Probiotic fermented traditional Chinese medicine composition for preventing and controlling canine distemper and preparation method thereof

Abstract

The invention discloses a probiotic fermented traditional Chinese medicine composition for preventing and controlling canine distemper and a preparation method of the probiotic fermented traditional Chinese medicine composition, and belongs to the technical field of fermented compositions. The feed additive is prepared from the following raw materials in parts by weight through fermentation: 100 to 200 parts of cortex phellodendri, 10 to 20 parts of radix aucklandiae, 100 to 200 parts of liquorice root, 200 to 400 parts of herba taraxaci, 100 to 200 parts of rhizoma coptidis and 10 to 40 parts of a microbial preparation. The microbial preparation comprises bacillus licheniformis SXDYYB15, the bacillus licheniformis SXDYYB15 is preserved in the China Center for Type Culture Collection on April 18, 2025, and the preservation number of the bacillus licheniformis SXDYYB15 is CCTCC (China Center for Type Culture Collection) NO: M2025814. The composition breaks through the bottleneck that the traditional Chinese medicine takes effect slowly through a triple synergistic mechanism of traditional Chinese medicine antivirus, probiotic immunoregulation and gastrointestinal function repair; on the premise of adapting to the canine intestinal environment, the pharmacodynamic bioavailability is remarkably improved through directional fermentation, and an efficient, safe and economical solution is provided for prevention and control of canine distemper.

Description

Technical Field

[0001] This invention belongs to the field of fermentation composition technology, specifically relating to a probiotic fermented traditional Chinese medicine composition for the prevention and control of canine distemper and its preparation method. Background Technology

[0002] Canine distemper is a highly contagious and fatal infectious disease caused by the canine distemper virus. Early symptoms include a biphasic fever pattern resembling a cold, followed by bronchitis, catarrhal pneumonia, and gastroenteritis. Later stages may present with neurological symptoms such as spasms and convulsions. Some cases may exhibit hyperkeratosis of the nose and paw pads (hard paw pad disease).

[0003] Canine distemper primarily affects puppies, and its pathogen is the canine distemper virus. Sick dogs exhibit a biphasic fever pattern, rhinitis, digestive disorders, and respiratory inflammation; encephalitis may also occur in a few cases. Various secretions, excrement, and blood of infected dogs contain large amounts of the virus, which can be excreted through respiratory secretions and urine. Healthy dogs can be infected through contact with infected dogs or through contaminated air or food via the respiratory or digestive tract. Besides puppies, fur-bearing animals such as foxes and minks are also highly susceptible. The disease can occur year-round, but is more prevalent in winter and spring. It has a certain periodicity, with a major epidemic occurring every three years. Dogs of all ages, sexes, and breeds can be infected, but young puppies are most susceptible. Purebred dogs and police dogs are more susceptible than mixed-breed dogs, and their symptoms are more severe, with a higher mortality rate. The most important sources of infection are nasal and ocular secretions and urine. Reports have indicated that dogs infected with canine distemper can still shed the virus in their urine 60-90 days later, making urine a dangerous source of infection. The primary route of transmission is direct contact between infected and healthy dogs, but it can also be transmitted through respiratory droplets. If a dog in a shared room is found to have canine distemper, no amount of preventative measures can completely prevent infection of other dogs living in the same room.

[0004] There is currently no specific chemical treatment for canine distemper; treatment is mainly symptomatic. For dogs exhibiting symptoms such as diarrhea, broad-spectrum antibiotics can be added to their feed or drinking water to control the infection. Prevention is paramount in treating canine distemper, but there are currently no effective preventative or curative drugs. Antibodies and chemical antibiotics are usually the first-line treatments for canine distemper. However, antibodies are extremely expensive and not suitable for ordinary dogs. Chemical antibiotics readily induce resistance; even with planned alternation or combination of multiple antibiotics for treatment or prevention, resistance cannot be completely avoided.

[0005] Traditional Chinese medicine (TCM) has shown good efficacy in preventing and treating viral infectious diseases. Selecting appropriate TCM herbs can effectively prevent and treat canine distemper. TCM's control of canine distemper primarily achieves this through enhancing the body's immunity, antiviral activity, and anti-inflammatory effects.

[0006] The mechanisms by which Traditional Chinese Medicine (TCM) controls canine distemper mainly include the following aspects: Enhancing Immunity: Many components of TCM, such as Astragalus and Codonopsis, can strengthen the pet's immune function and improve its resistance to viral infections. Antiviral Effects: Some TCM components have direct antiviral effects. For example, honeysuckle and Isatis root contain various effective ingredients that can inhibit the replication and spread of canine distemper virus. Regulating Immune Response: TCM can reduce viral inflammation and tissue damage by regulating the pet's immune response. For example, glycyrrhizic acid in licorice has anti-inflammatory effects and can relieve fever and pain symptoms caused by canine distemper. Improving Clinical Symptoms: TCM can also improve the clinical symptoms of canine distemper through symptomatic treatment. For example, using antitussive and antidiarrheal TCM components can relieve symptoms such as coughing and diarrhea in pets, improving their quality of life. Promoting Recovery: During the recovery period of canine distemper, TCM can help pets regain strength and promote the repair of damaged tissues. For example, using spleen-tonifying and qi-boosting TCM can strengthen the pet's constitution and accelerate its recovery process.

[0007] In summary, the mechanisms by which traditional Chinese medicine (TCM) prevents and controls canine distemper are multifaceted, including direct antiviral effects as well as indirect methods such as regulating immune responses and improving clinical symptoms to help pets resist viral infections. It is important to note that the use of TCM to treat canine distemper should be under the guidance of a professional veterinarian to ensure the safety and effectiveness of the treatment.

[0008] While traditional Chinese medicine has some effect on canine distemper, its efficacy remains limited, with slow-acting drugs failing to achieve the desired control effect. Therefore, there is a need for an effective traditional Chinese medicine combination with high efficiency and rapid efficacy. Summary of the Invention

[0009] In view of this, the purpose of this invention is to provide a probiotic fermented traditional Chinese medicine composition for the prevention and control of canine distemper and its preparation method. The composition of this invention breaks through the bottleneck of slow efficacy of traditional Chinese medicine through a triple synergistic mechanism of antiviral action of traditional Chinese medicine, immune regulation by probiotics, and repair of gastrointestinal function.

[0010] To achieve the above objectives, the present invention provides the following technical solution:

[0011] In a first aspect, the present invention provides a probiotic fermented traditional Chinese medicine composition for the prevention and control of canine distemper, which is obtained by fermentation from the following raw materials in parts by weight:

[0012] 100-200 parts of Phellodendron bark, 10-20 parts of Aucklandia lappa, 100-200 parts of Glycyrrhiza uralensis, 200-400 parts of Taraxacum mongolicum, 100-200 parts of Coptis chinensis, and 10-40 parts of microbial preparations;

[0013] The microbial preparation includes Bacillus licheniformis SXDYYB15, which was deposited at the China Center for Type Culture Collection on April 18, 2025, with accession number CCTCC NO:M2025814.

[0014] Phellodendron amurense (Latin name: Phellodendron amurense) has a slight odor and an extremely bitter taste, becoming sticky when chewed. It has the effects of clearing heat and drying dampness, purging fire and detoxifying, and relieving bone steaming fever. It is used to treat painful urination due to heat, damp-heat diarrhea, jaundice, damp-heat beriberi, flaccidity, bone steaming fever, night sweats, seminal emission, sores and carbuncles, and eczema with itching. Phellodendron amurense is bitter and cold in nature, and enters the kidney and bladder meridians.

[0015] Licorice: Latin name: Radix Glycyrrhizae; it is neutral in nature, sweet in taste, and has a slight odor. It is primarily used for clearing heat and detoxifying, resolving phlegm and relieving cough, and for abdominal ailments.

[0016] Dandelion: Latin name Herba Taraxaci; sweet, slightly bitter, cold. It clears heat and detoxifies, reduces swelling and dissipates nodules.

[0017] Costus root (Latin name: *Ligusticum chuanxiong Hort.*) is pungent, bitter, and warm in nature, with a fragrant and pungent aroma. It can unblock the flow of Qi throughout the body, especially effective in relieving discomfort caused by Qi stagnation in the spleen and stomach. Therefore, costus root is an excellent remedy for regulating Qi, harmonizing the middle Jiao, and relieving pain. It also strengthens the spleen, aids digestion, and stimulates appetite, thus it is often used to treat symptoms such as Qi stagnation in the spleen and stomach, abdominal distension and pain, food stagnation, indigestion, and loss of appetite. It can also treat tenesmus after diarrhea, damp-heat diarrhea, and other symptoms of incomplete defecation.

[0018] In this invention, Phellodendron bark, Coptis chinensis, and dandelion directly inhibit viral replication; licorice polysaccharides and probiotic metabolites synergistically activate macrophage and NK cell activity; Aucklandia lappa promotes gastrointestinal motility, and probiotics maintain flora balance to combat viral gastroenteritis; the anti-inflammatory components of licorice repair mucosal damage, and the fermentation process converts flavonoids, alkaloids, etc., into small molecules, improving bioavailability; probiotic metabolism produces novel active substances (such as antimicrobial peptides), enhancing anti-infection capabilities.

[0019] Secondly, the present invention also provides a method for preparing the above-mentioned probiotic fermented traditional Chinese medicine composition, comprising the following steps:

[0020] S1. Weigh the raw materials according to the stated weight proportions;

[0021] S2. Powder the following herbs: Phellodendron bark, Costus root, licorice root, dandelion, and Coptis root, then add water and decoct together to obtain a traditional Chinese medicine liquid.

[0022] S3. After the Chinese medicine liquid cools down, add microbial preparations for fermentation to obtain a probiotic fermented Chinese medicine composition.

[0023] Preferably, the material-to-liquid ratio in S2 is 1:(10~30).

[0024] Preferably, the simmering time in S2 is 25-35 minutes.

[0025] Preferably, the amount of microbial preparation added in step S3 is 1-5 wt%; the viable count of the microbial preparation is ≥1.80 × 10⁻⁶. 9 CFU / mL.

[0026] Preferably, the preparation method of the microbial preparation is as follows:

[0027] The seed culture of Bacillus licheniformis with accession number CCTCCNO.CCTCCM2025814 was inoculated into a culture medium and fermented.

[0028] Fermentation conditions: continuous culture at 30-37℃ for 24-72 hours, followed by static culture for 96-120 hours until the bacterial count is ≥1.80×10⁻⁶. 9 CFU / mL.

[0029] Preferably, the culture medium consists of the following components by weight: 1-5 parts peptone, 1-3 parts beef extract, 1-10 parts yeast extract, 5-15 parts sodium chloride, 10-100 parts fructooligosaccharide, and 1000 parts water.

[0030] Preferably, the fermentation conditions in S3 are: static closed fermentation at 0-37℃ for 72-240 hours.

[0031] It contains at least the following beneficial technical effects:

[0032] The composition of this invention overcomes the bottleneck of slow efficacy of traditional Chinese medicine through a triple synergistic mechanism of antiviral action by traditional Chinese medicine, immune regulation by probiotics, and repair of gastrointestinal function. The SXDYYB15 strain, adapted to the canine intestinal environment, significantly improves bioavailability through targeted fermentation, providing an efficient, safe, and economical solution for canine distemper prevention and control; simultaneously, it synergizes with the Chinese medicine components to exert a significant therapeutic effect and enhances the dog's own immunity.

[0033] Preservation information: Bacillus licheniformis SXDYYB15 was deposited at the China Center for Type Culture Collection on April 18, 2025, with accession number CCTCC NO:M2025814. Detailed Implementation

[0034] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0035] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0036] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0037] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be obvious to those skilled in the art. This application specification and embodiments are merely exemplary.

[0038] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0039] Unless otherwise specified, "room temperature" and "normal temperature" in this invention refer to 25±2℃.

[0040] Unless otherwise specified, all raw materials or instruments used in the following embodiments of the present invention are commercially available.

[0041] The Bacillus licheniformis SXDYYB15 was isolated from the intestines of healthy canine distemper patients and identified as Bacillus licheniformis by 16S RNA.

[0042] Preparation of microbial preparations:

[0043] It was prepared by inoculating a seed culture containing Bacillus licheniformis M 2025814 into a culture medium and fermenting it.

[0044] Fermentation conditions were: continuous culture at 35℃ for 48 hours followed by static culture for 96 hours, resulting in a bacterial count of 2.3 × 10⁻⁶. 9 CFU / mL;

[0045] The culture medium consists of the following components by weight: 3 parts peptone, 2 parts beef extract, 7 parts yeast extract, 11 parts sodium chloride, 75 parts fructooligosaccharide, and 1000 parts water.

[0046] Example 1

[0047] This embodiment provides a method for preparing a probiotic fermented traditional Chinese medicine composition for the prevention and control of canine distemper. The preparation steps are as follows:

[0048] S1. Weigh out 100 parts of Phellodendron bark, 10 parts of Aucklandia lappa, 100 parts of Glycyrrhiza uralensis, 200 parts of Taraxacum mongolicum, 100 parts of Coptis chinensis, and 10 parts of microbial preparation by weight.

[0049] S2. Pulverize the following herbs: Phellodendron bark, Costus root, licorice root, dandelion, and Coptis root, and pass them through an 80-mesh sieve. Add water equal to 10 times the total weight of the herbs and boil at normal pressure for 30 minutes to obtain a traditional Chinese medicine liquid.

[0050] S3. After the Chinese medicine liquid cools down, add 3wt% microbial preparation and ferment statically at 20℃ for 120h to obtain a probiotic fermented Chinese medicine composition.

[0051] Example 2

[0052] This embodiment provides a method for preparing a probiotic fermented traditional Chinese medicine composition for the prevention and control of canine distemper. The preparation steps are as follows:

[0053] S1. Weigh out 200 parts of Phellodendron bark, 20 parts of Aucklandia lappa, 200 parts of Glycyrrhiza uralensis, 400 parts of Taraxacum mongolicum, 200 parts of Coptis chinensis, and 20 parts of microbial preparation by weight.

[0054] S2. Pulverize the following herbs: Phellodendron bark, Costus root, licorice root, dandelion, and Coptis root, and pass them through an 80-mesh sieve. Add water equal to 20 times the total weight of the herbs and boil at normal pressure for 30 minutes to obtain a traditional Chinese medicine liquid.

[0055] S3. After the Chinese medicine liquid cools down, add 1wt% microbial preparation and ferment statically at 30℃ for 80 hours to obtain a probiotic fermented Chinese medicine composition.

[0056] Example 3

[0057] This embodiment provides a method for preparing a probiotic fermented traditional Chinese medicine composition for the prevention and control of canine distemper. The preparation steps are as follows:

[0058] S1. Weigh out 20 parts of Phellodendron bark, 2 parts of Aucklandia lappa, 10 parts of Glycyrrhiza uralensis, 20 parts of Taraxacum mongolicum, 10 parts of Coptis chinensis, and 1 part of microbial preparation by weight.

[0059] S2. Pulverize the following herbs: Phellodendron bark, Costus root, licorice root, dandelion, and Coptis root, and pass them through an 80-mesh sieve. Add water equal to 30 times the total weight of the herbs and boil at normal pressure for 30 minutes to obtain a traditional Chinese medicine liquid.

[0060] S3. After the Chinese medicine liquid cools down, add 5wt% microbial preparation and ferment statically at 15℃ for 180h to obtain a probiotic fermented Chinese medicine composition.

[0061] Example 4

[0062] This embodiment provides a method for preparing a probiotic fermented traditional Chinese medicine composition for the prevention and control of canine distemper. The preparation steps are as follows:

[0063] S1. Weigh out 30 parts of Phellodendron bark, 3 parts of Aucklandia lappa, 20 parts of Glycyrrhiza uralensis, 30 parts of Taraxacum mongolicum, 30 parts of Coptis chinensis, and 3 parts of microbial preparation by weight.

[0064] S2. Pulverize the following herbs: Phellodendron bark, Costus root, licorice root, dandelion, and Coptis root, and pass them through an 80-mesh sieve. Add water equal to 30 times the total weight of the herbs and boil at normal pressure for 30 minutes to obtain a traditional Chinese medicine liquid.

[0065] S3. After the Chinese medicine liquid cools down, add 2wt% microbial preparation and ferment statically at 5℃ for 240h to obtain a probiotic fermented Chinese medicine composition.

[0066] Comparative Example 1

[0067] This comparative example provides a method for preparing a traditional Chinese medicine composition for the prevention and control of canine distemper. The preparation steps are as follows:

[0068] S1. Weigh out 30 parts of Phellodendron bark, 3 parts of Aucklandia lappa, 20 parts of Glycyrrhiza uralensis, 30 parts of Taraxacum mongolicum, 30 parts of Coptis chinensis, and 3 parts of microbial preparation by weight.

[0069] S2. Pulverize Phellodendron bark, Costus root, licorice root, dandelion, and Coptis root through an 80-mesh sieve, add 30 times the total weight of the herbs to water, and boil at normal pressure for 30 minutes to obtain a traditional Chinese medicine composition.

[0070] Comparative Example 2

[0071] This comparative example provides a method for preparing probiotics for the prevention and control of canine distemper. The preparation steps are as follows:

[0072] Probiotics are obtained by fermenting microbial preparations.

[0073] Comparative Example 3

[0074] The preparation method of this comparative example is the same as that of Example 1, except that the bacterial agent used is commercially available Bacillus licheniformis.

[0075] Comparative Example 4

[0076] The preparation method of this comparative example is the same as that of Example 1, except that it does not contain dandelion and coptis.

[0077] Comparative Example 5

[0078] The preparation method of this comparative example is the same as that of Example 1, except that it does not contain Phellodendron bark and Costus root.

[0079] Experimental Example 1

[0080] Healing Experiment

[0081] Laboratory animals: 1-year-old dogs diagnosed with canine distemper.

[0082] 1-year-old dogs are divided into:

[0083] Group with infection: No treatment received;

[0084] Experimental group: Dogs were treated with the compositions of Examples 1-4 and Comparative Examples 1-5, respectively, at a feed intake of 50g / kg body weight / day per dog, mixed with 10wt% of the corresponding composition. The treatment was repeated for 3 consecutive days, followed by a 7-day interval before starting another course of treatment, for a total of 3 courses. The treatment results are shown in Table 1.

[0085] Table 1. Results of Treatment Efficacy

[0086] Quantity / piece Canine distemper / dog Composition Cure rate / % Infection group 20 20 none 0 Experimental group 1 20 20 Example 1 95 Experimental group 2 20 20 Example 2 95 Experimental group 3 20 20 Example 3 90 Experimental group 4 20 20 Example 4 90 Experimental group 5 20 20 Comparative Example 1 70 Experimental group 6 20 20 Comparative Example 2 75 Experimental group 7 20 20 Comparative Example 3 45 Experimental group 8 20 20 Comparative Example 4 70 Experimental group 9 20 20 Comparative Example 5 65

[0087] As can be seen from Table 1, the compositions of Examples 1-4 have a high cure rate for canine distemper, but their effectiveness is significantly reduced after the removal of the bactericidal agent. At the same time, compared with commercially available Bacillus licheniformis, the Bacillus licheniformis SXDYYB15 of this invention has a better therapeutic ability; and there is a significant synergistic effect between the traditional Chinese medicine formula and the strain SXDYYB15.

[0088] Example 2

[0089] Prevention experiment:

[0090] Laboratory animals: 1-year-old dogs.

[0091] Control group: No medication was administered;

[0092] Experimental group: The dogs were treated with the compositions of Examples 1-4 and Comparative Examples 1-5 respectively. Each dog was fed 50g / kg body weight / day of feed, which was mixed with 10wt% of the corresponding composition. The feeding was carried out for 3 consecutive days. Then, 2 1-year-old dogs diagnosed with canine distemper were added to each group and kept together. The number of infected dogs was calculated after 7 days.

[0093] The results are shown in Table 2.

[0094] Total quantity / piece Canine distemper / dog Composition Number of new cases / animals Blank group 20 2 none 15 Experimental group 1 20 2 Example 1 0 Experimental group 2 20 2 Example 2 0 Experimental group 3 20 2 Example 3 0 Experimental group 4 20 2 Example 4 0 Experimental group 5 20 2 Comparative Example 1 3 Experimental group 6 20 2 Comparative Example 2 2 Experimental group 7 20 2 Comparative Example 3 8 Experimental group 8 20 2 Comparative Example 4 4 Experimental group 9 20 2 Comparative Example 5 4

[0095] As shown in Table 2, the composition prepared in this invention has a good preventive effect against canine distemper and can improve the immunity of dogs.

[0096] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A probiotic fermented traditional Chinese medicine composition for the prevention and treatment of canine distemper, characterized in that, It is produced by fermentation of the following raw materials in parts by weight: 100-200 parts of Phellodendron bark, 10-20 parts of Aucklandia lappa, 100-200 parts of Glycyrrhiza uralensis, 200-400 parts of Taraxacum mongolicum, 100-200 parts of Coptis chinensis, and 10-40 parts of microbial preparations; The microbial preparation includes Bacillus licheniformis SXDYYB15, which was deposited at the China Center for Type Culture Collection on April 18, 2025, with accession number CCTCC NO:M2025814.

2. The method for preparing the probiotic fermented traditional Chinese medicine composition according to claim 1, characterized in that, Includes the following steps: S1. Weigh the raw materials according to the stated weight proportions; S2. Powder the following herbs: Phellodendron bark, Costus root, licorice root, dandelion, and Coptis root, then add water and decoct together to obtain a traditional Chinese medicine liquid. S3. After the Chinese medicine liquid cools down, add microbial preparations for fermentation to obtain a probiotic fermented Chinese medicine composition.

3. The preparation method according to claim 2, characterized in that, The material-to-liquid ratio in S2 is 1:(10~30).

4. The preparation method according to claim 2, characterized in that, The decocting time in S2 is 25-35 minutes.

5. The preparation method according to claim 2, characterized in that, The amount of microbial agent added in S3 is 1-5 wt%; the viable count of the microbial agent is ≥1.80 × 10⁻⁶. 9 CFU / mL.

6. The preparation method according to claim 5, characterized in that, The preparation method of the microbial agent is as follows: The seed culture of Bacillus licheniformis with accession number CCTCCNO.CCTCCM2025814 was inoculated into a culture medium and fermented. Fermentation conditions: continuous culture at 30-37℃ for 24-72 hours, followed by static culture for 96-120 hours until the bacterial count is ≥1.80×10⁻⁶. 9 CFU / mL.

7. The preparation method according to claim 6, characterized in that, The culture medium consists of the following components by weight: 1-5 parts peptone, 1-3 parts beef extract, 1-10 parts yeast extract, 5-15 parts sodium chloride, 10-100 parts fructooligosaccharide, and 1000 parts water.

8. The preparation method according to claim 2, characterized in that, The fermentation conditions in S3 are: static closed fermentation at 0-37℃ for 72-240 hours.