Functional pet fresh food and preparation method thereof

Abstract

The application discloses functional pet fresh food and a preparation method thereof, and belongs to the technical field of pet food. The functional pet fresh food is prepared from the following components in parts by mass: black soldier fly larva defatted powder 50-60 parts, medicinal fungus fermentation product 20-30 parts, Antarctic krill powder 8-10 parts, compound fruit and vegetable fermentation slurry 3.5-5 parts, prebiotic 3-4 parts, cold-pressed flaxseed oil 3-4 parts, ganoderma lucidum mycelium polysaccharide extract 2-3 parts and microencapsulated compound probiotic 0.5-1 part. The pet fresh food provided by the application can overcome the problems of high protein allergenicity, insufficient active ingredient activity retention and short shelf life of traditional pet fresh food in the prior art, has the functions of intestinal regulation and immune enhancement, and is suitable for industrialized production.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pet food, and particularly relates to a functional pet fresh food and a preparation method thereof. BACKGROUND

[0002] With the popularity of pet feeding, pet owners pay more and more attention to the health of pets. In recent years, pet fresh food has gradually been favored by the market, and its fresh ingredients and rich nutritional components are more in line with the natural dietary needs of pets. However, the current pet fresh food on the market has the following problems: 1. Single protein source: mainly relying on traditional animal proteins such as chicken and beef, which is easy to cause allergies and has a large environmental burden; 2. Serious nutritional loss: high-temperature sterilization (such as above 121℃) leads to the inactivation of heat-sensitive functional components (such as probiotics and active polysaccharides); 3. Dependence on synthetic additives: artificial feeding agents or preservatives need to be added, which affects the long-term health of pets; 4. Conflict between shelf life and nutrient retention: the shelf life of conventional fresh food is short (usually ≤14 days), and extending the shelf life requires sacrificing nutritional components. Although the existing technology attempts to use insect protein, it does not solve the problems of low protein bioavailability and functional component synergy; some technology discloses the application of fungal polysaccharides, but does not combine with low-temperature sterilization process. Therefore, there is an urgent need to develop a new type of pet fresh food with high digestibility, long shelf life and multiple health benefits. SUMMARY

[0003] In order to overcome the problems of high protein allergenicity, insufficient activity retention of functional components and short shelf life of traditional pet fresh food in the prior art, the present application provides a functional pet fresh food and a preparation method thereof.

[0004] In order to achieve the above-mentioned purposes, the present application provides the following technical solutions:

[0005] In one aspect, the present application provides a functional pet fresh food, which is prepared from the following components by mass fraction: black soldier fly larvae defatted powder 50-60 parts, medicinal fungus fermentation product 20-30 parts, Antarctic krill powder 8-10 parts, compound fruit and vegetable fermentation broth 3.5-5 parts, prebiotic 3-4 parts, cold-pressed flaxseed oil 3-4 parts, ganoderma lucidum mycelium polysaccharide extract 2-3 parts, and microencapsulated compound probiotics 0.5-1 part.

[0006] The medicinal fungus fermentation product is a co-fermentation product of Phellinus baumii and Hericium erinaceus;

[0007] The compound fruit and vegetable fermentation broth is prepared from fruits, vegetables, fermentation substrate and fermentation strains;

[0008] The fruits include at least one of apple residue, blueberry and pineapple peel;

[0009] The vegetables include at least one of carrot, purple cabbage and pumpkin;

[0010] the fermentation substrate comprises at least one of fructooligosaccharide, yeast extract;

[0011] the fermentation strain comprises at least one of lactobacillus plantarum, saccharomyces boulardii;

[0012] the prebiotic is a mixture of xylooligosaccharide and inulin;

[0013] the content of β-glucan in the ganoderma lucidum mycelium polysaccharide extract is ≥ 30wt%, and the content of polysaccharide is ≥ 80wt%;

[0014] the microencapsulated complex probiotic is prepared from complex probiotic and embedding material;

[0015] the complex probiotic comprises lactobacillus plantarum and bacillus subtilis;

[0016] the embedding material comprises at least one of sodium alginate and chitosan.

[0017] Optionally, the functional pet fresh food contains the following components by mass fraction: black soldier fly larva defatted powder 55 parts, medicinal fungus fermentation product 25 parts, antarctic krill powder 9 parts, complex fruit and vegetable fermentation broth 4 parts, prebiotic 3.5 parts, cold-pressed flaxseed oil 3.5 parts, ganoderma lucidum mycelium polysaccharide extract 2.5 parts, and microencapsulated complex probiotic 0.8 parts.

[0018] Optionally, the fat content of the black soldier fly larva defatted powder is ≤ 5wt%.

[0019] The particle size of the black soldier fly larva defatted powder is 150-180μm.

[0020] Optionally, the particle size of the black soldier fly larva defatted powder is independently selected from any value or range value between any two of 150μm, 160μm, 170μm and 180μm.

[0021] Optionally, the content of β-glucan in the phellinus-hericium co-fermentation product is ≥ 8wt%.

[0022] Optionally, the particle size of the antarctic krill powder is ≤ 50μm.

[0023] Optionally, in the complex fruit and vegetable fermentation broth, the mass ratio of fruit, vegetable and fermentation substrate is 2-3:1-2:1.

[0024] The inoculation amount of the fermentation strain is 3-5% of the total mass of fruit, vegetable and fermentation substrate.

[0025] Optionally, in the complex fruit and vegetable fermentation broth, the mass ratio of fruit, vegetable and fermentation substrate is 2.5:1.5:1.

[0026] Optionally, the inoculation amount of the fermentation strain is independently selected from any value or a range value between any two values of 3%, 4%, 5% of the total mass of the fruit, vegetable, fermentation substrate.

[0027] Optionally, the mixed mass ratio of xylo-oligosaccharide and inulin is 1:1-2.

[0028] Optionally, the mixed mass ratio of xylo-oligosaccharide and inulin is 1:1.

[0029] In a second aspect, the application provides a preparation method of the functional pet fresh food, comprising the following steps:

[0030] (1) mixing the black soldier fly larva defatted powder, medicinal fungus fermentation product and Antarctic krill powder, and performing enzymolysis by using exogenous enzymes to obtain a compound protein substrate;

[0031] (2) mixing the compound protein substrate obtained in step (1) with a compound fruit and vegetable fermentation slurry, prebiotics, cold-pressed flaxseed oil, ganoderma lucidum mycelium polysaccharide extract and microencapsulated compound probiotics, and sequentially performing homogenization, molding and sterilization to obtain the functional pet fresh food;

[0032] In step (1), the exogenous enzymes include alkaline protease and flavor protease.

[0033] Optionally, in step (1), the preparation step of the black soldier fly larva defatted powder comprises: performing ultrasonic-assisted enzymolysis on fresh black soldier fly larvae, centrifuging to obtain a precipitate with a fat content ≤5wt%, and drying and sieving the precipitate, wherein the undersize is the black soldier fly larva defatted powder;

[0034] The fresh black soldier fly larvae have a fat content ≤20wt%;

[0035] The ultrasonic-assisted enzymolysis has an ultrasonic frequency of 30-50 kHz;

[0036] The enzyme used in the ultrasonic-assisted enzymolysis is lipase;

[0037] The lipase has an enzyme activity ≥20000U / g;

[0038] The addition amount of the lipase is 0.4-0.6% of the mass of the fresh black soldier fly larvae;

[0039] The ultrasonic-assisted enzymolysis has a temperature of 40-50°C;

[0040] The ultrasonic-assisted enzymolysis has a time of 1-2h;

[0041] The ultrasonic-assisted enzymolysis has a pH of 6.7-7.2;

[0042] The centrifugal separation speed is 2500-3500 rpm.

[0043] Optionally, in step (1), the ultrasonic frequency of the ultrasonic-assisted enzymatic hydrolysis is independently selected from any value or a range between any two of 30 kHz, 35 kHz, 40 kHz, 45 kHz, 50 kHz.

[0044] Optionally, in step (1), the amount of the lipase added is independently selected from any value or a range between any two of 0.4%, 0.5%, 0.6% of the mass of the fresh black soldier fly larvae.

[0045] Optionally, in step (1), the temperature of the ultrasonic-assisted enzymatic hydrolysis is independently selected from any value or a range between any two of 40°C, 42°C, 45°C, 46°C, 48°C, 50°C.

[0046] Optionally, in step (1), the time of the ultrasonic-assisted enzymatic hydrolysis is independently selected from any value or a range between any two of 1 h, 1.2 h, 1.5 h, 1.8 h, 2 h.

[0047] Optionally, in step (1), the pH of the ultrasonic-assisted enzymatic hydrolysis is independently selected from any value or a range between any two of 6.7, 7, 7.2.

[0048] Optionally, in step (1), the centrifugal separation speed is independently selected from any value or a range between any two of 2500 rpm, 2800 rpm, 3000 rpm, 3200 rpm, 3500 rpm.

[0049] Optionally, in step (1), the preparation of the medicinal fungus fermentation product comprises: taking bran, soybean meal, and tea residue as the substrate, adjusting the water content to 53-57%, inoculating with a Phellinus baumii spore suspension and a Hericium erinaceus mycelium suspension, co-fermenting, and drying to obtain a Phellinus baumii-Hericium erinaceus co-fermentation product, i.e., the medicinal fungus fermentation product.

[0050] The mass ratio of the bran, soybean meal, and tea residue is 5-7:2-4:1.

[0051] The tea residue is green tea residue.

[0052] The number of spores in the Phellinus baumii spore suspension is ≥1×10 6 spores / mL.

[0053] The content of the Hericium erinaceus mycelium in the mycelium suspension is ≥1×10 6 CFU / mL.

[0054] The volume ratio of the Phellinus baumii spore suspension to the Hericium erinaceus mycelium suspension is 2-4:1.

[0055] the total inoculation amount of the Phellinus baumii spore suspension and the Hericium erinaceus mycelium suspension is 5-10%;

[0056] the temperature of the co-fermentation is 25-28℃;

[0057] the content of β-glucan in the fermentation product at the end of the co-fermentation is ≥8wt%;

[0058] the temperature of the drying is 35-45℃;

[0059] the time of the drying is 5-7h.

[0060] Optionally, the mass ratio of the wheat bran, soybean meal, and tea dregs is 6:3:1.

[0061] Optionally, the volume ratio of the Phellinus baumii spore suspension and the Hericium erinaceus mycelium suspension is 3:1.

[0062] Optionally, the total inoculation amount of the Phellinus baumii spore suspension and the Hericium erinaceus mycelium suspension is independently selected from any value in 5%, 6%, 7%, 8%, 9%, 10% or a range value between any two thereof.

[0063] Optionally, the temperature of the co-fermentation is independently selected from any value in 25℃, 26℃, 27℃, 28℃ or a range value between any two thereof.

[0064] Optionally, the temperature of the drying is independently selected from any value in 35℃, 38℃, 40℃, 43℃, 45℃ or a range value between any two thereof.

[0065] Optionally, the time of the drying is independently selected from any value in 5h, 6h, 7h or a range value between any two thereof.

[0066] Optionally, in step (1), the mass ratio of the alkaline protease and the flavor protease is 1-2:1;

[0067] the enzyme activity of the alkaline protease is ≥200000U / g;

[0068] the enzyme activity of the flavor protease is ≥30000U / g;

[0069] the total addition amount of the alkaline protease and the flavor protease is 4500-5500U per gram of protein;

[0070] the temperature of the enzymolysis is 45-50℃;

[0071] the time of the enzymolysis is 100-150min;

[0072] the pH during the enzymolysis is 6.8-7.2;

[0073] intermittent ultrasound is applied during the enzymatic hydrolysis process;

[0074] The frequency of the ultrasound is 30-50 kHz, and the working time of the ultrasound is 4-6 min every 30 min.

[0075] Optionally, the mass ratio of the alkaline protease and the flavor protease is 1:1.

[0076] Optionally, the total amount of the alkaline protease and the flavor protease added is independently selected from any value or a range value between any two of 4500 U, 4800 U, 5000 U, 5200 U, 5500 U per gram of protein.

[0077] Optionally, the temperature of the enzymatic hydrolysis is independently selected from any value or a range value between any two of 45℃, 46℃, 47℃, 48℃, 49℃, 50℃.

[0078] Optionally, the time of the enzymatic hydrolysis is independently selected from any value or a range value between any two of 100 min, 110 min, 120 min, 130 min, 140 min, 150 min.

[0079] Optionally, the pH during the enzymatic hydrolysis is independently selected from any value or a range value between any two of 6.8, 7, 7.2.

[0080] Optionally, the frequency of the ultrasound is independently selected from any value or a range value between any two of 30 kHz, 35 kHz, 40 kHz, 45 kHz, 50 kHz.

[0081] Optionally, the working time of the ultrasound is 5 min every 30 min.

[0082] Optionally, in step (2), the preparation step of the compound fruit and vegetable fermentation slurry comprises: mixing the fruits, vegetables, and fermentation substrate, inoculating fermentation bacteria, carrying out fermentation, and carrying out enzymatic hydrolysis on the fermentation product after the fermentation is completed to obtain the compound fruit and vegetable fermentation slurry.

[0083] The temperature of the fermentation is 30-37℃;

[0084] The time of the fermentation is 12-72 h;

[0085] The enzymatic hydrolysis is carried out using pectinase;

[0086] The enzyme activity of the pectinase is ≥100000 U / g;

[0087] The addition amount of the pectinase is 45-55 U per gram of fermentation product;

[0088] The temperature of the enzymatic hydrolysis is 35-45℃;

[0089] The enzymatic hydrolysis time is 1 to 2 hours.

[0090] Optionally, the fermentation temperature is independently selected from any value of 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, or a range between any two.

[0091] Optionally, the fermentation time is independently selected from any value of 12h, 24h, 36h, 48h, 60h, 72h or a range between any two.

[0092] Optionally, the amount of pectinase added is independently selected from any value among 45U, 48U, 50U, 52U, and 55U per gram of fermentation product, or a range between any two.

[0093] Optionally, the temperature of the enzymatic hydrolysis is independently selected from any value of 35°C, 38°C, 40°C, 42°C, 45°C, or a range between any two.

[0094] Optionally, the enzymatic hydrolysis time is independently selected from any value of 1h, 1.2h, 1.5h, 1.6h, 1.8h, 2h or any range between two of them.

[0095] Optionally, in step (2), the preparation step of the microencapsulated compound probiotics includes: encapsulating the compound probiotics with an encapsulation material to obtain microencapsulated compound probiotics;

[0096] The compound probiotics include Lactobacillus plantarum and Bacillus subtilis;

[0097] The embedding material includes at least one of sodium alginate and chitosan.

[0098] Optionally, the compound probiotic is a mixture of Lactobacillus plantarum suspension and Bacillus subtilis suspension;

[0099] The bacterial content of the *Lactobacillus plantarum* suspension is 1×10⁻⁶. 6 ~1×10 9 CFU / mL;

[0100] The bacterial content of the Bacillus subtilis suspension is ≥1×10⁻⁶. 9 CFU / mL;

[0101] The volume ratio of the *Lactobacillus plantarum* suspension to the *Bacillus subtilis* suspension is 1–2:1.

[0102] The embedding material is sodium alginate and chitosan;

[0103] The mass ratio of sodium alginate to chitosan is 3-5:1;

[0104] The embedding is mixing the composite probiotics with an aqueous solution of sodium alginate, electrostatically dropping into a CaCl2 solution to form a gel bead, and then coating the gel bead with an acetic acid solution containing chitosan.

[0105] Optionally, in step (2), the homogenization pressure is 45-55 MPa.

[0106] The homogenization rotation speed is 3000-4000 rpm.

[0107] The homogenization time is 25-35 min.

[0108] The sterilization uses high-voltage electrostatic field coupling ultraviolet pulse sterilization.

[0109] Optionally, in step (2), the homogenization pressure is independently selected from any value in 45 MPa, 48 MPa, 50 MPa, 52 MPa, 55 MPa or a range value between any two thereof.

[0110] Optionally, in step (2), the homogenization rotation speed is independently selected from any value in 3000 rpm, 3200 rpm, 3500 rpm, 3700 rpm, 4000 rpm or a range value between any two thereof.

[0111] Optionally, in step (2), the homogenization time is independently selected from any value in 25 min, 28 min, 30 min, 32 min, 35 min or a range value between any two thereof.

[0112] In a third aspect, the application provides the use of the above functional pet fresh food or the functional pet fresh food prepared by the above preparation method in the preparation of a product for improving the intestinal flora balance of pets and / or enhancing the immune function of pets.

[0113] The product includes health products and / or pharmaceutical products.

[0114] Compared with the prior art, the application has the following beneficial effects:

[0115] (1) The functional pet fresh food provided by the present application combines black soldier fly larva protein with medicinal fungus fermentation and Antarctic krill powder to form a complex protein system rich in full-spectrum amino acids, flavonoids (such as hispidin), and nerve growth factor (NGF) precursors. This not only solves the problems of traditional animal protein sources (chicken / beef) being easily allergenic and having high carbon emissions, but also achieves the synergistic effects of pet cognitive function and immune enhancement. The complex fruit and vegetable fermentation slurry added in the fresh food formula is rich in effective ingredients such as pectin, anthocyanins, bromelain, beta-carotene, and thioglucosides. This not only solves the problems of pet fresh food relying on synthetic additives and having a single source of fiber, but also enhances the functional value through biological transformation to achieve the purposes of prebiotic-probiotic synergistic regulation of intestinal health, enhancement of antioxidant and anti-inflammatory effects, and the like. The lactic acid bacteria fermentation also produces ethyl acetate (fruity aroma) and 2,3-butanedione (buttery aroma), and the yeast metabolism can reduce the bitterness of vegetables to improve the palatability of the pet fresh food. The microencapsulated complex probiotics added in the present application solve the problems of stability and bioavailability of functional components through microencapsulation, and break through the application bottleneck of probiotics in pet fresh food by using a sodium alginate-chitosan double-layer embedding method, which has the effects of targeting the intestinal tract, regulating intestinal flora, and enhancing immune function. In addition, the present application also adds components such as prebiotics, cold-pressed flaxseed oil, and ganoderma lucidum polysaccharide extract to further endow the pet fresh food with rich nutrition, which synergistically enhances the efficacy of the fresh food with the above-mentioned complex protein system, complex fruit and vegetable fermentation slurry, and microencapsulated complex probiotics.

[0116] (2) The preparation method of the functional pet fresh food provided by the present application performs mixed enzymolysis on the complex protein system to cut the peptide bonds (around hydrophobic amino acid residues) within the protein to generate polypeptide fragments, and further hydrolyze the peptide chain ends to release free amino acids, thereby improving the bioavailability of the protein. At the same time, the enzymolysis can destroy the allergenic epitopes of black soldier fly protein (such as the IgE binding region of Tropomyosin) and promote the generation of antibacterial peptides from black soldier fly protein. In addition, the present application uses low-temperature enzymolysis (up to 50°C) to protect heat-sensitive ingredients (such as ganoderma polysaccharides), and ultrasonic intermittent assistance can promote enzyme-substrate contact. The combination of low-temperature biological processing and physical field sterilization technology developed by the present application can ensure functional activity while enabling large-scale production using conventional food processing equipment, which is suitable for pet fresh food manufacturing.

[0117] (3) Experiments have proved that the protein digestion rate of the functional pet fresh food provided by the present application can be increased to 92.3%, the serum IgG and IgA levels are significantly improved, the shelf life is ≥12 months (4°C refrigeration) and no preservatives are needed. It can be seen that the pet fresh food provided by the present application has a broad application prospect in preparing products for improving the intestinal flora balance and / or enhancing the immune function of pets, and can effectively prolong the shelf life of fresh food, especially for companion animals such as dogs and cats. DETAILED DESCRIPTION

[0118] The present application is further described in conjunction with the following examples. The following examples are merely illustrative of the present application and are not intended to limit the present application in any way. Although the present application has been described in considerable detail with reference to certain preferred embodiments thereof, other versions are possible that are not expressly mentioned or shown. Therefore, certain changes and modifications are contemplated and, therefore, are intended to fall within the scope of the present application. It is understood that the examples and embodiments described herein are presented by way of example only and are not intended to limit the scope of the application.

[0119] Unless otherwise specified, the raw materials in the examples of the present application are purchased through commercial channels and are directly used without any special treatment.

[0120] Unless otherwise specified, the analytical methods in the examples all adopt the conventional settings of instruments or equipment and conventional analytical methods.

[0121] The sources of the materials in the following examples and comparative examples are shown in Tables 1 to 4.

[0122] Table 1 Raw material components

[0123]

[0124] Table 2 Strains and leavening agents

[0125]

[0126]

[0127] Table 3 Enzyme preparations

[0128]

[0129] Table 4 Functional additives

[0130]

[0131]

[0132] Example 1

[0133] A method for preparing a functional pet fresh food is as follows:

[0134] (1) Fresh black soldier fly larvae (fat content ≤20%) are subjected to ultrasonic-assisted lipase hydrolysis (30 kHz, 40°C, pH 6.7, 1 h, lipase enzyme activity 20,000 U / g, and lipase addition amount 0.4% of the mass of fresh black soldier fly larvae), centrifugal separation (2,500 rpm) to a fat content ≤5 wt% in the precipitate, and then hot air drying (50°C, 2 h) of the precipitate, sieving, and collection of undersize material, i.e., black soldier fly larvae defatted powder (particle size 150 μm);

[0135] (2) Using bran, soybean meal, and green tea residue as the substrate (bran: soybean meal: green tea residue = 5:2:1, mass ratio), the water content is adjusted to 53%, and a Phellinus baumii spore suspension and a Hericium erinaceus mycelium suspension are inoculated (wherein the Phellinus baumii spore suspension is scraped with 0.05% Tween 80 sterile water, filtered (200-mesh screen) to adjust to 1×10 6 spores / mL, and the Hericium erinaceus mycelium suspension is broken under sterile conditions (ultrasonic 3 min, 40 kHz) to adjust to a bacterial content of 1×10 6 CFU / mL, the volume ratio of the Phellinus baumii spore suspension to the Hericium erinaceus mycelium suspension is 2:1, and the total inoculation amount of the Phellinus baumii spore suspension and the Hericium erinaceus mycelium suspension is 5%), co-fermentation is carried out at 25°C, 70% humidity, and in the dark for 7 days, the fermentation end standard is a mycelium β-glucan content of 8.12 wt% (determined by the phenol-sulfuric acid method), and then the fermentation product is low-temperature dried at 35°C for 5 h to obtain a Phellinus baumii-Hericium erinaceus co-fermentation product, i.e., a medicinal fungus fermentation product;

[0136] (3) Lactobacillus plantarum and Bacillus subtilis suspensions (both adjusted to a bacterial concentration of 1×10 9 CFU / mL) are mixed in a volume ratio of 1:1 to obtain a mixed bacterial suspension; sodium alginate (2%, w / v) is dissolved in deionized water at 55°C and sterilized at 121°C for 15 min to obtain a sodium alginate solution; chitosan (degree of deacetylation ≥90%) is dissolved in 0.5% acetic acid solution (pH 5.0) to obtain a chitosan solution with a concentration of 0.5% (w / v); the mixed bacterial suspension and the sodium alginate solution are mixed in a volume ratio of 1:1, and electrostatically dropped into a CaCl2 solution (0.1M in itself, containing 0.05% Tween 80), with parameters set as voltage 8 kV, needle diameter 0.4 mm, needle dripping speed 15 drops / min, magnetic stirring (200 rpm), and solidification for 20 min to form gel beads with a diameter of 1.2-1.5 mm, then the gel beads are immersed in the chitosan solution, slowly oscillated (50 rpm) at 25°C for 15 min, washed with PBS buffer solution (pH 7.0) for 3 times after being taken out, and quickly frozen at -40°C for 2 h to obtain microencapsulated compound probiotics;

[0137] (4) mixing fruits (apple residue: blueberry: pineapple skin = 1:1:1, mass ratio), vegetables (carrot: purple cabbage: pumpkin = 1:1:1, mass ratio), fermentation substrate (fructooligosaccharide: yeast extract = 1:1) in a mass ratio of 2:1:1, inoculating Lactobacillus plantarum (without activation, direct-injection freeze-dried bacteria powder, viable bacterial count 1x10 10 CFU / g bacteria powder, 5% (w / w) inoculum) at 37°C for 24h, then inoculating Saccharomyces boulardii (inoculating in the form of bacterial suspension, bacterial content 1x10 10 CFU / mL suspension, 3% (v / w) inoculum) at 30°C for 48h, and then enzymatically hydrolyzing the fermentation product after fermentation ends, adding pectinase (45U per gram of fermentation product) at 35°C for 1h, sterilizing, and obtaining a compound fruit and vegetable fermentation slurry;

[0138] (5) weighing the following mass of raw materials: black soldier fly larva defatted powder 50 parts, medicinal fungus fermentation product 20 parts, Antarctic krill powder 8 parts, compound fruit and vegetable fermentation slurry 3.5 parts, prebiotic (xylo-oligosaccharide: inulin = 1:1, mass ratio) 3 parts, cold-pressed flaxseed oil 3 parts, ganoderma lucidum mycelium polysaccharide extract 2 parts, and microencapsulated compound probiotic 0.5 parts;

[0139] (6) mixing the black soldier fly larva defatted powder, medicinal fungus fermentation product, and Antarctic krill powder, and enzymatically hydrolyzing the mixture using alkaline protease and flavor protease (1:1, mass ratio) at 45°C and pH 6.8 for 100min, wherein the enzyme activity of the alkaline protease is 200000U / g, the enzyme activity of the flavor protease is 30000U / g, the total amount of enzyme added is 4500U per gram of protein, and intermittent ultrasonic waves (30kHz, acting for 4min every 30min) are applied during the enzymatic hydrolysis process, to obtain a compound protein substrate;

[0140] (7) mixing the compound protein substrate, microencapsulated compound probiotic, compound fruit and vegetable fermentation slurry, prebiotic, cold-pressed flaxseed oil, and ganoderma lucidum mycelium polysaccharide extract, homogenizing at 45MPa and 3000rpm for 25min, 3D printing extrusion molding (equipment parameters: nozzle diameter 1.2mm, extrusion pressure 0.3MPa, printing temperature 35°C (constant temperature nozzle), molding speed 20mm / s), and then high-voltage electrostatic field coupling ultraviolet pulse sterilization (high-voltage electrostatic field treatment: equipment is parallel plate electrodes (spacing 5cm), parameters are 15kV / cm, 10min, and ambient humidity 50%; ultraviolet pulse assistance: wavelength 254nm+185nm (ozone generation), intensity 50mJ / cm 2 , pulse frequency 5Hz), to obtain the functional pet fresh food.

[0141] Example 2

[0142] A method for preparing a functional pet fresh food, comprising the following steps:

[0143] (1) Fresh black soldier fly larvae (fat content ≤20%) were subjected to ultrasonic-assisted lipase hydrolysis (40 kHz, 45℃, pH 7, 1.5 h, lipase enzyme activity 20000 U / g, lipase addition amount 0.5% of the mass of fresh black soldier fly larvae), centrifugal separation (3000 rpm) to a fat content ≤5wt% in the precipitate, and then hot air drying (50℃, 2h) of the precipitate, sieving, and collection of undersize material, i.e. black soldier fly larvae defatted powder (particle size 160μm);

[0144] (2) Using bran, soybean meal, and green tea residue as the substrate (bran: soybean meal: green tea residue = 6:3:1, mass ratio), the moisture content was adjusted to 55%, and a Phellinus igniarius spore suspension and a Hericium erinaceus mycelium suspension were inoculated (wherein the Phellinus igniarius spore suspension: spores were scraped with 0.05% Tween 80 sterile water, filtered (200-mesh screen) to adjust to 1×10 6 spores / mL, the Hericium erinaceus mycelium suspension: the mycelium ball was broken under sterile conditions (ultrasonic 3 min, 40 kHz), and adjusted to a bacterial content of 1×10 6 CFU / mL, the volume ratio of the Phellinus igniarius spore suspension to the Hericium erinaceus mycelium suspension was 3:1, and the total inoculum of the Phellinus igniarius spore suspension and the Hericium erinaceus mycelium suspension was 8%), and co-fermentation was carried out at 27℃, 70% humidity, and in the dark for 7 days, with the mycelium β-glucan content of 8.97wt% (determined by the phenol-sulfuric acid method) as the fermentation end criterion, and then the fermentation product was dried at low temperature at 40℃ for 6h to obtain a Phellinus igniarius-Hericium erinaceus co-fermentation product, i.e. a medicinal fungal fermentation product;

[0145] (3) Lactobacillus plantarum and Bacillus subtilis suspensions (both adjusted to a bacterial concentration of 1×10 9 CFU / mL) were mixed in a volume ratio of 1:1 to obtain a mixed bacterial suspension; sodium alginate (2%, w / v) was dissolved in 55℃ deionized water, sterilized at 121℃ for 15 min to obtain a sodium alginate solution; chitosan (degree of deacetylation ≥90%) was dissolved in 0.5% acetic acid solution (pH 5.0) to obtain a chitosan solution with a concentration of 0.5% (w / v); the mixed bacterial suspension and the sodium alginate solution were mixed in a volume ratio of 1:1, and electrostatically dropped into a CaCl2 solution (0.1M in itself, containing 0.05% Tween 80), with parameters set as voltage 8kV, needle diameter 0.4mm, needle dropping speed 15 drops / min, magnetic stirring (200rpm), and solidification for 20 min to form gel beads with a diameter of 1.2-1.5mm, then the gel beads were immersed in the chitosan solution, slowly oscillated (50rpm) at 25℃ for 15 min, washed with PBS buffer solution (pH 7.0) for 3 times after taking out, and quickly frozen at -40℃ for 2h to obtain microencapsulated compound probiotics;

[0146] (4) Mix the fruit (apple residue: blueberry: pineapple skin = 1:1:1, mass ratio), the vegetable (carrot: purple cabbage: pumpkin = 1:1:1, mass ratio), the fermentation substrate (fructooligosaccharide: yeast extract = 1:1) according to the mass ratio of 2.5:1.5:1, inoculate Lactobacillus plantarum (without activation, direct-injection freeze-dried bacteria powder, viable bacterial count is 1x10 10 CFU / g bacteria powder, 5% (w / w) inoculum) at 37℃ for 24h, then inoculate Saccharomyces boulardii (inoculate in the form of bacteria suspension, bacteria content is 1x10 10 CFU / mL suspension, 3% (v / w) inoculum) at 30℃ for 48h, after fermentation, the fermentation product is subjected to enzymolysis, add pectinase (add 50U per gram of fermentation product) at 40℃ for 1.5h, sterilize, and obtain the compound fruit and vegetable fermentation slurry;

[0147] (5) Take the following mass parts of raw materials: black soldier fly larvae defatted powder 55 parts, medicinal fungus fermentation product 25 parts, Antarctic krill powder 9 parts, compound fruit and vegetable fermentation slurry 4 parts, prebiotic (xylo-oligosaccharide: inulin = 1:1, mass ratio) 3.5 parts, cold-pressed flaxseed oil 3.5 parts, ganoderma lucidum mycelium polysaccharide extract 2.5 parts, and microencapsulated compound probiotics 0.8 parts;

[0148] (6) Mix the black soldier fly larvae defatted powder, medicinal fungus fermentation product, and Antarctic krill powder, and subject to enzymolysis with alkaline protease and flavor protease (1:1, mass ratio) at 47℃ and pH 7 for 120min, wherein the enzyme activity of alkaline protease is 200000U / g, the enzyme activity of flavor protease is 30000U / g, the total amount of enzyme added is 5000U per gram of protein, and intermittent ultrasonic waves (40kHz, act for 5min every 30min) are applied during the enzymolysis process, to obtain a compound protein substrate;

[0149] (7) Mix the compound protein substrate, microencapsulated compound probiotics, compound fruit and vegetable fermentation slurry, prebiotic, cold-pressed flaxseed oil, and ganoderma lucidum mycelium polysaccharide extract, homogenize at 50MPa and 3500rpm for 30min, 3D print extrusion molding (equipment parameters: nozzle diameter 1.2mm, extrusion pressure 0.3MPa, printing temperature 35℃ (constant temperature nozzle), molding speed 20mm / s), and then high-voltage electrostatic field coupling ultraviolet pulse sterilization (high-voltage electrostatic field treatment: equipment is parallel plate electrodes (spacing 5cm), parameters are 15kV / cm, 10min, and ambient humidity 50%; ultraviolet pulse assistance: wavelength is 254nm+185nm (ozone generation), intensity is 50mJ / cm 2 , pulse frequency is 5Hz), to obtain the functional pet fresh food.

[0150] Example 3

[0151] A method for preparing a functional pet fresh food, comprising the following steps:

[0152] (1) Fresh black soldier fly larvae (fatty content ≤20%) are subjected to ultrasonic-assisted lipase hydrolysis (50 kHz, 50°C, pH 7.2, 2 h, lipase enzyme activity 20000 U / g, and the amount of lipase added is 0.6% of the mass of fresh black soldier fly larvae), centrifugal separation (3500 rpm) is performed until the fatty content in the precipitate is ≤5 wt%, then hot air drying (50°C, 2 h) is performed on the precipitate, sieving is performed, and the undersize is collected as black soldier fly larvae defatted powder (particle size 180 μm);

[0153] (2) Using bran, soybean meal and green tea residue as the substrate (bran: soybean meal: green tea residue = 7:4:1, mass ratio), the water content is adjusted to 57%, and a sporangium suspension of Phellinus baumii and a mycelium suspension of Hericium erinaceus are inoculated (wherein the sporangium suspension of Phellinus baumii is scraped with 0.05% Tween 80 sterile water, filtered (200-mesh sieve) to adjust to 1×10 6 spores / mL, the mycelium suspension of Hericium erinaceus is broken under sterile conditions (ultrasonic for 3 min, 40 kHz), and adjusted to 1×10 6 CFU / mL, the volume ratio of the sporangium suspension of Phellinus baumii to the mycelium suspension of Hericium erinaceus is 4:1, and the total inoculation amount of the sporangium suspension of Phellinus baumii and the mycelium suspension of Hericium erinaceus is 10%), and co-fermentation is carried out at 28°C, 70% humidity and in the dark for 7 days, the fermentation end standard is that the mycelium β-glucan content is 8.23 wt% (determined by the phenol-sulfuric acid method), then the fermentation product is low-temperature dried at 45°C for 7 h, and the Phellinus baumii-Hericium erinaceus co-fermentation product, i.e., the medicinal fungal fermentation product, is obtained;

[0154] (3) A Bacillus subtilis suspension and a Lactobacillus plantarum suspension (both adjusted to 1×10 9The mixed bacteria suspension was obtained by mixing the bacteria suspensions of Lactobacillus plantarum and Bifidobacterium longum in a volume ratio of 2:1; the sodium alginate solution was obtained by dissolving sodium alginate (2%, w / v) in deionized water at 55°C and sterilizing at 121°C for 15 min; the chitosan solution was obtained by dissolving chitosan (degree of deacetylation≥90%) in 0.5% acetic acid solution (pH 5.0) to a concentration of 0.5% (w / v); the mixed bacteria suspension was mixed with the sodium alginate solution in a volume ratio of 1:1, and was electrostatically dropped into a CaCl2 solution (0.1M in itself, containing 0.05% Tween 80) with the parameters set as follows: voltage 8kV, needle diameter 0.4mm, needle dropping speed 15 drops / min, magnetic stirring (200rpm), and solidification for 20 min, to form gel beads with a diameter of 1.2-1.5mm; then the gel beads were immersed in the chitosan solution, slowly oscillated (50rpm) at 25°C for 15 min, washed with PBS buffer solution (pH 7.0) for 3 times after being taken out, and quickly frozen at-40°C for 2h, to obtain the microencapsulated compound probiotics;

[0155] (4) The fruits (apple residue: blueberry: pineapple skin = 1:1:1, mass ratio), vegetables (carrot: purple cabbage: pumpkin = 1:1:1, mass ratio), and fermentation substrate (fructooligosaccharide: yeast extract = 1:1) were mixed in a mass ratio of 3:2:1, inoculated with Lactobacillus plantarum (without activation, directly used in the form of freeze-dried bacteria powder, with a viable bacterial count of 1×10 10 CFU / g of bacteria powder, at an inoculum of 5% (w / w)) for 24h at 37°C, and then inoculated with Bifidobacterium longum (in the form of bacteria suspension, with a bacteria content of 1×10 10 CFU / mL of suspension, at an inoculum of 3% (v / w)) for 48h at 30°C, and then subjected to enzymatic hydrolysis to obtain a compound fruit and vegetable fermentation slurry, by adding pectinase (55U per gram of fermentation product) and treating at 45°C for 2h, and sterilizing.

[0156] (5) The following raw materials were weighed: black soldier fly larvae defatted powder 60 parts, medicinal fungus fermentation product 30 parts, Antarctic krill powder 10 parts, compound fruit and vegetable fermentation slurry 5 parts, prebiotic (xylo-oligosaccharide: inulin = 1:2, mass ratio) 4 parts, cold-pressed flaxseed oil 4 parts, ganoderma lucidum mycelium polysaccharide extract 3 parts, and microencapsulated compound probiotics 1 part;

[0157] (6) The black soldier fly larvae defatted powder, medicinal fungus fermentation product, and Antarctic krill powder were mixed, and subjected to enzymatic hydrolysis with alkaline protease and flavor protease (1:1, mass ratio) at 50°C and pH 7.2 for 150min, wherein the enzyme activity of the alkaline protease was 200000U / g, the enzyme activity of the flavor protease was 30000U / g, the total amount of enzyme added was 5500U per gram of protein, and intermittent ultrasonic waves (50kHz, acting for 6min every 30min) were applied during the enzymatic hydrolysis, to obtain a compound protein substrate.

[0158] (7) mixing the complex protein matrix, microencapsulated complex probiotics, complex fruit and vegetable fermentation slurry with prebiotics, cold-pressed flaxseed oil, and ganoderma polysaccharide extract, homogenizing at 55 MPa and 4000 rpm for 35 min, 3D printing extrusion molding (device parameters: nozzle diameter 1.2 mm, extrusion pressure 0.3 MPa, printing temperature 35°C (constant temperature nozzle), molding speed 20 mm / s), and then high-voltage electrostatic field coupling ultraviolet pulse sterilization (high-voltage electrostatic field treatment: device is parallel plate electrodes (spacing 5 cm), parameters are 15 kV / cm, 10 min, ambient humidity 50%; ultraviolet pulse assistance: wavelength 254 nm + 185 nm (ozone generation), intensity 50 mJ / cm 2 , pulse frequency 5 Hz) to obtain the functional pet fresh food.

[0159] Comparative Example 1

[0160] A method for preparing a pet fresh food, comprising the following steps:

[0161] The difference from Example 2 is that the medicinal fungal fermentation product component is omitted:

[0162] (1) ultrasonic-assisted lipase hydrolysis of fresh black soldier fly larvae (fat content ≤20%) (40 kHz, 45°C, pH 7, 1.5 h, lipase enzyme activity 20000 U / g, lipase addition amount 0.5% of the mass of fresh black soldier fly larvae), centrifugal separation (3000 rpm) to a fat content of ≤5wt% in the precipitate, then hot air drying (50°C, 2 h) of the precipitate, sieving, and collecting the undersize material to obtain black soldier fly larvae defatted powder (particle size 160 μm);

[0163] (2) mixing the Lactobacillus plantarum suspension and Bacillus subtilis suspension (both adjusted to a bacterial concentration of 1×10 9The mixed bacteria suspension was obtained by mixing the mixed bacteria suspension and the sodium alginate solution at a volume ratio of 1:1; the chitosan solution was prepared by dissolving chitosan (degree of deacetylation ≥ 90%) in 0.5% acetic acid solution (pH 5.0) to a concentration of 0.5% (w / v); the mixed bacteria suspension was mixed with the sodium alginate solution at a volume ratio of 1:1, and electrostatically dropped into a CaCl2 solution (0.1 M in itself, containing 0.05% Tween 80), with the parameters set as follows: voltage 8 kV, needle diameter 0.4 mm, needle dropping speed 15 drops / min, magnetic stirring (200 rpm), and solidification time 20 min, to form gel beads with a diameter of 1.2-1.5 mm, which were then immersed in the chitosan solution, slowly oscillated (50 rpm) at 25°C for 15 min, washed with PBS buffer (pH 7.0) for 3 times after being taken out, and quickly frozen at -40°C for 2 h, to obtain the microencapsulated compound probiotics;

[0164] (3) The fruits (apple residue: blueberry: pineapple skin = 1:1:1, mass ratio), vegetables (carrot: purple cabbage: pumpkin = 1:1:1, mass ratio), and fermentation substrate (fructooligosaccharide: yeast extract = 1:1) were mixed at a mass ratio of 2.5:1.5:1, inoculated with Lactobacillus plantarum (without activation, directly used in the form of freeze-dried bacteria powder, with a viable bacterial count of 1×10 10 CFU / g of bacteria powder, at an inoculum of 5% (w / w)), inoculated with Saccharomyces boulardii (in the form of bacterial suspension, with a bacterial content of 1×10 10 CFU / mL of suspension, at an inoculum of 3% (v / w)), and fermented at 30°C for 48 h, and then the fermented product was subjected to enzymatic hydrolysis by adding pectinase (50 U per gram of fermented product) and treated at 40°C for 1.5 h, to obtain the compound fruit and vegetable fermented slurry;

[0165] (4) The following raw materials were weighed: black soldier fly larvae defatted powder 55 parts, medicinal fungus fermentation product 25 parts, Antarctic krill powder 9 parts, compound fruit and vegetable fermented slurry 4 parts, prebiotic (xylo-oligosaccharide: inulin = 1:1, mass ratio) 3.5 parts, cold-pressed flaxseed oil 3.5 parts, ganoderma lucidum mycelium polysaccharide extract 2.5 parts, and microencapsulated compound probiotics 0.8 part;

[0166] (5) The black soldier fly larvae defatted powder and the Antarctic krill powder were mixed, and subjected to enzymatic hydrolysis by alkaline protease and flavor protease (1:1, mass ratio) at 47°C and pH 7 for 120 min, wherein the enzyme activity of the alkaline protease was 200,000 U / g, the enzyme activity of the flavor protease was 30,000 U / g, the total amount of enzyme added was 5,000 U per gram of protein, and intermittent ultrasonic waves (40 kHz, acting for 5 min every 30 min) were applied during the enzymatic hydrolysis, to obtain the compound protein substrate;

[0167] (6) mixing the complex protein matrix, microencapsulated complex probiotics, complex fruit and vegetable fermentation slurry with prebiotics, cold-pressed flaxseed oil, and ganoderma lucidum mycelium polysaccharide extract, homogenizing at 50 MPa and 3500 rpm for 30 min, 3D printing extrusion molding (device parameters: nozzle diameter 1.2 mm, extrusion pressure 0.3 MPa, printing temperature 35°C (constant temperature nozzle), molding speed 20 mm / s), and then high-voltage electrostatic field coupling ultraviolet pulse sterilization (high-voltage electrostatic field treatment: device is parallel plate electrodes (spacing 5 cm), parameters are 15 kV / cm, 10 min, ambient humidity 50%; ultraviolet pulse assistance: wavelength 254 nm + 185 nm (ozone generation), intensity 50 mJ / cm 2 , pulse frequency 5 Hz) to obtain the pet fresh food.

[0168] Comparative Example 2

[0169] A preparation method of a pet fresh food, comprising the following steps:

[0170] The difference from Example 2 is that the process of enzymatic hydrolysis of black soldier fly larvae defatted powder, medicinal fungus fermentation, and Antarctic krill powder is omitted:

[0171] (1) ultrasonic-assisted lipase hydrolysis of fresh black soldier fly larvae (fatty acid content ≤20%) (40 kHz, 45°C, pH 7, 1.5 h, lipase enzyme activity 20000 U / g, lipase addition amount 0.5% of the mass of fresh black soldier fly larvae), centrifugal separation (3000 rpm) to a fatty acid content of ≤5 wt% in the precipitate, then hot air drying (50°C, 2 h) of the precipitate, sieving, and collection of undersize material as black soldier fly larvae defatted powder (particle size 160 μm);

[0172] (2) using wheat bran, soybean meal, and green tea dregs as the substrate (wheat bran: soybean meal: green tea dregs = 6:3:1, mass ratio), adjusting the moisture content to 55%, inoculating with a Phellinus igniarius spore suspension and a Hericium erinaceus mycelium suspension (wherein the Phellinus igniarius spore suspension: scraping spores with 0.05% Tween 80 sterile water, filtering (200 mesh sieve) to adjust to 1×10 6 spores / mL, the Hericium erinaceus mycelium suspension: sterilely crushing the mycelium ball (ultrasonic 3 min, 40 kHz) to adjust the mycelium content to 1×10 6CFU / mL, the volume ratio of the Phellinus spore suspension to the Hericium mycelium suspension was 3:1, the total inoculation amount of the Phellinus spore suspension and the Hericium mycelium suspension was 8%, and the co-fermentation was carried out at 27°C, 70% humidity and in the dark for 7 days, and the fermentation end standard was that the mycelium β-glucan content was 8.95wt% (determined by the phenol-sulfuric acid method), then the fermentation product was dried at low temperature at 40°C for 6h, and the Phellinus-Hericium co-fermentation product, i.e., the medicinal fungus fermentation product, was obtained;

[0173] (3) The Lactobacillus plantarum suspension and the Bacillus subtilis suspension (both of which were adjusted to a bacterial concentration of 1×10 9 CFU / mL) were mixed according to a volume ratio of 1:1 to obtain a mixed bacterial suspension; sodium alginate (2%, w / v) was dissolved in deionized water at 55°C and sterilized at 121°C for 15min to obtain a sodium alginate solution; chitosan (degree of deacetylation≥90%) was dissolved in 0.5% acetic acid solution (pH 5.0) to obtain a chitosan solution with a concentration of 0.5% (w / v); the mixed bacterial suspension and the sodium alginate solution were mixed according to a volume ratio of 1:1, and electrostatically dropped into a CaCl2 solution (0.1M in itself, containing 0.05% Tween 80), with parameters set as voltage 8kV, needle diameter 0.4mm, needle dropping speed 15 drops / min, magnetic stirring (200rpm), and solidification for 20min, to form gel beads with a diameter of 1.2-1.5mm, then the gel beads were immersed in the chitosan solution, slowly oscillated (50rpm) at 25°C for 15min, washed with PBS buffer solution (pH 7.0) for 3 times after being taken out, and quickly frozen at -40°C for 2h, to obtain the microencapsulated compound probiotics;

[0174] (4) Fruits (apple residue: blueberry: pineapple skin = 1:1:1, mass ratio), vegetables (carrot: purple cabbage: pumpkin = 1:1:1, mass ratio), and fermentation substrate (fructooligosaccharide: yeast extract = 1:1) were mixed according to a mass ratio of 2.5:1.5:1, inoculated with Lactobacillus plantarum (without activation, directly used in the form of freeze-dried bacterial powder, with a viable bacterial count of 1×10 10 CFU / g of bacterial powder, 5% (w / w) inoculation amount) at 37°C for 24h, and then inoculated with Saccharomyces boulardii (in the form of a bacterial suspension, with a bacterial content of 1×10 10 CFU / mL of suspension, 3% (v / w) inoculation amount) at 30°C for 48h, and then the fermentation product was subjected to enzymatic hydrolysis, pectinase (50U per gram of fermentation product) was added, and the mixture was treated at 40°C for 1.5h, and then sterilized, to obtain the compound fruit and vegetable fermentation slurry;

[0175] (5) Take the following mass parts of raw materials: black soldier fly larvae defatted powder 55 parts, medicinal fungus ferment 25 parts, Antarctic krill powder 9 parts, compound fruit and vegetable fermentation broth 4 parts, prebiotic (xylo-oligosaccharide: inulin = 1:1, mass ratio) 3.5 parts, cold-pressed flaxseed oil 3.5 parts, ganoderma lucidum mycelium polysaccharide extract 2.5 parts, and microencapsulated compound probiotics 0.8 parts;

[0176] (6) Mix the black soldier fly larvae defatted powder, medicinal fungus ferment, Antarctic krill powder, microencapsulated compound probiotics, compound fruit and vegetable fermentation broth, prebiotic, cold-pressed flaxseed oil, and ganoderma lucidum mycelium polysaccharide extract, and homogenize at 50 MPa and 3500 rpm for 30 min, 3D print extrusion molding (equipment parameters: nozzle diameter 1.2 mm, extrusion pressure 0.3 MPa, printing temperature 35°C (constant temperature nozzle), molding speed 20 mm / s), and then use high-voltage electrostatic field coupling ultraviolet pulse sterilization (high-voltage electrostatic field treatment: equipment is parallel plate electrodes (spacing 5 cm), parameters are 15 kV / cm, 10 min, and ambient humidity 50%; ultraviolet pulse assistance: wavelength 254 nm + 185 nm (ozone generation), intensity 50 mJ / cm 2 , pulse frequency 5 Hz), to obtain the pet fresh food.

[0177] Comparative Example 3

[0178] A preparation method of a pet fresh food, the steps being as follows:

[0179] The difference from Example 2 is only that the fermentation process of the compound fruit and vegetable is omitted:

[0180] (1) Ultrasonic-assisted lipase hydrolysis is performed on fresh black soldier fly larvae (fatty content ≤20%) (40 kHz, 45°C, pH 7, 1.5 h, lipase enzyme activity 20000 U / g, and lipase addition amount 0.5% of the mass of fresh black soldier fly larvae), centrifugal separation (3000 rpm) is performed until the fatty content in the precipitate is ≤5 wt%, then hot air drying (50°C, 2 h) is performed on the precipitate, sieving is performed, and the undersize is collected as black soldier fly larvae defatted powder (particle size 160 μm);

[0181] (2) Take bran, soybean meal, and green tea dregs as the substrate (bran: soybean meal: green tea dregs = 6:3:1, mass ratio), adjust the water content to 55%, inoculate the sporodochia suspension of the Phlebia sp. and the mycelium suspension of the Hericium erinaceus (among which, the sporodochia suspension: scrape the spores with 0.05% Tween 80 sterile water, filter (200-mesh sieve) to adjust to 1×10 6 spores / mL, and the mycelium suspension of the Hericium erinaceus: break the mycelium ball under sterile conditions (ultrasonic 3 min, 40 kHz), and adjust to a mycelium content of 1×10 6CFU / mL, the volume ratio of the Phellinus spore suspension to the Hericium erinaceus mycelium suspension was 3:1, the total inoculation amount of the Phellinus spore suspension and the Hericium erinaceus mycelium suspension was 8%, and co-fermentation was carried out at 27°C, 70% humidity and in the dark for 7 days, with the mycelium β-glucan content of 8.96wt% (determined by the phenol-sulfuric acid method) as the fermentation end standard, and then the fermentation product was dried at low temperature at 40°C for 6h to obtain the Phellinus-Hericium co-fermentation product, that is, the medicinal fungus fermentation product;

[0182] (3) Lactobacillus plantarum suspension and Bacillus subtilis suspension (both with a bacterial concentration of 1×10 9 CFU / mL) were mixed according to a volume ratio of 1:1 to obtain a mixed bacterial suspension; sodium alginate (2%, w / v) was dissolved in deionized water at 55°C and sterilized at 121°C for 15min to obtain a sodium alginate solution; chitosan (degree of deacetylation ≥90%) was dissolved in 0.5% acetic acid solution (pH 5.0) to obtain a chitosan solution with a concentration of 0.5% (w / v); the mixed bacterial suspension and the sodium alginate solution were mixed according to a volume ratio of 1:1, and electrostatically dropped into a CaCl2 solution (0.1M in itself, containing 0.05% Tween 80), with parameters set as voltage 8kV, needle diameter 0.4mm, needle dropping speed 15 drops / min, magnetic stirring (200rpm), and solidification for 20min to form gel beads with a diameter of 1.2-1.5mm, then the gel beads were immersed in the chitosan solution, slowly oscillated (50rpm) at 25°C for 15min, washed with PBS buffer solution (pH 7.0) for 3 times after taking out, and quickly frozen at -40°C for 2h to obtain the microencapsulated compound probiotics;

[0183] (4) Fruits (apple residue: blueberry: pineapple skin = 1:1:1, mass ratio), vegetables (carrot: purple cabbage: pumpkin = 1:1:1, mass ratio), and deionized water were mixed according to a mass ratio of 2.5:1.5:1, stirred, and sterilized to obtain a compound fruit and vegetable slurry;

[0184] (5) The following raw materials were weighed: black soldier fly larvae defatted powder 55 parts, medicinal fungus fermentation product 25 parts, Antarctic krill powder 9 parts, compound fruit and vegetable slurry 4 parts, prebiotic (xylo-oligosaccharide: inulin = 1:1, mass ratio) 3.5 parts, cold-pressed flaxseed oil 3.5 parts, Ganoderma lucidum mycelium polysaccharide extract 2.5 parts, and microencapsulated compound probiotics 0.8 part;

[0185] (6) The defatted powder of black soldier fly larvae, the fermented medicinal fungus and Antarctic krill powder were mixed and enzymatically hydrolyzed for 120 min at 47 °C and pH 7 using alkaline protease and flavor protease (1:1, mass ratio). The alkaline protease had an activity of 200,000 U / g and the flavor protease had an activity of 30,000 U / g. The total amount of enzyme added was 5,000 U per gram of protein. Intermittent ultrasound (40 kHz, 5 min every 30 min) was applied during the enzymatic hydrolysis process to obtain a composite protein matrix.

[0186] (7) The composite protein matrix, microencapsulated composite probiotics, composite fruit and vegetable pulp, prebiotics, cold-pressed flaxseed oil, and Ganoderma lucidum mycelium polysaccharide extract were mixed and homogenized at 50 MPa and 3500 rpm for 30 min. The mixture was then 3D printed and extruded (equipment parameters: nozzle diameter 1.2 mm, extrusion pressure 0.3 MPa, printing temperature 35℃ (constant temperature nozzle), forming speed 20 mm / s). The mixture was then sterilized using a high-voltage electrostatic field coupled with ultraviolet pulse (high-voltage electrostatic field treatment: equipment is a parallel plate electrode (5 cm spacing), parameters are 15 kV / cm, 10 min, ambient humidity 50%; ultraviolet pulse assistance: wavelength is 254 nm + 185 nm (ozone generation), intensity is 50 mJ / cm). 2 The fresh pet food was obtained by pulse frequency of 5Hz.

[0187] Comparative Example 4

[0188] A method for preparing fresh pet food, comprising the following steps:

[0189] The only difference from Example 2 is that the *Lactobacillus plantarum* in the microencapsulated probiotic complex is replaced with *Bifidobacterium adolescentis*.

[0190] (1) Fresh black soldier fly larvae (fat content ≤20%) were subjected to ultrasonic-assisted lipase hydrolysis (40kHz, 45℃, pH 7, 1.5h, lipase activity 20000U / g, lipase addition amount 0.5% of the mass of fresh black soldier fly larvae), centrifuged (3000rpm) until the fat content in the precipitate was ≤5wt%, and then the precipitate was dried by hot air (50℃, 2h), sieved, and the sieve-underfilled material was collected as black soldier fly larvae defatted powder (particle size 160μm).

[0191] (2) Using wheat bran, soybean meal, and green tea residue as a substrate (wheat bran: soybean meal: green tea residue = 6:3:1, mass ratio), adjust the moisture content to 55%, and inoculate with a suspension of Phellinus linteus spores and a suspension of Hericium erinaceus mycelium (for the Phellinus linteus spore suspension: scrape spores with 0.05% Tween 80 sterile water, filter (using a 200-mesh sieve), and adjust to 1×10⁻⁶. 6spores / mL, and the Hericium erinaceus mycelium suspension: breaking the mycelium ball under sterile conditions (ultrasonic 3 min, 40 kHz), adjusting to the mycelium content of 1 x 10 6 CFU / mL, the volume ratio of the Phellinus igniarius spore suspension to the Hericium erinaceus mycelium suspension was 3:1, and the total inoculation amount of the Phellinus igniarius spore suspension and the Hericium erinaceus mycelium suspension was 8%), and co-fermentation was carried out at 27°C, 70% humidity and in the dark for 7 days, the fermentation end standard was that the mycelium β-glucan content was 8.94 wt% (determined by the phenol-sulfuric acid method), and then the fermentation product was dried at low temperature at 40°C for 6 h to obtain the Phellinus igniarius-Hericium erinaceus co-fermentation product, that is, the medicinal fungus fermentation product;

[0192] (3) The Bifidobacterium adolescentis suspension and the Bacillus subtilis suspension (the mycelium concentration of both was adjusted to 1 x 10 9 CFU / mL) were mixed in a volume ratio of 1:1 to obtain a mixed bacteria suspension; sodium alginate (2%, w / v) was dissolved in deionized water at 55°C and sterilized at 121°C for 15 min to obtain a sodium alginate solution; chitosan (degree of deacetylation ≥90%) was dissolved in 0.5% acetic acid solution (pH 5.0) to obtain a chitosan solution with a concentration of 0.5% (w / v); the mixed bacteria suspension and the sodium alginate solution were mixed in a volume ratio of 1:1, and electrostatically dropped into a CaCl2 solution (its own concentration was 0.1M, containing 0.05% Tween 80), the parameters were set as follows: voltage 8kV, needle diameter 0.4mm, needle dropping speed 15 drops / min, magnetic stirring (200rpm), and solidification for 20 min to form gel beads with a diameter of 1.2-1.5mm, then the gel beads were immersed in the chitosan solution, slowly oscillated (50rpm) at 25°C for 15 min, washed with PBS buffer solution (pH 7.0) for 3 times after taking out, and frozen at -40°C for 2h to obtain the microencapsulated compound probiotics;

[0193] (4) Fruits (apple residue: blueberry: pineapple skin = 1:1:1, mass ratio), vegetables (carrot: purple cabbage: pumpkin = 1:1:1, mass ratio), and fermentation substrate (fructooligosaccharide: yeast extract = 1:1) were mixed in a mass ratio of 2.5:1.5:1, inoculated with Lactobacillus plantarum (without activation, directly used in the form of freeze-dried bacteria powder, the viable bacteria number was 1 x 10 10 CFU / g of bacteria powder, 5% (w / w) inoculation amount) at 37°C for 24h, and then inoculated with Saccharomyces boulardii (in the form of bacteria suspension, the bacteria content was 1 x 10 10 CFU / mL suspension, 3% (v / w) inoculation amount) at 30°C for 48h, and then the fermentation product was subjected to enzymatic hydrolysis by adding pectinase (50U per gram of fermentation product) at 40°C for 1.5h, and sterilized to obtain the compound fruit and vegetable fermentation slurry;

[0194] (5) take the following mass parts of raw materials: black soldier fly larvae defatted powder 55 parts, medicinal fungus ferment 25 parts, Antarctic krill powder 9 parts, compound fruit and vegetable fermentation broth 4 parts, prebiotic (xylo-oligosaccharide: inulin = 1:1, mass ratio) 3.5 parts, cold-pressed flaxseed oil 3.5 parts, ganoderma lucidum mycelium polysaccharide extract 2.5 parts, and microencapsulated compound probiotics 0.8 parts;

[0195] (6) The black soldier fly larvae defatted powder, medicinal fungus ferment, and Antarctic krill powder are mixed, and then subjected to enzymatic hydrolysis with alkaline protease and flavor protease (1:1, mass ratio) at 47°C and pH 7 for 120 min. The alkaline protease has an enzyme activity of 200,000 U / g, and the flavor protease has an enzyme activity of 30,000 U / g. The total amount of enzyme added is 5,000 U per gram of protein. Intermittent ultrasonic waves (40 kHz, 5 min of action every 30 min) are applied during the enzymatic hydrolysis process to obtain a compound protein matrix.

[0196] (7) The compound protein matrix, microencapsulated compound probiotics, compound fruit and vegetable fermentation broth, prebiotic, cold-pressed flaxseed oil, and ganoderma lucidum mycelium polysaccharide extract are mixed, and then subjected to homogenization at 50 MPa and 3,500 rpm for 30 min. 3D printing extrusion molding is performed (device parameters: nozzle diameter 1.2 mm, extrusion pressure 0.3 MPa, printing temperature 35°C (constant temperature nozzle), molding speed 20 mm / s). Then, high-voltage electrostatic field coupling ultraviolet pulse sterilization is performed (high-voltage electrostatic field treatment: device is a parallel plate electrode (spacing 5 cm), parameters are 15 kV / cm, 10 min, and ambient humidity 50%; ultraviolet pulse assistance: wavelength 254 nm + 185 nm (ozone generation), intensity 50 mJ / cm 2 , pulse frequency 5 Hz) to obtain the pet fresh food.

[0197] Comparative Example 5

[0198] A method for preparing a pet fresh food, comprising the following steps:

[0199] The difference from Example 2 is that the microencapsulation embedding process of probiotics is omitted.

[0200] (1) Fresh black soldier fly larvae (fatty content ≤20%) are subjected to ultrasonic-assisted lipase hydrolysis (40 kHz, 45°C, pH 7, 1.5 h, lipase enzyme activity 20,000 U / g, and lipase addition amount 0.5% of the mass of fresh black soldier fly larvae). Centrifugal separation (3,000 rpm) is performed until the fatty content in the precipitate is ≤5 wt%. Then, the precipitate is subjected to hot air drying (50°C, 2 h), sieving, and collection of undersize material to obtain black soldier fly larvae defatted powder (particle size 160 μm).

[0201] (2) with bran, soybean meal, green tea residue as substrate (bran: soybean meal: green tea residue = 6:3:1, mass ratio), the moisture content is adjusted to 55%, inoculated with Phellinus igniarius spore suspension and Hericium erinaceus mycelium suspension (in which, Phellinus igniarius spore suspension: spores are scraped with 0.05% Tween 80 sterile water, filtered (200 mesh screen) to adjust to 1×10 6 spores / mL, Hericium erinaceus mycelium suspension: mycelium ball is broken under sterile conditions (ultrasonic 3 min, 40 kHz), the content is adjusted to 1×10 6 CFU / mL, the volume ratio of the Phellinus igniarius spore suspension and the Hericium erinaceus mycelium suspension is 3:1, the total inoculation amount of the Phellinus igniarius spore suspension and the Hericium erinaceus mycelium suspension is 8%), co-fermented at 27°C, humidity 70% and light-avoiding conditions for 7 days, the fermentation end standard is that the mycelium β-glucan content is 8.93wt% (determined by phenol-sulfuric acid method), then the fermentation product is dried at low temperature at 40°C for 6h, to obtain Phellinus igniarius-Hericium erinaceus co-fermentation product, that is, medicinal fungus fermentation product;

[0202] (3) Lactobacillus plantarum suspension and Bacillus subtilis suspension (both of which are adjusted to 1×10 9 CFU / mL) are mixed according to a volume ratio of 1:1 to obtain a compound probiotic;

[0203] (4) fruits (apple residue: blueberry: pineapple skin = 1:1:1, mass ratio), vegetables (carrot: purple cabbage: pumpkin = 1:1:1, mass ratio), fermentation substrate (fructooligosaccharide: yeast extract = 1:1) are mixed according to a mass ratio of 2.5:1.5:1, first inoculated with Lactobacillus plantarum (without activation, direct injection type freeze-dried bacteria powder, viable bacteria number is 1×10 10 CFU / g bacteria powder, 5% (w / w) inoculation amount) at 37°C for 24h, then inoculated with Saccharomyces boulardii (inoculated in the form of bacteria suspension, bacteria content is 1×10 10 CFU / mL suspension, 3% (v / w) inoculation amount) at 30°C for 48h, after fermentation, the fermentation product is subjected to enzymatic hydrolysis, pectinase is added (50U per gram of fermentation product) at 40°C for 1.5h, sterilized to obtain a compound fruit and vegetable fermentation broth;

[0204] (5) the following mass parts of raw materials are weighed: black soldier fly larvae defatted powder 55 parts, medicinal fungus fermentation product 25 parts, Antarctic krill powder 9 parts, compound fruit and vegetable fermentation broth 4 parts, prebiotic (xylo-oligosaccharide: inulin = 1:1, mass ratio) 3.5 parts, cold-pressed flaxseed oil 3.5 parts, Ganoderma lucidum mycelium polysaccharide extract 2.5 parts, compound probiotic 0.8 part;

[0205] (6) mixing the black soldier fly larvae defatted powder, medicinal fungus fermentation product and Antarctic krill powder, and using alkaline protease and flavor protease (1:1, mass ratio) to hydrolyze for 120 min at 47°C and pH 7, wherein the enzyme activity of the alkaline protease is 200000 U / g, the enzyme activity of the flavor protease is 30000 U / g, the total amount of enzyme added is 5000 U per gram of protein, and intermittent ultrasonic waves (40 kHz, acting for 5 min every 30 min) are applied during the enzyme hydrolysis process to obtain a complex protein matrix;

[0206] (7) mixing the complex protein matrix, complex probiotics, complex fruit and vegetable fermentation slurry, prebiotics, cold-pressed flaxseed oil and ganoderma lucidum mycelium polysaccharide extract, homogenizing for 30 min at 50 MPa and 3500 rpm, 3D printing extrusion molding (equipment parameters: nozzle diameter 1.2 mm, extrusion pressure 0.3 MPa, printing temperature 35°C (constant temperature nozzle), molding speed 20 mm / s), and then using high-voltage electrostatic field coupling ultraviolet pulse sterilization (high-voltage electrostatic field treatment: equipment is parallel plate electrodes (spacing 5 cm), parameters are 15 kV / cm, 10 min, ambient humidity 50%; ultraviolet pulse assistance: wavelength is 254 nm + 185 nm (ozone generation), intensity is 50 mJ / cm 2 , pulse frequency 5 Hz) to obtain the pet fresh food.

[0207] Experimental Example 1

[0208] In-vitro Digestibility Determination Experiment:

[0209] Taking Examples 1-3, Comparative Examples 1-5 and a commercially available traditional pet fresh food (formula: chicken breast (boneless): 50%, vegetables: 30% (pumpkin 20%, carrot 10%), chicken liver: 10%, brown rice: 5%, supplements: 5% (fish oil 2%, calcium powder 2%, vitamin E 1%), high-temperature sterilization at 121°C) as examples, the protein digestibility of different pet fresh foods was verified, and the amino acid release characteristics were evaluated.

[0210] (1) Simulated Gastric-Intestinal Digestion Model

[0211] Gastric Phase:

[0212] 5 g of different samples (Examples 1-3, Comparative Examples 1-5 and commercially available traditional pet fresh food) were ground and then mixed with 10 mL of simulated gastric juice (containing 0.15 M NaCl, 0.006 M HCl, pH = 2.0) respectively;

[0213] 32 mg / 10 mL of pepsin (enzyme activity 10000 U / g) was added, and the mixture was shaken (100 rpm) at 37°C for 2 h.

[0214] Intestinal Phase:

[0215] Adjust pH to 6.8, add 10 mL simulated intestinal fluid (containing 0.1 M NaHCO3, 0.005 M trypsin);

[0216] 37℃ continue to shake digestion for 4h, sample every 30min to detect hydrolysis degree (DH).

[0217] (2) Key detection indicators

[0218] Protein digestibility: Kjeldahl determination of nitrogen content of undigested residue, calculate the digestibility:

[0219]

[0220] Amino acid release rate: HPLC determination of free amino acid content in the digestion solution (detect essential amino acids: lysine).

[0221] The results are shown in Table 5.

[0222] Table 5 Protein digestibility and amino acid release of different pet fresh foods

[0223] Sample Protein digestibility (%) Lysine release (mg / g) Example 1 91.6 57.2 Example 2 92.3 58.9 Example 3 90.8 56.4 Comparative Example 1 72.4 42.2 Comparative Example 2 73.8 44.8 Comparative Example 3 78.9 48.5 Comparative Example 4 75.1 46.4 Comparative Example 5 77.2 47.6 Commercial conventional pet fresh food 68.7 40.5

[0224] As can be seen from Table 5, compared with Comparative Examples 1-5 and commercially available conventional pet fresh foods, the protein digestibility of the pet fresh foods of the present application Examples 1-3 is significantly improved, and the essential amino acid release is more sufficient, especially the effect of Example 2 is the most significant, the protein digestibility is as high as 92.3%, and the lysine release amount is 58.9 mg / g.

[0225] Experimental Example 2

[0226] Immune performance determination experiment:

[0227] Verify the effect of different pet fresh foods on improving the immune function of pet dogs:

[0228] 1. Experimental design (see Table 6)

[0229] Table 6 Experimental design

[0230] Group Sample size Detection time point Example 1 n = 10 (dog) 0, 8 weeks Example 2 n = 10 (dog) 0, 8 weeks Example 3 n = 10 (dog) 0, 8 weeks Comparative Example 1 n = 10 (dog) 0, 8 weeks Comparative Example 2 n = 10 (dog) 0, 8 weeks Comparative Example 3 n = 10 (dog) 0, 8 weeks Comparative Example 4 n = 10 (dog) 0, 8 weeks Comparative Example 5 n = 10 (dog) 0, 8 weeks Commercial conventional pet fresh food n = 10 (dog) 0, 8 weeks Basal diet n = 10 (dog) 0, 8 weeks

[0231] Animal selection: healthy adult beagle dogs (body weight 10-15 kg, age 1-2 years old), randomly divided into groups, single cage feeding; exclude individuals using antibiotics or immunosuppressive agents in recent period.

[0232] 2. Feeding management

[0233] Daily ration formula:

[0234] Experimental group: fresh food of the present application Examples 1-3;

[0235] Control group: Comparative Example 1-5 fresh food and commercially available traditional pet fresh food (formula by mass percentage: chicken breast (boneless): 50%, vegetables: 30% (pumpkin 20%, carrot 10%), chicken liver: 10%, brown rice: 5%, supplements: 5% (fish oil 2%, calcium powder 2%, vitamin E 1%), 121°C high temperature sterilization);

[0236] Blank group: basal diet, containing only basal energy and protein (by mass percentage, beef powder 35%, soybean meal 20%, corn 25%, wheat bran 10%, spinach 5%, banana 5%, no functional ingredients).

[0237] Feeding scheme:

[0238] 2 times a day, 5% of body weight, free drinking water;

[0239] Record daily feed intake and health status.

[0240] 3, blood sampling and processing

[0241] Blood sampling time: after 12 hours of fasting, before 8 o'clock in the morning, blood sampling from the forelimb vein;

[0242] Sampling volume: 3mL each time, divided into anticoagulant-free vacuum tubes;

[0243] Serum preparation:

[0244] Room temperature for 30min→3000rpm centrifugation for 15min (4℃)→serum storage at-80℃.

[0245] 4, detection method (see Table 7)

[0246] Table 7 Detection method

[0247]

[0248] Operation process:

[0249] IgG detection:

[0250] Dilute serum according to kit instructions (1:1000);

[0251] Add 96-well plate, incubate at 37℃ for 1h→wash→add enzyme-labeled secondary antibody→TMB color development→measure OD value at 450nm.

[0252] IgA detection:

[0253] Mix serum with anti-dog IgA antibody, react at 37℃ for 10min;

[0254] Determine turbidity change, calculate concentration (mg / dL).

[0255] The results are shown in Table 8.

[0256] Table 8 Influence of different pet fresh foods on immune performance of pet dogs

[0257]

[0258]

[0259] From Table 8, it can be seen that the influence of the basic diet on the immune index is not large, excluding environmental interference. Compared with Comparative Examples 1-5 and commercially available conventional pet fresh foods, the IgG level and IgA level of the pet fresh foods of the present application Examples 1-3 are significantly improved, proving that the pet fresh foods of the present application can comprehensively activate the immune system of pets and enhance the immune performance of pets.

[0260] Experimental Example 3

[0261] Standardized fecal score experiment:

[0262] The improvement effect of different pet fresh foods on intestinal health of pet dogs is verified by a standardized fecal score system.

[0263] 1. Experimental design (see Table 9)

[0264] Table 9 Experimental design

[0265]

[0266]

[0267] Animal selection:

[0268] Healthy adult dogs Beagle (body weight 10-15 kg, age 1-2 years old), no recent history of digestive tract disease;

[0269] After adaptive feeding for 1 week, the formal experiment was started.

[0270] 2. Fecal score standard

[0271] A 5-point Bristol modified scale (pet special edition) was used, as shown in Table 10:

[0272] Table 10 Bristol modified scale (pet special edition)

[0273] Score Character description Health significance 1 point Hard particles, no mucus Constipation risk 2 points Good shape, slight crack Ideal state 3 points Soft stool but shaped, easy to pick up Mild indigestion 4 points Loose and unshaped, partially adhered Pre-diarrhea 5 points Water-like stool, unable to shape Severe diarrhea

[0274] Ideal range: 2-3 points.

[0275] 3. Feeding management

[0276] Diet formula:

[0277] Experimental group: fresh food of the present application Example 2;

[0278] Control group: Comparative Example 1-5 fresh food and commercially available traditional pet fresh food (formula: chicken breast (boneless): 50%, vegetables: 30% (pumpkin 20%, carrot 10%), chicken liver: 10%, brown rice: 5%, supplements: 5% (fish oil 2%, calcium powder 2%, vitamin E 1%), high temperature sterilization at 121℃);

[0279] Blank group: basal diet, containing only basal energy and protein (by mass percentage, beef powder 35%, soybean meal 20%, corn 25%, wheat bran 10%, spinach 5%, banana 5%, no functional ingredients).

[0280] Feeding scheme:

[0281] Feed 5% of body weight twice a day, and drink water freely;

[0282] Record daily feed intake and health status.

[0283] Fecal collection: collect fresh samples at a fixed time every day (first defecation in the morning).

[0284] 4, Fecal detection method (see Table 11)

[0285] Table 11 Fecal detection method

[0286] Detection item Method Tool / standard Appearance score Instant score according to Bristol scale Blindly scored by 3 experimenters independently Moisture content Drying method (105℃ to constant weight) Calculation: (wet weight - dry weight) / wet weight x 100% Mucus detection rate Visual + physiological saline smear microscopy Record the proportion of mucus coverage area

[0287] 5, Statistical analysis

[0288] Main indicators: weekly average fecal score (extreme values are excluded), fecal water content and mucus detection rate are counted in the 4th week.

[0289] The results are shown in Table 12.

[0290] Table 12 Improvement effect of different pet fresh food on intestinal health of pet dogs

[0291]

[0292] As shown in Table 12, the score of Example 2 of the present application decreased significantly to the ideal range (2.2 points) in the 4th week, while the scores of Comparative Examples 1-5 and commercially available traditional pet fresh food group were still ≥3.1 points; the water content of Example 2 of the present application decreased to below 70%, and the mucus detection rate was only 5%, while the water content of Comparative Examples 1-5 and commercially available traditional pet fresh food group was ≥72%, and the mucus detection rate was ≥10%, with a high risk of diarrhea. It can be seen that the pet fresh food of the present application has obvious improvement effect on the intestinal health of pets.

[0293] Experimental Example 4

[0294] Accelerated shelf life experiment:

[0295] Example 2 and commercially available conventional pet fresh food (formula: chicken breast (boneless): 50%, vegetables: 30% (pumpkin 20%, carrot 10%), chicken liver: 10%, brown rice: 5%, supplements: 5% (fish oil 2%, calcium powder 2%, vitamin E 1%), high-temperature sterilization at 121°C) were used as examples to verify the theoretical shelf life of different pet fresh food under 4°C refrigeration.

[0296] 1. Accelerated test conditions

[0297] (1) Accelerated model:

[0298] 37°C ± 1°C, RH 75% ± 5% (Arrhenius equation Q10 = 2.5, 1 day ≈ 7 days at 4°C)

[0299] (2) Detection time points (see Table 13):

[0300] Table 13 Detection time points

[0301] Conditions Detection time point Equivalent 4℃ time 37℃ 0, 14, 28, 42, 56 days 0, 3, 6, 9, 12 months

[0302] (3) 4°C real storage control:

[0303] Regular detection (0, 3, 6, 9, 12 months) to verify the accuracy of the accelerated model.

[0304] 2. Detection methods and qualification standards (see Table 14)

[0305] Table 14 Detection methods and qualification standards

[0306]

[0307] 3. Method steps

[0308] (1) Sample preparation:

[0309] Example 2 and commercially available conventional pet fresh food were divided into 50g per portion and numbered after vacuum sealing.

[0310] (2) Accelerated storage:

[0311] The samples were placed in layers in a constant temperature and humidity box to avoid the influence of heat transfer caused by stacking.

[0312] (3) Regular detection:

[0313] Total number of aerobic bacteria: sterile sampling → gradient dilution → coating PCA plate → 37°C incubation for 48h counting;

[0314] Peroxide value: Soxhlet extraction of fat → titration method calculation;

[0315] Water activity: 5g of sample was directly taken for determination, and the average value of three parallel determinations was taken.

[0316] Results are shown in Tables 15, 16.

[0317] Table 15 Accelerated test data (37°C)

[0318]

[0319] Table 16 4°C real storage validation

[0320] Time (months) Example 2 aerobic bacteria (CFU / g) Example 2 POV (g / 100g) Aw 0 1.5 x 10 2 ]]> 0.08 0.82 6 4.2 x 10 3 ]]> 0.16 0.83 12 9.1 x 10 3 ]]> 0.23 0.84

[0321] From Tables 15 and 16, it can be seen that the commercially available conventional pet fresh food has been spoiled (microorganisms exceed the standard + oil rancidity) at the equivalent of 6 months, while the pet fresh food of Example 2 of the present application still meets all the indicators at 37°C for 56 days (equivalent to 4°C for 12 months); after 12 months of real storage at 4°C, the POV (0.23) of the pet fresh food of Example 2 of the present application is close to but does not exceed the standard (≤0.25), verifying the reliability of the accelerated model. It can be seen that the pet fresh food of the present application has a shelf life of ≥12 months at 4°C, which is significantly better than commercially available products.

[0322] The above is only a few embodiments of the present application, and does not limit the present application in any form. Although the preferred embodiments of the present application are disclosed as above, they are not intended to limit the present application. Any skilled person in the art can make some changes or modifications to the disclosed technical content without departing from the scope of the technical solutions of the present application, and such changes or modifications are equivalent to equivalent embodiments, and are within the scope of the technical solutions.

Claims

1. A functional fresh pet food, characterized in that, It is prepared from the following components in parts by weight: 50-60 parts of defatted black soldier fly larvae powder, 20-30 parts of medicinal fungal fermentation product, 8-10 parts of Antarctic krill powder, 3.5-5 parts of compound fruit and vegetable fermentation paste, 3-4 parts of prebiotics, 3-4 parts of cold-pressed flaxseed oil, 2-3 parts of Ganoderma lucidum mycelium polysaccharide extract, and 0.5-1 part of microencapsulated compound probiotics; The medicinal fungal ferment is a co-fermentation product of Phellinus linteus and Hericium erinaceus; The preparation steps of the medicinal fungal fermentation product include: using wheat bran, soybean meal, and tea residue as substrates, adjusting the moisture content to 53-57%, inoculating with Sanghuang spore suspension and Hericium erinaceus mycelial suspension, co-fermenting, drying, and obtaining Sanghuang-Hericium erinaceus co-fermentation product, i.e., medicinal fungal fermentation product; The compound fruit and vegetable fermentation slurry is prepared from fruits, vegetables, fermentation substrate, and fermentation strains. The fruit includes at least one of apple pomace, blueberries, and pineapple peel; The vegetables include at least one of carrots, purple cabbage, and pumpkin; The fermentation substrate includes fructooligosaccharides and yeast extract; The fermentation strains include Lactobacillus plantarum and Saccharomyces boulardii; The prebiotic is a mixture of xylooligosaccharides and inulin; The Ganoderma lucidum mycelium polysaccharide extract contains ≥30wt% β-glucan and ≥80wt% polysaccharide. The microencapsulated compound probiotics are prepared from compound probiotics and encapsulation materials; The compound probiotics include Lactobacillus plantarum and Bacillus subtilis; The embedding materials include sodium alginate and chitosan; The fat content of the black soldier fly larvae defatting powder is ≤5 wt%; The particle size of the defatted powder of black soldier fly larvae is 150~180μm; The β-glucan content in the co-fermented product of Phellinus linteus and Hericium erinaceus is ≥8 wt%. The particle size of the Antarctic krill powder is ≤50μm; In the compound fruit and vegetable fermentation slurry, the mass ratio of the fruit, vegetables, and fermentation substrate is 2~3:1~2:1; The inoculation amount of the fermentation strain is 3-5% of the total mass of the fruit, vegetables, and fermentation substrate; The mass ratio of xylooligosaccharide to inulin is 1:1~2.

2. The method for preparing a functional fresh pet food according to claim 1, characterized in that, Includes the following steps: (1) The defatted powder of black soldier fly larvae, the fermented medicinal fungus and Antarctic krill powder were mixed and enzymatically hydrolyzed with exogenous enzymes to obtain a composite protein matrix; (2) The composite protein matrix obtained in step (1) is mixed with composite fruit and vegetable fermentation pulp, prebiotics, cold-pressed flaxseed oil, Ganoderma lucidum mycelium polysaccharide extract and microencapsulated composite probiotics, and then homogenized, shaped and sterilized in sequence to obtain the functional pet food. In step (1), the exogenous enzymes include alkaline protease and flavor protease.

3. The method for preparing a functional fresh pet food according to claim 2, characterized in that, In step (1), the preparation steps of the black soldier fly larvae defatting powder include: subjecting fresh black soldier fly larvae to ultrasonic-assisted enzymatic hydrolysis, centrifuging to separate the fat content in the precipitate to ≤5wt%, drying the precipitate, sieving, and the sieved material is black soldier fly larvae defatting powder; The fat content of the fresh black soldier fly larvae is ≤20wt%; The ultrasonic frequency for the ultrasonic-assisted enzymatic hydrolysis is 30~50kHz. The enzyme used in the ultrasound-assisted enzymatic hydrolysis is lipase. The lipase activity is ≥20000 U / g; The amount of lipase added is 0.4-0.6% of the weight of fresh black soldier fly larvae; The temperature for ultrasound-assisted enzymatic hydrolysis is 40~50℃; The time for ultrasound-assisted enzymatic hydrolysis is 1-2 hours; The pH of the ultrasound-assisted enzymatic hydrolysis is 6.7~7.2; The centrifugal separation speed is 2500~3500 rpm.

4. The method for preparing a functional fresh pet food according to claim 2, characterized in that, In step (1), the preparation steps of the medicinal fungal fermentation product include: using wheat bran, soybean meal, and tea residue as substrates, adjusting the moisture content to 53-57%, inoculating the spore suspension of Phellinus linteus and the mycelial suspension of Hericium erinaceus, co-fermenting, drying, and obtaining the Phellinus linteus-Hericium erinaceus co-fermentation product, i.e., the medicinal fungal fermentation product; The mass ratio of wheat bran, soybean meal, and tea residue is 5~7:2~4:1; The tea residue is green tea residue; The number of spores in the *Phellinus linteus* spore suspension is ≥1×10 6 spores / mL; The bacterial content of the Hericium erinaceus mycelial suspension is ≥1×10⁻⁶. 6 CFU / mL; The volume ratio of the *Phellinus linteus* spore suspension to the *Hericium erinaceus* mycelial suspension is 2-4:1; The total inoculum amount of Phellinus linteus spore suspension and Hericium erinaceus mycelial suspension is 5-10%; The temperature for co-fermentation is 25~28℃; The content of β-glucan in the fermentation product at the time of termination of co-fermentation is ≥8 wt%; The drying temperature is 35~45℃; The drying time is 5-7 hours.

5. The method for preparing a functional fresh pet food according to claim 2, characterized in that, In step (1), the mass ratio of alkaline protease to flavor protease is 1~2:1; The alkaline protease has an enzyme activity ≥200,000 U / g; The enzyme activity of the flavor protease is ≥30000 U / g; The total amount of alkaline protease and flavor protease added is 4500~5500U per gram of protein; The enzymatic hydrolysis temperature is 45~50℃; The enzymatic hydrolysis time is 100-150 min; The pH during the enzymatic hydrolysis process is 6.8~7.2; Intermittent ultrasound is applied during the enzymatic hydrolysis process; The frequency of the ultrasound is 30~50kHz, and the working time of the ultrasound is 4~6 minutes every 30 minutes.

6. The method for preparing a functional fresh pet food according to claim 2, characterized in that, In step (2), the preparation steps of the compound fruit and vegetable fermentation slurry include: mixing the fruit, vegetables and fermentation substrate, inoculating with fermentation bacteria, fermenting, and enzymatically hydrolyzing the fermentation product after fermentation to obtain the compound fruit and vegetable fermentation slurry. The fermentation temperature is 30~37℃; The fermentation time is 12-72 hours; The enzymatic hydrolysis was performed using pectinase; The pectinase activity is ≥100,000 U / g; The amount of pectinase added is 45-55 U per gram of fermentation product; The enzymatic hydrolysis temperature is 35~45℃; The enzymatic hydrolysis time is 1-2 hours.

7. The method for preparing a functional fresh pet food according to claim 2, characterized in that, In step (2), the preparation steps of the microencapsulated compound probiotics include: encapsulating the compound probiotics with an encapsulation material to obtain microencapsulated compound probiotics; The compound probiotics include Lactobacillus plantarum and Bacillus subtilis; The embedding materials include sodium alginate and chitosan.

8. The method for preparing a functional fresh pet food according to claim 7, characterized in that, The compound probiotics are a mixture of Lactobacillus plantarum suspension and Bacillus subtilis suspension; The bacterial content of the *Lactobacillus plantarum* suspension is 1×10⁻⁶. 6 ~1×10 9 CFU / mL; The bacterial content of the Bacillus subtilis suspension is ≥1×10⁻⁶. 9 CFU / mL; The volume ratio of the *Lactobacillus plantarum* suspension to the *Bacillus subtilis* suspension is 1~2:1; The embedding material is sodium alginate and chitosan; The mass ratio of sodium alginate to chitosan is 3~5:1; The encapsulation process involves mixing the compound probiotics with an aqueous solution of sodium alginate, electrostatically dropping the mixture into a CaCl2 solution to form gel beads, and then coating the gel beads with an acetic acid solution containing chitosan.

9. The method for preparing a functional fresh pet food according to claim 2, characterized in that, In step (2), the pressure of the homogenizer is 45~55MPa; The rotational speed of the homogenizer is 3000~4000 rpm; The homogenization time is 25-35 minutes; The sterilization process employs high-voltage electrostatic field coupled ultraviolet pulse sterilization.

10. The application of the functional fresh pet food according to claim 1, or the functional fresh pet food prepared by the preparation method according to any one of claims 2 to 9, in the preparation of products that enhance the immune function of pets; The products include health supplements and / or pharmaceuticals.

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