Preparation method and application of composite lactic acid bacteria dairy cow hoof bath solution

Abstract

The present invention relates to a preparation method and application of a composite lactic acid bacteria dairy cow hoof bath liquid, and belongs to the technical field of livestock and poultry drug preparations. The present invention is based on the main pathogenic bacteria of dairy cow hoof disease, and through the compounding of specific Bacillus subtilis and Lactobacillus pentosus, the antibacterial effect is synergistically exerted, and the selection of auxiliary materials can further improve the performance of the hoof bath liquid. The hoof bath liquid of the present invention can significantly reduce the incidence of dairy cow hoof disease, improve the economic benefits of dairy cow breeding, and has broad market prospects.

Description

Technical Field

[0001] The invention relates to the technical field of livestock and poultry pharmaceutical preparations, and in particular to a preparation method and application of a composite lactic acid bacteria cow hoof bath liquid. Background Art

[0002] Hoof disease in dairy cows is one of the diseases with a higher incidence rate in dairy cow breeding at home and abroad, and is an important cause of dairy cows being forced to be eliminated. Hoof disease not only affects the normal walking of dairy cows, but also reduces the milk production and fertility of dairy cows, reduces the economic benefits of dairy cows, and leads to huge economic losses. The main factors causing hoof disease in dairy cows include environmental factors, such as untimely drainage of cowsheds and cleaning of manure; ground factors, such as the presence of stones and sharp objects; hoof trimming factors and other nutritional factors. The main types of hoof disease in dairy cows include hoof rot, hoof warts, interdigital dermatitis, hoof sole ulcers, etc. The treatment methods for different hoof diseases are slightly different.

[0003] The prevention and control of hoof diseases in dairy cows includes hoof trimming management, such as regular hoof trimming and timely treatment of sick cows; it also includes hoof bathing management. Generally, the length of the hoof bath trough is 3-5m, the depth is about 15cm, and the liquid level is greater than 10cm, so that the cow's hooves are completely immersed in the hoof bath liquid; during the feeding process, attention should be paid to the hygiene of the cowshed to ensure timely cleaning of feces and reduce the chance of hoof contamination; in addition, the flatness of the ground in the cowshed can be improved, and relatively soft rubber mats can be laid; in the feeding process, attention should also be paid to the balanced nutrition of dairy cows to reduce the probability of disease.

[0004] Among the above-mentioned preventive measures, hoof bathing is the most effective method for treating and preventing hoof diseases in dairy cows. The hoof bathing liquid works by cleaning and disinfecting the cow's hooves. This method not only has a therapeutic effect on early hoof warts, but also can control the recurrence of late hoof warts. At the same time, it can effectively reduce the incidence of interdigital dermatitis and foot rot, and has a good improvement effect on common types of hoof diseases.

[0005] The main ingredients of the known hoof baths generally include copper sulfate, which has high permeability and improves the hardness of the hoof, widely used formalin, zinc sulfate, which is usually used in combination with copper sulfate, lime powder and other chemical ingredients. Natural ingredients such as thymol, Chinese herbal extracts, and plant extracts can also be used. Hoof baths have a variety of compositions and their ingredients vary greatly. There are many problems in the use of the above ingredients. For example, the effect of hoof bathing is unclear, and there are usually problems of environmental pollution. For example, the use of chemical ingredients such as formalin and copper sulfate in the composition of commonly used hoof baths will increase harmful emissions from farms and put pressure on the environment. In addition, the use of drug ingredients will inevitably lead to drug resistance problems. The use of chemical ingredients is also highly irritating, which will irritate the animal's hooves after being applied to animals. Therefore, seeking safer and less irritating alternatives is the focus of hoof baths.

[0006] In the development of existing hoof baths, people usually focus on developing hoof baths that are pollution-free, safe, effective, and residue-free. There are studies on the use of safe hoof baths without toxic side effects in the prior art. For example, a related study with publication number CN110507748A provides a cow hoof bath and a preparation method thereof, wherein the raw materials include Chinese herbal extracts, antibacterial agents, antifreeze agents, and water; the Chinese herbal extracts are made of the following raw materials in parts by weight: indigo, frankincense, myrrh, safflower, sophora flavescens, phellodendron, aloe, etc., and the corresponding effects are achieved by using a large amount of Chinese herbal ingredients. The document with publication number CN110248670A discloses the use of a composition comprising a bacterial mixture for preparing a medicament for preventing and improving the symptoms of ungulates suffering from diseases involving foot parasites or bacterial infections. The composition comprises: at least one strain of Bacillus bacteria, especially at least one Bacillus subtilis, and at least one lactic acid bacteria strain, especially at least one Lactococcus lactis or at least one Pediococcus pentosaceus. The above bacteria are combined or compounded with a disinfectant to treat related diseases of ungulates, such as treating symptoms such as lameness by spraying antibacterial agents.

[0007] In summary, although the research and development of existing hoof baths pays attention to the safety and effectiveness of use, the research is not in-depth, especially there are no reports on the development of bacterial compounding specifically for cow hoof diseases. Based on this, the present invention provides a preparation method and application of a composite lactic acid bacteria cow hoof bath. Summary of the invention

[0008] Based on the problems of the prior art, the present invention provides a composite lactic acid bacteria dairy cow hoof bath and a preparation method thereof. Based on the main pathogenic bacteria of dairy cow hoof diseases, by compounding specific Bacillus subtilis and Lactobacillus pentosus, the antibacterial effect is synergistically exerted, and the selection of auxiliary materials can further improve the performance of the hoof bath. The hoof bath of the present invention can significantly reduce the incidence of dairy cow hoof diseases, improve the economic benefits of dairy cow farming, and has broad market prospects.

[0009] In a first aspect of the present invention, a composite lactic acid bacteria cow hoof bath solution is provided, wherein the composite lactic acid bacteria cow hoof bath solution is prepared from composite lactic acid bacteria, auxiliary materials and water;

[0010] The composite lactic acid bacteria are: Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ;

[0011] Bacillus subtilis LB-KC is deposited in the General Microbiological Center of China Microorganism Culture Collection Administration with the deposit number of CGMCC NO.25562.

[0012] Lactobacillus pentosus MRS-RGJ was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration with the deposit number of CGMCC NO.25561;

[0013] The excipients are: trehalose, carrageenan, polyvinyl pyrrolidone;

[0014] And water.

[0015] When selecting compound lactic acid bacteria, first of all, according to existing studies, the main pathogens that cause hoof diseases in dairy cows vary according to the type of hoof disease. For example, hoof rot in dairy cows is mainly caused by Fusobacterium necroticum and Bacteroides melanogenes, and also includes Bacteroides nodosa and Anaerobic Bacillus necroticus. Among them, Bacteroides nodosa can produce protease, digest keratin, and make the surface and base of the hoof vulnerable to damage, thus causing hoof rot. For another type of wart dermatitis, it is generally believed that its pathogens are Treponema and Fusobacterium necroticum, and may also include pathogenic bacteria such as Bacteroides, Mycoplasma, Campylobacter, etc.; the main pathogens of hoof ulcer disease include Corynebacterium pyogenes and Anaerobic Bacillus necroticus. There are differences in the main pathogens of different types of hoof diseases. For the above pathogens, the inventors selected the main pathogens of hoof diseases, Fusobacterium necroticum, Corynebacterium pyogenes and Bacteroides nodosa, as the pathogens for investigation. By screening the types and dosages of bacteria, they determined to select Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ for compounding.

[0016] In terms of the selection of raw material dosage, the composite lactic acid bacteria cow hoof bath liquid is prepared by: 0.5-1.1 parts of composite lactic acid bacteria, 6-10 parts of trehalose, 10-15 parts of carrageenan, 12-20 parts of polyvinyl pyrrolidone and 200-300 parts of water, and the ratio of the effective live bacteria count of Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ in the composite lactic acid bacteria agent is 2-4:1.

[0017] Preferably, calculated by weight: the composite lactic acid bacteria cow hoof bath liquid is prepared by 0.8 parts of composite lactic acid bacteria, 8 parts of trehalose, 13 parts of carrageenan, 16 parts of polyvinyl pyrrolidone and 250 parts of water, and the ratio of the effective live bacteria count of Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ in the composite lactic acid bacteria agent is 3:1.

[0018] Further preferably, the pH of the composite lactic acid bacteria cow hoof bath solution is 6.5-7.2.

[0019] The pH of the composite lactic acid bacteria cow hoof bath liquid of the present invention is controlled within a range suitable for the bacterial strains to exert their efficacy, thereby ensuring that the bacterial strains can exert their activity.

[0020] Another aspect of the present invention provides a method for preparing a composite lactic acid bacteria cow hoof bath liquid, which is characterized by comprising the following steps: mixing the composite lactic acid bacteria, auxiliary materials and water, and stirring for 3-5 minutes to obtain the cow hoof bath liquid.

[0021] The preparation method of the invention is simple, and the raw materials can be used after being fully mixed, and the industrial use is convenient.

[0022] The third aspect of the present invention also provides an application of a composite lactic acid bacteria cow hoof bath in preventing and treating cow hoof diseases.

[0023] The composite lactic acid bacteria cow hoof bathing liquid of the present invention has good effect of preventing and treating cow hoof diseases and has definite curative effect.

[0024] As a preferred embodiment, the hoof disease of dairy cows includes one or more of hoof rot, verrucous dermatitis, and hoof sole ulcer.

[0025] Further preferably, the cow hoof disease is mainly caused by Fusobacterium necroticum, Corynebacterium pyogenes and Bacteroides nodosa.

[0026] Compared with the prior art, the technical solution adopted by the present invention can achieve at least one of the following beneficial effects:

[0027] (1) The composite lactic acid bacteria hoof bath of the present invention uses lactic acid bacteria with outstanding antibacterial effect as the active ingredient. Compared with the chemical ingredients in the prior art, it has the advantages of high safety and outstanding efficacy, filling the gap of the lack of composite probiotic combination with definite efficacy in the preparation of dairy cow hoof bath. The product of the present invention has broad market prospects.

[0028] (2) The inventors identified two lactic acid bacteria through screening research, namely Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ; based on the previous research, their biological characteristics and physiological forms were determined, and further starting from the main pathogenic bacteria of dairy cow hoof disease, through experimental screening research, it was clarified that the composite lactic acid bacteria prepared by compounding the above two bacteria has synergistic effects in inhibiting dairy cow hoof disease-related bacteria species and has outstanding antibacterial properties.

[0029] (3) In terms of auxiliary material selection, the inventors adjusted the composition of auxiliary materials based on the growth characteristics of lactic acid bacteria in order to fully exert their activity. The addition of trehalose can maintain the stability of the cell structure and function of lactic acid bacteria. Carrageenan, as a stabilizer, can improve the adhesion of hoof bath liquid, promote the adhesion and absorption of bacteria during hoof bathing, and improve the effect. Polyvinyl pyrrolidone can promote film formation and cooperate with carrageenan to improve the durability of the antibacterial performance of the composite lactic acid bacteria.

[0030] (4) The composite lactic acid bacteria hoof bath of the present invention has an outstanding effect in preventing and treating hoof diseases of dairy cows, and there is a clear synergistic effect between the two bacteria. The experimental research results show that Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ have a synergistic effect in inhibiting the main pathogenic bacteria of dairy cow hoof diseases; the industrial application results show that when the dairy cow hoof bath under the conditions of the present invention is used to treat sick dairy cows, the treatment efficiency is as high as 95%, the curative effect is definite, and it has a broad market application prospect. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 It is a photo of the hoof bath during the experimental study of the effect of the present invention;

[0032] Figure 2 It is a photo of the scene of the cow hoof bath of the present invention. DETAILED DESCRIPTION

[0033] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. Obviously, the described embodiments are only part of the embodiments of the present invention, not all embodiments.

[0034] The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0035] Unless otherwise specified, the experimental methods used in the examples of the present invention are all conventional methods; the materials and reagents used are all commercially available materials and reagents unless otherwise specified.

[0036] Example 1 Study on the antibacterial properties of different strains against the main pathogenic bacteria of dairy cow hoof diseases

[0037] 1. Source of experimental strains

[0038] (1) Screening experiment of Bacillus subtilis LB-KC

[0039] Screening of strains

[0040] Bacillus subtilis LB-KC was collected and isolated from the soil in Hohhot, Inner Mongolia Autonomous Region. The separation and purification method is as follows:

[0041] a) Weigh 5 g of soil sample from a breeding farm in Tumotezuoqi, Hohhot, and place it in a 100 mL Erlenmeyer flask containing 50 mL of sterile water. Shake for 30 min and mix thoroughly to obtain a soil suspension with a concentration of 1 / 10.

[0042] b) Pipette 0.5 mL of the above soil suspension into a sterile centrifuge tube containing 4.5 mL of sterile water, mix well to make the solution concentration 1 / 100, and then pipette 100 μL of the diluted soil suspension and spread it on LB solid culture medium. Purify and culture at 25°C for 3-5 days, pick the bacterial colony with the largest number of colonies for separation and purification, obtain a single colony, and obtain Bacillus subtilis.

[0043] Identification of Bacillus subtilis LB-KC

[0044] After sequencing analysis and identification, its taxonomic characteristics are confirmed as: Bacillus, Gram-positive bacteria, slightly yellow colonies, rough surface forming wrinkles, shape: long oval column, with strong acid resistance, bile salt resistance, oxidation resistance, high temperature resistance, stress resistance and strong stability, etc., which can be identified as Bacillus subtilis. The strain is named Bacillus subtilis LB-KC, the deposit number is CGMCC NO.25562, the deposit address is: General Microbiology Center of China Microbiological Culture Collection Administration, and the deposit date is: August 19, 2022.

[0045] (2) Screening experiment of Lactobacillus pentosus MRS-RGJ

[0046] Screening of strains

[0047] The Lactobacillus pentosus MRS-RGJ used in the present invention is isolated from Hohhot kimchi, and the isolation and identification process is as follows:

[0048] 3 mL of Hohhot kimchi juice was selected and added to 100 mL of MRS liquid culture medium. The culture was carried out at a constant temperature of 35° C., during which the rotation speed was controlled at 200 r / min and the culture was carried out for 36 hours to obtain a culture solution rich in lactic acid bacteria. 0.2 mL of the culture solution was drawn from the culture solution for ten-fold gradient dilution, and the culture was spread on a culture dish. The culture was continued at 35° C. After 48 hours, single colonies of different morphologies were separated, and the single colonies were picked for purification culture in MRS liquid culture medium to obtain Lactobacillus pentosus MRS-RGJ.

[0049] Identification of Lactobacillus pentosus MRS-RGJ

[0050] After sequencing analysis and identification, it was confirmed that the taxonomic characteristics of Lactobacillus pentosus were a lactic acid bacterium, which belongs to the Firmicutes, Bacillus, Lactobacilales, Lactobacilaceae, and Lactobacillus in taxonomy. It is a Gram-positive bacterium, facultative anaerobic, without spores, and with yellowish rough ring-shaped precipitation. The bacteria are medium-long rod-shaped; they are salt-resistant, acid-resistant or freeze-drying-resistant, and can use lactose, mannitol, maltose, cellobiose, galactose, xylose, and sorbitol to produce acid. The strain was named Lactobacillus pentosus MRS-RGJ, which is deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration, with the deposit number CGMCC NO.25561; the deposit address is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing; the deposit date is August 19, 2022.

[0051] 2. Experimental methods

[0052] Antibacterial research method: The main pathogens of dairy cow hoof disease, Fusobacterium necroticum, Corynebacterium pyogenes and Bacteroides nodosa, were selected as experimental strains. The conventional Oxford cup method was used in the antibacterial experiment. First, the selected experimental strains were prepared into a bacterial suspension with a concentration of 106cfμ / ml, and then the above bacterial suspensions were added to the culture medium for coating culture. Different lactic acid bacteria were added to the Oxford cup to measure the diameter of the antibacterial zone; each treatment was repeated 3 times during the experimental determination.

[0053] The treatments with different lactic acid bacteria were as follows: Treatment 1 was 200 μl of Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ, and the ratio of the effective live bacteria counts of Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ was controlled to be 3:1; Treatment 2 was 200 μl of Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ, and the ratio of the effective live bacteria counts of Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ was controlled to be 1:1; Treatment 3 was 200 μl of Bacillus subtilis LB-KC; Treatment 4 was 200 μl of Lactobacillus pentosus MRS-RGJ.

[0054] 3. Experimental results

[0055] The antibacterial properties of different lactic acid bacteria combinations were determined according to the experimental method, and the diameters of the antibacterial circles of different treatment groups against the three bacteria were obtained. The results are shown in Table 1:

[0056] Table 1 Comparison of antibacterial effects of different treatment groups against different pathogenic bacteria

[0057]

[0058] From the results in Table 1, it can be seen that there are great differences in the antibacterial effect results of different treatment groups against different pathogenic bacteria. The antibacterial effect of treatments 1 and 2 after the combination of the two strains (Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ) of the present invention is significantly better than that of treatments 3 and 4 of a single strain, and the overall effect under the conditions of treatment group 1 is the best, and it has a good effect in inhibiting the main pathogenic bacteria, which can reflect the synergistic effect of the above strain combination. The above antibacterial experiment can show that the synergy of the specific strains of the present invention has a clear synergistic effect in inhibiting the main pathogenic bacteria of dairy cow hoof diseases, and can be used for the further development of dairy cow hoof bath products.

[0059] Example 2

[0060] A composite lactic acid bacteria cow hoof bath liquid, which is prepared from composite lactic acid bacteria, auxiliary materials and water; the composite lactic acid bacteria are: Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ, and the auxiliary materials are: trehalose, carrageenan, polyvinyl pyrrolidone;

[0061] and water;

[0062] The raw material dosage is calculated by weight fraction as follows: 0.8 parts of composite lactic acid bacteria, 8 parts of trehalose, 13 parts of carrageenan, 16 parts of polyvinyl pyrrolidone and 250 parts of water, and the ratio of the effective live bacteria count of Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ in the composite lactic acid bacteria agent is 3:1;

[0063] The preparation process adopts the following steps: mixing the composite lactic acid bacteria, auxiliary materials and water, and stirring for 3-5 minutes to obtain the cow hoof bath liquid; and controlling the pH of the hoof bath liquid to be 6.5-7.2.

[0064] Effect Experimental Research

[0065] 1. The selection criteria for sick dairy cows are as follows:

[0066] Hoof rot: lameness of one or more limbs, prone to lying down, red or dark purple skin between toes and hoof crowns, congestion, redness and swelling of the skin between hooves and hoof crowns, and ulceration between hooves.

[0067] Verrucous dermatitis: lameness of one or more limbs, prone to lying down, cauliflower-like true fibropapilloma on the dorsal or palmoplantar skin between toes, redness, swelling and erosion of the hoof; lack of energy, poor appetite, and decreased milk production;

[0068] Hoof ulcer: lameness of one or more limbs, prone to lying down, loss of keratin in the hoof sole, leading to hyperplasia of granulation tissue or even suppuration, infection of hoof tissue; decreased feed intake and milk production;

[0069] 2. Experimental methods: A total of 180 dairy cows with hoof diseases in a large breeding farm in Inner Mongolia were selected as the research subjects, including 60 cows with hoof rot, verrucous dermatitis and hoof sole ulcer. In the study of each type of hoof disease, 20 cows were studied in the experimental group, control group 1 and control group 2. The treatment methods of different treatment groups are as follows:

[0070] Experimental group: using the dairy cow hoof bath obtained in Example 2;

[0071] Control group 1: The composite lactic acid bacteria in Example 2 were replaced with the lactic acid bacteria in treatment 3, that is, only Bacillus subtilis LB-KC was used to replace the composite lactic acid bacteria combination in Example 2, and the other treatments were the same as in Example 2;

[0072] Control group 2: The composite lactic acid bacteria in Example 2 were replaced by the lactic acid bacteria in treatment 4, that is, only Lactobacillus pentosus MRS-RGJ was used to replace the composite lactic acid bacteria combination in Example 2, and other treatments were the same as in Example 2.

[0073] During use, the sick cows were given a hoof bath once in the morning and evening. Figure 1 Perform the footbath in the manner shown in the figure. Figure 2 Hoof bathing was performed in the manner shown; the treatment was continued for 8 days, and the experimental results were recorded according to the judgment criteria after the end, as shown in Tables 2 and 3.

[0074] 3. Judgment criteria

[0075] Table 2 Criteria for judging different hoof diseases in dairy cows after treatment

[0076]

[0077] 4. Experimental results

[0078] According to the criteria in Table 2, the treatment status of dairy cows with hoof disease was recorded, and the number of effective, markedly effective and ineffective dairy cows in each treatment group under each hoof disease category was recorded, and the effective rate was calculated, where the effective rate = (effective + markedly effective) / number of dairy cows * 100%. The specific results are shown in Table 3:

[0079] Table 3 Comparison of therapeutic effects in different treatment groups

[0080]

[0081] From the results in Table 3, it can be seen that under the conditions of the embodiments of the present invention, the obtained cow hoof bath product has a significantly better therapeutic effect on the sick cows than the control group 1 and the control group 2. The effects of the control group 1 and the control group 2 are not as good as the experimental group, and the lowest therapeutic effect on the hoof disease of the cow is only 65%. The experimental research results on sick cows can show that the combined use of Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ has a significant synergistic effect on the hoof disease of the cow relative to a single lactic acid bacteria. The cow hoof bath using Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ as composite lactic acid bacteria shows a good industrial application effect.

[0082] The above description is only a specific implementation mode of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily conceived by a technician familiar with the technical field within the technical scope disclosed by the present invention should be covered within the protection scope of the present invention.

Claims

1. A composite lactic acid bacteria cow hoof bath, characterized in that: The composite lactic acid bacteria cow hoof bath solution is prepared from composite lactic acid bacteria, auxiliary materials and water; The compound lactic acid bacteria is a mixture of Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ; Among them, Bacillus subtilis LB-KC is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number CGMCC NO.25562; the deposit address is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing; the deposit date is August 19, 2022; Lactobacillus pentosus MRS-RGJ is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number CGMCC NO.25561; the deposit address is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing; the deposit date is August 19, 2022; The excipients are: trehalose, carrageenan, polyvinyl pyrrolidone; And water.

2. The composite lactic acid bacteria cow hoof bath according to claim 1, characterized in that: The composite lactic acid bacteria cow hoof bath liquid is prepared by the following weight fractions: 0.5-1.1 parts of composite lactic acid bacteria, 6-10 parts of trehalose, 10-15 parts of carrageenan, 12-20 parts of polyvinyl pyrrolidone and 200-300 parts of water; the ratio of the effective live bacteria counts of Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ in the composite lactic acid bacteria agent is 2-4:

1.

3. The composite lactic acid bacteria cow hoof bath according to claim 2, characterized in that: The composite lactic acid bacteria cow hoof bath liquid is prepared by: 0.8 parts of composite lactic acid bacteria, 8 parts of trehalose, 13 parts of carrageenan, 16 parts of polyvinyl pyrrolidone and 250 parts of water, and the ratio of the effective live bacteria count of Bacillus subtilis LB-KC and Lactobacillus pentosus MRS-RGJ in the composite lactic acid bacteria agent is 3:

1.

4. The composite lactic acid bacteria cow hoof bath liquid according to claim 3, characterized in that: The pH value of the composite lactic acid bacteria cow hoof bath liquid is 6.5-7.

2.

5. A method for preparing the composite lactic acid bacteria cow hoof bath liquid as claimed in any one of claims 1 to 4, characterized in that: The method comprises the following steps: mixing composite lactic acid bacteria, auxiliary materials and water, and stirring for 3-5 minutes to obtain the cow hoof bath liquid.

6. Use of the composite lactic acid bacteria cow hoof bath liquid according to any one of claims 1 to 4 in the preparation of a pharmaceutical preparation for preventing and treating cow hoof diseases, wherein the cow hoof diseases are one or more of hoof rot, verrucous dermatitis, and hoof sole ulcers.

Images