inclined spore suspension, pick 1 ring of spores to 5mL liquid seed culture medium in a 25mL test tube, and incubate at 37℃ 180 r·min -1 Culture on an air shaker for 2-3 days and measure the OD 600 The value was between 0.6 and 0.8, and the liquid seeds were used after microscopic examination. MRS medium (g·L -1 ): Peptone 10.0, Beef Extract Powder 10.0, Yeast Extract Powder 5.0, Glucose 20.0, Tween 80 1.0, Sodium Acetate 5.0, Triammonium Citrate 2.0, K 2 HPO 4 2.0, MgSO 4 7H 2 O 0.2 、MnSO 4 ·H 2 O 0.05, 121℃, sterilize for 15 min.

[0034] Preparation of Bacillus licheniformis fermentation broth: Use distilled water slant spore suspension, pick 1 ring of spores to 5mL liquid seed culture medium in a 25mL test tube, and incubate at 37℃ 180 r·min -1 Incubate on an air shaker for 1-2 days and measure the OD 600 The value was between 0.6 and 0.8, and the liquid seeds were used after microscopic examination. LB medium (g·L -1 ): Tryptone 10, yeast extract 5, sodium chloride 10, 121℃, sterilize for 15 min.

[0035] Preparation of Bacillus Velez fermentation broth: Use distilled water slant spore suspension, pick 1 ring of spores to 5mL liquid seed culture medium in a 25mL test tube, and incubate at 37℃ 180 r·min -1 Incubate on an air shaker for 1-2 days and measure the OD 600 The value is between 0.6-0.8, and it is used as liquid seed after microscopic examination. LB medium is used.

[0036] Preparation method of Pediococcus pentosaceus fermentation broth: Use distilled water slant spore suspension, pick 1 ring of spores to 5mL liquid seed culture medium in a 25mL test tube, and incubate at 37℃ 180 r·min -1Culture on an air shaker for 2-3 days and measure the OD 600 The value is between 0.6-0.8, and it is used as liquid seed after microscopic examination. MRS medium is used.

[0037] In the present invention, Trichoderma reesei was purchased from China Industrial Microbiological Culture Collection Center (CICC), with strain number: CICC 13052.

[0038] In the present invention, the yeast was purchased from China Industrial Microorganism Culture Collection Center (CICC), strain number: CICC1445.

[0039] In the present invention, Lactobacillus plantarum was purchased from China Industrial Culture Collection Center (CICC), strain number: CICC 24936.

[0040] In the present invention, Bacillus licheniformis was purchased from China Industrial Microbiological Culture Collection Center (CICC), strain number: CICC 10291.

[0041] In the present invention, Bacillus velezensis was purchased from China Industrial Microbiological Culture Collection Center (CICC), with strain number: CICC 24693.

[0042] In the present invention, Pediococcus pentosaceus was purchased from China Industrial Microbiological Culture Collection Center (CICC), with strain number: CICC 22738.

[0043] In the present invention, Candida tropicalis was purchased from China Industrial Culture Collection Center (CICC), strain number: CICC 1253.

[0044] In the present invention, Lactococcus lactis was purchased from China Industrial Culture Collection Center (CICC), strain number: CICC 20408.

[0045] In the present invention, Geotrichum candidum was purchased from China Industrial Microbiological Culture Collection Center (CICC), with strain number: CICC1315.

[0046] In the present invention, the content of β-glucan is detected according to the kit. The kit is purchased from Megazyme Company, with the item number: AOAC995.16. It should be noted that the terms used herein are for describing specific embodiments only and are not intended to be limiting of exemplary embodiments according to the present invention.

[0047] Embodiment 1 The present embodiment discloses a potato residue fermented feed and a preparation method thereof. The feed comprises 180 parts of potato residue, 140 parts of auxiliary materials, 120 parts of purified water, 3 parts of Trichoderma reesei fermentation liquid, 10 parts of composite bacteria fermentation liquid, 3 parts of plant extracts, 2 parts of vitamins and 2 parts of trace element additives; wherein the auxiliary materials comprise 100 parts of bran, 50 parts of yam and 40 parts of purslane; the composite bacteria fermentation liquid comprises 4 parts of yeast, 3 parts of Lactobacillus plantarum, 3 parts of Bacillus licheniformis, 1 part of Bacillus velezensis and 2 parts of Pediococcus pentosaceus; the plant extract comprises 2 parts of curcumin and 2 parts of quercetin; the vitamins comprise 0.5 parts of vitamin A, 0.5 parts of vitamin E and 2 parts of vitamin C; the trace element additives comprise 2 parts of ammonium sulfate, 0.8 parts of dipotassium hydrogen phosphate, 1 part of sodium chloride and 1 part of magnesium sulfate.

[0048] The method for preparing potato residue fermented feed comprises: (1) preparing materials: drying auxiliary materials (yam and purslane) until they contain 70-80% moisture, and then crushing them into 0.5 cm size by machine; and compressing the potato residue until the moisture content is 70-80%.

[0049] (2): Prepare bacterial fermentation broth: Inoculate the frozen bacterial strains (Trichoderma reesei, yeast, Lactobacillus plantarum, Bacillus licheniformis, Bacillus velezensis, Pediococcus pentosaceus) into appropriate culture medium for activation and culture until OD 600 The first generation activation solution was inoculated into the liquid culture medium at a rate of 5% (v / v) and the culture was expanded under the same culture conditions until the OD 600 is 0.8, and the second generation activation liquid is obtained, which is the strain fermentation liquid.

[0050] The preparation method of the composite bacteria fermentation liquid is as follows: the second generation activation liquid is mixed according to 4 parts of yeast, 3 parts of Lactobacillus plantarum, 3 parts of Bacillus licheniformis, 1 part of Bacillus velezaceus and 2 parts of Pediococcus pentosaceus.

[0051] (3): Primary fermentation: Mix the prepared potato residue, auxiliary materials, purified water and trace elements, steam at 100°C for 30 minutes, and cool to room temperature to obtain a fermentation material; add the obtained Trichoderma reesei fermentation liquid to the fermentation material and stir evenly; ferment at 25°C and 0.2 vvm for 72 hours until the primary fermentation is completed with white mycelium; (4): Secondary fermentation: Pour the prepared composite bacteria fermentation liquid into the material that has completed the primary fermentation, mix evenly, ferment at 37℃ with ventilation of 0.2 vvm for 24 hours, and then ferment in a sealed chamber for 48 hours to complete the secondary fermentation; (5): Mixing and packaging: Mix the product from the secondary fermentation with 3 parts of plant extract and 2 parts of vitamins, dry in an oven at 60°C to a moisture content of less than 15%, crush and package.

[0052] Embodiment 2 The present embodiment discloses a potato residue fermented feed and a preparation method thereof. Different from the first embodiment, the feed comprises 200 parts of potato residue, 150 parts of auxiliary materials, 120 parts of purified water, 3 parts of Trichoderma reesei fermentation liquid, 12 parts of composite bacteria fermentation liquid, 2 parts of plant extract, 1 part of vitamins and 1 part of trace element additives; wherein the auxiliary materials comprise 100 parts of bran, 60 parts of yam and 50 parts of purslane; the composite bacteria fermentation liquid comprises 4 parts of yeast, 2 parts of Lactobacillus plantarum, 5 parts of Bacillus licheniformis, 2 parts of Bacillus velezensis and 3 parts of Pediococcus pentosaceus; the plant extract comprises 2 parts of curcumin and 2 parts of quercetin; the vitamins comprise 0.3 parts of vitamin A, 0.8 parts of vitamin E and 3 parts of vitamin C; the trace element additives comprise 2 parts of ammonium sulfate, 1 part of dipotassium hydrogen phosphate, 1 part of sodium chloride and 0.8 parts of magnesium sulfate.

[0053] Comparative Example 1 The comparative example discloses a potato residue fermented feed and a preparation method thereof. The preparation method of the potato residue fermented feed comprises: (1) preparing materials: drying auxiliary materials (yam and purslane) until they contain 70-80% moisture, and then crushing them into 0.5 cm size by machine; and compressing the potato residue to a moisture content of 70-80%.

[0054] (2): Prepare bacterial fermentation broth: Inoculate the frozen bacterial strains (yeast, Lactobacillus plantarum, Bacillus licheniformis, Bacillus velezensis, Pediococcus pentosaceus) into appropriate culture medium for activation and culture until OD 600 The first generation activation solution was inoculated into the liquid culture medium at a rate of 5% (v / v) and the culture was expanded under the same culture conditions until the OD 600 is 0.8, and the second generation activation liquid is obtained, which is the strain fermentation liquid.

[0055] The preparation method of the composite bacteria fermentation liquid is as follows: the second generation activation liquid is mixed according to 4 parts of yeast, 3 parts of Lactobacillus plantarum, 3 parts of Bacillus licheniformis, 1 part of Bacillus velezaceus and 2 parts of Pediococcus pentosaceus.

[0056] (3): Fermentation: Mix the prepared potato residue, auxiliary materials, purified water and trace elements, steam at 100°C for 30 minutes, and cool to room temperature to obtain fermented material; pour the prepared composite bacteria fermentation liquid into the material that has completed the primary fermentation, mix evenly, ferment at 37°C with ventilation of 0.2 vvm, ferment for 24 hours, and then ferment in a sealed manner for 48 hours to complete the fermentation; (4): Mixing and packaging: Mix the fermentation product with 3 parts of plant extract and 2 parts of vitamins, dry in an oven at 60°C to a moisture content of less than 15%, crush and package.

[0057] Different from Example 1, Trichoderma reesei was not used in this comparative example, and there was no primary fermentation process.

[0058] Comparative Example 2 The comparative example discloses a potato residue fermented feed and a preparation method thereof. The preparation method of the potato residue fermented feed comprises: (1) preparing materials: drying auxiliary materials (yam and purslane) until they contain 70-80% moisture, and then crushing them into 0.5 cm size by machine; and compressing the potato residue to a moisture content of 70-80%.

[0059] (2): Prepare bacterial fermentation broth: Inoculate the frozen bacterial strains (Trichoderma reesei, yeast, Lactobacillus plantarum, Bacillus licheniformis, Bacillus velezensis, Pediococcus pentosaceus) into appropriate culture medium for activation and culture until OD 600 The first generation activation solution was inoculated into the liquid culture medium at a rate of 5% (v / v) and the culture was expanded under the same culture conditions until the OD 600 is 0.8, and the second generation activation liquid is obtained, which is the strain fermentation liquid.

[0060] The preparation method of the composite bacteria fermentation liquid is as follows: the second generation activation liquid is mixed according to 4 parts of yeast, 3 parts of Lactobacillus plantarum, 3 parts of Bacillus licheniformis, 1 part of Bacillus velezaceus and 2 parts of Pediococcus pentosaceus.

[0061] (3): Fermentation: Mix the prepared potato residue, auxiliary materials, purified water and trace elements, steam at 100°C for 30 minutes, and cool to room temperature to obtain a fermentation material; pour 10 parts of the prepared composite bacteria fermentation liquid and 3 parts of the Trichoderma reesei fermentation liquid into the fermentation material, mix evenly, ferment at 37°C with ventilation of 0.2 vvm, ferment for 24 hours, and then ferment in a sealed manner for 48 hours to complete the fermentation; (4): Mixing and packaging: Mix the fermentation product with 3 parts of plant extract and 2 parts of vitamins, dry in an oven at 60°C to a moisture content of less than 15%, crush and package.

[0062] Different from Example 1, in this comparative example, Trichoderma reesei and other strains were fermented simultaneously.

[0063] Comparative Example 3 The comparative example discloses a potato residue fermented feed and a preparation method thereof. The preparation method of the potato residue fermented feed comprises: (1) preparing materials: drying auxiliary materials (yam and purslane) until they contain 70-80% moisture, and then crushing them into 0.5 cm size by machine; and compressing the potato residue to a moisture content of 70-80%.

[0064] (2): Prepare bacterial fermentation broth: Inoculate the frozen bacterial strains (Trichoderma reesei, yeast, Lactobacillus plantarum, Bacillus licheniformis, Bacillus velezensis, Pediococcus pentosaceus) into appropriate culture medium for activation and culture until OD 600 The first generation activation solution was inoculated into the liquid culture medium at a rate of 5% (v / v) and the culture was expanded under the same culture conditions until the OD 600 is 0.8, and the second generation activation liquid is obtained, which is the strain fermentation liquid.

[0065] The preparation method of the composite bacteria fermentation liquid is as follows: the second generation activation liquid is mixed according to 4 parts of yeast, 3 parts of Lactobacillus plantarum, 3 parts of Bacillus licheniformis, 1 part of Bacillus velezaceus and 2 parts of Pediococcus pentosaceus.

[0066] (3): Primary fermentation: Mix the prepared potato residue, auxiliary materials, purified water and trace elements to obtain fermentation material; add the obtained Trichoderma reesei fermentation liquid to the fermentation material and stir evenly; ferment at 25°C and ventilation of 0.2 vvm for 72 hours until the primary fermentation is completed with white mycelium; (4): Secondary fermentation: Pour the prepared composite bacteria fermentation liquid into the material that has completed the primary fermentation, mix evenly, ferment at 37℃ with ventilation of 0.2 vvm for 24 hours, and then ferment in a sealed chamber for 48 hours to complete the secondary fermentation; (5): Mixing and packaging: Mix the product from the secondary fermentation with 3 parts of plant extract and 2 parts of vitamins, dry in an oven at 60°C to a moisture content of less than 15%, crush and package.

[0067] Different from Example 1, in step (3), the potato residue and auxiliary materials are not steamed at 100° C. for 30 min.

[0068] Comparative Example 4 The difference between this comparative example and Example 1 is that the mixed bacterial strains in this example are 4 parts of Candida tropicalis, 3 parts of Geotrichum candidum, 3 parts of Lactobacillus plantarum, 1 part of Lactococcus lactis, and 3 parts of Pediococcus pentosaceus.

[0069] Comparative Example 5 The difference between this comparative example and Example 1 is that no trace elements are added in this comparative example.

[0070] Comparative Example 6 The difference between this comparative example and Example 1 is that no plant additive is added in this comparative example.

[0071] Comparative Example 7 The difference between this comparative example and Example 1 is that in this comparative example, ammonium sulfate in the trace elements is replaced by ammonium dihydrogen phosphate.

[0072] Comparative Example 8 The difference between this comparative example and Example 1 is that no vitamins are added in this comparative example.

[0073] Experimental Example 1 Nutritional changes before and after fermentation After the feed is prepared, take 2 g of feed and add it to 10 mL of sterilized distilled water for the detection of yeast and Lactobacillus plantarum, etc., and measure the pH. In addition, weigh 400 g of fermented material and put it in a refrigerator at 20°C to test its nutritional components. The crude protein content was determined in accordance with GB5009.5-2010 using the Kjeldahl method. The crude fiber content was determined in accordance with GB / T6434-94. Yeast, Bacillus licheniformis, Bacillus velezensii, Pediococcus pentosaceus and Lactobacillus plantarum were determined in accordance with GB 4789.15-2016, GB / T 34224-2017, GB / T 26428-2010, GB 4789.35-2023 and GB 4789.35-2023, respectively. The changes in crude protein and crude fiber content are shown in Table 1, and the bacterial content is shown in Table 1. Figure 1 , Figure 2 shown.

[0074] The results in Table 1 show that the potato residue in the present invention effectively reduces the cellulose content and increases the protein content after fermentation, while not adding Trichoderma reesei, using only one fermentation, not adding trace elements or replacing the types of trace elements will affect the fermentation results and are not conducive to protein accumulation and cellulose degradation.

[0075] By testing the number of live bacteria in the finished feed, it can be found that the total number of live bacteria in the potato residue feed provided by the present invention is greater than 4×10 8 CFU / g, whether Trichoderma reesei was added and the way of addition also had a great influence on the final total bacterial count ( Figure 1 ). Using Trichoderma reesei to achieve the first fermentation is more conducive to promoting the growth and reproduction of the mixed strains. Without adding Trichoderma reesei or mixing Trichoderma reesei with other strains for fermentation, the final number of bacteria decreased compared with the comparative example 1 ( Figure 2 ), the possible reason is that Trichoderma reesei produces more enzymes and small molecules after the first fermentation, which can promote the decomposition of fermentation materials to produce substances that are beneficial to the reproduction of composite strains, so the number of live bacteria is greater. Although Trichoderma reesei was used for the first fermentation in Comparative Example 4, after the composite strains were replaced, the growth promotion effect between them was weakened, especially the growth of Pediococcus pentosaceus and Geotrichum candidum was greatly affected ( Figure 2). Interestingly, in Comparative Example 7, ammonium sulfate was more conducive to the total bacterial count and the reproduction of each strain than diammonium phosphate. The reason may be that ammonium sulfate and other trace elements are more conducive to bacterial reproduction. Trace elements may also cooperate with the subsequent fermentation process to promote the release of enzymes or other small molecules in Trichoderma reesei, thereby promoting the reproduction of the composite strain ( Figure 1 , Figure 2 The pH results showed that there was no significant difference between the pH of the feed obtained in the embodiment and the comparative example, and the pH decreased by an average of 0.6 compared with the initial pH (initial pH = 6.5).

[0076] Polypeptides and polysaccharides can regulate animal immunity, improve animal immune resistance and production performance, especially oligopeptides and oligo-β-glucans are excellent in animal immunity. In order to detect the effect of the fermented potato residue feed of the present invention on the animal immune regulation performance, the polypeptides and β-glucans in the feed obtained in Example 1 were detected by HPLC, and the results are shown in Tables 2 and 3.

[0077] Table 1 Effects of different ratios and preparation steps on nutritional components

[0078] Table 2 Molecular weight distribution of peptides before and after fermentation

[0079] Table 3 β-glucan content and molecular weight distribution before and after fermentation

[0080] Note: Differential detection cannot distinguish protein peptides, dextrin and small molecular amylose in the sample. After the total protein of each component was measured by Bradford method, it can be seen that the protein peptides are mainly distributed in ≤10 4 The protein in the other molecular weight range can be ignored. Starch can generate ≤10 4 The solubility of starch in other molecular weight ranges is low and can be ignored.

[0081] Through testing, it was found that the feed obtained by the method provided in Example 1 of the present invention can effectively increase the content of polypeptides with a molecular weight <1000 and oligosaccharides, which is beneficial to promoting animal intestinal absorption, promoting animal growth and improving animal immunity.

[0082] Experimental Example 2 Potato residue is easily contaminated by Aspergillus flavus, which can produce aflatoxin. Aflatoxin can exist in the process of feed processing, transportation and storage, and its toxicity is extremely strong, posing a threat to animal life safety. The present invention verifies the inhibitory effect of the potato residue fermented feed and its preparation method provided by the present invention on Aspergillus flavus through antibacterial experiments.

[0083] Take the mixed fermentation broth obtained before drying at 60°C in the embodiment and the comparative example, 100 mL of the fermentation broth, centrifuge at 12000 r / min for 10 min, discard the precipitate, and filter with a 0.22 μm membrane to obtain a sterile supernatant; take 100 mL of the fermentation broth, centrifuge at 4°C and 12000 r / min for 10 min, add solid ammonium sulfate to the supernatant to 70% saturation, let stand at 4°C for 12 h, centrifuge at 4°C and 10000 r / min for 20 min, discard the supernatant, add 4 mL of 0.02 mol / L phosphate buffer with a pH value of 6.5 to the precipitate and dissolve it, which is the crude protein extract.

[0084] In a mixture with a suspension of Aspergillus flavus (spore concentration 1×10 6 CFU / mL) PDA medium plate was placed in an Oxford cup, and 1 mL of the above metabolites was added to the cup. The control was 1 mL of sterile water. Each treatment was repeated 3 times. The plate was incubated at 28°C until the Aspergillus flavus in the control covered the plate. The radius of the inhibition zone of each treatment was measured, and the inhibition rate was calculated. The results are shown in Table 4.

[0085] Table 4 Antibacterial effect of metabolites on Aspergillus flavus Unit: %

[0086] As shown in Table 4, the fermented liquid produced by the fermented feed preparation method provided by the present invention has a good inhibitory effect on Aspergillus flavus. The inhibition rate of the crude protein extract on Aspergillus flavus can reach more than 83%, and the inhibition rate of the supernatant can reach 45.9%, indicating that Aspergillus flavus is sensitive to active substances and plant extracts produced by mixed strain fermentation; compared with comparative example 4 (different strain types), the inhibition rates of its protein extract and supernatant on Aspergillus flavus are 46.4% and 16.5% respectively, which are weaker, indicating that the active substances produced by the first fermentation and the second fermentation of Trichoderma reesei, yeast, Lactobacillus plantarum, Bacillus licheniformis, Bacillus velez, and Pediococcus pentosaceus provided by the present invention can effectively inhibit the growth of Aspergillus flavus.

[0087] Experimental Example 3 Room temperature storage stability test The potato residue fermented feed provided in Example 1, Example 2, and Comparative Examples 1 to 7 was subjected to a room temperature storage stability test.

[0088] The test method for room temperature storage stability is: store the feed at room temperature, and then use the plate count method to determine the total number of live bacteria and calculate the live bacteria retention rate at 2, 4, 6, 8, 10 and 12 months of storage.

[0089] The results of the room temperature storage stability test are shown in Table 5.

[0090] Table 5 Results of room temperature storage test of feeds obtained by different treatment methods

[0091] As can be seen from the above table, the retention rate of live bacteria of the fermented potato residue feed provided by the present application is greater than 95% after being stored at room temperature for 6 months; and the retention rate of live bacteria is greater than 80% after being stored for 12 months. However, the retention rates of live bacteria of Comparative Example 3, in which the material is not heated, and Comparative Example 6 and Comparative Example 8, in which plant extracts and vitamins are not added, are less than 75% after being stored for 12 months, indicating that vitamins and plant additives can cooperate with the active substances produced by microbial fermentation to prolong the storage time of the feed, and heating the potato residue and auxiliary materials before fermentation can also prolong the storage time of the potato residue feed to a certain extent. This should be because the growth of some miscellaneous bacteria is inhibited during the heating process, and this operation can also partially inhibit Aspergillus flavus. Therefore, the fermented potato residue feed provided in the embodiment of the present invention can be stored for a long time.

[0092] Experimental Example 4 Animal growth effect experiment 1. Impact on the growth and development of piglets Feeding experiments were conducted on pigs using the feeds obtained in Example 1, Comparative Example 1, Comparative Examples 4 to 6, and Comparative Example 8.

[0093] A total of 180 30-day-old weaned piglets weighing about 8 kg were randomly divided into 6 groups, 30 piglets in each group (half male and half female), and placed in a pig house disinfected according to epidemic prevention requirements. During the trial, all conditions were basically the same, and according to the routine immunization program, immunization and deworming, etc., they were fed 4 times a day, free to eat and drink, and cleaned feces once a day. The pre-trial period was 7 days, and the formal trial period was 30 days. The production performance and diarrhea rate of the piglets during the formal trial period were measured, and the results are shown in Table 6.

[0094] (1) Production performance measurement method: On the morning of the first day of the experiment, the weaned piglets were weighed on an empty stomach; the daily feed intake of the weaned piglets was recorded during the experiment; feeding was stopped after the experiment, and the weight of the weaned piglets was recorded 24 hours later. The average daily weight gain, average daily feed intake and feed-to-meat ratio of the weaned piglets were calculated respectively.

[0095] (2) Method for determining diarrhea rate: During the test recording period, the number of diarrheal animals was recorded daily. Diarrhea was considered to occur if there was feces near the anus. Finally, the total number of diarrheal animals during the test period was counted.

[0096] The calculation formula of diarrhea rate: Diarrhea rate = total number of diarrhea pigs / (total number of weaned piglets × number of test days) × 100% Table 6 Results of production performance and diarrhea rate of piglets

[0097] As can be seen from the above table, the average daily feed intake and average daily weight gain of the piglets fed with the feed in comparative example 1 are significantly higher than those of the other control groups. Therefore, it is shown that when weaned piglets eat the fermented potato residue feed provided by the present invention, the appetite of weaned piglets can be significantly improved, and more of the pig feed fed can be converted into pork.

[0098] According to the test results of diarrhea rate, the diarrhea rate of piglets fed with the feed in Example 1 is 4%, which is significantly lower than the diarrhea rate of piglets fed with the feed in Comparative Examples 1, 4, 5, 6, 7, and 8. This indicates that the fermented potato residue feed provided by the present invention can significantly reduce the diarrhea rate of weaned piglets; in particular, the diarrhea rate can be effectively reduced after adding plant additives and vitamins, and the active substances produced by the synergistic microbial fermentation of the two can effectively regulate animal immunity, improve animal immunity and resistance, and reduce the incidence of diarrhea.

[0099] 2. Experiment on the effect on duck egg quality A feeding test was conducted at a duck farm of a farmer in Jining, Shandong. The experimental animals were 4,000 20-week-old Jinding ducks, which were randomly divided into 4 groups and fed with the feed obtained in Example 1, Comparative Example 1, Comparative Example 4 and Comparative Example 6, respectively. Each group had 4 replicates, and each replicate had 250 ducks. The average daily feed intake of each duck was controlled at 225-230 grams. The feeding period was 20 weeks to 32 weeks of age. The method was based on GB 2749-2015, and the test results are shown in Table 7.

[0100] Table 7 Effects of different feeds on egg quality

[0101] It can be seen from the statistical results in the above table that the egg-laying quality and egg-laying performance of Jinding ducks can be effectively improved after feeding the feed provided in Example 1 of the present invention.

[0102] Although the above describes the specific implementation mode of the present invention in conjunction with the accompanying drawings, it is not intended to limit the scope of protection of the present invention. Those skilled in the art should understand that various modifications or variations that can be made by those skilled in the art on the basis of the technical solution of the present invention without creative work are still within the scope of protection of the present invention.

Claims

1. A potato residue fermented feed, characterized in that: The invention comprises 150-200 parts of potato residue, 120-150 parts of auxiliary materials, 100-120 parts of purified water, 2-4 parts of Trichoderma reesei fermentation liquid, 8-15 parts of compound bacteria fermentation liquid, 2-4 parts of plant extract, 1-3 parts of vitamins and 1-3 parts of trace element additives; The auxiliary materials include 90-120 parts of bran, 40-60 parts of yam, and 30-50 parts of purslane; The composite bacteria fermentation liquid comprises 3-5 parts of yeast, 2-5 parts of Lactobacillus plantarum, 3-5 parts of Bacillus licheniformis, 1-2 parts of Bacillus velezaceus, and 1-3 parts of Pediococcus pentosaceus; The plant extract comprises 1-3 parts of curcumin and 1-3 parts of quercetin.

2. A potato residue fermented feed as claimed in claim 1, characterized in that: The vitamins include 0.3-0.8 parts of vitamin A, 0.3-0.8 parts of vitamin E, and 1.5-3 parts of vitamin C.

3. The potato residue fermented feed according to claim 1, characterized in that: The trace element additive includes 1-2 parts of ammonium sulfate, 0.5-1 parts of dipotassium hydrogen phosphate, 0.5-1 parts of sodium chloride, and 0.5-1 parts of magnesium sulfate.

4. The potato residue fermented feed according to claim 1, characterized in that: The invention comprises 180-200 parts of potato residue, 140-150 parts of auxiliary materials, 100-120 parts of purified water, 2.5-3.5 parts of Trichoderma reesei fermentation liquid, 8-13 parts of compound bacteria fermentation liquid, 2.5-3.5 parts of plant extract, 1.5-2 parts of vitamins and 1.5-2.5 parts of trace element additives; The auxiliary materials include 100-120 parts of bran, 45-60 parts of yam, and 40-50 parts of purslane; The composite bacteria fermentation liquid comprises 3-4 parts of yeast, 2-4 parts of Lactobacillus plantarum, 2-4 parts of Bacillus licheniformis, 1-2 parts of Bacillus velezaceus, and 1-3 parts of Pediococcus pentosaceus; The plant extract comprises 2-3 parts of curcumin and 1.5-2 parts of quercetin.

5. The potato residue fermented feed according to claim 4, characterized in that: The invention comprises 180 parts of potato residue, 140 parts of auxiliary materials, 120 parts of purified water, 3 parts of Trichoderma reesei fermentation liquid, 10 parts of compound bacteria fermentation liquid, 3 parts of plant extract, 2 parts of vitamins and 2 parts of trace element additives; The auxiliary materials include 100 parts of bran, 50 parts of yam and 40 parts of purslane; The composite bacteria fermentation liquid comprises 4 parts of yeast, 3 parts of Lactobacillus plantarum, 3 parts of Bacillus licheniformis, 1 part of Bacillus velezaceus, and 2 parts of Pediococcus pentosaceus; The plant extracts are 2 parts of curcumin and 2 parts of quercetin.

6. The method for preparing potato residue fermented feed according to claim 1, characterized in that: The following steps are involved: 1) Add 2-4 parts of Trichoderma reesei fermentation liquid to 150-200 parts of potato residue, 120-150 parts of auxiliary materials, 100-120 parts of purified water, and 1-3 parts of trace element additives, and ferment at 24-28℃ with ventilation for 48-72 hours until the primary fermentation is completed with white mycelium; 2) Stir 8-15 portions of the compound bacteria fermentation liquid into the product of the primary fermentation, ferment at 30-37°C with ventilation for 24-48 hours, and then ferment in a sealed environment for 24-72 hours to complete the secondary fermentation; 3) Mix the secondary fermentation product with 2-4 parts of plant extract and 1-3 parts of vitamins, dry until the water content is below 15%, crush and package.

7. The method for preparing potato residue fermented feed according to claim 6, characterized in that: The auxiliary materials are crushed to 0.5-1 cm.

8. The method for preparing potato residue fermented feed according to claim 6, characterized in that: In the step 1), the potato residue and auxiliary materials are first steamed at 100° C. for 20-30 minutes, and then cooled to room temperature for later use.

9. The method for preparing potato residue fermented feed according to claim 6, characterized in that: In step 1) and step 2), the ventilation volume is 0.1-0.3 vvm.

10. The method for preparing potato residue fermented feed according to claim 6, characterized in that: In the step 3), the drying temperature is 40-60°C.

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