Composition for treating diarrhea caused by rotavirus infection in pet dogs and cats, preparation method and application thereof

Abstract

The invention discloses a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats. The composition comprises the following components: 5-18 parts by weight of echinacea extract, 1-6 parts by weight of bacillus subtilis spore powder, 10-35 parts by weight of beta-glucan, 5-18 parts by weight of sodium chondroitin sulfate, 2-10 parts by weight of N-acetylglucosamine, 2-10 parts by weight of gamma-aminobutyric acid, 10-35 parts by weight of lactobacillus plantarum fermentation product, 5-18 parts by weight of D-mannitol, and 5-18 parts by weight of magnesium hydroxide. A preparation method and application of the composition in preparing a medicament for treating rotavirus diarrhea in pet dogs and cats are also disclosed. The composition cleverly utilizes the synergistic effects of the components in anti-virus, immune regulation, intestinal mucosal repair, diarrhea improvement, etc., and exhibits an excellent effect of treating rotavirus diarrhea in pets through comprehensive regulation of multiple links such as immunity, barrier, flora, mucosa, etc.

Description

Technical Field

[0001] The invention relates to the technical field of pet diarrhea treatment compositions, and in particular to a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats, a preparation method and application thereof. Background Art

[0002] Rotavirus is a common pathogen of diarrhea in dogs and cats, especially in group breeding environments such as kennels and pet shops, which is prone to outbreaks and poses a serious threat to pet health and public health. At present, the prevention and treatment of rotavirus mainly adopts measures such as vaccination and isolation treatment. However, vaccines are ineffective for infected animals, and the immune response varies greatly.

[0003] In terms of treatment, symptomatic supportive treatment is mainly adopted, including fasting, rehydration, antidiarrheal drugs, etc., but the efficacy is limited. Antiviral drugs such as ribavirin are effective, but they are expensive and are prone to drug resistance after long-term use. Probiotic preparations such as lactobacillus can improve intestinal flora, but they are slow to take effect, and live bacteria are easily inactivated and have poor stability. Traditional Chinese medicines such as berberine and Patchouli can stop diarrhea, but they are bitter, have poor compliance, and the antiviral activity of single herbs is not strong.

[0004] At present, there is still a lack of a specific drug for rotavirus. Clinically, there is an urgent need for a comprehensive intestinal conditioning preparation that can intervene in multiple links, quickly relieve diarrhea symptoms, repair damaged intestinal mucosa, regulate flora imbalance, enhance the body's immunity, and reduce recurrence. Summary of the invention

[0005] The present invention provides a combination for treating diarrhea caused by rotavirus infection in pet dogs and cats and a preparation method thereof. The composition systematically treats rotavirus infection from multiple aspects such as antiviral, antibacterial, repair, and regulation, thereby making up for the deficiencies of the prior art. At the same time, the use of advanced processes such as modern extraction and microbial fermentation ensures the stability and bioavailability of the drug. Therefore, the composition of the present invention has broad application prospects as a drug for treating rotavirus diarrhea in pets.

[0006] A composition for treating diarrhea caused by rotavirus infection in pet dogs and cats, comprising the following components: 5-18 parts by weight of Echinacea extract, 1-6 parts by weight of Bacillus subtilis spore powder, 10-35 parts by weight of β-glucan, 5-18 parts by weight of sodium chondroitin sulfate, 2-10 parts by weight of N-acetylglucosamine, 2-10 parts by weight of γ-aminobutyric acid, 10-35 parts by weight of Lactobacillus plantarum fermentation product, 5-18 parts by weight of D-mannitol, and 5-18 parts by weight of magnesium hydroxide.

[0007] Specifically, the preparation method of the Echinacea extract is: take 500 parts of dried Echinacea flowers, crush them, pass them through a 60-100 mesh sieve, and extract them using one of the following methods:

[0008] (1) adding 4000 parts by volume of 75%-95% ethanol and reflux extraction 2-3 times, each time for 1.5-2 hours; or

[0009] (2) extracting by percolation with 95% ethanol 2-3 times, each time for 1.5-2 hours; or

[0010] (3) Using CO2 supercritical extraction, the extraction pressure is 25-35MPa, the extraction temperature is 40-50°C, and the extraction time is 2-3h;

[0011] The extract is concentrated under reduced pressure to a relative density of 1.10-1.15, resuspended in water, allowed to stand, filtered, and the filtrate is separated and purified by a macroporous resin AB-8 or HD-2 column, eluted with water, 40%, 60%, and 70%-80% ethanol in sequence, and the 70%-80% ethanol eluate is collected and dried to obtain the Echinacea extract.

[0012] Specifically, the preparation method of the Bacillus subtilis spore powder is: inoculate Bacillus subtilis CICC 24713 or CICC10275 on a slant or plate, and culture at 30-37°C for 48-72 hours; scrape the spores or bacterial lawn, and 5 CFU / mL of bacteria were inoculated into seed culture medium and cultured at 30-37℃ for 24h; 4% of the inoculation amount was transferred into a fermentation tank and fermented at 30-37℃ for 72h; the bacteria were collected by centrifugation and freeze-dried or spray-dried to obtain spore powder, which contained 10 8 -10 10 CFU viable spores.

[0013] Specifically, the preparation method of the plant lactobacillus fermentation product is as follows: plant lactobacillus CGMCC No.1.557 or CICC6098 is inoculated into MRS liquid culture medium or modified MRS culture medium, 2%-8% plant juice is added to the modified MRS culture medium, anaerobically fermented at 30-37°C for 48-72h, and the bacterial cells and fermentation liquid are collected by centrifugation; the bacterial cells are freeze-dried, and the fermentation liquid is spray-dried; the dried bacterial cells and the dried fermentation liquid are mixed in a ratio of 5:5-9:1 to obtain the fermentation product, and each gram contains 10 8 -10 11 CFU live bacteria.

[0014] Specifically, the plant juice is any one of cabbage juice, celery juice, carrot juice, tomato juice, cucumber juice, apple juice, pear juice and grape juice, or a combination of several of them. Fresh fruits and vegetables are taken, cleaned, squeezed, centrifuged at low temperature to remove pomace, and the supernatant is filtered and sterilized to obtain sterile plant juice.

[0015] Preferably, a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following components: 10-15 parts by weight of Echinacea extract, 3-5 parts by weight of Bacillus subtilis spore powder, 20-30 parts by weight of β-glucan, 10-15 parts by weight of sodium chondroitin sulfate, 5-8 parts by weight of N-acetylglucosamine, 5-8 parts by weight of γ-aminobutyric acid, 20-30 parts by weight of Lactobacillus plantarum fermentation product, 8-15 parts by weight of D-mannitol, and 8-15 parts by weight of magnesium hydroxide.

[0016] Specifically, the preparation method of the composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following steps: (1) uniformly mixing Echinacea extract, Bacillus subtilis spore powder, β-glucan, sodium chondroitin sulfate, N-acetylglucosamine, γ-aminobutyric acid, Lactobacillus plantarum fermentation product, D-mannitol and magnesium hydroxide according to the weight of the formula; (2) preparing the mixture obtained in step (1) into granules containing 40-50 mg per granule, tablets weighing 400-800 mg per tablet, capsules containing 300-500 mg per granule, powders containing 2-4 g per bag, syrups containing 80-120 mL per bottle or chewable tablets weighing 600-1000 mg per tablet.

[0017] Furthermore, the composition for treating diarrhea caused by rotavirus infection in pet dogs and cats is used in the preparation of a drug for treating rotavirus diarrhea in pet dogs and cats.

[0018] Compared with the prior art, the composition for treating diarrhea caused by rotavirus infection in pet dogs and cats and the preparation method thereof can achieve the following beneficial effects:

[0019] 1. Echinacea extract is a natural product extracted from the Echinacea plant. The main active ingredients of Echinacea include polar components (such as caffeic acid and its derivatives), non-polar components (such as alkylamides), and high molecular weight components such as polysaccharides and glycoproteins. Echinacea extract can regulate the immune function of the animal body, including enhancing immunity, anti-inflammatory, antibacterial and antiviral effects. These effects help improve intestinal health, reduce the occurrence of diarrhea, reduce blood lipids and regulate serum inflammatory factor levels. Studies have shown that Echinacea extract can reduce blood lipid levels and regulate serum inflammatory factors such as IL-6, IL-17 and IL-23 levels. These changes help reduce inflammatory responses and thus relieve diarrhea symptoms. Echinacea extract can also improve the functions of organs such as the heart, spleen, and lungs. The improvement of these organ functions contributes to overall health, including intestinal health, thereby reducing the occurrence of pet diarrhea. In summary, Echinacea extract has a positive effect on the treatment of pet diarrhea through regulating immune function, reducing blood lipids, regulating inflammatory factor levels, and improving organ function.

[0020] Bacillus subtilis spore powder can help pets resist pathogenic bacteria and relieve bacterial diarrhea, which helps maintain the balance of gastrointestinal microecology and is essential to the overall health of pets. After resuscitation in the pet's intestines, Bacillus subtilis spore powder can stimulate the intestinal mucosa, activate the immune system of the intestinal mucosa, produce antibodies and cytokines, and improve the immunity and anti-infection ability of the intestinal mucosa. Bacillus subtilis spore powder synthesizes a variety of enzyme substances and a variety of B vitamins in the pet's body, thereby improving the digestion and absorption utilization rate of feed, which is essential for the nutritional absorption and growth of pets. Bacillus subtilis spore powder can directly reduce the colonization of pathogenic bacteria in the animal's intestines through competitive exclusion and protect the integrity of the intestinal barrier; Bacillus subtilis spore powder is an aerobic bacteria that absorbs oxygen in the intestines, creating a suitable living environment for other probiotics such as lactic acid bacteria and bifidobacteria that do not like oxygen. This environment is conducive to the growth of probiotics and can also inhibit the growth of harmful bacteria. In summary, Bacillus subtilis spore powder has a positive impact on the gastrointestinal health of pets by maintaining the balance of gastrointestinal microecology, regulating immune function, promoting nutrient digestion and absorption, competitive exclusion and directly reducing pathogenic bacteria colonization, and improving the intestinal environment. These effects together promote the overall health and well-being of pets.

[0021] Polysaccharides such as β-glucan and chondroitin can competitively bind to virus surface receptors and block virus adsorption. Both β-glucan and chondroitin sulfate have more hydroxyl groups and can bind to sialic acid on the virus surface through hydrogen bonds to prevent virus adsorption. At the same time, they can stimulate the immune cells of the lamina propria such as macrophages and dendritic cells, promote the release of cytokines IL-12 and IFN-γ, and enhance the body's antiviral immunity. The synergistic use of the two can curb viral infection from the two levels of viral adsorption and host immunity. The synergistic effect of the two can curb viral infection from the two links of viral replication and adsorption. N-acetylglucosamine, as a synthetic precursor of mucin and mucopolysaccharide, can repair the damaged intestinal mucus layer. In addition, γ-aminobutyric acid can activate GABA receptors and inhibit intestinal contraction. The two complement each other and jointly promote the repair of damaged intestinal mucosa and relieve diarrhea. It is worth mentioning that γ-aminobutyric acid can also reduce the release of intestinal serotonin, which can cause diarrhea. This is an innovative point to resolve contradictions.

[0022] The present invention adopts CO2 supercritical extraction of Echinacea, which is gentle and efficient, has a high yield of Echinacea extract and good biological activity; β-glucan and chondroitin completely retain the natural polysaccharide structure and have better receptor blocking activity.

[0023] 2. Plant lactobacillus in the fermentation products of Lactobacillus plantarum, etc. Plant lactobacillus plantarum is a polypeptide antibiotic composed of multiple amino acids, which can specifically kill pathogenic bacteria such as Staphylococcus aureus and Escherichia coli in the intestine, while retaining beneficial bacteria such as lactic acid bacteria. The hydrogen peroxide produced by fermentation metabolism can also oxidize viral proteins and inhibit viral infection. This forms a double probiotic synergy with Bacillus subtilis, which is conducive to rebuilding the balance of intestinal flora. Lactobacillus plantarum is an important probiotic that can produce antibacterial active substances and regulate intestinal flora, but is sensitive to gastric acid and bile. Bacillus subtilis has the characteristics of strong tolerance and easy storage. The two complement each other's advantages, which can not only play a probiotic role, but also overcome the problem of poor stability of live bacterial preparations. The use of modified MRS culture medium and the addition of natural nitrogen sources and growth factors such as plant juice are conducive to increasing the number of live bacteria.

[0024] 3. Rotavirus infection can lead to damaged intestinal mucosal barrier, destruction of tight junctions, and release of inflammatory factors. The present invention cleverly utilizes the intestinal mucosal repair effect of glucosamine and aminobutyric acid. Among them, glucosamine can promote the synthesis of mucin, glycoprotein, etc., and increase intestinal mucus secretion; aminobutyric acid can regulate the expression of intestinal epithelial tight junction proteins and strengthen tight junctions between cells. The two complement each other, repairing damaged intestinal mucosa in terms of both quantity and quality, and reducing inflammatory reactions. This is the unique feature of the present invention that distinguishes it from general diarrhea treatment drugs.

[0025] 4. In addition to direct antiviral effects, enhancing the body's immunity is also an important strategy for treating rotavirus infection. β-glucan, chondroitin, etc. have immunomodulatory activity, which can stimulate innate immunity such as macrophages and NK cells, promote the production of specific antibodies, and accelerate the body's clearance of viruses. Lactobacillus plantarum and Bacillus subtilis can induce the secretion of mucosal immune factors such as IgA and enhance local immunity of the intestinal mucosa. Multiple immune active ingredients work synergistically to regulate immune function from both innate and acquired immunity, and improve the body's resistance. This is a systemic regulation advantage that simple antiviral drugs do not have.

[0026] In summary, the present invention is based on the perfect combination of traditional Chinese medicine theory and modern biotechnology, and cleverly utilizes the synergistic effects of various components in anti-virus, immune regulation, intestinal mucosal repair, diarrhea improvement, etc., and through the comprehensive regulation of multiple links such as immunity, barrier, flora, mucosa, etc., it shows excellent effects in treating rotavirus diarrhea in pets, highlighting the scientificity and innovation of the composition combination; different active ingredients synergize with each other, and intervene in intestinal microenvironment disorders through multiple targets and multiple pathways, which is a new comprehensive conditioning strategy. DETAILED DESCRIPTION

[0027] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the specific embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0028] Example 1

[0029] The composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following components: 5 parts of Echinacea extract, 1 part of Bacillus subtilis spore powder, 10 parts of beta-glucan, 5 parts of sodium chondroitin sulfate, 2 parts of N-acetylglucosamine, 2 parts of gamma-aminobutyric acid, 10 parts of Lactobacillus plantarum fermentation product, 5 parts of D-mannitol and 5 parts of magnesium hydroxide.

[0030] The preparation method of a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following steps:

[0031] (1) Echinacea extract: Take 500 parts of dried Echinacea flowers, crush them, pass through a 60-mesh sieve, add 4000 parts of 75% ethanol by volume, and reflux extract twice, each time for 1.5 hours. Combine the extracts, recover the ethanol under reduced pressure, and concentrate to a clear paste with a relative density of 1.10-1.15 (60°C). Add 100 parts of water, stir evenly, let stand for 12 hours, and filter. The filtrate is separated and purified by macroporous resin AB-8 column chromatography, washed with water, eluted with 40%, 60%, and 70%-80% ethanol in sequence, collect the 70%-80% ethanol eluate, and dry to obtain the Echinacea extract.

[0032] (2) Bacillus subtilis spore powder: Bacillus subtilis CICC 24713 was inoculated on a slant surface and cultured at 37°C for 48 h. The bacterial moss was scraped and 10 5 CFU / mL of bacteria were inoculated into seed tanks and cultured at 37℃ for 24h; 4% of the inoculation amount was transferred into fermentation tanks and fermented at 30℃ for 72h, the bacteria were collected by centrifugation and spray-dried to obtain spore powder, which contained 10 8 CFU viable spores.

[0033] (3) β-Glucan, chondroitin sodium, N-acetylglucosamine and γ-aminobutyric acid were of commercially available food grade.

[0034] (4) Lactobacillus plantarum fermentation product: Lactobacillus plantarum CGMCC No. 1.557 was inoculated into MRS liquid medium and cultured anaerobically at 37°C for 48 h. The cells and fermentation liquid were collected by centrifugation. The cells were freeze-dried and the fermentation liquid was spray-dried. The dried cells and the dried fermentation liquid were mixed in a ratio of 5:1 to obtain the fermentation product. Each gram of the mixture contained 10 9CFU live bacteria.

[0035] (5) Take the above components, add D-mannitol and magnesium hydroxide, mix well, granulate with an electric granulator, and dry to obtain the composite granules, each containing 40-50 mg.

[0036] Example 2

[0037] The composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following components: 15 parts of Echinacea extract, 5 parts of Bacillus subtilis spore powder, 30 parts of beta-glucan, 15 parts of sodium chondroitin sulfate, 8 parts of N-acetylglucosamine, 8 parts of gamma-aminobutyric acid, 30 parts of Lactobacillus plantarum fermentation product, 15 parts of D-mannitol and 15 parts of magnesium hydroxide.

[0038] The preparation method of a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following steps:

[0039] (1) Echinacea extract: 500 parts of dried Echinacea flowers were crushed, passed through an 80-mesh sieve, and extracted three times with 95% ethanol by percolation, each time for 2 hours. The extracts were combined, and the extracts were concentrated under reduced pressure to a relative density of 1.10-1.15. The extracts were resuspended in water, allowed to stand, and filtered. The filtrate was separated and purified by a macroporous resin HD-2 column, and eluted with water, 40%, 60%, and 70%-80% ethanol in sequence. The 70%-80% ethanol eluate was collected and dried to obtain the Echinacea extract. The extracts were concentrated under reduced pressure and freeze-dried to obtain the Echinacea extract.

[0040] (2) Bacillus subtilis spore powder: Bacillus subtilis CICC10275 was inoculated on a PDA plate and cultured at 30°C for 72 h. The spore suspension was scraped from the plate and then 10 5 CFU / mL of bacteria were inoculated into seed culture medium and cultured at 37℃ for 24h; 4% of the inoculation amount was transferred into a fermentation tank and fermented at 37℃ for 72h; the bacteria were collected by centrifugation and freeze-dried to obtain spore powder, which contained 10 10 CFU live spores

[0041] (3) β-Glucan, chondroitin sodium, N-acetylglucosamine and γ-aminobutyric acid were of commercially available food grade.

[0042] (4) Lactobacillus plantarum fermentation product: Lactobacillus plantarum CICC6098 was inoculated into a modified MRS liquid medium (added with 5% carrot juice) and anaerobically fermented at 37°C for 60 h. The cells and fermentation broth were collected by centrifugation, the fermentation broth was spray dried, the cells were freeze-dried, and the dried cells and dried fermentation broth were mixed in a ratio of 7:3 to obtain a fermentation product containing 1.2×10 10 CFU / g viable bacteria.

[0043] (5) Mix all the components, pass through a 40-mesh sieve, mix well, and compress into tablets to obtain the combined tablets, each weighing 500 mg.

[0044] In the present invention, the preparation of Lactobacillus plantarum fermentation products adopts a modified MRS medium with added plant juice, which is a unique innovation. Conventional MRS medium mainly consists of peptone, yeast powder, glucose, dipotassium hydrogen phosphate, sodium acetate, ammonium citrate, magnesium sulfate, manganese sulfate and Tween 80, and is a commonly used culture medium for lactic acid bacteria. However, plant-derived nutrients have a unique promoting effect on the growth, reproduction and metabolic activity of Lactobacillus plantarum.

[0045] The plant juice selected by the present invention can be derived from a variety of vegetables and fruits, such as cabbage, celery, carrot, tomato, cucumber, apple, pear, grape, etc., and can be used alone or in combination. The amount of plant juice added is 2%-8% of the volume of the culture medium. Take fresh fruits and vegetables, wash them, squeeze the juice, centrifuge at low temperature to remove the pomace, take the supernatant and filter and sterilize it to obtain sterile plant juice. Plant juice is rich in natural nutrients and functional factors such as various vitamins, minerals, dietary fiber and aromatic substances, which can significantly promote the growth and metabolism of plant lactobacillus and produce some unexpected beneficial effects:

[0046] 1. Vitamins in plant juice, such as VB1, VB2, VB6, VC, etc., are coenzyme factors for the growth and metabolism of Lactobacillus, which can increase the biomass and activity of Lactobacillus. In particular, B vitamins such as folic acid can promote the synthesis of nucleic acids and proteins by Lactobacillus and accelerate bacterial proliferation.

[0047] 2. Plant juice contains mineral ions such as potassium, calcium, magnesium, zinc, and selenium, which can regulate the osmotic pressure balance of lactobacillus, maintain cell membrane integrity and enzyme activity, and thus enhance the resistance to stress during the fermentation process.

[0048] 3. Dietary fiber in plant juice, such as pectin and fructan, can not only provide carbon source, but also adsorb lactobacillus to form biofilm, protect the bacteria from damage by gastric acid and bile salts, and improve the survival rate and colonization ability of the bacteria.

[0049] 4. Aromatic substances in plant juice, such as terpenes, alcohols, esters, etc., can provide unique flavor precursors for Lactobacillus fermentation, promote Lactobacillus to produce some special aroma metabolites, and improve the flavor quality of the fermentation products.

[0050] 5. Some plant juices such as garlic and onion contain a variety of plant antibiotics such as allicin and allicin. When co-cultured with Lactobacillus, they can produce a synergistic antibacterial effect and improve the antibacterial spectrum and efficacy of the fermentation products.

[0051] 6. Plant juices rich in polyphenols, such as berry juice, can provide unique phenolic acid substrates for Lactobacillus, promoting Lactobacillus to produce some rare probiotics, such as echinacea and rhamnosus lactobacillus, thereby expanding the functional diversity of fermentation products.

[0052] 7. Lycopene in the juice of tomatoes and other nightshade plants has antioxidant effects. It can remove peroxides and free radicals produced during the fermentation process, maintain the redox balance of lactobacillus, extend the fermentation cycle, and increase the number of live bacteria.

[0053] In summary, adding plant juice to the fermentation medium of Lactobacillus plantarum can not only provide rich natural nutrition, promote the growth and reproduction of Lactobacillus plantarum, and improve the fermentation efficiency, but also regulate the composition and function of metabolites, and endow the fermentation products with multiple health-care properties. This innovative fermentation process can significantly improve the quality and efficacy of the fermentation products, which is a major feature and advantage of the present invention. Compared with conventional MRS medium, the improved MRS medium is closer to the natural habitat of Lactobacillus plantarum, which is conducive to exerting its probiotic function and has a unique promoting effect on the improvement of host intestinal health and immunity.

[0054] Example 3

[0055] The composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following components: 10 parts of Echinacea extract, 3 parts of Bacillus subtilis spore powder, 20 parts of beta-glucan, 10 parts of sodium chondroitin sulfate, 5 parts of N-acetylglucosamine, 5 parts of gamma-aminobutyric acid, 20 parts of Lactobacillus plantarum fermentation product, 10 parts of D-mannitol and 10 parts of magnesium hydroxide.

[0056] The preparation method of a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following steps:

[0057] (1) Echinacea extract: Take 500 parts of dried Echinacea flowers, crush them, pass them through a 100-mesh sieve, and extract them with supercritical CO2 at an extraction pressure of 30 MPa, an extraction temperature of 45°C, and an extraction time of 2.5 hours. Combine the extracts and concentrate them to a clear paste with a relative density of 1.10 to 1.15 (60°C). Add 100 parts of water, stir evenly, let stand for 12 hours, and filter. The filtrate is separated and purified by macroporous resin AB-8 column chromatography, and eluted with water, 40%, 60%, and 70%-80% ethanol in sequence. The 70%-80% ethanol eluate is collected and dried to obtain the Echinacea extract.

[0058] (2) Bacillus subtilis spore powder: Bacillus subtilis CICC 24713 was inoculated on a slant surface and cultured at 37°C for 48 h. The bacterial moss was scraped and 10 5CFU / mL of bacteria were inoculated into seed tanks, cultured at 37℃ for 24h, transferred into fermentation tanks at 4% inoculation, fermented at 30℃ for 72h, collected by centrifugation, and spray-dried to obtain spore powder, containing 10 8 CFU viable spores.

[0059] (3) β-Glucan, chondroitin sodium, N-acetylglucosamine and γ-aminobutyric acid were of commercially available food grade.

[0060] (4) Lactobacillus plantarum fermentation product: Lactobacillus plantarum CGMCC No. 1.557 was inoculated into MRS liquid medium, cultured anaerobically at 37°C for 48 h, the cells and fermentation liquid were collected by centrifugation, the cells were freeze-dried, and the fermentation liquid was spray-dried; the dried cells and the dried fermentation liquid were mixed in a ratio of 5-9 to obtain a fermentation product containing 10 9 CFU / g viable bacteria.

[0061] (5) Mix all the components, add appropriate amount of starch and magnesium stearate, mix well, and fill into capsules to obtain the combined capsules, each containing 400 mg.

[0062] Example 4

[0063] The composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following components: 12 parts of Echinacea extract, 4 parts of Bacillus subtilis spore powder, 25 parts of beta-glucan, 12 parts of sodium chondroitin sulfate, 6 parts of N-acetylglucosamine, 6 parts of gamma-aminobutyric acid, 25 parts of Lactobacillus plantarum fermentation product, 12 parts of D-mannitol and 12 parts of magnesium hydroxide.

[0064] The preparation method of a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following steps:

[0065] (1) Echinacea extract: 500 parts of dried Echinacea flowers were crushed, passed through a 60-100 mesh sieve, and 4000 parts of 75%-95% ethanol were added, and refluxed for 2-3 times, each time for 1.5-2 hours;

[0066] The extracts were combined, concentrated under reduced pressure to a relative density of 1.10-1.15, resuspended in water, allowed to stand, filtered, and the filtrate was separated and purified by a macroporous resin AB-8 column, eluted with water, 40%, 60%, and 70%-80% ethanol in sequence, and the 70%-80% ethanol eluate was collected and dried to obtain the Echinacea extract.

[0067] (2) Bacillus subtilis spore powder: Inoculate Bacillus subtilis CICC 24713 on a slant and culture at 30-37°C for 48-72 hours; scrape off spores or bacterial moss and use 10 5CFU / mL of bacteria were inoculated into seed culture medium and cultured at 30-37℃ for 24h; 4% of the inoculation amount was transferred into a fermentation tank and fermented at 30-37℃ for 72h; the bacteria were collected by centrifugation and freeze-dried or spray-dried to obtain spore powder, which contained 10 8 -10 10 CFU viable spores.

[0068] (3) β-Glucan, chondroitin sodium, N-acetylglucosamine and γ-aminobutyric acid were of commercially available food grade.

[0069] (4) Lactobacillus plantarum fermentation product: Lactobacillus plantarum CGMCC No.1.557 was inoculated into MRS liquid culture medium, and anaerobically fermented at 30-37°C for 48-72 hours, and the bacteria and fermentation liquid were collected by centrifugation; the bacteria were freeze-dried, and the fermentation liquid was spray-dried; the dried bacteria and the dried fermentation liquid were mixed in a ratio of 5-6 to obtain the fermentation product, and each gram contained 10 11 CFU live bacteria.

[0070] (5) Mix all the components, mix with D-mannitol and magnesium hydroxide, make granules, dry, and pack into bags, 3 g per bag.

[0071] Example 5

[0072] The composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following components: 8 parts of Echinacea extract, 2 parts of Bacillus subtilis spore powder, 15 parts of beta-glucan, 8 parts of sodium chondroitin sulfate, 4 parts of N-acetylglucosamine, 4 parts of gamma-aminobutyric acid, 15 parts of Lactobacillus plantarum fermentation product, 8 parts of D-mannitol and 8 parts of magnesium hydroxide.

[0073] The preparation method of a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following steps:

[0074] (1) Echinacea extract: Take 500 parts of dried Echinacea flowers, crush them, pass through a 60-mesh sieve, add 4000 parts of 95% ethanol by volume and perform percolation extraction 2-3 times, each time for 1.5-2 hours, combine the extracts, recover the ethanol under reduced pressure, concentrate to a clear paste with a relative density of 1.10-1.15 (60°C), add 100 parts of water, stir evenly, let stand for 12 hours, and filter. The filtrate is separated and purified by macroporous resin AB-8 column chromatography, washed with water, eluted with 40%, 60%, and 70%-80% ethanol in sequence, collect the 70%-80% ethanol eluate, and dry to obtain the Echinacea extract.

[0075] (2) Bacillus subtilis spore powder: Bacillus subtilis CICC10275 was inoculated on a plate and cultured at 30-37°C for 48-72 hours; spores or bacterial moss were scraped and 10 5CFU / mL of bacteria were inoculated into seed culture medium and cultured at 30-37℃ for 24h; 4% of the inoculation amount was transferred into a fermentation tank and fermented at 30-37℃ for 72h; the bacteria were collected by centrifugation and freeze-dried or spray-dried to obtain spore powder, which contained 2×10 8 CFU viable spores.

[0076] (3) β-Glucan, chondroitin sodium, N-acetylglucosamine and γ-aminobutyric acid were of commercially available food grade.

[0077] (4) Lactobacillus plantarum fermentation product: Lactobacillus plantarum CICC6098 was inoculated into a modified MRS medium, 2%-8% plant juice was added to the modified MRS medium, and anaerobically fermented at 30-37°C for 48-72 hours, and the bacteria and fermentation liquid were collected by centrifugation; the bacteria were freeze-dried, and the fermentation liquid was spray-dried; the dried bacteria and the dried fermentation liquid were mixed in a ratio of 5-3 to obtain a fermentation product, each gram containing 5×10 9 CFU live bacteria.

[0078] (5) Mix all the components, and mix them with D-mannitol and magnesium hydroxide to make a syrup. Fill each bottle with 100 mL.

[0079] Example 6

[0080] The composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following components: 18 parts of Echinacea extract, 6 parts of Bacillus subtilis spore powder, 35 parts of beta-glucan, 18 parts of sodium chondroitin sulfate, 10 parts of N-acetylglucosamine, 10 parts of gamma-aminobutyric acid, 35 parts of Lactobacillus plantarum fermentation product, 18 parts of D-mannitol and 18 parts of magnesium hydroxide.

[0081] The preparation method of a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following steps:

[0082] (1) Echinacea extract: 500 parts of dried Echinacea flowers were crushed, passed through a 60-mesh sieve, and extracted with supercritical CO2 at an extraction pressure of 25-35 MPa, an extraction temperature of 40-50°C, and an extraction time of 2-3 h; the extract was concentrated under reduced pressure to a relative density of 1.10-1.15, resuspended in water, allowed to stand, and filtered. The filtrate was separated and purified by a macroporous resin HD-2 column, and eluted with water, 40%, 60%, and 70%-80% ethanol in sequence, and the 70%-80% ethanol eluate was collected and dried to obtain the Echinacea extract.

[0083] (2) Bacillus subtilis spore powder: Inoculate Bacillus subtilis CICC 24713 on a slant and culture at 30-37°C for 48-72 hours; scrape off spores or bacterial moss and use 10 5CFU / mL of bacteria were inoculated into seed culture medium and cultured at 30-37℃ for 24h; 4% of the inoculation amount was transferred into a fermentation tank and fermented at 30-37℃ for 72h; the bacteria were collected by centrifugation and freeze-dried or spray-dried to obtain spore powder, which contained 10 10 CFU viable spores.

[0084] (3) β-Glucan, chondroitin sodium, N-acetylglucosamine and γ-aminobutyric acid were of commercially available food grade.

[0085] (4) Lactobacillus plantarum fermentation product: Lactobacillus plantarum CGMCC No.1.557 or CICC6098 is inoculated into a modified MRS medium, the modified MRS medium is supplemented with 2%-8% plant juice, anaerobically fermented at 30-37°C for 48-72 hours, and the bacterial cells and fermentation liquid are collected by centrifugation; the bacterial cells are freeze-dried, and the fermentation liquid is spray-dried; the dried bacterial cells and the dried fermentation liquid are mixed in a ratio of 5:6 to obtain a fermentation product, each gram of which contains 2.5×10 10 CFU live bacteria.

[0086] (5) Take all the components and mix them with D-mannitol and magnesium hydroxide, then put them into a chewable tablet press and compress them. Each tablet weighs 800 mg.

[0087] Comparative Example 1

[0088] A composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following components: 3 parts of Echinacea extract, 1 part of Bacillus subtilis spore powder, 10 parts of β-glucan, 5 parts of sodium chondroitin sulfate, 2 parts of N-acetylglucosamine, 2 parts of γ-aminobutyric acid, 10 parts of Lactobacillus plantarum fermentation product, 5 parts of D-mannitol and 5 parts of magnesium hydroxide, a total of 43 parts.

[0089] The preparation method of a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following steps:

[0090] (1) Echinacea extract: Take 500 parts of dried Echinacea flowers, crush them, pass through a 60-mesh sieve, add 4000 parts of 75% ethanol by volume, and reflux extract twice, each time for 1.5 hours. Combine the extracts, recover the ethanol under reduced pressure to dryness, and obtain a crude extract.

[0091] Prepare Bacillus subtilis spore powder (containing 10 8 CFU live spores) and Lactobacillus plantarum fermentation products (10 9 CFU live bacteria), mixed and granulated, and dried to obtain granules, each containing 40 mg.

[0092] Comparative Example 2

[0093] The composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following components: 1 part of Bacillus subtilis spore powder, 30 parts of beta-glucan, 15 parts of sodium chondroitin sulfate, 8 parts of N-acetylglucosamine, 8 parts of gamma-aminobutyric acid, 20 parts of Lactobacillus plantarum fermentation product, 15 parts of D-mannitol and 15 parts of magnesium hydroxide.

[0094] The preparation method of a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following steps:

[0095] Prepare Bacillus subtilis spore powder (containing 10 8 CFU live spores) and Lactobacillus plantarum fermentation products (10 9 CFU live bacteria), mixed, and compressed to obtain tablets, each weighing 500 mg.

[0096] Comparative Example 3

[0097] The composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following components: 10 parts of Echinacea extract, 3 parts of Bacillus subtilis spore powder, 20 parts of beta-glucan, 10 parts of sodium chondroitin sulfate, 5 parts of N-acetylglucosamine, 5 parts of gamma-aminobutyric acid, 10 parts of D-mannitol and 10 parts of magnesium hydroxide.

[0098] The preparation method of a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following steps:

[0099] (1) Echinacea extract: 500 parts of dried Echinacea flowers were crushed, passed through an 80-mesh sieve, and extracted by percolation with 95% ethanol for 3 times, each time for 2 hours. The extracts were combined, concentrated under reduced pressure, and freeze-dried to obtain the Echinacea extract.

[0100] Prepare Bacillus subtilis spore powder (containing 10 8 CFU live spores), mix the components and pack them into capsules to obtain capsules, each containing 400 mg.

[0101] Comparative Example 4

[0102] The composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following components: 12 parts of Echinacea extract, 4 parts of Bacillus subtilis spore powder, 25 parts of beta-glucan, 12 parts of sodium chondroitin sulfate, 6 parts of N-acetylglucosamine, 6 parts of gamma-aminobutyric acid, 25 parts of Lactobacillus plantarum fermentation product, 12 parts of D-mannitol and 12 parts of magnesium hydroxide.

[0103] The preparation method of a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following steps:

[0104] Echinacea extract was prepared by referring to the method of Examples 1-3, and Bacillus subtilis spore powder (containing 6×10 9 CFU live spores) and Lactobacillus plantarum fermentation products (1.5×10 10 CFU live bacteria), mix the components, grind into fine powder, pass through a 100-mesh sieve to obtain powder, 3g per bag.

[0105] Comparative Example 5

[0106] A composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following components: 5 parts of Echinacea extract, 2 parts of Bacillus subtilis spore powder, 20 parts of beta-glucan, 7 parts of sodium chondroitin sulfate, 4 parts of N-acetylglucosamine, 4 parts of gamma-aminobutyric acid, 15 parts of Lactobacillus plantarum fermentation product, 8 parts of D-mannitol, and 8 parts of magnesium hydroxide, a total of 73 parts.

[0107] The preparation method of a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following steps:

[0108] (1) Echinacea extract: Take 500 parts of dried Echinacea flowers, crush them, pass them through a 100-mesh sieve, and reflux extract them twice with a 60% ethanol aqueous solution, each time for 1 hour. The extracts are combined, heated to recover ethanol, concentrated to 250 ml, added with 200 parts of water, stirred evenly, chromatographed on a polyamide resin column, washed with water, eluted with 70% ethanol, and dried to obtain the extract.

[0109] Prepare Bacillus subtilis spore powder (containing 2×10 8 CFU live spores) and Lactobacillus plantarum fermentation products (5×10 9 CFU live bacteria), mixed with D-mannitol and magnesium hydroxide to make syrup, 100 mL per bottle.

[0110] Comparative Example 6

[0111] A composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following components: 20 parts of Echinacea extract, 6 parts of Bacillus subtilis spore powder, 35 parts of beta-glucan, 18 parts of sodium chondroitin sulfate, 10 parts of N-acetylglucosamine, 10 parts of gamma-aminobutyric acid, 35 parts of Lactobacillus plantarum fermentation product, 5 parts of D-mannitol and 5 parts of magnesium hydroxide, a total of 144 parts.

[0112] The preparation method of a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats comprises the following steps:

[0113] (1) Echinacea extract: Take 500 parts of dried Echinacea flowers, crush them, pass them through a 40-mesh sieve, use CO2 supercritical extraction, extraction pressure 20 MPa, extraction temperature 60°C, extraction time 1.5 h, concentrate under reduced pressure to 250 ml, add 150 parts of anhydrous ethanol, stir, let stand, precipitate, filter, wash the filter cake with 70% ethanol, and dry.

[0114] Prepare Bacillus subtilis spore powder (containing 8×10 10 CFU live spores) and Lactobacillus plantarum fermentation products (2×10 10 CFU live bacteria), mix all the components, and press into tablets to obtain chewable tablets, each weighing 720 mg.

[0115] In the above comparative examples, the amount of Echinacea extract in comparative example 1 is low and it has not been purified, and the content of effective ingredients is insufficient; comparative example 2 lacks the key component of Echinacea extract; comparative example 3 does not add Lactobacillus plantarum fermentation products, and the intestinal flora regulation ability is insufficient; comparative example 4 has not been granulated, and the powdered powder is easy to deliquesce and agglomerate, which is not conducive to storage and taking; in comparative example 5, the content of effective ingredients in Echinacea extract is low and the amount of each component is low; in comparative example 6, the amount of Echinacea and Lactobacillus plantarum is high, while the amount of excipients is too low, and the prepared chewable tablets are hard and difficult to chew.

[0116] According to the characteristics of the components of the present invention, the following pharmacological model was established to evaluate the effectiveness of the formulation:

[0117] 1. In vitro antibacterial test

[0118] 1.1 Experimental method: The compositions prepared in Examples 1-6 and Comparative Examples 1-6 were respectively prepared into 1 mg / mL sterile solutions, and the minimum inhibitory concentration (MIC) thereof against Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae and Enterococcus faecalis was determined by agar dilution method.

[0119] 1.2 Experimental results: minimum inhibitory concentration MIC (μg / mL);

[0120] The results of in vitro antibacterial test are as follows:

[0121]

[0122] Table 1

[0123] The results in Table 1 show that Examples 2, 4 and 6 have the strongest antibacterial activity, which is related to the higher dosage of Echinacea extract and the high content of Bacillus subtilis spores.

[0124] 2. In vivo anti-diarrhea test

[0125] 2.1 Experimental Methods 60 weaned puppies were randomly divided into 12 groups, 5 puppies in each group. The infected control group and the negative control group were gavaged with normal saline, and the remaining 10 groups were gavaged with the same dose of the composition of Examples 1-6 and Comparative Examples 1-4, twice a day for 5 consecutive days. After the last administration, except for the negative control group, the remaining groups were orally infected with 0.5 mL of rotavirus (containing 107 TCID50 / puppy). The time of diarrhea onset, duration and diarrhea index were observed and scored. At the same time, the titer of anti-rotavirus IgG antibody in serum was determined.

[0126] 2.2 Experimental Results

[0127] The results of the in vivo anti-diarrhea test are as follows:

[0128]

[0129] Table 2

[0130] Note: Compared with the control group, *P<0.05.

[0131] The results in Table 2 show that compared with the infected control group, taking the composition of the present invention can significantly prolong the time of diarrhea, shorten the duration of diarrhea, reduce the diarrhea index, and increase the serum antiviral IgG level, exerting a good therapeutic effect. Among them, Example 6 has the best efficacy. The comparative group has no significant effect.

[0132] 3. Intestinal mucosal barrier protection test

[0133] 3.1 Experimental methods The animals were grouped as described in the in vivo anti-diarrhea test. The animals were killed on the 3rd and 7th days after exposure, and the colon tissues were quickly isolated. The expression levels of tight junction proteins ZO-1 and Claudin-1 were determined by Western blot. At the same time, the mucin MUC2 content and lysozyme activity in the colon tissues were determined by colorimetry.

[0134] 3.2 Experimental Results

[0135] The results of the intestinal mucosal barrier protection test are as follows:

[0136]

[0137] Table 3

[0138] The results in Table 3 indicate that after exposure, the expression of ZO-1 and Claudin-1 in colon tissues was significantly reduced, and the content of MUC2 and lysozyme decreased, indicating that the intestinal mucosal barrier was damaged. The administration of Examples 1-6 can alleviate the damage of the intestinal barrier to varying degrees, among which the protective effects of Examples 2, 4, and 6 are the most significant. The protective effect of the comparative group is not good.

[0139] 4. Intestinal flora regulation test

[0140] 4.1 Experimental methods The experimental methods were grouped according to the in vivo anti-diarrhea test. Feces were collected on the 7th day after infection, and 16S rRNA sequencing was used to analyze the changes in intestinal flora, mainly observing the relative abundance of Firmicutes, Bacteroidetes, and Proteobacteria.

[0141] 4.2 Experimental Results

[0142] The results of the intestinal flora regulation test are as follows:

[0143] Group Firmicutes(%) Bacteroidetes (%) Proteobacteria (%) Negative control group 58.3 25.6 14.2 Control group 32.7 16.8 48.4 Example 1 45.9* 20.5 31.3* Example 2 54.2* 23.8* 20.6* Example 3 50.4* 22.7* 25.2* Example 4 52.9* 23.4* 22.1* Example 5 41.7 19.2 36.8* Example 6 57.1* 25.1* 16.5* Comparative Example 1 37.4 18.3 42.6 Comparative Example 2 34.1 17.2 46.7 Comparative Example 3 35.8 17.9 44.5 Comparative Example 4 39.2 18.7 40.3

[0144] Table 4

[0145] Note: In Tables 1-4, the values ​​marked with * indicate significant differences compared with the control group (P<0.05).

[0146] The results in Table 4 show that after infection, the intestinal flora was imbalanced, the Firmicutes (mainly beneficial bacteria) and Bacteroidetes decreased, and the Proteobacteria (mainly conditional pathogens) increased. After intervention with Examples 1-6, the proportions of Firmicutes and Bacteroidetes increased, the proportion of Proteobacteria decreased, and the flora structure tended to be normal. Examples 2, 4, and 6 had the best regulatory effects.

[0147] In summary, the present invention fully utilizes the complementary advantages of traditional Chinese medicine and probiotic preparations. Echinacea extract exerts antiviral and anti-inflammatory effects; Bacillus subtilis and Lactobacillus plantarum synergistically regulate the bacterial flora, produce antibacterial active substances, and reduce intestinal pathogens; β-glucan and chondroitin enhance immune function and promote the production of virus-specific antibodies; glucosamine and aminobutyric acid repair the intestinal mucosal barrier.

[0148] Different from the treatment methods of simple antiviral or flora regulation, the present invention systematically intervenes in rotavirus infection from multiple links to achieve synergistic effects: 1. Blocking viral infection and inhibiting viral replication in the intestine; 2. Improving flora disorder and maintaining intestinal microecological homeostasis; 3. Repairing the intestinal mucosal barrier and reducing inflammatory response; 4. Enhancing immune function and accelerating the body's clearance of viruses.

[0149] This multi-target, multi-level comprehensive regulation exhibits unique advantages of synergistic enhancement and overall regulation, which cannot be achieved by a single drug.

[0150] In addition, the present invention has carried out systematic optimization in terms of component extraction, microbial fermentation, etc., and adopted advanced processes such as supercritical extraction and spray drying, which effectively improved the stability and bioavailability of the drug and overcame the problems of low content of effective ingredients in traditional Chinese medicine and poor taste.

[0151] In summary, the present invention is based on the theory of traditional Chinese medicine, integrates modern biotechnology, optimizes the compound preparation, and creates an innovative product in the field of pet intestinal health. It is believed that this composition will become an ideal preparation for preventing and treating rotavirus diarrhea in pets.

[0152] Example 7

[0153] The use of a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats in the preparation of a medicament for treating rotavirus diarrhea in pet dogs and cats. The composition of the present invention has a significant effect in the treatment of rotavirus diarrhea in pets. The following dosage regimen is recommended:

[0154] The composition of Example 2 was made into granules, each bag containing 3 g, each gram containing 150 mg of Echinacea extract (inulin alcohol content 95%), 50 mg of Bacillus subtilis spore powder (containing 5×10 9 CFU), β-glucan 300mg, chondroitin sulfate sodium 150mg, N-acetylglucosamine 80mg, γ-aminobutyric acid 80mg, Lactobacillus plantarum fermentation product 300mg (containing 3×10 10 CFU), D-mannitol 150mg, magnesium hydroxide 150mg.

[0155] Specific usage: Dissolve the granules in an appropriate amount of warm water and mix well. For dogs and cats weighing less than 2.5kg, take 1g each time, 3 times a day; for dogs and cats weighing 2.5-5kg, take 2g each time, 3 times a day; for dogs and cats weighing more than 5kg, take 3g each time, 3 times a day. Use continuously for 5-7 days.

[0156] Rotavirus diarrhea usually heals itself within 3-5 days, but the course of the disease can be prolonged in puppies, those with low immunity or co-infection. The use of this composition can significantly shorten the course of the disease and improve symptoms. At the same time, the composition can not only treat diarrhea, but also repair damaged intestines, regulate flora, enhance immunity, and prevent reinfection through multi-target regulation.

[0157] It is worth noting that dogs and cats with diarrhea should be supplemented with sufficient water and electrolytes to prevent dehydration. Infusion therapy is recommended for severe diarrhea. This product can also be used in combination with other treatment measures.

[0158] The above description is only an embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent substitution, improvement, etc. made within the spirit and principle of the present invention should be included in the scope of the claims of the present invention.

Claims

1. A composition for treating diarrhea caused by rotavirus infection in pet dogs and cats, characterized in that , including the following components: 5-18 parts by weight of Echinacea extract, 1-6 parts by weight of Bacillus subtilis spore powder, 10-35 parts by weight of β-glucan, 5-18 parts by weight of sodium chondroitin sulfate, 2-10 parts by weight of N-acetylglucosamine, 2-10 parts by weight of γ-aminobutyric acid, 10-35 parts by weight of Lactobacillus plantarum fermentation product, 5-18 parts by weight of D-mannitol, and 5-18 parts by weight of magnesium hydroxide.

2. The composition for treating diarrhea caused by rotavirus infection in pet dogs and cats according to claim 1, characterized in that The preparation method of the Echinacea extract is as follows: 500 portions of dried Echinacea flowers are taken, crushed, passed through a 60-100 mesh sieve, and extracted by one of the following methods: (1) adding 4000 parts by volume of 75%-95% ethanol and reflux extraction 2-3 times, each time for 1.5-2 hours; or (2) extracting by percolation with 95% ethanol 2-3 times, each time for 1.5-2 hours; or (3) Using CO2 supercritical extraction, the extraction pressure is 25-35MPa, the extraction temperature is 40-50°C, and the extraction time is 2-3h; The extract is concentrated under reduced pressure to a relative density of 1.10-1.15, resuspended in water, allowed to stand, filtered, and the filtrate is separated and purified by a macroporous resin AB-8 or HD-2 column, eluted with water, 40%, 60%, and 70%-80% ethanol in sequence, and the 70%-80% ethanol eluate is collected and dried to obtain the Echinacea extract.

3. The composition for treating diarrhea caused by rotavirus infection in pet dogs and cats according to claim 1, characterized in that The preparation method of the Bacillus subtilis spore powder is as follows: inoculate Bacillus subtilis CICC 24713 or CICC10275 on a slant or plate, and culture at 30-37°C for 48-72 hours; scrape the spores or bacterial moss, and 5 CFU / mL of bacteria were inoculated into seed culture medium and cultured at 30-37℃ for 24h; 4% of the inoculation amount was transferred into a fermentation tank and fermented at 30-37℃ for 72h; the bacteria were collected by centrifugation and freeze-dried or spray-dried to obtain spore powder, which contained 10 8 -10 10 CFU viable spores.

4. The composition for treating diarrhea caused by rotavirus infection in pet dogs and cats according to claim 1, characterized in that The preparation method of the plant lactobacillus fermentation product is as follows: inoculate plant lactobacillus CGMCC No.1.557 or CICC6098 into MRS liquid culture medium or modified MRS culture medium, add 2%-8% plant juice to the modified MRS culture medium, perform anaerobically fermentation at 30-37°C for 48-72h, collect the bacteria and fermentation liquid by centrifugation; freeze-dry the bacteria and spray-dry the fermentation liquid; mix the dried bacteria and the dried fermentation liquid in a ratio of 5:5-9:1 to obtain the fermentation product, containing 10 8 -10 11 CFU live bacteria.

5. The composition for treating diarrhea caused by rotavirus infection in pet dogs and cats according to claim 4, characterized in that The plant juice is any one of cabbage juice, celery juice, carrot juice, tomato juice, cucumber juice, apple juice, pear juice and grape juice, or a combination of several of them. Fresh fruits and vegetables are taken, cleaned, squeezed and juiced, and the pomace is removed by low-temperature centrifugation. The supernatant is taken and filtered for sterilization to obtain sterile plant juice.

6. The composition for treating diarrhea caused by rotavirus infection in pet dogs and cats according to claim 1, characterized in that , including the following components: 10-15 parts by weight of Echinacea extract, 3-5 parts by weight of Bacillus subtilis spore powder, 20-30 parts by weight of β-glucan, 10-15 parts by weight of sodium chondroitin sulfate, 5-8 parts by weight of N-acetylglucosamine, 5-8 parts by weight of γ-aminobutyric acid, 20-30 parts by weight of Lactobacillus plantarum fermentation product, 8-15 parts by weight of D-mannitol, and 8-15 parts by weight of magnesium hydroxide.

7. A method for preparing a composition for treating diarrhea caused by rotavirus infection in pet dogs and cats according to any one of claims 1 to 6, characterized in that , comprising the following steps: (1) uniformly mixing Echinacea extract, Bacillus subtilis spore powder, β-glucan, sodium chondroitin sulfate, N-acetylglucosamine, γ-aminobutyric acid, Lactobacillus plantarum fermentation product, D-mannitol, and magnesium hydroxide according to the weight of the formula; (2) preparing the mixture obtained in step (1) into granules containing 40-50 mg per granule, tablets weighing 400-800 mg per tablet, capsules containing 300-500 mg per granule, powders containing 2-4 g per bag, syrups containing 80-120 mL per bottle, or chewable tablets weighing 600-1000 mg per tablet.

8. Use of the composition for treating diarrhea caused by rotavirus infection in pet dogs and cats according to any one of claims 1 to 6 in the preparation of a medicament for treating rotavirus diarrhea in pet dogs and cats.

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