Functional pet fresh food and preparation method thereof

Abstract

The invention discloses functional pet fresh food and a preparation method thereof, and belongs to the technical field of pet food. The functional fresh food for pets provided by the invention is prepared from the following components in parts by mass: 50-60 parts of hermetia illucens larva degreased powder, 20-30 parts of a medicinal fungus fermented product, 8-10 parts of euphausia superba powder, 3.5-5 parts of compound fruit and vegetable fermented pulp, 3-4 parts of prebiotics, 3-4 parts of cold-pressed linseed oil, 2-3 parts of a ganoderma lucidum mycelium polysaccharide extract and 0.5-1 part of microencapsulated compound probiotics. The pet fresh food provided by the invention can overcome the problems of high protein sensitization, insufficient functional component activity retention, short shelf life and the like of the traditional pet fresh food in the prior art, has the functions of regulating intestinal tracts, enhancing immunity and the like, and is suitable for industrial production.

Description

Technical Field

[0001] This application belongs to the technical field of pet food, and particularly relates to a functional fresh pet food and a preparation method thereof. Background Art

[0002] With the popularization of pet raising, pet owners' attention to pet health has been continuously increasing. In recent years, fresh pet food has gradually gained market favor, and its fresh ingredients and rich nutritional components are more in line with the natural dietary needs of pets. However, currently, the commercially available fresh pet food generally has the following problems: First, the protein source is single: mainly relying on traditional animal proteins such as chicken and beef, which is prone to cause allergies and has a large environmental burden; Second, serious nutritional loss: high-temperature sterilization (above 121°C) leads to the inactivation of heat-sensitive functional components (such as probiotics and active polysaccharides); Third, dependence on synthetic additives: artificial attractants or preservatives need to be added, which affects the long-term health of pets; Fourth, the contradiction between shelf life and nutritional retention: the refrigerated shelf life of conventional fresh pet food is short (usually ≤14 days), and extending the shelf life requires sacrificing nutritional components. Although the prior art has tried to use insect protein, it has not solved the problems of low protein bioavailability and the synergy of functional components; there is also a technology that discloses the application of fungal polysaccharides, but it does not combine with the low-temperature sterilization process. Therefore, there is an urgent need to develop a new type of fresh pet food with high digestibility, long shelf life, and multiple health effects. Summary of the Invention

[0003] To overcome the problems in the prior art such as high protein allergenicity, insufficient retention of functional component activity, and short shelf life of traditional fresh pet food, this application provides a functional fresh pet food and a preparation method thereof.

[0004] To achieve the above-mentioned invention purpose, this application provides the following technical solutions:

[0005] On the one hand, this application provides a functional fresh pet food, which is prepared from components comprising the following parts by mass: 50 - 60 parts of defatted black soldier fly larvae powder, 20 - 30 parts of medicinal fungus fermentate, 8 - 10 parts of Antarctic krill powder, 3.5 - 5 parts of compound fruit and vegetable fermentation pulp, 3 - 4 parts of prebiotics, 3 - 4 parts of cold-pressed linseed oil, 2 - 3 parts of ganoderma mycelium polysaccharide extract, 0.5 - 1 part of microencapsulated compound probiotics;

[0006] The medicinal fungus fermentate is a co-fermentate of Phellinus linteus and Hericium erinaceus;

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

[0008] The fruits include at least one of apple pomace, blueberries, and pineapple peels;

[0009] The vegetables include at least one of carrots, purple cabbages, and pumpkins;

[0010] The fermentation substrate includes at least one of fructooligosaccharide and yeast extract;

[0011] The fermentation strains include at least one of Lactobacillus plantarum and Saccharomyces boulardii;

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

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

[0014] The microencapsulated compound probiotics are prepared from compound probiotics and embedding materials;

[0015] The compound probiotics include Lactobacillus plantarum and Bacillus subtilis;

[0016] The embedding materials include at least one of sodium alginate and chitosan.

[0017] Optionally, the functional fresh pet food contains the following components in parts by mass: 55 parts of defatted black soldier fly larvae powder, 25 parts of medicinal fungus fermentate, 9 parts of Antarctic krill powder, 4 parts of compound fruit and vegetable fermentation slurry, 3.5 parts of prebiotic, 3.5 parts of cold-pressed linseed oil, 2.5 parts of Ganoderma lucidum mycelium polysaccharide extract, and 0.8 parts of microencapsulated compound probiotics.

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

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

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

[0021] Optionally, in the Sanghuang-Hericium erinaceus co-fermentate, the content of β-glucan is ≥8 wt%.

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

[0023] Optionally, in the compound fruit and vegetable fermentation slurry, the mass ratio of the fruit, vegetable, and fermentation substrate is 2 - 3:1 - 2:1;

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

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

[0026] Optionally, the inoculation amount of the fermentation strains is independently selected from any value of 3%, 4%, 5% of the total mass of fruits, vegetables, and fermentation substrates, or a range value between any two of them.

[0027] Optionally, the mixing mass ratio of xylooligosaccharide to inulin is 1:1 to 2.

[0028] Optionally, the mixing mass ratio of xylooligosaccharide to inulin is 1:1.

[0029] In a second aspect, the present application provides a method for preparing the above-mentioned functional fresh pet food, including the following steps:

[0030] (1) Mix the defatted black soldier fly larvae powder, medicinal fungus fermentate, and Antarctic krill powder, and perform enzymatic hydrolysis with exogenous enzymes to obtain a composite protein matrix;

[0031] (2) Mix the composite protein matrix obtained in step (1) with the composite fruit and vegetable fermentation pulp, prebiotics, cold-pressed linseed oil, Ganoderma lucidum mycelium polysaccharide extract, and microencapsulated composite probiotics, and perform homogenization, forming, and sterilization in sequence to obtain the functional fresh pet food;

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

[0033] Optionally, in step (1), the preparation steps of the defatted black soldier fly larvae powder include: subjecting fresh black soldier fly larvae to ultrasonic-assisted enzymatic hydrolysis, centrifugally separating until the fat content in the precipitate is ≤5 wt%, drying and sieving the precipitate, and the material passing through the sieve is the defatted black soldier fly larvae powder;

[0034] The fat content of the fresh black soldier fly larvae is ≤20 wt%;

[0035] The ultrasonic frequency of the ultrasonic-assisted enzymatic hydrolysis is 30 - 50 kHz;

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

[0037] The enzyme activity of the lipase is ≥20000 U / 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 temperature of the ultrasonic-assisted enzymatic hydrolysis is 40 - 50 °C;

[0040] The time of the ultrasonic-assisted enzymatic hydrolysis is 1 - 2 h;

[0041] The pH of the ultrasonic-assisted enzymatic hydrolysis is 6.7 - 7.2;

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

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

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

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

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

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

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

[0049] Optionally, in step (1), the preparation steps of the medicinal fungal ferment include: using wheat bran, soybean meal, and green tea residue as substrates, adjusting the water content to 53 - 57%, inoculating the Phellinus igniarius spore suspension and the Hericium erinaceus mycelium suspension, performing co-fermentation, and drying to obtain the Phellinus igniarius-Hericium erinaceus co-ferment, namely the medicinal fungal ferment;

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

[0051] The green tea residue is green tea residue;

[0052] The number of spores in the Phellinus igniarius spore suspension ≥ 1×10 6 spores / mL;

[0053] The bacterial content of the Hericium erinaceus mycelium suspension ≥ 1×10 6 CFU / mL;

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

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

[0056] The temperature of the co-fermentation is 25-28 °C;

[0057] When the co-fermentation ends, the content of β-glucan in the fermented product is ≥8 wt%;

[0058] The temperature of the drying is 35-45 °C;

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

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

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

[0062] Optionally, the total inoculation amount of the Sanghuang spore suspension and the Hericium erinaceus mycelium suspension independently selects any value from 5%, 6%, 7%, 8%, 9%, 10% or the range value between any two of them.

[0063] Optionally, the temperature of the co-fermentation independently selects any value from 25 °C, 26 °C, 27 °C, 28 °C or the range value between any two of them.

[0064] Optionally, the temperature of the drying independently selects any value from 35 °C, 38 °C, 40 °C, 43 °C, 45 °C or the range value between any two of them

[0065] Optionally, the time of the drying independently selects any value from 5 h, 6 h, 7 h or the range value between any two of them.

[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 ≥200000 U / g;

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

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

[0070] The temperature of the enzymatic hydrolysis is 45-50 °C;

[0071] The time of the enzymatic hydrolysis is 100-150 min;

[0072] The pH during the enzymatic hydrolysis is 6.8-7.2;

[0073] Intermittent ultrasonic waves are applied during the enzymatic hydrolysis process;

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

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

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

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

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

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

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

[0081] Optionally, the working time of the ultrasonic waves is 5 minutes every 30 minutes.

[0082] Optionally, in step (2), the preparation steps of the composite fruit and vegetable fermentation slurry include: mixing the fruits, vegetables, and fermentation matrix, inoculating fermentation bacteria, performing fermentation, and enzymatically hydrolyzing the fermentation product after fermentation to obtain the composite fruit and vegetable fermentation slurry;

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

[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 ≥ 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°C;

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

[0104] The embedding is to mix the compound probiotics with an aqueous solution of sodium alginate and electrostatically drop it into a CaCl 2 solution to form gel beads, and then coat the gel beads with an acetic acid solution containing chitosan.

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

[0106] The rotation speed of the homogenization is 3000-4000 rpm;

[0107] The time of the homogenization is 25-35 min;

[0108] The sterilization adopts high-voltage electrostatic field coupled with ultraviolet pulse sterilization.

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

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

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

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

[0113] The products include health products and / or drugs.

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

[0115] (1) The functional fresh pet food provided by this application scientifically combines black soldier fly larvae protein with medicinal fungus ferment and Antarctic krill powder to form a composite protein system with full-spectrum amino acids and rich in flavonoids (such as hispidin) and precursors of nerve growth factor (NGF). It not only solves the problems of easy sensitization and high carbon emissions of traditional animal protein sources (chicken / beef), but also realizes the dual synergistic effects of pet cognitive function + immune enhancement. The composite fruit and vegetable fermentation slurry added to the fresh pet food formula of this application is rich in effective ingredients such as pectin, anthocyanin, bromelain, β-carotene, and glucosinolate. It not only solves the problems of dependence on synthetic additives and single fiber source in fresh pet food, but also enhances the functional value through biotransformation, achieving the purposes of prebiotic-probiotic synergistic regulation of intestinal health, enhancing antioxidant and anti-inflammatory effects, etc. The lactic acid bacteria fermentation used can also produce ethyl acetate (fruity aroma) and 2,3-butanedione (buttery aroma), and yeast metabolism can also reduce the astringency of vegetables, so as to improve the palatability of fresh pet food. The microencapsulated composite probiotics added in this application solve the problems of functional component stability and bioavailability through microencapsulation. Using the sodium alginate-chitosan double-layer embedding method, it breaks through the application bottleneck of probiotics in fresh pet food and has the effects of targeted intestinal release, regulation of intestinal flora, and enhancement of body immunity. In addition, this application also adds components such as prebiotics, cold-pressed linseed oil, and Ganoderma lucidum mycelium polysaccharide extract to further endow the fresh pet food with rich nutrition and synergistically improve the efficacy of the fresh pet food with the above-mentioned composite protein system, composite fruit and vegetable fermentation slurry, and microencapsulated composite probiotics.

[0116] (2) The preparation method of the functional fresh pet food provided by this application performs mixed enzymatic hydrolysis on the composite protein system, which can cut the peptide bonds inside the protein (near hydrophobic amino acid residues), generate polypeptide fragments, and further hydrolyze the peptide chain ends to release free amino acids, thereby improving the protein bioavailability. At the same time, enzymatic hydrolysis 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 by enzymatic hydrolysis of black soldier fly. In addition, this application uses low-temperature enzymatic hydrolysis (up to 50 °C), which can also protect heat-sensitive components (such as Ganoderma lucidum polysaccharide), and intermittent ultrasonic assistance can promote the contact between enzyme and substrate. The combination of low-temperature biological processing + physical field sterilization technology developed in this application can achieve large-scale production using conventional food processing equipment while ensuring functional activity, and is applicable to the fresh pet food manufacturing industry.

[0117] (3) Experimental results have shown that when using the functional fresh pet food provided by this application, the protein digestibility can be increased to 92.3%, the serum IgG and IgA levels are significantly increased, the shelf life ≥ 12 months (refrigerated at 4 °C) and no preservatives are required. It can be seen that the fresh pet food provided by this application has broad application prospects in the preparation of products for improving the balance of pet intestinal flora and / or enhancing pet immune function, and can effectively extend the shelf life of fresh pet food, especially suitable for companion animals such as dogs and cats. Detailed implementation manners

[0118] The following further elaborates on this application in conjunction with specific embodiments. The following descriptions are merely several embodiments of this application and do not impose any form of limitation on this application. Although this application is disclosed below with preferred embodiments, it is not intended to limit this application. Any person skilled in the relevant art, without departing from the scope of the technical solution of this application, making some changes or modifications using the disclosed technical content is equivalent to equivalent implementation cases and all fall within the scope of the technical solution.

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

[0120] Unless otherwise specified, the analysis methods in the embodiments adopt the conventional settings and conventional analysis methods of the instruments or equipment.

[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 fermenting agents

[0125]

[0126]

[0127] Table 3 Enzyme preparations

[0128]

[0129] Table 4 Functional additives

[0130]

[0131]

[0132] Example 1

[0133] A preparation method of a functional fresh pet food, the steps are as follows:

[0134] (1) Fresh black soldier fly larvae (fat content ≤ 20%) were subjected to ultrasonic-assisted lipase hydrolysis (30 kHz, 40 °C, pH 6.7, 1 h, lipase activity 20000 U / g, lipase addition amount 0.4% of the mass of fresh black soldier fly larvae), centrifuged (2500 rpm) until the fat content in the precipitate was ≤ 5 wt%, then the precipitate was dried by hot air (50 °C, 2 h), sieved, and the material passing through the sieve was the defatted powder of black soldier fly larvae (particle size 150 μm);

[0135] (2) Using wheat bran, soybean meal, and green tea residue as substrates (wheat bran:soybean meal:green tea residue = 5:2:1, mass ratio), adjusting the water content to 53%, inoculating with a suspension of Sanghuang spores and a suspension of Hericium erinaceus mycelia (wherein, the Sanghuang spore suspension: scraping spores with 0.05% Tween 80 sterile water, filtering (200-mesh sieve) and adjusting to 1×10 6 spores / mL, the Hericium erinaceus mycelia suspension: crushing mycelial pellets under sterile conditions (ultrasonic for 3 min, 40 kHz), adjusting to a bacterial content of 1×10 6 CFU / mL, the volume ratio of the Sanghuang spore suspension to the Hericium erinaceus mycelia suspension is 2:1, and the total inoculation amount of the Sanghuang spore suspension and the Hericium erinaceus mycelia suspension is 5%), co-fermenting for 7 days at 25 °C, a humidity of 70% and in the dark. The end standard of fermentation is that the β-glucan content in the mycelium is 8.12 wt% (determined by the phenol-sulfuric acid method), and then the fermentation product is dried at a low temperature of 35 °C for 5 h to obtain the Sanghuang-Hericium erinaceus co-fermented product, that is, the medicinal fungus fermented product;

[0136] (3) Mixing a suspension of Lactobacillus plantarum and a suspension of Bacillus subtilis (both with the bacterial concentration adjusted to 1×10 9 CFU / mL) in a volume ratio of 1:1 to obtain a mixed bacterial suspension; dissolving sodium alginate (2%, w / v) in deionized water at 55 °C, sterilizing at 121 °C for 15 min to obtain a sodium alginate solution; dissolving chitosan (degree of deacetylation ≥ 90%) in 0.5% acetic acid solution (pH 5.0) at a concentration of 0.5% (w / v) to obtain a chitosan solution; mixing the mixed bacterial suspension and the sodium alginate solution in a volume ratio of 1:1, electrostatically dripping into a CaCl 2 solution (its own concentration 0.1 M, containing 0.05% Tween 80), with the parameters set as voltage 8 kV, needle diameter 0.4 mm, needle dropping speed 15 drops / min, magnetic stirring (200 rpm), curing for 20 min to form gel beads in the range of 1.2 - 1.5 mm in diameter, then immersing the gel beads in the chitosan solution, slowly oscillating at 25 °C (50 rpm) for 15 min, taking them out and rinsing 3 times with PBS buffer at pH 7.0, and quick-freezing at -40 °C for 2 h to obtain microencapsulated composite probiotics;

[0137] (4) Mix fruits (apple pomace: blueberry: pineapple peel = 1:1:1, by mass), vegetables (carrot: red cabbage: pumpkin = 1:1:1, by mass), and a fermentation substrate (fructooligosaccharide: yeast extract = 1:1) in a mass ratio of 2:1:1. First, inoculate Lactobacillus plantarum (direct-inoculation freeze-dried bacterial powder without activation, viable cell count of 1×10 10 CFU / g of bacterial powder, inoculation amount of 5% (w / w)) at 37°C for 24 h. Then, inoculate Saccharomyces boulardii (inoculated in the form of a bacterial suspension, bacterial content of 1×10 10 CFU / mL of suspension, inoculation amount of 3% (v / w)) at 30°C for 48 h. After fermentation, enzymatically hydrolyze the fermentation product, add pectinase (45 U per gram of fermentation product) and treat at 35°C for 1 h, and then sterilize to obtain a composite fruit and vegetable fermentation slurry;

[0138] (5) Weigh the following raw materials by mass: 50 parts of defatted black soldier fly larvae powder, 20 parts of medicinal fungus fermentation product, 8 parts of Antarctic krill powder, 3.5 parts of composite fruit and vegetable fermentation slurry, 3 parts of prebiotics (xylooligosaccharide: inulin = 1:1, by mass), 3 parts of cold-pressed linseed oil, 2 parts of Ganoderma lucidum mycelium polysaccharide extract, and 0.5 part of microencapsulated composite probiotics;

[0139] (6) Mix the defatted black soldier fly larvae powder, medicinal fungus fermentation product, and Antarctic krill powder, and enzymatically hydrolyze with alkaline protease and flavor protease (1:1, by mass) at 45°C and pH 6.8 for 100 min. Among them, the enzyme activity of alkaline protease is 200000 U / g, the enzyme activity of flavor protease is 30000 U / g, the total enzyme addition amount is 4500 U per gram of protein, and intermittent ultrasonic waves (30 kHz, acting for 4 min every 30 min) are applied during the enzymatic hydrolysis process to obtain a composite protein matrix;

[0140] (7) Mix the composite protein matrix, microencapsulated composite probiotics, composite fruit and vegetable fermentation slurry with prebiotics, cold-pressed linseed oil, and Ganoderma lucidum mycelium polysaccharide extract, homogenize at 45 MPa and 3000 rpm for 25 min, and perform 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). Then, sterilize with high-voltage electrostatic field coupled with ultraviolet pulse (high-voltage electrostatic field treatment: the equipment is a parallel plate electrode (spacing 5 cm), parameters are 15 kV / cm, 10 min, environmental 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 functional pet fresh food.

[0141] Example 2

[0142] A method for preparing a functional pet fresh food, the steps are as follows:

[0143] (1) Fresh black soldier fly larvae (fat content ≤ 20%) are subjected to ultrasonic-assisted lipase hydrolysis (40 kHz, 45 °C, pH 7, 1.5 h, lipase activity 20000 U / g, lipase addition amount 0.5% of the mass of fresh black soldier fly larvae), centrifuged (3000 rpm) until the fat content in the precipitate is ≤ 5 wt%, then the precipitate is dried by hot air (50 °C, 2 h), sieved, and the material passing through the sieve is the defatted powder of black soldier fly larvae (particle size 160 μm);

[0144] (2) Using wheat bran, soybean meal, and green tea residue as substrates (wheat bran:soybean meal:green tea residue = 6:3:1, mass ratio), adjusting the water content to 55%, inoculating with a suspension of Inonotus sanghuang spores and a suspension of Hericium erinaceus mycelia (wherein, the suspension of Inonotus sanghuang spores: scraping spores with 0.05% Tween 80 sterile water, filtering (200-mesh sieve) and adjusting to 1×10 6 spores / mL, the suspension of Hericium erinaceus mycelia: crushing mycelial balls under sterile conditions (ultrasonic for 3 min, 40 kHz), adjusting to a bacterial content of 1×10 6 CFU / mL, the volume ratio of the suspension of Inonotus sanghuang spores to the suspension of Hericium erinaceus mycelia is 3:1, and the total inoculation amount of the suspension of Inonotus sanghuang spores and the suspension of Hericium erinaceus mycelia is 8%), co-fermenting for 7 days under the conditions of 27 °C, humidity 70% and in the dark. The end standard of fermentation is that the β-glucan content in the mycelium is 8.97 wt% (determined by the phenol-sulfuric acid method), and then the fermentation product is dried at a low temperature of 40 °C for 6 h to obtain the co-fermented product of Inonotus sanghuang and Hericium erinaceus, that is, the medicinal fungal fermented product;

[0145] (3) Mixing a suspension of Lactobacillus plantarum and a suspension of Bacillus subtilis (the bacterial concentrations of both are adjusted to 1×10 9 CFU / mL) according to a volume ratio of 1:1 to obtain a mixed bacterial suspension; dissolving sodium alginate (2%, w / v) in deionized water at 55 °C, sterilizing at 121 °C for 15 min to obtain a sodium alginate solution; dissolving chitosan (degree of deacetylation ≥ 90%) in 0.5% acetic acid solution (pH 5.0) with a concentration of 0.5% (w / v) to obtain a chitosan solution; mixing the mixed bacterial suspension and the sodium alginate solution according to a volume ratio of 1:1, electrostatically dripping into a CaCl 2 solution (its own concentration 0.1 M, containing 0.05% Tween 80), with parameters set as voltage 8 kV, needle diameter 0.4 mm, needle dropping speed 15 drops / min, magnetic stirring (200 rpm), and curing for 20 min to form gel beads with a diameter in the range 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, taken out and rinsed 3 times with PBS buffer at pH 7.0, and quick-frozen at -40 °C for 2 h to obtain microencapsulated composite probiotics;

[0146] (4) Mix fruits (apple pomace: blueberries: pineapple peels = 1:1:1, by mass), vegetables (carrots: purple cabbage: pumpkin = 1:1:1, by mass), and a fermentation substrate (fructooligosaccharides: yeast extract = 1:1) in a mass ratio of 2.5:1.5:1. First, inoculate Lactobacillus plantarum (no activation required, direct-inoculation freeze-dried bacterial powder, viable cell count of 1×10 10 CFU / g of bacterial powder, inoculation amount of 5% (w / w)) at 37°C for 24 h, then inoculate Saccharomyces boulardii (inoculated in the form of a bacterial suspension, bacterial content of 1×10 10 CFU / mL of suspension, inoculation amount of 3% (v / w)) at 30°C for 48 h. After fermentation, enzymatically hydrolyze the fermentation product, add pectinase (50 U per gram of fermentation product) and treat at 40°C for 1.5 h, then sterilize to obtain a composite fruit and vegetable fermentation slurry;

[0147] (5) Weigh the following raw materials by mass: 55 parts of defatted black soldier fly larvae powder, 25 parts of medicinal fungus fermentation product, 9 parts of Antarctic krill powder, 4 parts of composite fruit and vegetable fermentation slurry, 3.5 parts of prebiotics (xylooligosaccharides: inulin = 1:1, by mass), 3.5 parts of cold-pressed linseed oil, 2.5 parts of Ganoderma lucidum mycelium polysaccharide extract, and 0.8 part of microencapsulated composite probiotics;

[0148] (6) Mix the defatted black soldier fly larvae powder, medicinal fungus fermentation product, and Antarctic krill powder, and enzymatically hydrolyze at 47°C and pH 7 for 120 min using alkaline protease and flavor protease (1:1, by mass). Among them, the enzyme activity of alkaline protease is 200000 U / g, the enzyme activity of flavor protease is 30000 U / g, and the total enzyme addition amount is 5000 U per gram of protein. Apply intermittent ultrasonic waves (40 kHz, acting for 5 min every 30 min) during the enzymatic hydrolysis process to obtain a composite protein matrix;

[0149] (7) Mix the composite protein matrix, microencapsulated composite probiotics, composite fruit and vegetable fermentation slurry with prebiotics, cold-pressed linseed oil, and Ganoderma lucidum mycelium polysaccharide extract, and homogenize at 50 MPa and 3500 rpm for 30 min, then perform 3D printing and 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). Then, use high-voltage electrostatic field coupled with ultraviolet pulse sterilization (high-voltage electrostatic field treatment: the equipment is a parallel plate electrode (spacing 5 cm), parameters are 15 kV / cm, 10 min, environmental 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 functional fresh pet food.

[0150] Example 3

[0151] A preparation method of a functional fresh pet food, the steps are as follows:

[0152] (1) Perform ultrasonic-assisted lipase hydrolysis on fresh black soldier fly larvae (fat content ≤ 20%) (50 kHz, 50 °C, pH 7.2, 2 h, lipase activity 20000 U / g, lipase addition amount 0.6% of the mass of fresh black soldier fly larvae), centrifuge (3500 rpm) until the fat content in the precipitate ≤ 5 wt%, then perform hot air drying on the precipitate (50 °C, 2 h), sieve, and collect the material under the sieve to obtain black soldier fly larvae defatted powder (particle size 180 μm);

[0153] (2) Use wheat bran, soybean meal, and green tea residue as substrates (wheat bran:soybean meal:green tea residue = 7:4:1, mass ratio), adjust the water content to 57%, inoculate with Sanghuang spore suspension and Hericium erinaceus mycelium suspension (wherein, Sanghuang spore suspension: scrape spores with 0.05% Tween 80 sterile water, filter (200-mesh sieve) and adjust to 1×10 6 spores / mL, Hericium erinaceus mycelium suspension: break mycelial balls under sterile conditions (ultrasonic for 3 min, 40 kHz), adjust to a bacterial content of 1×10 6 CFU / mL, the volume ratio of the Sanghuang spore suspension to the Hericium erinaceus mycelium suspension is 4:1, and the total inoculation amount of the Sanghuang spore suspension and the Hericium erinaceus mycelium suspension is 10%), and carry out co-fermentation for 7 days at 28 °C, humidity 70% and in the dark. The end standard of fermentation is that the β-glucan content in the mycelium is 8.23 wt% (determined by the phenol-sulfuric acid method), and then the fermentation product is dried at low temperature at 45 °C for 7 h to obtain a Sanghuang-Hericium erinaceus co-fermentation product, that is, a medicinal fungus fermentation product;

[0154] (3) Mix the Lactobacillus plantarum suspension and the Bacillus subtilis suspension (the bacterial concentrations of both are adjusted to 1×10 9 CFU / mL) according to a volume ratio of 2:1 to obtain a mixed bacterial suspension; dissolve sodium alginate (2%, w / v) in deionized water at 55 °C, sterilize at 121 °C for 15 min to obtain a sodium alginate solution; dissolve chitosan (deacetylation degree ≥ 90%) in 0.5% acetic acid solution (pH 5.0) with a concentration of 0.5% (w / v) to obtain a chitosan solution; mix the mixed bacterial suspension and the sodium alginate solution according to a volume ratio of 1:1, and electrostatically drip into CaCl 2Solution (self - concentration 0.1 M, containing 0.05% Tween 80), parameters set as voltage 8 kV, needle diameter 0.4 mm, needle dropping speed 15 drops / min, magnetic stirring (200 rpm), curing for 20 min to form gel beads with a diameter in the range of 1.2 - 1.5 mm. Then immerse the gel beads in chitosan solution, slowly oscillate at 25°C (50 rpm) for 15 min, take out and rinse 3 times with PBS buffer at pH 7.0, and quick - freeze at - 40°C for 2 h to obtain microencapsulated composite probiotics;

[0155] (4) Mix fruits (apple pomace: blueberry: pineapple peel = 1:1:1, mass ratio), vegetables (carrot: purple cabbage: pumpkin = 1:1:1, mass ratio), and fermentation substrate (fructooligosaccharide: yeast extract = 1:1) in a mass ratio of 3:2:1. First, inoculate Lactobacillus plantarum (no need to activate, direct - injection freeze - dried powder, viable cell count 1×10 10 CFU / g of powder, 5% (w / w) inoculation amount) at 37°C for 24 h, then inoculate Saccharomyces boulardii (inoculated in the form of cell suspension, cell content 1×10 10 CFU / mL of suspension, 3% (v / w) inoculation amount) at 30°C for 48 h. After fermentation, enzymatically hydrolyze the fermentation product, add pectinase (55 U per gram of fermentation product) and treat at 45°C for 2 h, and sterilize to obtain composite fruit and vegetable fermentation pulp;

[0156] (5) Weigh the following raw materials in parts by mass: 60 parts of defatted black soldier fly larvae powder, 30 parts of medicinal fungus fermentation product, 10 parts of Antarctic krill powder, 5 parts of composite fruit and vegetable fermentation pulp, 4 parts of prebiotics (xylo - oligosaccharide: inulin = 1:2, mass ratio), 4 parts of cold - pressed linseed oil, 3 parts of Ganoderma lucidum mycelium polysaccharide extract, and 1 part of microencapsulated composite probiotics;

[0157] (6) Mix the defatted black soldier fly larvae powder, medicinal fungus fermentation product and Antarctic krill powder, and enzymatically hydrolyze with alkaline protease and flavor protease (1:1, mass ratio) at 50°C and pH 7.2 for 150 min. Among them, the enzyme activity of alkaline protease is 200000 U / g, the enzyme activity of flavor protease is 30000 U / g, the total enzyme addition amount is 5500 U per gram of protein, and intermittent ultrasonic waves (50 kHz, acting for 6 min every 30 min) are applied during the enzymatic hydrolysis process to obtain a composite protein matrix;

[0158] (7) Mix the composite protein matrix, microencapsulated composite probiotics, composite fruit and vegetable fermentation pulp with prebiotics, cold-pressed linseed oil, and Ganoderma lucidum mycelium polysaccharide extract, homogenize at 55 MPa and 4000 rpm for 35 min, and perform 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). Then, use high-voltage electrostatic field coupled with ultraviolet pulse sterilization (high-voltage electrostatic field treatment: the equipment is parallel plate electrodes (spacing 5 cm), parameters are 15 kV / cm, 10 min, environmental humidity 50%; ultraviolet pulse assistance: wavelength is 254 nm + 185 nm (ozone generation), intensity is 50 mJ / cm 2 , and the pulse frequency is 5 Hz) to obtain the functional fresh pet food.

[0159] Comparative Example 1

[0160] A method for preparing fresh pet food, the steps are as follows:

[0161] The difference from Example 2 is only that the component of the medicinal fungus fermentate is omitted:

[0162] (1) Perform ultrasonic-assisted lipase hydrolysis on fresh black soldier fly larvae (fat content ≤ 20%) (40 kHz, 45 °C, pH 7, 1.5 h, lipase activity is 20000 U / g, lipase addition amount is 0.5% of the mass of fresh black soldier fly larvae), centrifuge (3000 rpm) until the fat content in the precipitate ≤ 5 wt%, then perform hot air drying on the precipitate (50 °C, 2 h), sieve, and collect the material under the sieve, which is the defatted powder of black soldier fly larvae (particle size is 160 μm);

[0163] (2) Mix the suspension of Lactobacillus plantarum and the suspension of Bacillus subtilis (the bacterial concentrations of both are adjusted to 1×10 9 CFU / mL) in a volume ratio of 1:1 to obtain a mixed bacterial suspension; dissolve sodium alginate (2%, w / v) in deionized water at 55 °C, sterilize at 121 °C for 15 min to obtain a sodium alginate solution; dissolve chitosan (degree of deacetylation ≥ 90%) in 0.5% acetic acid solution (pH 5.0) with a concentration of 0.5% (w / v) to obtain a chitosan solution; mix the mixed bacterial suspension and the sodium alginate solution in a volume ratio of 1:1, and electrostatically drop into CaCl 2Solution (self - concentration 0.1 M, containing 0.05% Tween 80), parameters are set as voltage 8 kV, needle diameter 0.4 mm, needle dropping speed 15 drops / min, magnetic stirring (200 rpm), curing for 20 min to form gel beads in the range of 1.2 - 1.5 mm in diameter, then the gel beads are immersed in chitosan solution, slowly oscillated at 25°C (50 rpm) for 15 min, taken out and rinsed 3 times with PBS buffer at pH 7.0, and quick - frozen at - 40°C for 2 h to obtain microencapsulated composite probiotics;

[0164] (3) Mix fruits (apple pomace: blueberry: pineapple peel = 1:1:1, by mass), vegetables (carrot: purple cabbage: pumpkin = 1:1:1, by mass), and fermentation substrate (fructooligosaccharide: yeast extract = 1:1) in a mass ratio of 2.5:1.5:1. First, inoculate Lactobacillus plantarum (no need to activate, direct - inoculation freeze - dried powder, viable count 1×10 10 CFU / g of powder, 5% (w / w) inoculation amount) at 37°C for 24 h, then inoculate Saccharomyces boulardii (inoculated in the form of bacterial suspension, bacterial content 1×10 10 CFU / mL of suspension, 3% (v / w) inoculation amount) at 30°C for 48 h. After fermentation, enzymatically hydrolyze the fermentation product, add pectinase (50 U per gram of fermentation product) and treat at 40°C for 1.5 h, then sterilize to obtain composite fruit and vegetable fermentation pulp;

[0165] (4) Weigh the following raw materials in parts by mass: 55 parts of defatted black soldier fly larvae powder, 25 parts of medicinal fungus fermentation product, 9 parts of Antarctic krill powder, 4 parts of composite fruit and vegetable fermentation pulp, 3.5 parts of prebiotics (xylooligosaccharide: inulin = 1:1, by mass), 3.5 parts of cold - pressed linseed oil, 2.5 parts of Ganoderma lucidum mycelium polysaccharide extract, 0.8 parts of microencapsulated composite probiotics;

[0166] (5) Mix the defatted black soldier fly larvae powder and Antarctic krill powder, and enzymatically hydrolyze with alkaline protease and flavor protease (1:1, by mass) at 47°C and pH 7 for 120 min. Among them, the enzyme activity of alkaline protease is 200000 U / g, the enzyme activity of flavor protease is 30000 U / g, the total enzyme addition amount is 5000 U per gram of protein, and intermittent ultrasonic waves (40 kHz, acting for 5 min every 30 min) are applied during the enzymatic hydrolysis process to obtain a composite protein matrix;

[0167] (6) Mix the composite protein matrix, microencapsulated composite probiotics, composite fruit and vegetable fermentation broth with prebiotics, cold-pressed linseed oil, and Ganoderma lucidum mycelium polysaccharide extract, and homogenize at 50 MPa and 3500 rpm for 30 min, then perform 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 use high-voltage electrostatic field coupled with ultraviolet pulse sterilization (high-voltage electrostatic field treatment: the equipment is a parallel plate electrode (spacing 5 cm), parameters are 15 kV / cm, 10 min, environmental 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.

[0168] Comparative Example 2

[0169] A method for preparing pet fresh food, the steps are as follows:

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

[0171] (1) Perform ultrasonic-assisted lipase hydrolysis on fresh black soldier fly larvae (fat content ≤ 20%) (40 kHz, 45 °C, pH 7, 1.5 h, lipase activity 20000 U / g, lipase addition amount is 0.5% of the mass of fresh black soldier fly larvae), centrifuge (3000 rpm) until the fat content in the precipitate ≤ 5 wt%, then perform hot air drying on the precipitate (50 °C, 2 h), sieve, and collect the material under the sieve as defatted black soldier fly larvae powder (particle size 160 μm);

[0172] (2) Use wheat bran, soybean meal, and green tea residue as the matrix (wheat bran:soybean meal:green tea residue = 6:3:1, mass ratio), adjust the water content to 55%, inoculate with Sanghuang spore suspension and Hericium erinaceus mycelium suspension (wherein, Sanghuang spore suspension: scrape spores with 0.05% Tween 80 sterile water, filter (200-mesh sieve) and adjust to 1×10 6 spores / mL, Hericium erinaceus mycelium suspension: break mycelial balls under sterile conditions (ultrasonic for 3 min, 40 kHz), adjust to a bacterial content of 1×10 6 CFU / mL, the volume ratio of the Sanghuang spore suspension to the Hericium erinaceus mycelium suspension is 3:1, and the total inoculation amount of the Sanghuang spore suspension and the Hericium erinaceus mycelium suspension is 8%), and perform co-fermentation for 7 days at 27 °C, humidity 70% and in the dark. The end standard of fermentation is that the β-glucan content of the mycelium is 8.95 wt% (determined by the phenol-sulfuric acid method), and then the fermentation product is dried at low temperature at 40 °C for 6 h to obtain the Sanghuang-Hericium erinaceus co-fermented product, that is, the medicinal fungus fermented product;

[0173] (3) Mix the Lactobacillus plantarum suspension and the Bacillus subtilis suspension (both with the bacterial concentration adjusted to 1×10 9 CFU / mL) in a volume ratio of 1:1 to obtain a mixed bacterial suspension; dissolve sodium alginate (2%, w / v) in deionized water at 55°C and sterilize it at 121°C for 15 min to obtain a sodium alginate solution; dissolve chitosan (degree of deacetylation ≥90%) in a 0.5% acetic acid solution (pH 5.0) at a concentration of 0.5% (w / v) to obtain a chitosan solution; mix the mixed bacterial suspension and the sodium alginate solution in a volume ratio of 1:1, and electrostatically drip it into a CaCl 2 solution (its own concentration is 0.1 M, containing 0.05% Tween 80), with the parameters set as voltage 8 kV, needle diameter 0.4 mm, needle dropping speed of 15 drops / min, magnetic stirring (200 rpm), and curing for 20 min to form gel beads with a diameter in the range of 1.2 - 1.5 mm. Then immerse the gel beads in the chitosan solution, slowly oscillate (50 rpm) at 25°C for 15 min, take them out and rinse them 3 times with PBS buffer at pH 7.0, and quick-freeze at -40°C for 2 h to obtain microencapsulated composite probiotics;

[0174] (4) Mix fruits (apple pomace: blueberry: pineapple peel = 1:1:1, mass ratio), vegetables (carrot: purple cabbage: pumpkin = 1:1:1, mass ratio), and fermentation substrates (fructooligosaccharide: yeast extract = 1:1) in a mass ratio of 2.5:1.5:1. First, inoculate Lactobacillus plantarum (no need to activate, direct-fed freeze-dried bacterial powder, viable bacteria count is 1×10 10 CFU / g bacterial powder, 5% (w / w) inoculation amount) at 37°C for 24 h, then inoculate Saccharomyces boulardii (inoculated in the form of a bacterial suspension, bacterial content is 1×10 10 CFU / mL suspension, 3% (v / w) inoculation amount) at 30°C for 48 h. After fermentation, enzymatically hydrolyze the fermentation product, add pectinase (50 U per gram of fermentation product) and treat it at 40°C for 1.5 h, and then sterilize it to obtain a composite fruit and vegetable fermentation slurry;

[0175] (5) Weigh the following raw materials in parts by mass: 55 parts of defatted black soldier fly larvae powder, 25 parts of medicinal fungus fermentation product, 9 parts of Antarctic krill powder, 4 parts of composite fruit and vegetable fermentation slurry, 3.5 parts of prebiotics (xylooligosaccharide: inulin = 1:1, mass ratio), 3.5 parts of cold-pressed linseed oil, 2.5 parts of Ganoderma lucidum mycelium polysaccharide extract, and 0.8 part of microencapsulated composite probiotics;

[0176] (6) Mix the defatted black soldier fly larvae powder, medicinal fungus fermentate, Antarctic krill powder, microencapsulated compound probiotics, compound fruit and vegetable fermented pulp, prebiotics, cold-pressed linseed oil, and Ganoderma lucidum mycelium polysaccharide extract, and homogenize at 50 MPa and 3500 rpm for 30 min. Then, perform 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). Subsequently, use high-voltage electrostatic field coupled with ultraviolet pulse sterilization (high-voltage electrostatic field treatment: the equipment is parallel plate electrodes (spacing 5 cm), parameters are 15 kV / cm, 10 min, environmental humidity 50%; ultraviolet pulse assistance: wavelength is 254 nm + 185 nm (ozone generation), intensity is 50 mJ / cm 2 , and pulse frequency is 5 Hz) to obtain the pet fresh food.

[0177] Comparative Example 3

[0178] A preparation method of pet fresh food is as follows:

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

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

[0181] (2) Use wheat bran, soybean meal, and green tea residue as the matrix (wheat bran:soybean meal:green tea residue = 6:3:1, mass ratio), adjust the water content to 55%, inoculate the Sanghuang spore suspension and Hericium erinaceus mycelium suspension (wherein, Sanghuang spore suspension: scrape spores with 0.05% Tween 80 sterile water, filter (200-mesh sieve) and adjust to 1×10 6 spores / mL, Hericium erinaceus mycelium suspension: break the mycelial balls under sterile conditions (ultrasound for 3 min, 40 kHz), and adjust to a bacterial content of 1×10 6 CFU / mL, the volume ratio of the Sanghuang spore suspension to the Hericium erinaceus mycelium suspension is 3:1, and the total inoculation amount of the Sanghuang spore suspension and the Hericium erinaceus mycelium suspension is 8%). Ferment for 7 days at 27 °C, humidity 70%, and in the dark. The fermentation end standard is that the β-glucan content of the mycelium is 8.96 wt% (determined by the phenol-sulfuric acid method). Then, low-temperature dry the fermentation product at 40 °C for 6 h to obtain the Sanghuang-Hericium erinaceus co-fermentate, that is, the medicinal fungus fermentate;

[0182] (3) Mix the Lactobacillus plantarum suspension and the Bacillus subtilis suspension (the bacterial concentrations of both are adjusted to 1×10 9 CFU / mL) in a volume ratio of 1:1 to obtain a mixed bacterial suspension; dissolve sodium alginate (2%, w / v) in deionized water at 55°C and sterilize it at 121°C for 15 min to obtain a sodium alginate solution; dissolve chitosan (degree of deacetylation ≥90%) in 0.5% acetic acid solution (pH 5.0) at a concentration of 0.5% (w / v) to obtain a chitosan solution; mix the mixed bacterial suspension and the sodium alginate solution in a volume ratio of 1:1 and electrostatically drip them into CaCl 2 solution (its own concentration is 0.1 M, containing 0.05% Tween 80), with the parameters set as voltage 8 kV, needle diameter 0.4 mm, needle dropping speed of 15 drops / min, magnetic stirring (200 rpm), and curing for 20 min to form gel beads with a diameter in the range of 1.2 - 1.5 mm. Then immerse the gel beads in the chitosan solution, slowly oscillate (50 rpm) at 25°C for 15 min, take them out, rinse them 3 times with PBS buffer at pH 7.0, and quickly freeze them at -40°C for 2 h to obtain microencapsulated composite probiotics;

[0183] (4) Mix fruits (apple pomace: blueberry: pineapple peel = 1:1:1, mass ratio), vegetables (carrot: purple cabbage: pumpkin = 1:1:1, mass ratio), and deionized water in a mass ratio of 2.5:1.5:1, beat them, and sterilize them to obtain a composite fruit and vegetable pulp;

[0184] (5) Weigh the following raw materials in parts by mass: 55 parts of defatted black soldier fly larvae powder, 25 parts of medicinal fungus fermentate, 9 parts of Antarctic krill powder, 4 parts of composite fruit and vegetable pulp, 3.5 parts of prebiotics (xylooligosaccharide: inulin = 1:1, mass ratio), 3.5 parts of cold-pressed linseed oil, 2.5 parts of Ganoderma lucidum mycelium polysaccharide extract, and 0.8 part of microencapsulated composite probiotics;

[0185] (6) Mix the defatted black soldier fly larvae powder, the medicinal fungus fermentate, and the Antarctic krill powder, and enzymatically hydrolyze them with alkaline protease and flavor protease (1:1, mass ratio) at 47°C and pH 7 for 120 min. Among them, the enzyme activity of alkaline protease is 200000 U / g, the enzyme activity of flavor protease is 30000 U / g, the total enzyme addition amount is 5000 U per gram of protein, and intermittent ultrasonic waves (40 kHz, acting for 5 min every 30 min) are applied during the enzymatic hydrolysis process to obtain a composite protein matrix;

[0186] (7) Mix the composite protein matrix, microencapsulated composite probiotics, composite fruit and vegetable pulp with prebiotics, cold-pressed linseed oil, and Ganoderma lucidum mycelium polysaccharide extract, homogenize at 50 MPa and 3500 rpm for 30 min, and perform 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). Then, perform high-voltage electrostatic field coupled with ultraviolet pulse sterilization (high-voltage electrostatic field treatment: the 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 , and the pulse frequency is 5 Hz) to obtain the fresh pet food.

[0187] Comparative Example 4

[0188] A method for preparing fresh pet food, the steps are as follows:

[0189] The difference from Example 2 is only that Lactobacillus plantarum in the microencapsulated composite probiotics is replaced with Bifidobacterium adolescentis:

[0190] (1) Perform ultrasonic-assisted lipase hydrolysis on fresh black soldier fly larvae (fat content ≤ 20%) (40 kHz, 45 °C, pH 7, 1.5 h, lipase activity is 20000 U / g, lipase addition amount is 0.5% of the mass of fresh black soldier fly larvae), centrifuge (3000 rpm) until the fat content in the precipitate is ≤ 5 wt%, then perform hot air drying on the precipitate (50 °C, 2 h), sieve, and collect the material under the sieve as black soldier fly larvae defatted powder (particle size is 160 μm);

[0191] (2) Use wheat bran, soybean meal, and green tea residue as the matrix (wheat bran:soybean meal:green tea residue = 6:3:1, mass ratio), adjust the water content to 55%, inoculate with Sanghuang spore suspension and Hericium erinaceus mycelium suspension (wherein, Sanghuang spore suspension: scrape spores with 0.05% Tween 80 sterile water, filter (200-mesh sieve) and adjust to 1×10 6 spores / mL, Hericium erinaceus mycelium suspension: break mycelial balls under sterile conditions (ultrasound for 3 min, 40 kHz), adjust to a bacterial content of 1×10 6 CFU / mL, the volume ratio of the Sanghuang spore suspension to the Hericium erinaceus mycelium suspension is 3:1, and the total inoculation amount of the Sanghuang spore suspension and the Hericium erinaceus mycelium suspension is 8%). Carry out co-fermentation for 7 days at 27 °C, humidity 70% and in the dark. The end standard of fermentation is that the β-glucan content of the mycelium is 8.94 wt% (determined by the phenol-sulfuric acid method). Then, low-temperature dry the fermentation product at 40 °C for 6 h to obtain the Sanghuang-Hericium erinaceus co-fermentation product, that is, the medicinal fungus fermentation product;

[0192] (3) Mix the Bifidobacterium adolescentis suspension and the Bacillus subtilis suspension (the bacterial concentrations of both are adjusted to 1×10 9 CFU / mL) in a volume ratio of 1:1 to obtain a mixed bacterial suspension; dissolve sodium alginate (2%, w / v) in deionized water at 55°C and sterilize it at 121°C for 15 min to obtain a sodium alginate solution; dissolve chitosan (degree of deacetylation ≥90%) in a 0.5% acetic acid solution (pH 5.0) at a concentration of 0.5% (w / v) to obtain a chitosan solution; mix the mixed bacterial suspension and the sodium alginate solution in a volume ratio of 1:1, and electrostatically drip them into a CaCl 2 solution (its own concentration is 0.1 M, containing 0.05% Tween 80), with the parameters set as voltage 8 kV, needle diameter 0.4 mm, needle dropping speed of 15 drops / min, magnetic stirring (200 rpm), and curing for 20 min to form gel beads with a diameter in the range of 1.2 - 1.5 mm. Then immerse the gel beads in the chitosan solution, slowly oscillate (50 rpm) at 25°C for 15 min, take them out, rinse them 3 times with PBS buffer at pH 7.0, and quickly freeze them at -40°C for 2 h to obtain microencapsulated composite probiotics;

[0193] (4) Mix fruits (apple pomace: blueberry: pineapple peel = 1:1:1, mass ratio), vegetables (carrot: purple cabbage: pumpkin = 1:1:1, mass ratio), and fermentation substrates (fructooligosaccharide: yeast extract = 1:1) in a mass ratio of 2.5:1.5:1. First, inoculate Lactobacillus plantarum (no need to activate, direct - injection freeze - dried bacterial powder, viable bacteria count is 1×10 10 CFU / g bacterial powder, 5% (w / w) inoculation amount) at 37°C for 24 h, then inoculate Saccharomyces boulardii (inoculated in the form of a bacterial suspension, bacterial content is 1×10 10 CFU / mL suspension, 3% (v / w) inoculation amount) at 30°C for 48 h. After fermentation, enzymatically hydrolyze the fermentation product, add pectinase (50 U per gram of fermentation product) and treat it at 40°C for 1.5 h, and then sterilize it to obtain a composite fruit and vegetable fermentation slurry;

[0194] (5) Weigh the following raw materials in parts by mass: 55 parts of defatted black soldier fly larvae powder, 25 parts of medicinal fungus fermentate, 9 parts of Antarctic krill powder, 4 parts of composite fruit and vegetable fermentation slurry, 3.5 parts of prebiotics (xylooligosaccharide: inulin = 1:1, mass ratio), 3.5 parts of cold - pressed linseed oil, 2.5 parts of Ganoderma lucidum mycelium polysaccharide extract, and 0.8 part of microencapsulated composite probiotics;

[0195] (6) Mix the defatted black soldier fly larvae powder, the medicinal fungus fermentate, and Antarctic krill powder, and enzymatically hydrolyze them with alkaline protease and flavor protease (1:1, mass ratio) at 47 °C and pH 7 for 120 min. Among them, the enzyme activity of alkaline protease is 200,000 U / g, the enzyme activity of flavor protease is 30,000 U / g, the total enzyme addition amount is 5000 U per gram of protein, and intermittent ultrasonic waves (40 kHz, acting for 5 min every 30 min) are applied during the enzymatic hydrolysis process to obtain a composite protein matrix;

[0196] (7) Mix the composite protein matrix, microencapsulated composite probiotics, composite fruit and vegetable fermented pulp, prebiotics, cold-pressed linseed oil, and Ganoderma lucidum mycelium polysaccharide extract, homogenize at 50 MPa and 3500 rpm for 30 min, and perform 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). Then, use high-voltage electrostatic field coupled with ultraviolet pulse sterilization (high-voltage electrostatic field treatment: the equipment is parallel plate electrodes (spacing 5 cm), the parameters are 15 kV / cm, 10 min, and environmental humidity 50%; ultraviolet pulse assistance: wavelength is 254 nm + 185 nm (ozone generation), intensity is 50 mJ / cm 2 , and pulse frequency 5 Hz) to obtain the pet fresh food.

[0197] Comparative Example 5

[0198] A preparation method of pet fresh food, the steps are as follows:

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

[0200] (1) Perform ultrasonic-assisted lipase hydrolysis on fresh black soldier fly larvae (fat content ≤ 20%) (40 kHz, 45 °C, pH 7, 1.5 h, lipase enzyme activity is 20,000 U / g, and the lipase addition amount is 0.5% of the mass of fresh black soldier fly larvae), centrifuge (3000 rpm) until the fat content in the precipitate ≤ 5 wt%, then perform hot air drying (50 °C, 2 h) on the precipitate, sieve, and collect the undersize as defatted black soldier fly larvae powder (particle size is 160 μm);

[0201] (2) Use wheat bran, soybean meal, and green tea residue as the matrix (wheat bran:soybean meal:green tea residue = 6:3:1, mass ratio), adjust the water content to 55%, and inoculate the Sanghuang spore suspension and Hericium erinaceus mycelium suspension (among them, the Sanghuang spore suspension: scrape the spores with 0.05% Tween 80 sterile water, filter (200-mesh sieve) and adjust to 1×10 6spores / mL, Hericium erinaceus mycelium suspension: The mycelial pellets were broken under sterile conditions (ultrasonic treatment for 3 min at 40 kHz), and adjusted to a bacterial content of 1×10 6 CFU / mL. The volume ratio of the Sanghuang spore suspension to the Hericium erinaceus mycelium suspension was 3:1, and the total inoculation amount of the Sanghuang spore suspension and the Hericium erinaceus mycelium suspension was 8%). Co-fermentation was carried out for 7 days at 27°C, 70% humidity and in the dark. The end standard of fermentation was that the β-glucan content of the mycelium was 8.93 wt% (determined by the phenol-sulfuric acid method). Then, the fermentation product was dried at a low temperature of 40°C for 6 h to obtain the Sanghuang-Hericium erinaceus co-fermentation product, that is, the medicinal fungus fermentation product;

[0202] (3) Mix the Lactobacillus plantarum suspension and the Bacillus subtilis suspension (the bacterial concentrations of both were adjusted to 1×10 9 CFU / mL) in a volume ratio of 1:1 to obtain a composite probiotic;

[0203] (4) Mix fruits (apple pomace: blueberry: pineapple peel = 1:1:1, mass ratio), vegetables (carrot: purple cabbage: pumpkin = 1:1:1, mass ratio), and fermentation substrate (fructooligosaccharide: yeast extract = 1:1) in a mass ratio of 2.5:1.5:1. First, inoculate Lactobacillus plantarum (no activation required, direct-inoculation freeze-dried bacterial powder, viable bacteria count of 1×10 10 CFU / g bacterial powder, 5% (w / w) inoculation amount) at 37°C for 24 h, and then inoculate Saccharomyces boulardii (inoculated in the form of a bacterial suspension, bacterial content of 1×10 10 CFU / mL suspension, 3% (v / w) inoculation amount) at 30°C for 48 h. After fermentation, the fermentation product was enzymatically hydrolyzed, adding pectinase (50 U per gram of fermentation product) and treating at 40°C for 1.5 h, and then sterilized to obtain a composite fruit and vegetable fermentation slurry;

[0204] (5) Weigh the following raw materials in parts by mass: 55 parts of defatted black soldier fly larvae powder, 25 parts of medicinal fungus fermentation product, 9 parts of Antarctic krill powder, 4 parts of composite fruit and vegetable fermentation slurry, 3.5 parts of prebiotics (xylooligosaccharide: inulin = 1:1, mass ratio), 3.5 parts of cold-pressed linseed oil, 2.5 parts of Ganoderma lucidum mycelium polysaccharide extract, 0.8 part of composite probiotic;

[0205] (6) Mix the defatted black soldier fly larvae powder, the medicinal fungus fermentation product and the Antarctic krill powder, and enzymatically hydrolyze them with alkaline protease and flavor protease (1:1, mass ratio) at 47°C and pH 7 for 120 min. Among them, the enzyme activity of alkaline protease is 200000 U / g, the enzyme activity of flavor protease is 30000 U / g, and the total enzyme addition amount is 5000 U per gram of protein. Intermittent ultrasonic waves (40 kHz, acting for 5 min every 30 min) were applied during the enzymatic hydrolysis process to obtain a composite protein matrix;

[0206] (7) Mix the composite protein matrix, composite probiotics, composite fruit and vegetable fermentation slurry with prebiotics, cold-pressed linseed oil, and Ganoderma lucidum mycelium polysaccharide extract, homogenize at 50 MPa and 3500 rpm for 30 min, and perform 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). Then, use high-voltage electrostatic field coupled with ultraviolet pulse sterilization (high-voltage electrostatic field treatment: the equipment is parallel plate electrodes (spacing 5 cm), parameters are 15 kV / cm, 10 min, environmental humidity 50%; ultraviolet pulse assistance: wavelength is 254 nm + 185 nm (ozone generation), intensity is 50 mJ / cm 2 , and pulse frequency 5 Hz) to obtain the pet fresh food.

[0207] Experimental Example 1

[0208] In vitro digestibility determination experiment:

[0209] Taking Examples 1 to 3, Comparative Examples 1 to 5, and commercially available traditional pet fresh food (formula by mass percentage: deboned chicken breast: 50%, vegetables: 30% (20% pumpkin, 10% carrot), chicken liver: 10%, brown rice: 5%, supplement: 5% (2% fish oil, 2% calcium powder, 1% vitamin E), sterilized at 121 °C) as examples, verify the protein digestibility of different pet fresh foods and evaluate their amino acid release characteristics.

[0210] (1) Simulate the gastric-intestinal digestion model

[0211] Gastric phase:

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

[0213] Add pepsin (enzyme activity 10000 U / g) at 32 mg / 10 mL gastric juice, and digest at 37 °C with shaking (100 rpm) for 2 h.

[0214] Intestinal phase:

[0215] Adjust the pH to 6.8, and add 10 mL of simulated intestinal juice (containing 0.1 M NaHCO 3 , 0.005 M trypsin);

[0216] Continue to digest at 37 °C with shaking for 4 h, and take samples every 30 min to detect the degree of hydrolysis (DH).

[0217] (2) Key detection indicators

[0218] Protein digestibility: The nitrogen content of the undigested residue was determined by the Kjeldahl method, and the digestibility was calculated as follows:

[0219]

[0220] Amino acid release rate: The content of free amino acids in the digestive fluid was determined by HPLC (detecting essential amino acid: lysine).

[0221] The results are shown in Table 5.

[0222] Table 5 Protein digestibility and amino acid release of different fresh pet 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 Commercially available traditional fresh pet food 68.7 40.5

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

[0225] Experimental Example 2

[0226] Immune performance determination experiment:

[0227] Verify the improvement effect of different fresh pet foods on 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 (dogs) Weeks 0 and 8 Example 2 n = 10 (dogs) Weeks 0 and 8 Example 3 n = 10 (dogs) Weeks 0 and 8 Comparative Example 1 n = 10 (dogs) Weeks 0 and 8 Comparative Example 2 n = 10 (dogs) Weeks 0 and 8 Comparative Example 3 n = 10 (dogs) Weeks 0 and 8 Comparative Example 4 n = 10 (dogs) Weeks 0 and 8 Comparative Example 5 n = 10 (dogs) Weeks 0 and 8 Commercially available traditional fresh pet food n = 10 (dogs) Weeks 0 and 8 Basal diet n = 10 (dogs) Weeks 0 and 8

[0231] Animal selection: Healthy adult beagle dogs (weight 10-15 kg, age 1-2 years), randomly grouped and individually caged; individuals who had recently used antibiotics or immunosuppressants were excluded.

[0232] 2. Feeding management

[0233] Diet formula:

[0234] Experimental group: Fresh pet foods of Examples 1-3 of the present application;

[0235] Control group: Fresh pet foods of Comparative Examples 1-5 and commercially available traditional fresh pet foods (formula by mass percentage: boneless chicken breast: 50%, vegetables: 30% (20% pumpkin, 10% carrot), chicken liver: 10%, brown rice: 5%, supplement: 5% (2% fish oil, 2% calcium powder, 1% vitamin E), sterilized at 121 °C);

[0236] Blank group: Basal diet, containing only basic 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] Twice a day, feed at 5% of body weight, with free access to water;

[0239] Record the daily feed intake and health status.

[0240] 3. Blood sample collection and processing

[0241] Blood collection time: After fasting for 12 h, collect blood from the anterior limb vein at 8 am;

[0242] Sampling volume: 3 mL each time, aliquoted into a vacuum tube without anticoagulant;

[0243] Serum preparation:

[0244] Let stand at room temperature for 30 min → centrifuge at 3000 rpm for 15 min (4 °C) → aliquot the serum and store at -80 °C.

[0245] 4. Detection method (see Table 7)

[0246] Table 7 Detection method

[0247]

[0248] Operation process:

[0249] IgG detection:

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

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

[0252] IgA detection:

[0253] Mix the serum with anti-canine IgA antibody and react at 37 °C for 10 min;

[0254] Measure the turbidity change and calculate the concentration (mg / dL).

[0255] The results are shown in Table 8.

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

[0257]

[0258]

[0259] As can be seen from Table 8, the impact of the basal diet on immune indices fluctuates little, excluding environmental interference. Compared with Comparative Examples 1-5 and commercially available traditional fresh pet foods, the IgG and IgA levels in Examples 1-3 of this application are significantly increased, proving that the fresh pet food of this application can comprehensively activate the immune system of pets and enhance their immune performance.

[0260] Experimental Example 3

[0261] Standardized fecal score experiment:

[0262] Through the standardized fecal score system, verify the improvement effect of different fresh pet foods on the intestinal health of pet dogs.

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

[0264] Table 9 Experimental design

[0265]

[0266]

[0267] Animal selection:

[0268] Healthy adult beagle dogs (weight 10-15 kg, age 1-2 years), without a recent history of digestive tract diseases;

[0269] After 1 week of adaptive feeding, the formal experiment began.

[0270] 2. Fecal score criteria

[0271] Adopt the 5-point Bristol modified scale (pet-specific version), see Table 10:

[0272] Table 10 Bristol modified scale (pet-specific version)

[0273] Score Trait description Health significance 1 point Hard pellets, no mucus Risk of constipation 2 points Well-formed, slight cracks Ideal state 3 points Soft but formed, easy to pick up Mild indigestion 4 points Loose and amorphous, partially adherent Pre-diarrhea 5 points Watery stool, unable to form Severe diarrhea

[0274] Ideal range: 2-3 points.

[0275] 3. Feeding management

[0276] Diet formula:

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

[0278] Control group: the fresh foods of Comparative Examples 1-5 and commercially available traditional fresh pet foods (the formula is by mass percentage: boneless chicken breast: 50%, vegetables: 30% (20% pumpkin, 10% carrot), chicken liver: 10%, brown rice: 5%, supplement: 5% (2% fish oil, 2% calcium powder, 1% vitamin E), sterilized at 121 °C);

[0279] Blank group: Basal diet, containing only basic 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 plan:

[0281] Twice a day, feeding at 5% of body weight, with free access to water;

[0282] Record the daily feed intake and health status.

[0283] Fecal collection: Collect fresh samples at a fixed time every day (the first bowel movement 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 Immediate scoring according to the Bristol scale Independently scored blindly by 3 experimenters Water content Drying method (105 °C to constant weight) Calculation: (wet weight - dry weight) / wet weight × 100% Mucus detection rate Visual inspection + microscopic examination with physiological saline smear Record the proportion of the mucus-covered area

[0287] 5. Statistical analysis

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

[0289] The results are shown in Table 12.

[0290] Table 12 Improvement effects of different fresh pet foods on the intestinal health of pet dogs

[0291]

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

[0293] Experimental Example 4

[0294] Accelerated shelf life experiment:

[0295] Taking Example 2 and commercially available traditional fresh pet food (formula by mass percentage: boneless chicken breast: 50%, vegetables: 30% (20% pumpkin, 10% carrot), chicken liver: 10%, brown rice: 5%, supplement: 5% (2% fish oil, 2% calcium powder, 1% vitamin E), sterilized at 121 °C) as examples, verify the theoretical shelf life of different fresh pet foods under 4 °C refrigeration.

[0296] 1. Accelerated test conditions

[0297] (1)Acceleration 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] Condition Detection time point Equivalent time at 4 °C 37℃ Days 0, 14, 28, 42, 56 Months 0, 3, 6, 9, 12

[0302] (3)4°C real storage control:

[0303] Regularly detect (0, 3, 6, 9, 12 months) to verify the accuracy of the acceleration model.

[0304] 2. Detection methods and acceptance criteria (see Table 14)

[0305] Table 14 Detection methods and acceptance criteria

[0306]

[0307] 3. Method steps

[0308] (1)Sample preparation:

[0309] The samples of Example 2 and commercially available traditional fresh pet food are divided into 50 g portions, vacuum-sealed and numbered.

[0310] (2)Accelerated storage:

[0311] Place the samples in layers in a thermostatic and humidistatic chamber, avoiding stacking that may affect heat transfer.

[0312] (3)Regular detection:

[0313] Total aerobic bacteria count: Sterile sampling → Serial dilution → Spreading on PCA plates → Incubation at 37°C for 48 h for counting;

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

[0315] Water activity: Directly take 5 g of the sample for determination, and take the average value in parallel three times.

[0316] The results are shown in Tables 15 and 16.

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

[0318]

[0319] Table 16 4°C real storage verification

[0320] Time (months) Aerobic bacteria in Example 2 (CFU / g) POV in Example 2 (g / 100g) Aw 0 <![CDATA[1.5×10 2 > 0.08 0.82 6 <![CDATA[4.2×10 3 > 0.16 0.83 12 <![CDATA[9.1×10 3 > 0.23 0.84

[0321] As can be seen from Table 15 and Table 16, the commercially available traditional fresh pet food has deteriorated (excessive microorganisms + rancid oil) at the equivalent of 6 months, while Example 2 of this application still meets all the indicators at 56 days (equivalent to 12 months at 4°C) at 37°C; after real storage at 4°C for 12 months, the POV (0.23) of Example 2 of this application is close to but does not exceed the standard (≤0.25), verifying the reliability of the acceleration model. It can be seen that the shelf life of the fresh pet food of this application is ≥12 months at 4°C, which is significantly better than the commercially available products.

[0322] The above are only several embodiments of this application and do not impose any form of limitation on this application. Although this application is disclosed above with preferred embodiments, it is not intended to limit this application. Any person skilled in the art, without departing from the scope of the technical solution of this application, makes some changes or modifications using the technical content disclosed above, which are all equivalent to equivalent implementation cases and fall within the scope of the technical solution.

Claims

1. A functional fresh pet food, characterized in that: The invention is prepared from the following components in parts by weight: 50-60 parts of defatted black soldier fly larvae powder, 20-30 parts of fermented medicinal fungi, 8-10 parts of Antarctic krill powder, 3.5-5 parts of compound fruit and vegetable fermented pulp, 3-4 parts of prebiotics, 3-4 parts of cold-pressed linseed oil, 2-3 parts of polysaccharide extract from mycelium of Ganoderma lucidum, and 0.5-1 part of microencapsulated compound probiotics; The medicinal fungus fermentation product is a co-fermentation product of Phellinus linteus and Hericium erinaceus; The composite fruit and vegetable fermented pulp is prepared from fruits, vegetables, fermentation substrates and fermentation bacteria; The fruit includes at least one of apple pomace, blueberry, and pineapple peel; The vegetables include at least one of carrots, purple cabbage, and pumpkin; The fermentation substrate includes at least one of oligofructose and yeast extract; The fermentation bacteria include at least one of Lactobacillus plantarum and Saccharomyces boulardii; The prebiotic is a mixture of xylooligosaccharides and inulin; In the Ganoderma lucidum mycelium polysaccharide extract, the content of β-glucan is ≥30wt%, and the content of polysaccharide is ≥80wt%; The microencapsulated composite probiotics are prepared from composite probiotics and embedding materials; The composite probiotics include Lactobacillus plantarum and Bacillus subtilis; The embedding material includes at least one of sodium alginate and chitosan.

2. The functional fresh pet food according to claim 1, characterized in that: The fat content of the black soldier fly larvae defatted powder is ≤5wt%; The particle size of the black soldier fly larvae defatted powder is 150 to 180 μm; Preferably, the content of β-glucan in the co-fermentation product of Phellinus linteus and Hericium erinaceus is ≥ 8wt%; Preferably, the particle size of the Antarctic krill powder is ≤50 μm.

3. The functional fresh pet food according to claim 1, characterized in that: In the composite fruit and vegetable fermentation slurry, the mass ratio of the fruit, the vegetable, and the fermentation substrate is 2 to 3:1 to 2:1; The inoculation amount of the fermentation bacteria is 3-5% of the total mass of the fruits, vegetables and fermentation substrates.

4. The functional fresh pet food according to claim 1, characterized in that: The mass ratio of the xylo-oligosaccharide to inulin is 1:1-2.

5. The method for preparing a functional fresh pet food according to any one of claims 1 to 4, characterized in that: The steps include: (1) mixing the black soldier fly larvae defatted powder, the fermented medicinal fungus and the Antarctic krill powder, and performing enzymolysis with exogenous enzymes to obtain a composite protein matrix; (2) mixing the composite protein matrix obtained in step (1) with composite fruit and vegetable fermented pulp, prebiotics, cold-pressed linseed oil, Ganoderma lucidum mycelium polysaccharide extract, and microencapsulated composite probiotics, and sequentially homogenizing, molding, and sterilizing to obtain the functional pet fresh food; In step (1), the exogenous enzymes include alkaline protease and flavor protease.

6. The method for preparing functional fresh pet food according to claim 5, characterized in that: In step (1), the preparation step of the black soldier fly larvae defatted powder comprises: subjecting fresh black soldier fly larvae to ultrasonic-assisted enzymatic hydrolysis, centrifuging until the fat content in the precipitate is ≤5wt%, drying and sieving the precipitate, and the sieved material is the black soldier fly larvae defatted powder; The fat content of the fresh black soldier fly larvae is ≤20wt%; The ultrasonic frequency of the ultrasonic-assisted enzymatic hydrolysis is 30 to 50 kHz; The enzyme used in the ultrasonic-assisted enzymatic hydrolysis is lipase; The lipase activity is ≥20000U / g; The amount of lipase added is 0.4-0.6% of the mass of fresh black soldier fly larvae; The temperature of the ultrasonic-assisted enzymatic hydrolysis is 40-50°C; The ultrasound-assisted enzymatic hydrolysis time is 1 to 2 hours; The pH of the ultrasound-assisted enzymatic hydrolysis is 6.7 to 7.2; The rotation speed of the centrifugal separation is 2500-3500 rpm; Preferably, in step (1), the preparation step of the medicinal fungus fermentation product comprises: using wheat bran, soybean meal and tea residue as a matrix, adjusting the water content to 53-57%, inoculating a spore suspension of Phellinus linteus and a mycelium suspension of Hericium erinaceus, co-fermenting, and drying to obtain a co-fermentation product of Phellinus linteus and Hericium erinaceus, i.e., a medicinal fungus fermentation product; The mass ratio of the 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 spore suspension of Phellinus linteus is ≥ 1×10 6 spores / mL; The bacterial content of the Hericium erinaceus mycelium suspension is ≥1×10 6 CFU / mL; The volume ratio of the mulberry linterus spore suspension to the Hericium erinaceus mycelium suspension is 2 to 4:1; The total inoculation amount of the phellus linterus spore suspension and the Hericium erinaceus mycelium suspension is 5-10%; The co-fermentation temperature is 25-28°C; When the co-fermentation is terminated, the content of β-glucan in the fermentation product is ≥ 8wt%; The drying temperature is 35-45°C; The drying time is 5 to 7 hours.

7. The method for preparing functional fresh pet food according to claim 5, characterized in that: In step (1), the mass ratio of alkaline protease to flavor protease is 1 to 2:1; The enzymatic activity of the alkaline protease is ≥ 200000 U / g; The enzyme activity of the flavor protease is ≥30000U / g; The total amount of alkaline protease and flavor protease added is 4500-5500U per gram of protein; The temperature of the enzymatic hydrolysis is 45-50°C; The enzymatic hydrolysis time is 100 to 150 minutes; The pH during the enzymatic hydrolysis process is 6.8 to 7.2; Applying intermittent ultrasound during the enzymatic hydrolysis process; The frequency of the ultrasonic wave is 30 to 50 kHz, and the working time of the ultrasonic wave is 4 to 6 minutes every 30 minutes.

8. The method for preparing functional fresh pet food according to claim 5, characterized in that: In step (2), the preparation steps of the composite fruit and vegetable fermentation slurry include: mixing the fruits, vegetables and fermentation substrate, inoculating fermentation bacteria, fermenting, and enzymolyzing the fermentation product after the fermentation is completed to obtain the composite fruit and vegetable fermentation slurry; The fermentation temperature is 30-37°C; The fermentation time is 12 to 72 hours; The enzymatic hydrolysis is carried out using pectinase; The enzymatic activity of the pectinase is ≥100000 U / g; The amount of pectinase added is 45-55U per gram of fermentation product; The temperature of the enzymatic hydrolysis is 35-45°C; The enzymatic hydrolysis time is 1 to 2 hours.

9. The method for preparing functional fresh pet food according to claim 5, characterized in that: In step (2), the preparation step of the microencapsulated composite probiotics includes: embedding the composite probiotics with an embedding material to obtain the microencapsulated composite probiotics; The composite probiotics include Lactobacillus plantarum and Bacillus subtilis; The embedding material comprises at least one of sodium alginate and chitosan; Preferably, the composite probiotic is a mixture of a Lactobacillus plantarum suspension and a Bacillus subtilis suspension; The bacterial content of the plant lactobacillus suspension is 1×10 6 ~1×10 9 CFU / mL; The bacterial content of the Bacillus subtilis suspension is ≥ 1×10 9 CFU / mL; The volume ratio of the Lactobacillus plantarum suspension to the Bacillus subtilis suspension is 1 to 2:1; The embedding materials are sodium alginate and chitosan; The mass ratio of sodium alginate to chitosan is 3 to 5:1; The embedding is to mix the composite probiotics with an aqueous solution of sodium alginate, electrostatically drip a CaCl2 solution into it to form gel beads, and then coat the gel beads with an acetic acid solution containing chitosan; Preferably, in step (2), the homogenization pressure is 45-55 MPa; The homogenizing speed is 3000-4000 rpm; The homogenization time is 25 to 35 minutes; The sterilization adopts high-voltage electrostatic field coupled ultraviolet pulse sterilization.

10. Use of the functional fresh pet food according to any one of claims 1 to 4, or the functional fresh pet food prepared by the preparation method according to claim 5, in preparing products for improving the balance of pet intestinal flora and / or enhancing the immune function of pets; The products include health products and / or medicines.

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