Microecological preparation liquid-state fermentation compound traditional Chinese medicine for preventing cat parvovirus disease and preparation method of microecological preparation liquid-state fermentation compound traditional Chinese medicine

Abstract

The invention discloses a microecological preparation liquid-state fermentation compound traditional Chinese medicine for preventing cat parvovirus diseases and a preparation method thereof, and belongs to the technical field of traditional Chinese medicines. The preparation method comprises the following steps: crushing radix isatidis, hemarthria hemarthria, oldenlandia diffusa, herba patriniae, liquorice, radix sophorae flavescentis, coptis chinensis, purslane and sedum lineare, mixing and decocting to obtain traditional Chinese medicine liquid; and mixing the traditional Chinese medicine liquid with a culture medium, and sequentially carrying out aerobic fermentation and anaerobic fermentation to obtain the microecological preparation liquid-state fermentation compound traditional Chinese medicine. Compared with a common traditional Chinese medicine mixture or common micro-ecological bacterium liquid, the fermented traditional Chinese medicine liquid obtained by the invention can achieve the purposes of improving the immunity of group cats and preventing the cat parvovirus disease, strengthen the body of the group cats and effectively prevent the cat parvovirus disease.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine fermentation, and particularly relates to a liquid fermentation compound traditional Chinese medicine of a microecological preparation for preventing feline parvovirus disease and a preparation method thereof. Background Art

[0002] Feline parvovirus (CPV) is a highly contagious disease caused by the feline parvovirus (Canine Parvovirus, CPV). It primarily affects kittens, but also adult cats. Cats between weaning and 90 days of age are more likely to be infected, and the condition is more severe. Some kittens may develop symptoms of myocarditis and die suddenly. The incidence is higher in purebred and exotic cats than in indigenous cats. The disease can occur year-round, but is more common in cold weather. The feces of infected cats contain the highest levels of toxins. Cats infected with parvovirus are clinically divided into enteritis and myocarditis types. Initial symptoms include fever (above 40°C), depression, loss of appetite, vomiting, and diarrhea. Initially, the stool is watery; as the disease progresses, it becomes coffee-colored or ketchup-colored, bloody, and has a distinctive fishy odor. Severe dehydration occurs in later stages. The myocarditis type is more common in cats around 40 days of age, and some cats may die after experiencing mild diarrhea.

[0003] There is currently no specific treatment for feline parvovirus, and treatment is primarily symptomatic. For cats exhibiting symptoms such as diarrhea, broad-spectrum antibiotics can be added to their feed or drinking water to control the infection. While prevention is paramount for feline parvovirus, there are currently no effective preventive or therapeutic medications. Antibodies and chemical antibiotics are generally the preferred treatments for parvovirus. However, antibodies are extremely expensive and unsuitable for use in the average cat. Chemical antibiotics are highly susceptible to developing resistance, and even systematic alternating or combined use of multiple antibiotics for treatment or prevention cannot completely prevent the development of resistance.

[0004] Feline parvovirus has become a common occurrence in my country, posing an extremely challenging prevention and control situation. Traditional Chinese Medicine (TCM) has demonstrated effective prevention and treatment of similar viral infections. Appropriately selected TCM herbs can play a significant role in preventing and treating feline parvovirus. Herbs such as Astragalus membranaceus can enhance the body's immune function and improve resistance to viruses. Astragalus polysaccharide, one of Astragalus membranaceus's main active ingredients, promotes the phagocytic function of macrophages and enhances the function of T and B lymphocytes, thereby enhancing the body's immune response. Some TCM herbs, such as Isatis root and Honeysuckle, have antiviral properties, inhibiting viral replication and spread. For example, active ingredients in Isatis root, such as indirubin and indigo, can inhibit the synthesis of viral RNA and DNA, thereby exerting their antiviral effects. Herbs such as Honeysuckle and Folium Isatidis have anti-inflammatory properties, alleviating the inflammatory response caused by viral infection. Active ingredients in these herbs, such as chlorogenic acid and flavonoids, can inhibit the release of inflammatory mediators, reduce inflammation, and protect tissues from damage. Traditional Chinese medicine can also enhance the body's resistance to disease by regulating the body's yin and yang balance, as well as the balance of qi and blood. For example, astragalus root can replenish qi and blood, regulating the body's qi and blood balance; while isatis root can clear heat and detoxify, regulating the body's yin and yang balance.

[0005] Traditional Chinese medicine has certain potential in preventing and controlling feline parvovirus disease, which is mainly achieved through enhancing immunity, antiviral, and anti-inflammatory effects.

[0006] Probiotics, also known as microecological preparations, are live microorganisms that benefit the human body by improving the balance of intestinal flora. These preparations are made by adding probiotic strains to a sterilized liquid culture medium under controlled conditions, producing a variety of nutritious small molecules and beneficial microorganisms. This makes the liquid more nutritious while also improving animal health, combating disease, promoting growth, and saving feed costs. Probiotics have experienced rapid growth in the past decade and are a research hotspot in nutrition. Their primary function is to help establish a favorable gastrointestinal microbial flora for the host through competitive exclusion of digestive tract organisms, preventing bacterial, viral, and coccidial diarrhea, and promoting growth.

[0007] Although Chinese herbal medicine or probiotics can improve the intestinal microecological environment of kittens and enhance their immunity, for serious infections that have already occurred, especially those caused by feline parvovirus, the use of Chinese herbal medicine or probiotics alone may not achieve the ideal prevention and control effect. Sometimes the kittens die from severe diarrhea before the medicine takes effect, and the medicine loses its intended effect. Summary of the Invention

[0008] In view of this, the purpose of the present invention is to provide a liquid fermentation compound Chinese medicine of a probiotic preparation for preventing feline parvovirus disease and a preparation method thereof. The compound Chinese medicine of the present invention improves the immunity of cat populations, effectively treats and / or prevents feline parvovirus disease, and is of high quality and more nutritious.

[0009] In order to achieve the above object, the present invention provides the following technical solutions:

[0010] In a first aspect, the present invention provides a method for preparing a liquid fermentation compound Chinese medicine of a probiotic preparation for preventing feline parvovirus disease, comprising the following steps:

[0011] S1. According to the mass parts weighed isatis root, bullwhip, white flower tongue grass, Patrinia grass, licorice, Sophora flavescens, Coptis chinensis, Portulaca oleracea, Sedum grass after crushing, mixing, decocting to obtain a Chinese medicine liquid;

[0012] S2. The Chinese medicine liquid is mixed with the culture medium and then subjected to aerobic fermentation and anaerobic fermentation to obtain a microecological liquid fermentation compound Chinese medicine;

[0013] The fermentation bacteria used in the aerobic fermentation are composed of Bacillus licheniformis SXDYYB11, Bacillus cereus, and Hansenula in a mass ratio of (0.5-3): (0.5-3): (0.5-4);

[0014] The Bacillus licheniformis SXDYYB11 was deposited in the China Center for Type Culture Collection on April 18, 2025, with the deposit number CCTCCNO: M2025815;

[0015] The fermentation bacteria used in the anaerobic fermentation are composed of Lactobacillus reuteri and Bacillus butyricum in a mass ratio of (0.5-5): (0.5-5).

[0016] Preferably, the mass ratio of Radix Isatidis, Herba Verbenae, Herba Hedyotis Diffusae, Herba Patriniae, Radix Glycyrrhizae, Sophora flavescens, Rhizoma Coptidis, Herba Portulacae, and Herba Sedum in S1 is (10-15): (5-10): (20-25): (20-25): (20-25): (5-15): (5-15): (5-15): (5-15).

[0017] Preferably, the crushing mesh size in S1 is 50-80 mesh.

[0018] Preferably, the decoction time in S1 is 3-6 hours; and the mass ratio of material to liquid during decoction is 1:(5-10).

[0019] Preferably, the culture medium in S2 comprises the following components in parts by weight:

[0020] 65 parts of corn flour, 20 parts of fish meal, 12 parts of wheat bran powder, 0.5 parts of trace elements, 0.5 parts of vitamins, 1 part of potassium carbonate, 1 part of ammonium chloride, and 2000 parts of water.

[0021] Preferably, the corn flour, fish meal and wheat bran flour have a mesh size of 40-60 mesh.

[0022] Preferably, the trace elements are composed of cobalt chloride, chromium picolinate and strontium chloride in a mass ratio of 1:1:1.

[0023] Preferably, the vitamins are composed of vitamin B6, vitamin B12, and vitamin E in a mass ratio of 1:1:1.

[0024] Preferably, the mass ratio of the Chinese medicinal solution to the culture medium in S2 is (5-100):(1-10).

[0025] Preferably, the amount of bacteria used for aerobic fermentation and anaerobic fermentation in S2 is 0.1-10% of the mass of the culture medium;

[0026] The number of strains is 10 6 -10 10 CFU / g.

[0027] Preferably, the temperature of the aerobic fermentation in S2 is 20-35° C., and the fermentation time is 24-120 h; oxygen is introduced during the fermentation, and the fermentation is allowed to stand for 12-24 h after the aerobic fermentation is completed.

[0028] Preferably, the temperature of the anaerobic fermentation in S2 is 20-35° C., and the fermentation time is 24-120 h.

[0029] In a second aspect, the present invention provides a liquid fermentation compound Chinese medicine of a probiotic preparation for preventing feline parvovirus disease, which is prepared by the above method.

[0030] Contains at least the following beneficial technical effects:

[0031] The present invention adds boiled Chinese medicinal liquid to a liquid culture medium to adjust the water content of the fermentation culture medium for microbial fermentation. Compared with common Chinese medicinal mixtures or common microecological bacterial liquids, the present invention can achieve the purpose of improving the immunity of cats and preventing feline parvovirus disease, and strengthen the physical fitness of cats. Compared with adding Chinese medicinal additives and common microecological bacterial liquids, the present invention saves costs. The present invention combines aerobic and anaerobic fermentation methods, so that the microecological bacterial liquid prepared by the present invention takes into account the advantages of anaerobic fermentation and aerobic fermentation, and produces a variety of high-density nutritious small molecular substances and beneficial microorganisms. In particular, aerobic fermentation is first performed, and then the fermentation is allowed to stand for a period of time after completion to exhaust the dissolved oxygen in the bacterial liquid before anaerobic fermentation is performed, which can achieve the effect of reducing fermentation costs. Compared with common microecological bacterial liquids, the microecological bacterial liquid prepared by the present invention is of higher quality and more nutritious. At the same time, the Chinese medicinal components contained in the microecological bacterial liquid can improve animal health, play the role of disease resistance, growth promotion and feed cost saving. The use of microecological bacterial liquid and traditional Chinese medicine has a dual-effect enhancing effect on improving the disease resistance of cats. The results show that for prevention, the feeding time can be halved for 7-10 days, which can effectively control the occurrence of feline parvovirus in the epidemic area or reduce the infection rate, with an efficiency of 75-95%.

[0032] Deposit information: Bacillus licheniformis SXDYYB11 was deposited in the China Center for Type Culture Collection on April 18, 2025, with the deposit number CCTCCNO: M2025815. DETAILED DESCRIPTION

[0033] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0034] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each smaller range between any intermediate value within a stated value or stated range and any other stated value or intermediate value within the stated range is also encompassed by the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0035] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.

[0036] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the present invention. The present description and examples are intended to be illustrative only.

[0037] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0038] Unless otherwise specified, the "room temperature" and "normal temperature" mentioned in the present invention are all calculated as 25±2°C.

[0039] Unless otherwise specified, all raw materials and instruments used in the following examples of the present invention are commercially available.

[0040] Bacillus licheniformis SXDYYB11 was isolated from the intestine of a healthy kitten.

[0041] Culture medium: 65 parts of corn flour, 20 parts of fish meal, 12 parts of wheat bran powder, 0.5 parts of trace elements, 0.5 parts of vitamins, 1 part of potassium carbonate, 1 part of ammonium chloride, and 2000 parts of water; the trace elements are composed of cobalt chloride, chromium picolinate, and strontium chloride in a mass ratio of 1:1:1.

[0042] The vitamins are composed of vitamin B6, vitamin B12, and vitamin E in a mass ratio of 1:1:1.

[0043] Example 1

[0044] The present invention provides a method for preparing a liquid fermentation compound Chinese medicine of a probiotic preparation for preventing feline parvovirus disease, comprising the following steps:

[0045] S1. According to the mass ratio of 10:10:20:16:12:12:10:8:2, Isatis root, Herba Hedyotis diffusae, Herba Patriniae, Glycyrrhizae, Sophora flavescens, Coptis chinensis, Herba Portulacae, and Herba Sedum were weighed and ground into 50 mesh, and then 5 times the mass of water was added and decocted for 5h to obtain a Chinese medicine solution;

[0046] S2. The Chinese medicine liquid and the culture medium were mixed in a mass ratio of 2:1 and sterilized at 115°C for 60 minutes. After cooling, 0.1wt% of aerobic fermentation bacteria was added and then stirred continuously at 25°C for 72 hours. During the stirring process, sterile air was introduced into the liquid fermentation tank for aerobic fermentation to obtain an intermediate product, which was allowed to stand for 12 hours. Then, 0.1wt% of anaerobic fermentation bacteria was added and anaerobically fermented at 25°C for 48 hours to obtain a microecological preparation liquid fermentation compound Chinese medicine; wherein the aerobic fermentation bacteria consisted of Bacillus licheniformis SXDYYB11, Bacillus cereus, and Hansenula in a mass ratio of 0.5:0.5:0.5, and each strain was 10 8 CFU / g; the fermentation bacteria used in anaerobic fermentation were composed of Lactobacillus reuteri and Bacillus butyricum in a mass ratio of 0.5:0.5, and each strain was 10 8 CFU / g.

[0047] Example 2

[0048] The present invention provides a method for preparing a liquid fermentation compound Chinese medicine of a probiotic preparation for preventing feline parvovirus disease, comprising the following steps:

[0049] S1. According to the mass ratio of 10::12:22:15:15:10:8:6:2, weighed isatis root, bullwhip, white flower tongue grass, Patrinia, licorice, sophora flavescens, coptis chinensis, purslane, and sedum were crushed to 80 mesh and then added with 10 times the mass of water and boiled for 4h to obtain a Chinese medicine solution;

[0050] S2. The Chinese medicine liquid and the culture medium were mixed in a mass ratio of 1:15, and then 0.1wt% of aerobic fermentation bacteria was added and then stirred continuously at 25°C for 72h. During the stirring process, sterile air was introduced into the liquid fermentation tank for aerobic fermentation to obtain an intermediate product, which was allowed to stand for 12h; then 0.15wt% of anaerobic fermentation bacteria was added and anaerobically fermented at 25°C for 48h to obtain a microecological preparation liquid fermentation compound Chinese medicine; wherein the aerobic fermentation bacteria consisted of Bacillus licheniformis SXDYYB11, Bacillus cereus, and Hansenula in a mass ratio of 0.5:0.7:1, and each strain was 10 8 CFU / g; the fermentation bacteria used in anaerobic fermentation were composed of Lactobacillus reuteri and Bacillus butyricum in a mass ratio of 0.5:1, and each strain was 10 8 CFU / g.

[0051] Example 3

[0052] The present invention provides a method for preparing a liquid fermentation compound Chinese medicine of a probiotic preparation for preventing feline parvovirus disease, comprising the following steps:

[0053] S1. According to the mass ratio of 10:5:20:20:20:5:5:5:5, Isatis root, Herba Hedyotis diffusae, Herba Patriniae, Glycyrrhizae, Sophora flavescens, Coptis chinensis, Herba Portulacae, and Herba Sedum were weighed and crushed to 60 mesh, and then 8 times the mass of water was added and decocted for 3h to obtain a Chinese medicine solution;

[0054] S2. The Chinese medicine liquid and the culture medium were mixed in a mass ratio of 10:1, and 5 wt% of aerobic fermentation bacteria were added, and then the mixture was stirred continuously at 20°C for 120 h. During the stirring process, sterile air was introduced into the liquid fermentation tank for aerobic fermentation to obtain an intermediate product, which was allowed to stand for 24 h. Then, 5 wt% of anaerobic fermentation bacteria was added and anaerobically fermented at 20°C for 120 h to obtain a microecological preparation liquid fermentation compound Chinese medicine; wherein the aerobic fermentation bacteria consisted of Bacillus licheniformis SXDYYB11, Bacillus cereus, and Hansenula in a mass ratio of 3:3:4, and each strain was 10 8 CFU / g; the fermentation bacteria used in anaerobic fermentation were composed of Lactobacillus reuteri and Bacillus butyricum in a mass ratio of 0.5:5, and each strain was 10 8 CFU / g.

[0055] Comparative Example 1

[0056] The preparation method of this comparative example is the same as that of Example 1, except that the Chinese medicine liquid is not fermented.

[0057] Comparative Example 2

[0058] The preparation method of this comparative example is the same as that of Example 1, except that no culture medium components are added during fermentation.

[0059] Comparative Example 3

[0060] The preparation method of this comparative example is the same as that of Example 1, except that commercially available Bacillus licheniformis is used instead of Bacillus licheniformis SXDYYB11 during fermentation.

[0061] Comparative Example 4

[0062] The preparation method of this comparative example is the same as that of Example 1, except that only aerobic fermentation is used during fermentation.

[0063] Experimental Example 1

[0064] Preventive effect experiment

[0065] Prevention for weaned cats (50 days old, weaned at 35 days old);

[0066] The groups were divided into a blank group (no prevention) and a prevention group (prevention). The prevention group was treated with the compositions of Examples 2-4 and Comparative Examples 1-6, respectively, at a rate of 10 grams per kitten mixed into cat food for 5 consecutive days. The kittens were then placed with kittens infected with parvovirus and the onset of disease was observed within 30 days. The prevention effect results are shown in the following table:

[0067] Prevention effect results table

[0068]

[0069] As shown in Table 1, the composition prepared by the present invention can effectively prevent parvovirus disease, and the effect is significantly weakened when a certain component or strain is replaced.

[0070] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A method for preparing a liquid fermentation compound Chinese medicine of a probiotic preparation for preventing feline parvovirus disease, characterized in that: The following steps are involved: S1. According to the mass parts, weighed isatis root, bullwhip, white flower tongue grass, Patrinia grass, licorice, Sophora flavescens, Coptis chinensis, Portulaca oleracea, Sedum grass were crushed, mixed, and decocted to obtain a Chinese medicine liquid; S2. The Chinese medicine liquid is mixed with the culture medium and then subjected to aerobic fermentation and anaerobic fermentation to obtain a microecological liquid fermentation compound Chinese medicine; The fermentation bacteria used in the aerobic fermentation are composed of Bacillus licheniformis SXDYYB11, Bacillus cereus, and Hansenula in a mass ratio of (0.5-3): (0.5-3): (0.5-4); The Bacillus licheniformis SXDYYB11 was deposited in the China Center for Type Culture Collection on April 18, 2025, with the deposit number CCTCCNO: M2025815; The fermentation bacteria used in the anaerobic fermentation are composed of Lactobacillus reuteri and Bacillus butyricum in a mass ratio of (0.5-5): (0.5-5).

2. The preparation method according to claim 1, characterized in that The mass ratio of Radix Isatidis, Herba Verbenae, Herba Hedyotis Diffusae, Herba Patriniae, Radix Glycyrrhizae, Radix Sophorae Flavescentis, Rhizoma Coptidis, Herba Portulacae, and Herba Sedum in S1 is (10-15): (5-10): (20-25): (20-25): (20-25): (5-15): (5-15): (5-15): (5-15).

3. The preparation method according to claim 1, characterized in that The culture medium in S2 includes the following components in parts by weight: 65 parts of corn flour, 20 parts of fish meal, 12 parts of wheat bran powder, 0.5 parts of trace elements, 0.5 parts of vitamins, 1 part of potassium carbonate, 1 part of ammonium chloride, and 2000 parts of water.

4. The preparation method according to claim 5, characterized in that The trace elements are composed of cobalt chloride, chromium picolinate and strontium chloride in a mass ratio of 1:1:

1.

5. The preparation method according to claim 5, characterized in that The vitamins are composed of vitamin B6, vitamin B12, and vitamin E in a mass ratio of 1:1:

1.

6. The preparation method according to claim 1, characterized in that The mass ratio of the Chinese medicinal solution in S2 to the culture medium is (5-100): (1-10).

7. The preparation method according to claim 1, characterized in that The amount of bacteria used for aerobic and anaerobic fermentation in S2 is 0.1-10% of the mass of the culture medium; The number of strains is 10 6 -10 10 CFU / g.

8. The preparation method according to claim 1, characterized in that The temperature of the aerobic fermentation in S2 is 20-35° C., and the fermentation time is 24-120 hours. Oxygen is introduced during the fermentation, and the fermentation is allowed to stand for 12-24 hours after the aerobic fermentation is completed.

9. The preparation method according to claim 1, characterized in that The temperature of the anaerobic fermentation in S2 is 20-35° C., and the fermentation time is 24-120 hours.

10. A liquid fermentation compound Chinese medicine of a probiotic preparation for preventing feline parvovirus disease, characterized in that: The method is prepared by any one of claims 1 to 9.

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