A composition for improving immunity and its use in pet food

Abstract

The present application relates to a kind of composition for improving immunity and its application in pet food, belong to pet food technical field.The composition for improving immunity of the present application includes the following raw materials: chrysanthemum powder, medlar powder, pear powder, magnolia powder, the plant powder described above is uniformly mixed and then fermented using red yeast with preservation number CCTCC NO:M2015296, to obtain the composition for improving immunity.The composition for improving immunity described in the present application can significantly improve the immunity of pets, enhance the physical quality of pets, especially for pets recovering from serious illness or young pets, and the effect is very good.

Description

Technical Field

[0001] This invention relates to the field of pet food technology, and more particularly to a composition for enhancing immunity and its application in pet food. Background Technology

[0002] Pets have become important members of many families, and their health and welfare are receiving increasing attention. Immunity is the core of maintaining a pet's health and resisting disease. For pets whose immune systems are not yet fully developed in their early years, those in their old age whose functions are declining, or those recovering from serious illnesses and whose bodies are weak, how to safely and effectively enhance their immunity is a continuous focus of the pet nutrition and food technology field.

[0003] Currently, commercially available pet foods or additives with immune-enhancing functions mainly fall into the following technical categories: Directly adding nutrients: such as vitamins (e.g., vitamin C, vitamin E), minerals (e.g., zinc, selenium), and amino acids (e.g., taurine, arginine) to the formula. These substances are essential for maintaining the normal function of the immune system, but their mechanisms of action are relatively basic, and their effects on the overall regulation and enhancement of the immune system are limited. Moreover, relatively high amounts are usually required to show results.

[0004] Traditional Chinese medicine extracts are commonly used, such as water or alcohol extraction of single or compound Chinese herbs with immune-enhancing effects like Astragalus membranaceus, Codonopsis pilosula, and Poria cocos, and then adding the extracts to pet food. However, this method has significant drawbacks: First, the plant cell walls are not destroyed, resulting in low dissolution rates of active ingredients and low bioavailability; second, the extracts may have a strong medicinal taste, affecting the palatability for pets; third, simple physical mixing or solvent extraction fails to transform the macromolecules in the raw materials, potentially generating new active ingredients, and the immune-enhancing effect often falls short of expectations.

[0005] Adding probiotics or prebiotics can indirectly affect the immune system by regulating the gut microbiota. This is a worthwhile approach, but its mechanism of action is indirect, its effects are slow, and the results are inconsistent depending on the strain and individual differences.

[0006] Fermentation technology, as a biotransformation method, has been applied in the food and health product industries. It can degrade macromolecules into smaller, more easily absorbed molecules and may generate new active ingredients. However, research on fermenting specific combinations of plant raw materials with specific probiotic strains and applying them to pet food to synergistically enhance immunity is still limited. In particular, how to maximize the immune-enhancing potential of raw materials through fermentation processes and apply them safely and effectively in pet food is a direction worth exploring in the current pet food technology field.

[0007] Therefore, there is an urgent need to develop a new, safe, and efficient composition and its application scheme that can significantly enhance the immunity of pets for the market to choose from. Summary of the Invention

[0008] The purpose of this invention is to overcome the shortcomings of the prior art and provide a composition for enhancing immunity and its application in pet food.

[0009] To achieve the above objectives, the technical solution adopted by the present invention is as follows: In a first aspect, the present invention provides a composition for enhancing immunity, comprising the following raw materials in parts by weight: Chrysanthemum powder: 6-9 parts; Goji berry powder: 1-3 parts; Pear powder: 5-7 parts; Magnolia officinalis powder: 12-15 parts; The preparation method of the above composition includes the following steps: S1: Mix chrysanthemum powder, wolfberry powder, pear powder, and magnolia bark powder evenly to obtain a mixed powder; S2: Mix deionized water and mixed powder evenly to obtain a mixed solution, wherein the ratio of mixed powder to deionized water is 1:20-40 g / mL; S3: Sterilize the mixture, then inoculate with the inoculum, ferment for 20-30 hours, sterilize after fermentation, filter to obtain fermentation filtrate, freeze-dry the fermentation filtrate to obtain the composition.

[0010] Preferably, the inoculum used in the culture is Rhodotorula glutinis with preservation number CCTCC NO: M2015296; the viable count of the inoculum is 1×10⁻⁶. 7 -1×10 9 CFU / mL; the volume ratio of inoculum to mixed solution is 1:19-24.

[0011] Preferably, the fermentation temperature in step S3 is 25-30℃ and the fermentation pH is 4.0-6.0.

[0012] In a second aspect, the present invention provides the use of the immune-enhancing composition described in the first aspect in the preparation of pet food.

[0013] Thirdly, the present invention provides the use of the immune-enhancing composition described in the first aspect in the preparation of a pet nutritional fortifier.

[0014] Fourthly, the present invention provides the use of the immune-enhancing composition described in the first aspect in the preparation of pet food additives.

[0015] Fifthly, the present invention provides a pet food that enhances immunity, the pet food comprising the immunity-enhancing composition described in the first aspect.

[0016] Preferably, the aforementioned immune-boosting pet foods also include chondroitin sulfate, glucosamine hydrochloride, animal protein sources, grains, vitamins, and minerals.

[0017] Preferably, the animal protein source includes at least one of poultry meat meal, fish meal, and meat and bone meal; the grains include at least one of corn, wheat, rice, millet, peas, chickpeas, and oats; the vitamins include at least one of vitamin A, vitamin B, vitamin C, and vitamin E; and the minerals include at least one of copper salts, iron salts, manganese salts, zinc salts, calcium salts, and phosphates.

[0018] Sixthly, the present invention provides a pet food for enhancing immunity, the pet food comprising the following components by weight percentage: Composition to enhance immunity: 1-3 wt% Duck meat meal: 25-31 wt% Duck liver powder: 3-5 wt%; Chicken meal: 25-35 wt% Chicken liver powder: 4-7 wt% Chicken raw meat and bones: 1-3 wt% Duck meat and bones: 1-3wt% Taurine: 0.1-0.3 wt%; DL-methionine: 0.05-0.2 wt%; L-Lysine: 0.05-0.3 wt%; Chondroitin sulfate: 0.05-0.15 wt%; Glucosamine hydrochloride: 0.05-0.1 wt%; dicalcium phosphate: 1-5 wt% Vitamin A: 0.01-0.1 wt% Vitamin B: 0.01-0.1 wt% Vitamin C: 0.01-0.1 wt% Zinc citrate: 0.01-0.05 wt%; Ferric citrate: 0.01-0.05 wt%; Drinking water was replenished to 100 wt%.

[0019] Compared with the prior art, the beneficial effects of the present invention are as follows: The immune-enhancing composition provided by this invention uses chrysanthemum powder, wolfberry powder, pear powder, and magnolia bark powder in a specific ratio as a fermentation substrate, and employs Rhodotorula glutinis with preservation number CCTCC NO: M2015296 for directional fermentation. Significant synergistic effects are observed among the components, specifically in the following aspects: High Synergy Between Raw Material Formulation and Fermentation Strain: The chrysanthemum powder, wolfberry powder, pear powder, and magnolia bark powder used in this invention are not simply physical mixtures. Their specific mass ratio (6-9 parts chrysanthemum powder, 1-3 parts wolfberry powder, 5-7 parts pear powder, and 12-15 parts magnolia bark powder) forms a composite matrix that is highly compatible with the metabolic characteristics of a specific strain (Rhodotorula glutinis CCTCC NO: M2015296). This strain can efficiently utilize this specific ratio of plant components as a nutrient source, and its produced enzyme system can specifically degrade macromolecular active substances (such as polysaccharides and flavonoids) in the raw materials into small molecule active peptides, oligosaccharides, and novel metabolites that are more easily absorbed and utilized by the pet's body. This greatly improves the bioavailability and immune activity of the final composition, achieving a synergistic effect of "strain-substrate" compatibility.

[0020] Synergistic enhancement of functional activity through fermentation: By controlling specific fermentation conditions, not only was the growth and metabolism of Rhodotorula glutinis optimized, but more importantly, it promoted complex biotransformation reactions between the strain and the plant substrate. Compared to simple physical mixing or solvent extraction, this fermentation process produces components with greater immune-enhancing activity, resulting in a final composition that exhibits a synergistic effect in improving pet immunity that goes beyond the simple additive effect of individual components.

[0021] The comprehensive advantages of this composition in pet food: Adding this composition to the basic formula of pet food in a specific ratio not only significantly enhances the product's immunomodulatory function but also effectively improves the palatability of the pet food. This indicates that the flavor compounds in the composition prepared by the method of this invention are also optimized during the synergistic fermentation process, solving the industry problem of poor palatability that functional additives may cause, and achieving a synergistic improvement in efficacy and taste.

[0022] In summary, this invention, through innovative raw material compatibility, specific strain screening, and precise fermentation process control, has successfully prepared an immune-enhancing composition with significant synergistic effects among its components, high safety and efficiency, and good palatability, providing a new and effective solution for developing high-quality functional pet foods. Detailed Implementation

[0023] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0024] The sources of some of the raw materials used in this invention are as follows: Chrysanthemum powder: Guangzhou Kang Sheng Pharmaceutical Co., Ltd., batch number 250601; Goji berry powder: Guangzhou Kang Sheng Pharmaceutical Co., Ltd., batch number 250301; Pear powder: Guangzhou Kang Sheng Pharmaceutical Co., Ltd., batch number 250801; Magnolia officinalis powder: Guangzhou Kang Sheng Pharmaceutical Co., Ltd., batch number 250601; Red yeast rice: Preservation number: CCTCCNO M2015296, kindly donated by Guangdong Ocean University; All other raw materials and reagents used in this invention are commercially available.

[0025] Preparation of compositions for enhancing immunity: Composition 1 It is prepared from the following parts by weight of raw materials: Chrysanthemum powder: 8 parts; Goji berry powder: 2 parts; Pear powder: 6 parts; Magnolia officinalis powder: 13 parts; The method for preparing the composition includes the following steps: S1: Mix chrysanthemum powder, wolfberry powder, pear powder, and magnolia bark powder evenly to obtain a mixed powder; S2: Mix deionized water and mixed powder evenly to obtain a mixed solution, wherein the ratio of mixed powder to deionized water is 1:30 g / mL; S3: Sterilize the mixture, then inoculate with the inoculum, ferment for 24 hours, sterilize after fermentation, filter to obtain fermentation filtrate, freeze-dry the fermentation filtrate to obtain the composition; The inoculum used in the culture solution was Rhodotorula glutinis with accession number CCTCC NO M2015296; the viable count of the inoculum solution was 1×10⁻⁶. 8 CFU / mL; the volume ratio of inoculum to mixed solution is 1:23; The fermentation temperature in step S3 is 28°C; the initial fermentation pH is 5.0.

[0026] Composition 2 It is prepared from the following parts by weight of raw materials: Chrysanthemum powder: 6 parts; Goji berry powder: 1 part; Pear powder: 5 portions; Magnolia officinalis powder: 12 parts; The method for preparing the composition includes the following steps: S1: Mix chrysanthemum powder, wolfberry powder, pear powder, and magnolia bark powder evenly to obtain a mixed powder; S2: Mix deionized water and mixed powder evenly to obtain a mixed solution, wherein the ratio of mixed powder to deionized water is 1:20 g / mL; S3: Sterilize the mixture, then inoculate with the inoculum, ferment for 20 hours, sterilize after fermentation, filter to obtain fermentation filtrate, freeze-dry the fermentation filtrate to obtain the composition; The inoculum used in the culture solution was Rhodotorula glutinis with preservation number CCTCC NO: M2015296; the viable count of the inoculum solution was 1×10⁻⁶. 7 CFU / mL; the volume ratio of inoculum to mixed solution is 1:19; The fermentation temperature in step S3 is 25°C; the initial fermentation pH is 4.0.

[0027] Composition 3 It is prepared from the following parts by weight of raw materials: Chrysanthemum powder: 9 parts; Goji berry powder: 3 parts; Pear powder: 7 parts; Magnolia officinalis powder: 15 parts; The method for preparing the composition includes the following steps: S1: Mix chrysanthemum powder, wolfberry powder, pear powder, and magnolia bark powder evenly to obtain a mixed powder; S2: Mix deionized water and mixed powder evenly to obtain a mixed solution, wherein the ratio of mixed powder to deionized water is 1:40 g / mL; S3: Sterilize the mixture, then inoculate with the inoculum, ferment for 30 hours, sterilize after fermentation, filter to obtain fermentation filtrate, freeze-dry the fermentation filtrate to obtain the composition; The inoculum used in the culture solution was Rhodotorula glutinis with preservation number CCTCC NO: M2015296; the viable count of the inoculum solution was 1×10⁻⁶. 9 CFU / mL; the volume ratio of inoculum to mixed solution is 1:24; In step S3, the fermentation temperature is 30°C and the initial fermentation pH is 6.0.

[0028] Composition ① Unlike composition 1, this composition lacks chrysanthemum powder. The missing mass fraction is made up by wolfberry powder, pear powder, and magnolia bark powder in a mass ratio of 2:6:13. The other raw materials, the mass fractions of the raw materials, the preparation method of the composition, and the preparation parameters are the same as those of composition 1.

[0029] Composition ② Unlike composition 1, this composition lacks wolfberry powder. The missing mass is made up by chrysanthemum powder, pear powder, and magnolia bark powder in a mass ratio of 8:6:13. The other raw materials, the mass proportions of the raw materials, the preparation method of the composition, and the preparation parameters are the same as those of composition 1.

[0030] Composition ③ Unlike composition 1, this composition lacks pear powder. The missing mass is made up by chrysanthemum powder, wolfberry powder, and magnolia bark powder in a mass ratio of 8:2:13. The other raw materials, the mass proportions of the raw materials, the preparation method of the composition, and the preparation parameters are the same as those of composition 1.

[0031] Composition ④ Unlike composition 1, this composition lacks Magnolia officinalis powder. The missing mass fraction is made up by chrysanthemum powder, wolfberry powder, and pear powder in a mass ratio of 8:2:6. The other raw materials, the mass fraction of the raw materials, the preparation method of the composition, and the preparation parameters are the same as those of composition 1.

[0032] Composition ⑤ Unlike composition 1, the strain used is different; the strain used in the inoculum is Rhodotorula glutinis with preservation number CGMCC No. 22900; the viable count of the inoculum is 1 × 10⁻⁶. 8 CFU / mL; the volume ratio of inoculum to mixed liquid is 1:23; the other raw materials, raw material mass parts, preparation method of composition, and preparation parameters are the same as those of composition 1.

[0033] Composition ⑥ The raw materials and their mass fractions are the same as those in Composition 1, but the preparation method is different. The preparation method includes the following steps: S1: Mix chrysanthemum powder, wolfberry powder, pear powder, and magnolia bark powder evenly to obtain a mixed powder; S2: Mix 60wt% ethanol-water solution with the mixed powder evenly to obtain a mixed solution, wherein the ratio of the mixed powder to the 60wt% ethanol-water solution is 1:30 g / mL; S3: Heat the mixture to 65°C, reflux for 2 hours, and then filter to obtain the extract; S4: The extract is freeze-dried to obtain the composition.

[0034] Composition ⑦ The preparation method is the same as that of composition 1, but the raw materials are different. The specific raw materials and preparation method include the following steps: Raw material: YEPD liquid culture medium (Chinnok, CN230463) S1: Prepare YEPD liquid culture medium according to the YEPD liquid culture medium instructions; S2: Sterilize YEPD liquid culture medium, then inoculate with inoculum, ferment for 24 hours, sterilize after fermentation, filter to obtain fermentation filtrate, freeze-dry the fermentation filtrate to obtain the composition; The inoculum used in the culture solution was Rhodotorula glutinis with accession number CCTCC NO M2015296; the viable count of the inoculum solution was 1×10⁻⁶. 8 CFU / mL; the volume ratio of inoculum to YEPD liquid culture medium was 1:23; The fermentation temperature in step S2 is 28°C; the initial fermentation pH is 5.0.

[0035] Composition ⑧ It is prepared from the following parts by weight of raw materials: Chrysanthemum powder: 2 parts; Goji berry powder: 13 parts; Pear powder: 8 portions; Magnolia officinalis powder: 6 parts; The method for preparing the composition includes the following steps: S1: Mix chrysanthemum powder, wolfberry powder, pear powder, and magnolia bark powder evenly to obtain a mixed powder; S2: Mix deionized water and mixed powder evenly to obtain a mixed solution, wherein the ratio of mixed powder to deionized water is 1:30 g / mL; S3: Sterilize the mixture, then inoculate with the inoculum, ferment for 24 hours, sterilize after fermentation, filter to obtain fermentation filtrate, freeze-dry the fermentation filtrate to obtain the composition; The inoculum used in the culture solution was Rhodotorula glutinis with accession number CCTCC NO M2015296; the viable count of the inoculum solution was 1×10⁻⁶. 8 CFU / mL; the volume ratio of inoculum to mixed solution is 1:23; In step S3, the fermentation temperature is 28°C and the fermentation pH is 5.0.

[0036] The ingredients and their percentage by weight in pet food designed to boost immunity are as follows: Pet food 1 Composed of the following components by mass percentage: Composition to enhance immunity: 1:2wt% Duck meat meal: 29wt% Duck liver powder: 4wt% Chicken meal: 30wt% Chicken liver powder: 6wt% Chicken meat and bones: 2wt% Duck meat and bones: 2wt% Taurine: 0.2 wt%; DL-methionine: 0.1 wt% L-Lysine: 0.15 wt% Chondroitin sulfate: 0.1 wt% Glucosamine hydrochloride: 0.07 wt% Calcium hydrogen phosphate: 3wt% Vitamin A: 0.05 wt% Vitamin B: 0.05 wt% Vitamin C: 0.05 wt% Zinc citrate: 0.01 wt% Ferric citrate: 0.04 wt% Drinking water replenished to 100 wt%; Preparation method: Mix all components evenly, fill into containers, and obtain pet food.

[0037] Pet food 2 Composed of the following components by mass percentage: Composition to enhance immunity: 1:1wt% Duck meat meal: 25wt% Duck liver powder: 3wt%; Chicken meal: 25wt% Chicken liver powder: 4wt% Chicken raw meat and bones: 1 wt% Duck meat and bones: 1 wt% Taurine: 0.1 wt% DL-methionine: 0.05 wt% L-Lysine: 0.05 wt% Chondroitin sulfate: 0.05 wt% Glucosamine hydrochloride: 0.05 wt% Calcium hydrogen phosphate: 1 wt% Vitamin A: 0.01 wt% Vitamin B: 0.01 wt% Vitamin C: 0.01 wt% Zinc citrate: 0.05 wt% Ferric citrate: 0.05 wt% Drinking water replenished to 100 wt%; The preparation method is the same as that for pet food 1.

[0038] Pet food 3 Composed of the following components by mass percentage: Composition to enhance immunity: 1:3wt% Duck meat meal: 31 wt% Duck liver powder: 5wt%; Chicken meal: 35wt% Chicken liver powder: 7wt% Chicken meat and bones: 3wt% Duck meat and bones: 3wt% Taurine: 0.3 wt%; DL-methionine: 0.2wt% L-Lysine: 0.3 wt% Chondroitin sulfate: 0.15 wt%; Glucosamine hydrochloride: 0.1 wt% Calcium hydrogen phosphate: 5 wt% Vitamin A: 0.1 wt% Vitamin B: 0.1 wt% Vitamin C: 0.1 wt% Zinc citrate: 0.01 wt% Ferric citrate: 0.01 wt% Drinking water replenished to 100 wt%; The preparation method is the same as that for pet food 1.

[0039] Pet food 4 Unlike pet food 1, composition 2 is used to replace composition 1 by the same mass percentage, while the remaining components, component mass percentages, and preparation methods are the same as pet food 1.

[0040] Pet food 5 Unlike pet food 1, composition 3 is used to replace composition 1 by the same mass percentage, while the remaining components, component mass percentages, and preparation methods are the same as pet food 1.

[0041] Pet food ① Unlike pet food 1, composition ① is used to replace composition 1 by the same mass percentage, while the remaining components, component mass percentages, and preparation methods are the same as pet food 1.

[0042] Pet food ② Unlike pet food 1, composition 2 is used to replace composition 1 by the same mass percentage, while the remaining components, component mass percentages, and preparation methods are the same as pet food 1.

[0043] Pet food ③ Unlike pet food 1, composition 3 is used to replace composition 1 by the same mass percentage, while the remaining components, component mass percentages, and preparation methods are the same as pet food 1.

[0044] Pet food ④ Unlike pet food 1, composition 4 is used to replace composition 1 by a certain percentage by mass, while the remaining components, component mass percentages, and preparation methods are the same as those for pet food 1.

[0045] Pet food ⑤ Unlike pet food 1, composition 5 is used to replace composition 1 by a mass percentage, while the remaining components, component mass percentages, and preparation methods are the same as those in pet food 1.

[0046] Pet food ⑥ Unlike pet food 1, composition 6 is used to replace composition 1 by a certain percentage by mass, while the remaining components, component mass percentages, and preparation methods are the same as those in pet food 1.

[0047] Pet food ⑦ Unlike pet food 1, composition ⑦ is used to replace composition 1 by the same mass percentage, while the remaining components, component mass percentages, and preparation methods are the same as pet food 1.

[0048] Pet food ⑧ Unlike pet food 1, composition ⑧ is used to replace composition 1 by a certain percentage by mass, while the remaining components, component mass percentages, and preparation methods are the same as those for pet food 1.

[0049] Blank pet food Unlike pet food 1, this product lacks the composition, and the missing mass is made up with drinking water. The remaining components, their mass percentages, and preparation methods are the same as those in pet food 1.

[0050] Test Example 1: Effect of the Composition on Immunity Test samples: Compositions 1-3, Compositions ①-⑧; Test sample preparation: The composition was prepared into a 2wt% mixture using drinking water.

[0051] Blank test sample: The blank test sample is drinking water.

[0052] Spayed dogs (12-15 months old, healthy) were randomly divided into 11 experimental groups and 1 control group, with 10 dogs in each group. They were fed for 4 weeks, receiving the experimental sample via gavage once daily at a dose of 50 mL / 10 kg body weight. The experimental groups received the experimental sample via gavage, while the control group received the blank experimental sample via gavage. After 4 weeks, blood samples were collected from each dog for complete blood count (CBC) examination. Serum IgG, IgA, IgM, and T-AOC levels were recorded, and all data were averaged. The results are shown in Table 1.

[0053] Table 1. Effects of the composition on serum IgG, IgA, IgM, and T-AOC levels in dogs. blank control 6.74* 1.45* 0.81* 9.13* composition 1 7.93 1.82 0.97 10.72 composition 2 7.91 1.78 0.94 10.68 composition 3 7.88 1.81 0.96 10.65 composition 1 7.42* 1.72* 0.89* 10.36* composition 2 7.58* 1.73* 0.91* 10.41* composition 3 7.66* 1.68* 0.88* 10.39* composition 4 7.15* 1.59* 0.87* 9.52* composition 5 7.34* 1.57* 0.85* 10.08* composition 6 6.81* 1.48* 0.82* 9.34* composition 7 6.98* 1.51* 0.84* 9.92* composition 8 7.47* 1.72* 0.90* 10.09* Note: "*" indicates that p < 0.05 compared to composition 1.

[0054] According to the results in Table 1, comparing the results of compositions 1-3 with the blank control group, it can be seen that the compositions can significantly increase the content of IgG, IgA, IgM, and T-AOC in canine serum. Comparing the results of composition 1 with compositions ①-④ and composition ⑧, it can be seen that the compositions obtained by fermenting chrysanthemum powder, wolfberry powder, pear powder, and magnolia bark powder as substrates with specific strains have the effect of significantly increasing the content of IgG, IgA, IgM, and T-AOC in canine serum, indicating that they can significantly improve canine immunity. Comparing the results of composition 1 with composition ⑤, it can be seen that even among Rhodotorula glutinis, Rhodotorula glutinis with different preservation numbers have different physiological characteristics. The compositions obtained using Rhodotorula glutinis with the preservation number specified in this invention and the raw materials and preparation methods specified in this invention can significantly improve canine immunity. Comparing the results of composition 1 with compositions ⑥ and ⑦, it can be seen that there is a significant synergistic effect between the fermentation raw materials and fermentation strains in this invention. The compositions prepared using the raw materials, strains, and preparation methods specified in this invention have the effect of significantly improving canine immunity.

[0055] Test Example 2: Toxicity Test Test samples: Pet food 1-5, blank pet food; Experimental procedure: Healthy sterilized dogs (6 months old) were selected. The experimental period was 4 weeks. Necessary immunization and deworming treatments were performed before the experiment. The dogs were randomly divided into 5 experimental groups and 1 blank control group, with 5 dogs in each group. The breeding environment was 25℃ and 55% relative humidity. The experimental results are shown in Table 2.

[0056] Table 2 Acute toxicity test of oral liquids that help improve gut microbiota

[0057] The acute toxicity test results showed that no dogs showed obvious signs of poisoning during the test, and no animals died, indicating that the test sample prepared by this invention is non-toxic.

[0058] Test Example 3: Palatability Test Test substances: Pet food 1-5, Pet food ①-⑧.

[0059] Blank test subject: Blank pet food.

[0060] Test subjects: Spayed / neutered dogs (12-15 months old), divided into 14 groups of 6 dogs each.

[0061] Test Method: Test subjects were fed either the test substance or a blank test substance. All subjects were kept in the same diet and at the same time, except for the test substance or blank test substance. The time of feeding the test substance was also consistent, such as 9:00 AM or 4:00 PM daily. Subjects were fed in a controlled environment, with free movement within this controlled environment (closed environment at 20-25℃), maintaining a diurnal rhythm to ensure a stress-free palatability test. The health of the subjects was strictly monitored by a licensed veterinarian. No forced or aggressive testing was conducted, and no cage holding or long-term isolation was implemented. All personnel involved in the test underwent rigorous training. All test subjects received excellent care and socialization outside of the feeding test. The test subjects met the palatability test requirements.

[0062] Single feed intake test: Provide a feeding bowl to the test subjects at the same time every day during the test period. Put the corresponding test substance or blank test substance in the feeding bowl, weigh the amount of feed consumed by each test subject in one feeding and calculate the feed intake rate (%). Calculate the average feed intake rate of the group with a test period of 5 days. It should be noted that the amount of test substance or blank test substance provided to the test subjects should be higher than their average amount of feed consumed in one feeding.

[0063] First bite preference test: Provide two feeding bowls to the test subjects at the same time each day during the test period. One feeding bowl contains the test substance, and the other contains a blank test substance. For example, for the test subjects participating in the pet food 1 test, provide two feeding bowls, one containing pet food 1 and the other containing blank pet food. The quality of pet food in each feeding bowl is the same. Record the first bite choice of each test subject. Count the number of test subjects who chose the test substance in the first bite of the group in a 5-day test cycle. It should be noted that the positions of the feeding bowls should be reversed every other day.

[0064] Selective feeding rate test: Two feeding bowls were provided to the test subjects at the same time each day during the test period. One feeding bowl contained the test substance, and the other contained a blank test substance. The mass of the test substance or blank test substance in each feeding bowl was the same. The amount of food consumed by each test subject in one feeding process was measured. The average selective feeding rate of the group was calculated with a test period of 5 days. The selective feeding rate of a test subject in one feeding process was the amount of food consumed in the single bowl in that feeding process / the total amount of food consumed in the two bowls x 100%. For example, if the amount of food consumed in the test substance bowl was 1g and the amount of food consumed in the blank test substance bowl was 9g, then the selective feeding rate was 10%. It should be noted that the positions of the feeding bowls were reversed every other day.

[0065] Table 3 Palatability Test blank pet food - 75.4 - pet food 1 97.6 93.1 28 pet food 2 83.3 87.2 24 pet food 3 94.5 92.4 27 pet food 4 93.4 90.6 27 pet food 5 90.6 91.9 25 pet food 1 85.6 84.6 19 pet food 2 80.3 87.7 17 pet food 3 83.2 85.3 15 pet food 4 87.1 82.4 18 pet food 5 43.6 69.5 6 pet food 6 52.5 72.1 11 pet food 7 69.3 78.2 13 pet food 8 79.4 88.5 16 As shown in Table 3, comparing the selective feeding rate, single feeding rate, and number of test subjects selected for the first bite of pet food 1-5 with the blank pet food, it can be seen that the composition can significantly improve the palatability of pet food and increase the intake of pet food by the test subjects. Comparing the results of pet food 1 with pet food ①-⑧, it can be seen that the raw materials, fermentation strains, and preparation methods of the composition significantly improve the palatability of pet food and increase the intake of pet food by the test subjects.

[0066] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A composition for enhancing immunity, characterized in that, The composition comprises the following raw materials in parts by weight: Chrysanthemum powder: 6-9 parts; Goji berry powder: 1-3 parts; Pear powder: 5-7 parts; Magnolia officinalis powder: 12-15 parts; The method for preparing the composition includes the following steps: S1: Mix chrysanthemum powder, wolfberry powder, pear powder, and magnolia bark powder evenly to obtain a mixed powder; S2: Mix deionized water and mixed powder evenly to obtain a mixed solution, wherein the ratio of mixed powder to deionized water is 1:20-40 g / mL; S3: Sterilize the mixture, then inoculate with the inoculum, ferment for 20-30 hours, sterilize after fermentation, filter to obtain fermentation filtrate, freeze-dry the fermentation filtrate to obtain the composition.

2. The composition according to claim 1, characterized in that, The inoculum used in the culture solution was Rhodotorula glutinis with preservation number CCTCCNO: M2015296; the viable count of the inoculum solution was 1×10⁻⁶. 7 -1×10 9 CFU / mL; the volume ratio of inoculum to mixed solution is 1:19-24.

3. The composition according to claim 1, characterized in that, In step S3, the fermentation temperature is 25-30℃; the initial fermentation pH is 4.0-6.

0.

4. The use of an immune-enhancing composition as described in any one of claims 1-3 in the preparation of pet food.

5. The use of an immune-enhancing composition as described in any one of claims 1-3 in the preparation of a pet nutritional fortifier.

6. The use of an immune-enhancing composition as described in any one of claims 1-3 in the preparation of a pet food additive.

7. A pet food for enhancing immunity, characterized in that, The pet food includes the immune-enhancing composition according to any one of claims 1-3.

8. The pet food as described in claim 7, characterized in that, The food also includes animal protein sources, grains, vitamins, minerals, chondroitin sulfate, and glucosamine hydrochloride.

9. The pet food as described in claim 8, characterized in that, The animal protein source includes at least one of poultry meat meal, fish meal, and meat and bone meal; the grains include at least one of corn, wheat, rice, millet, peas, chickpeas, and oats; the vitamins include at least one of vitamin A, vitamin B, vitamin C, and vitamin E; and the minerals include at least one of copper salts, iron salts, manganese salts, zinc salts, calcium salts, and phosphates.

10. A pet food for enhancing immunity as described in any one of claims 7-9, characterized in that, Includes the following components by mass percentage: Composition to enhance immunity: 1-3 wt% Duck meat meal: 25-31 wt% Duck liver powder: 3-5 wt%; Chicken meal: 25-35 wt% Chicken liver powder: 4-7 wt% Chicken raw meat and bones: 1-3 wt% Duck meat and bones: 1-3 wt% Taurine: 0.1-0.3 wt%; DL-methionine: 0.05-0.2 wt%; L-Lysine: 0.05-0.3 wt%; Chondroitin sulfate: 0.05-0.15 wt%; Glucosamine hydrochloride: 0.05-0.1 wt%; dicalcium phosphate: 1-5 wt% Vitamin A: 0.01-0.1 wt% Vitamin C: 0.01-0.1 wt% Vitamin B: 0.01-0.1 wt% Zinc citrate: 0.01-0.05 wt%; Ferric citrate: 0.01-0.05 wt%; Add water to 100wt%.

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