A Lysostaphin Milk Film Preparation and Its Preparation Process

Abstract

This application relates to the technical field of dairy cow pharmaceutical preparations, and specifically discloses a lysostaphin milk protection film preparation and its preparation process. A lysostaphin milk protection film preparation comprises raw materials in the following mass percentages: 0.0001 - 2% of lysostaphin, 0.0001 - 1% of polyhexamethylene biguanide, 5 - 20% of lubricant, 0.5 - 2% of film-forming agent, 1 - 3% of thickening agent, 1 - 3% of moisturizing agent, 0.1 - 0.5% of vitamin B group, and the balance being purified water; the film-forming agent is a mixture composed of sodium carboxymethyl cellulose and plant polysaccharide in a mass ratio of 1:(2.3 - 4.1). The milk protection film preparation obtained by this application has excellent antibacterial and bactericidal effects, is mild in nature, non-irritating to the skin, and has good repair and nursing effects at the same time.

Description

Technical Field

[0001] The present application relates to the technical field of dairy cow pharmaceutical preparations, and more specifically, to a lysostaphin milk protection film preparation and its preparation process. Background Art

[0002] Mastitis in dairy cows refers to a disease in which the mammary glands of dairy cows become inflamed. It is one of the most harmful diseases in the dairy cow breeding industry and causes the most serious economic losses to the dairy cow industry. After a dairy cow contracts mastitis, the milk yield, milk quality, and nutritional value will all decline, and it may even easily lead to a shortening of the milk production years of the dairy cow.

[0003] Mastitis is mainly caused by pathogenic bacteria invading the mammary ducts or mammary tissues. It has been verified that immediately medicating the teats with a bactericide after each milking can effectively reduce bacterial infection. However, the commonly used teat medicaments in clinical practice are mainly iodophor, chlorhexidine, peracetic acid, sodium hypochlorite, etc., which have a certain irritating or corrosive effect on the teat skin. Long-term use is likely to cause conditions such as dryness and chapping of the teat skin, seriously affecting teat health and thus normal production capacity. In addition, long-term use of chemical or antibiotic disinfectants is likely to cause bacterial drug resistance, which is not conducive to the management and healthy development of the farm. Moreover, if the teats are disinfected with iodine or chlorhexidine after milking, and the teats are not thoroughly cleaned and wiped before milking, it may lead to an increase in the iodine or chlorhexidine content in the milk, seriously affecting milk quality and endangering human health.

[0004] Therefore, developing a new type of milk protection film preparation with excellent antibacterial and bactericidal effects, mild and non-irritating properties, and at the same time having a repair and care effect is of great significance for the health of dairy cow teats and the prevention and treatment of mastitis. Summary of the Invention

[0005] In order to develop a new type of milk protection film preparation with excellent antibacterial and bactericidal effects, mild and non-irritating properties, and a repair and care effect, the present application provides a lysostaphin milk protection film preparation and its preparation process.

[0006] In the first aspect, the present application provides a lysostaphin milk protection film preparation, adopting the following technical solution:

[0007] A lysostaphin milk protection film preparation, comprising the following raw materials in mass percentages: lysostaphin 0.0001 - 2%, polyhexamethylene biguanide 0.0001 - 1%, lubricant 5 - 20%, film-forming agent 0.5 - 2%, thickening agent 1 - 3%, moisturizing agent 1 - 3%, vitamin B group 0.1 - 0.5%, and the balance being purified water; the film-forming agent is a mixture composed of sodium carboxymethyl cellulose and plant polysaccharide in a mass ratio of 1:(2.3 - 4.1).

[0008] By adopting the above technical solution, lysostaphin and polyhexamethylene biguanide are used as antibacterial and bactericidal active ingredients. Lysostaphin is a broad-spectrum, highly efficient and mild metal protease. The mechanism of its bactericidal action is to act on the pentaglycine peptide bridge of peptidoglycan in the cell wall, destroy the cell wall, and cause the leakage of cell contents and death. While lysostaphin efficiently lyses bacteria, it also has the effect of promoting wound healing, which can significantly shorten the wound healing time. And the core component of lysostaphin contains Zn2+, which can increase the zinc content in the locally damaged skin and mucosal tissues. Zinc ions can promote the biosynthesis of nucleic acids and proteins, increase the activity of alkaline phosphatase (AKP) in the local tissues, increase the content of collagen in the tissues, and are beneficial to the growth and division of epidermal cells, thus promoting the rapid healing of the wounds of damaged skin and mucosal tissues and significantly shortening the mucosal healing time.

[0009] Polyhexamethylene biguanide is a safe and highly efficient broad-spectrum antibacterial agent, which has good antibacterial and bactericidal effects on Gram-positive bacteria, Gram-negative bacteria, fungi and viruses. At the same time, polyhexamethylene biguanide has the characteristics of low bacteriostatic concentration, broad-spectrum low toxicity, fast action speed and low foam volume; and after applying the medicated nursing film, polyhexamethylene biguanide can form a cationic protective film and exist relatively stably on the bovine udder, so as to achieve the effect of long-term antibacterial and bactericidal. In addition, the bactericidal process of polyhexamethylene biguanide mainly relies on physical action, that is, it quickly adsorbs and captures bacteria or viruses, forms polymers on the surface of microorganisms to inhibit the replication of bacteria and viruses, so the problem of bacterial drug resistance can be effectively reduced.

[0010] In summary, the organic combination of lysostaphin and polyhexamethylene biguanide has a significantly higher synergistic effect than the original separate use of bactericidal effects. Specifically, lysostaphin and polyhexamethylene biguanide form a high-molecular complex. On the one hand, lysostaphin is protected, which can effectively ensure the activity of lysostaphin in the system; on the other hand, due to the action of polyhexamethylene biguanide, microorganisms are quickly aggregated and captured, and the binding efficiency of lysostaphin to bacterial cells is increased exponentially, which is more conducive to lysostaphin to further act on the cells. The bactericidal rate of the antibacterial and bactericidal active ingredients of the present invention is significantly increased, and the effective concentration used can be lower, the biological safety is higher, and the use cost is also lower.

[0011] In addition, sodium carboxymethylcellulose and plant polysaccharides are further compounded in a specified ratio as a film-forming agent. Since sodium carboxymethylcellulose has a good dispersion effect in the system, the film-forming rate is relatively fast after application, and the formed film is relatively uniform. At the same time, it has good hygroscopicity, and the formed film can provide a certain moisturizing effect. Plant polysaccharides belong to natural extracts, and their structures contain a large number of hydroxyl groups, which can bind to water in the form of hydrogen bonds. At the same time, they have good film-forming properties and can form a "water-locking film" on the skin surface to prevent the loss of skin moisture and achieve a long-lasting moisturizing effect. The two cooperate with each other synergistically, which can effectively improve the film-forming efficiency, film-forming effect and moisturizing effect of the nursing film preparation.

[0012] A certain amount of vitamin B group is further added. First of all, the vitamin B group has a certain positive significance for the antibacterial and repair effects of the system; in addition, the vitamin B group can be well dispersed in the system, so that it is evenly dispersed in the film layer after application. And the vitamin B group is prone to decompose and lose its side chain under the action of visible light, that is, photodegradation occurs, and then fine channels are left on the film layer, making the air permeability of the film layer better.

[0013] Combining the above effects, the nursing film preparation prepared in this application has the advantages of high-efficiency sterilization, repair and promotion of healing, not easy to develop drug resistance, mild nature, moisturizing and nourishing, and good air permeability of the film layer, and the practical effect is relatively excellent.

[0014] In a specific feasible implementation scheme, the nursing film comprises the following raw materials in mass percentage: lysostaphin 0.4 - 1%, polyhexamethylene biguanide 0.04 - 0.2%, lubricant 10 - 20%, film-forming agent 1 - 2%, thickening agent 1 - 2%, moisturizing agent 1.5 - 2.4%, vitamin B group 0.2 - 0.4%, and the balance is purified water; the film-forming agent is a mixture composed of sodium carboxymethylcellulose and plant polysaccharides in a mass ratio of 1:(2.8 - 3.5).

[0015] By adopting the above technical scheme, the ratio of each component is further optimized, thereby further optimizing the comprehensive performance of the nursing film preparation.

[0016] In a specific feasible implementation scheme, the moisturizing agent is a mixture composed of hyaluronic acid and vegetable oil in a mass ratio of 1:(1.4 - 1.9).

[0017] By adopting the above technical scheme, hyaluronic acid and vegetable oil are compounded as a moisturizing agent according to the above mass ratio. Among them, hyaluronic acid can not only have an excellent moisturizing effect, but also promote the effect of wound healing. Vegetable oil can form a water-lipid film on the skin surface, which not only has a moisturizing effect, but also enables the nursing film layer to have good hydrophobic properties, that is, makes the milk show a distant state during the process of contacting the nursing film layer, so that the pollution situation of the nursing film layer contacting and dissolving in the milk during the subsequent milking operation can be effectively reduced.

[0018] In a specific feasible embodiment, the vegetable oil is a mixture composed of coconut oil, grape seed oil and wheat germ oil according to a mass ratio of 1:(2.6 - 3.2):(1.5 - 2.0).

[0019] By adopting the above technical solution, coconut oil, grape seed oil and wheat germ oil are compounded as vegetable oil according to the above mass ratio, which is safe, environmentally friendly, mild in nature and has a good moisturizing effect. At the same time, coconut oil has moisturizing, anti-inflammatory and antibacterial effects, grape seed oil has moisturizing, anti-inflammatory, antibacterial and wound-healing promoting effects, and wheat germ oil has moisturizing and wound-healing promoting effects; when the three are compounded and used according to the above ratio, better moisturizing and nourishing, anti-inflammatory and antibacterial as well as wound-healing promoting effects can be achieved, that is, the synergistic cooperation effect is better.

[0020] In a specific feasible embodiment, the plant polysaccharide includes at least one of aloe polysaccharide, astragalus polysaccharide and cactus polysaccharide.

[0021] By adopting the above technical solution, aloe polysaccharide, astragalus polysaccharide and cactus polysaccharide all belong to natural plant extracts, which are safe, environmentally friendly and mild in nature, and the "water-locking film" formed has good moisturizing effects.

[0022] In a specific feasible embodiment, the vitamin B group is a mixture composed of vitamin B5 and vitamin B12 according to a mass ratio of (3.5 - 4.2):1.

[0023] By adopting the above technical solution, vitamin B5 has good dispersibility in the system and has a certain hygroscopicity, which is of positive significance for improving the moisturizing and nourishing performance of the nursing film preparation. In addition, vitamin B12 contains cobalt ions and has certain antibacterial properties, which is of positive significance for improving the antibacterial performance of the nursing film preparation. At the same time, both are prone to photodegradation reactions. When compounded and used according to the above ratio, the air permeability of the nursing film layer can be effectively improved, and the comprehensive performance of the nursing film preparation can be further optimized.

[0024] In a specific feasible embodiment, the lubricant includes at least one of glycerol and lanolin.

[0025] In a specific feasible embodiment, the thickener includes at least one of sodium alginate, carrageenan and gum arabic.

[0026] In the second aspect, the present application provides a preparation process of a staphylolytic enzyme nursing film preparation, adopting the following technical solution:

[0027] A preparation process of a staphylolytic enzyme nursing film preparation includes the following steps:

[0028] Primary mixing: Mix the film-forming agent, thickening agent, humectant, vitamin B group and partial purified water, and stir well to obtain solution A;

[0029] Secondary mixing: Dissolve staphylococcal enzyme and polyhexamethylene biguanide in partial purified water, stir well and let stand, then add the lubricant, stir well to obtain solution B;

[0030] Tertiary mixing: Mix solution A and solution B, stir well to obtain the finished product of the nursing film preparation, and store it in the dark.

[0031] By adopting the above technical solution, solution A and solution B are separately prepared, and staphylococcal enzyme and polyhexamethylene biguanide are pre-formed into a polymer complex, so that staphylococcal enzyme can maintain a high activity in the system.

[0032] In a specific feasible embodiment, in the primary mixing and secondary mixing steps, the purified water is obtained by heating and boiling and then keeping warm at 30 - 45 °C.

[0033] By adopting the above technical solution, the germs in the purified water can be effectively killed during the boiling process of the purified water, and by using the purified water kept warm at 30 - 45 °C to prepare solution A and solution B, the dissolution and dispersion effect of each component in the system can be effectively improved.

[0034] In summary, the present application has the following beneficial effects:

[0035] 1. The present application uses staphylococcal enzyme and polyhexamethylene biguanide as antibacterial and bactericidal active ingredients. When used in combination, they can achieve a bactericidal effect significantly higher than that of using them alone; and after the two are combined, they exist in the form of a polymer complex, which can effectively guarantee the activity of staphylococcal enzyme in the system; in addition, polyhexamethylene biguanide can quickly aggregate and capture pathogenic microorganisms, so that the binding efficiency of staphylococcal enzyme and bacterial cells is increased exponentially, thus achieving a high-efficiency bactericidal effect.

[0036] 2. In the present application, staphylococcal enzyme, hyaluronic acid and vegetable oil all have a certain degree of promoting wound healing effect on the skin, so as to achieve the effect of repair and care while efficiently killing bacteria.

[0037] 3. The properties of each component of the nursing film preparation in the present application are relatively mild, without irritation, corrosion, etc., and the drug application safety is relatively high.

[0038] 4. In the present application, the bactericidal process of polyhexamethylene biguanide is mainly a physical effect, while staphylococcal enzyme plays a bactericidal role by directly lysing the cell wall of pathogenic bacteria. The bactericidal mechanisms of both are not likely to cause the problem of drug resistance. Detailed implementation mode

[0039] The present application will be further described in detail below in combination with examples and comparative examples. All raw materials involved in the present application can be obtained commercially.

[0040] Example

[0041] Example 1

[0042] A staphylolytic enzyme breast care film preparation comprises raw materials with the following masses: 1 kg of staphylolytic enzyme, 0.2 kg of polyhexamethylene biguanide, 10 kg of lubricant, 2 kg of film-forming agent, 1 kg of thickening agent, 2 kg of moisturizing agent, 0.3 kg of vitamin B complex, and purified water to make up to 100 kg;

[0043] The film-forming agent is a mixture composed of sodium carboxymethyl cellulose and plant polysaccharide in a mass ratio of 1:3.5, and the plant polysaccharide is aloe polysaccharide;

[0044] The lubricant is glycerol;

[0045] The thickening agent is carrageenan;

[0046] The moisturizing agent is a mixture composed of hyaluronic acid and vegetable oil in a mass ratio of 1:1.6, and the vegetable oil is a mixture composed of coconut oil, grape seed oil and wheat germ oil in a mass ratio of 1:2.8:1.7;

[0047] The vitamin B complex is a mixture composed of vitamin B5 and vitamin B12 in a mass ratio of 3.9:1.

[0048] The preparation process of the above staphylolytic enzyme breast care film preparation comprises the following steps:

[0049] S1 Pretreatment of purified water: Heat the purified water to boiling and keep it at 40 °C for standby;

[0050] S2 Primary mixing: Put the film-forming agent, thickening agent, moisturizing agent, vitamin B complex and 80% of the total amount of purified water into a container, stir evenly for 2 hours, and keep it in the dark during the stirring process to make it a uniform aqueous slurry to obtain solution A;

[0051] S3 Secondary mixing: Take another container, add staphylolytic enzyme and polyhexamethylene biguanide, mix and dissolve them in 15% of the total amount of pure water at 40 °C and pH 6.5, stir evenly at a stirring speed of 150 rpm1, let it stand for 1 h, then add the lubricant and stir evenly to obtain solution B;

[0052] S4 Tertiary mixing: Pour solution B into solution A, wash the container that has held solution B with the remaining purified water, and add the washing liquid to solution A as well. Stir evenly in the dark to obtain the finished breast care film preparation, and store the finished breast care film preparation in the dark.

[0053] Examples 2 to 5 are different from Example 1 in that the ratios of the raw materials are different, as shown in the following table.

[0054] Table 1 Raw Material Ratio Table

[0055]

[0056]

[0057] Example 6

[0058] The difference between this example and Example 1 is that the film-forming agent is a mixture composed of sodium carboxymethyl cellulose and plant polysaccharide in a mass ratio of 1:2.3.

[0059] Example 7

[0060] The difference between this example and Example 1 is that the film-forming agent is a mixture composed of sodium carboxymethyl cellulose and plant polysaccharide in a mass ratio of 1:4.1.

[0061] Example 8

[0062] The difference between this example and Example 1 is that the film-forming agent is a mixture composed of sodium carboxymethyl cellulose and plant polysaccharide in a mass ratio of 1:2.8.

[0063] Example 9

[0064] The difference between this example and Example 1 is that the moisturizer is a mixture composed of hyaluronic acid and vegetable oil in a mass ratio of 1:1.4.

[0065] Example 10

[0066] The difference between this example and Example 1 is that the moisturizer is a mixture composed of hyaluronic acid and vegetable oil in a mass ratio of 1:1.9.

[0067] Example 11

[0068] The difference between this example and Example 1 is that the moisturizer is a mixture composed of hyaluronic acid and vegetable oil in a mass ratio of 1:0.8.

[0069] Example 12

[0070] The difference between this example and Example 1 is that the moisturizer is a mixture composed of hyaluronic acid and vegetable oil in a mass ratio of 1:3.

[0071] Example 13

[0072] The difference between this example and Example 1 is that the moisturizer is hyaluronic acid.

[0073] Example 14

[0074] The difference between this embodiment and Embodiment 1 is that the moisturizer is vegetable oil.

[0075] Embodiment 15

[0076] The difference between this embodiment and Embodiment 1 is that the vegetable oil is a mixture composed of coconut oil, grape seed oil and wheat germ oil in a mass ratio of 1:2.6:1.5.

[0077] Embodiment 16

[0078] The difference between this embodiment and Embodiment 1 is that the vegetable oil is a mixture composed of coconut oil, grape seed oil and wheat germ oil in a mass ratio of 1:3.2:1.5.

[0079] Embodiment 17

[0080] The difference between this embodiment and Embodiment 1 is that the vegetable oil is a mixture composed of coconut oil, grape seed oil and wheat germ oil in a mass ratio of 1:3.2:2.

[0081] Embodiment 18

[0082] The difference between this embodiment and Embodiment 1 is that the vegetable oil is a mixture composed of coconut oil, grape seed oil and wheat germ oil in a mass ratio of 1:2.6:2.

[0083] Embodiment 19

[0084] The difference between this embodiment and Embodiment 1 is that the vegetable oil is a mixture composed of coconut oil and grape seed oil in a mass ratio of 1:2.8.

[0085] Embodiment 20

[0086] The difference between this embodiment and Embodiment 1 is that the vegetable oil is a mixture composed of grape seed oil and wheat germ oil in a mass ratio of 2.8:1.7.

[0087] Embodiment 21

[0088] The difference between this embodiment and Embodiment 1 is that the B vitamins are a mixture composed of vitamin B5 and vitamin B12 in a mass ratio of 3.5:1.

[0089] Embodiment 22

[0090] The difference between this embodiment and Embodiment 1 is that the B vitamins are a mixture composed of vitamin B5 and vitamin B12 in a mass ratio of 4.2:1.

[0091] Embodiment 23

[0092] The difference between this embodiment and Embodiment 1 is that the B vitamins are vitamin B5.

[0093] Example 24

[0094] The difference between this example and Example 1 is that the B vitamins are vitamin B12.

[0095] Example 25

[0096] The difference between this example and Example 1 is that the B vitamins are vitamin B1.

[0097] Comparative Example

[0098] Comparative Example 1

[0099] The difference between this comparative example and Example 1 is that lysostaphin is not added, and the balance is made up with polyhexamethylene biguanide.

[0100] Comparative Example 2

[0101] The difference between this comparative example and Example 1 is that polyhexamethylene biguanide is not added, and the balance is made up with lysostaphin.

[0102] Comparative Example 3

[0103] The difference between this comparative example and Example 1 is that the film-forming agent is not added, and the balance is made up with purified water.

[0104] Comparative Example 4

[0105] The difference between this comparative example and Example 1 is that the film-forming agent is sodium carboxymethyl cellulose.

[0106] Comparative Example 5

[0107] The difference between this comparative example and Example 1 is that the film-forming agent is vegetable oil.

[0108] Comparative Example 6

[0109] The difference between this comparative example and Example 1 is that the film-forming agent is a mixture composed of sodium carboxymethyl cellulose and plant polysaccharide in a mass ratio of 1:1.

[0110] Comparative Example 7

[0111] The difference between this comparative example and Example 1 is that the film-forming agent is a mixture composed of sodium carboxymethyl cellulose and plant polysaccharide in a mass ratio of 1:6.

[0112] Comparative Example 8

[0113] The difference between this comparative example and Example 1 is that the humectant is not added, and the balance is made up with purified water.

[0114] Comparative Example 9

[0115] The difference between this comparative example and Example 1 is that B vitamins are not added, and the balance is made up with purified water.

[0116] Comparative Example 10

[0117] A breast care film preparation, comprising the following raw materials in mass percentages: lysostaphin 10%, chitosan 2%, surface tension regulator 5%, pH regulator 2%, and the balance being medical purified water;

[0118] Wherein the surface tension regulator is polysiloxanes and their derivatives;

[0119] The pH regulator is glacial acetic acid.

[0120] The preparation method of the above breast care film preparation is as follows:

[0121] Add chitosan to purified water, and obtain a chitosan mixture after sufficient stirring; adjust the pH of the chitosan mixture to 5 under sterile conditions, perform ultrasonic defoaming and then filter; then add lysostaphin and the surface tension regulator, and stir evenly to obtain the finished breast care film preparation.

[0122] Performance detection test method

[0123] In vitro bactericidal experiment results: Using the breast care film preparations in Examples 1-25 and Comparative Examples 1-10 as experimental samples, respectively divide the experimental samples into 6 sterilized test tubes, 3 ml in each tube, inoculate each kind of bacteria into the above-mentioned test tubes, culture until the required action time, and then take out three inoculation loops from each tube and inoculate them into ordinary broth test tubes (streptococcus hemolyticus is inoculated into 10% serum broth), and observe the results after culturing at 37°C for 24 hours.

[0124] Table 2 Bactericidal and antibacterial experiment results of breast care film preparation (action for 5 minutes)

[0125]

[0126]

[0127] Note: + represents bacterial growth; - represents no bacterial growth.

[0128] Stability experiment: Using the breast care film preparation in Example 1 as an experimental sample, conduct the following product stability experiment. As shown in the results, after three cycles of freezing and thawing at room temperature and -20°C, its properties, pH value, enzyme activity, and killing rate against common bacteria were not significantly changed when detected in the laboratory.

[0129] Table 3 Stability of breast care film preparation at room temperature (25°C)

[0130]

[0131] Table 4 Stability results of the breast care film preparation after being repeatedly frozen and thawed three times at -20°C

[0132]

[0133]

[0134] Safety experiment: The breast care film preparation prepared in Example 1 was subjected to acute, subacute and chronic toxicity tests and mucosal irritation tests by the National Institutes for Food and Drug Control of China according to the Disinfection Technical Specification. The results showed that the product formula of the present invention is non-toxic, residue-free and has no irritation to the skin and mucosa.

[0135] Table 5 Safety identification results of the breast care film preparation

[0136] Test Items Safety Evaluation Acute Oral Toxicity Test Belongs to the practically non-toxic class Skin Irritation Test Belongs to non-irritating Vaginal Mucosa Irritation Test Belongs to non-irritating Micronucleus Test No micronucleus-inducing effect Oral Mucosa Irritation Test No oral mucosa irritation Acute Eye Irritation Test Belongs to non-irritating

[0137] The efficacy of the staphylolytic enzyme breast care film preparation of the present invention is illustrated by the clinical trial report. Specifically, the breast care film preparation prepared in Example 1 was clinically tested on dairy cows in a dairy farm in Shanghai, and the results are as follows.

[0138] 1. Test group and experimental method

[0139] Test group: The number of test dairy cows is not less than 50.

[0140] Before the start of the test, the specific conditions of the dairy cows were registered, including: cattle number, gestational age, days in milk, nipple skin trait score (by naked eye observation), results of CMT tests on each mammary gland area before the test (continuous 2 times), presence or absence of clinical mastitis and medical history.

[0141] 1.1 Usage method and test period:

[0142] Usage method of the nipple film-forming care agent: Pour this product into the medicated bath cup and directly dip the entire nipple of the dairy cow without dilution.

[0143] 1.2 Test period: Use for 30 - 35 days.

[0144] 1.3 Scoring standard for the skin condition of dairy cow nipples

[0145] The internationally common 5-point system standard is adopted, and the higher the score, the worse the skin condition. 1 point: The nipple skin is smooth, without peeling, cracks or chapping; 2 points: Some peeling or scaly phenomena appear on the nipple skin; 3 points: The nipple skin is chapped, and there may be some small warts on the nipple skin; 4 points: The nipple skin has chapping, bleeding points, obvious inflammation and many warts; 5 points: The nipple skin is severely damaged and crusted, and there are many large-area warts affecting the function of the nipple.

[0146] 2. Monitoring of test data:

[0147] 2.1 Nipple skin scoring: The nipple skin traits were scored (by naked-eye observation) 1 day before the start of the trial and 1 day after the end of the trial.

[0148] 2.2 Number of clinical mastitis incidences: The number of mammary gland regions with clinical mastitis during the trial period was counted.

[0149] 2.3 CMT detection: The udders of the trial dairy cows were subjected to subclinical mastitis detection (CMT) (twice consecutively) before the start of the trial and after the end of the trial, and the number of mammary gland regions with a score of "+" or above was counted.

[0150] 3. Statistical results of the clinical application effect of the breast protection film preparation

[0151] Table 6 Statistical results of the clinical application effect of the breast protection film preparation

[0152]

[0153]

[0154] 4. Changes in nipple skin

[0155] Through naked-eye observation of the changes in the nipple skin of each mammary gland region during the trial, it was statistically found that the average nipple skin score of the dairy cows decreased by 26.87% before and after the trial. That is, the use of the breast protection film preparation can significantly improve the nipple skin condition of dairy cows.

[0156] 5. Changes in subclinical mastitis (CMT) detection

[0157] The somatic cell counts of the milk of the trial dairy cows were measured using the CMT diagnostic solution, and then the positive mammary gland regions with a detection result of "+" or above were counted. The test results showed that after 4 weeks of using the breast protection film preparation series medicated bath solution, the number of positive CMT detections in the mammary gland regions of dairy cows decreased from 25 before the trial to 13, and the reduction rate was 48%. There was no change in the number of incidences of clinical mastitis in each mammary gland region of the trial dairy cows before and after the trial.

[0158] 6. Conclusion

[0159] The test results showed that the use of the breast protection film preparation prepared in Example 1 can significantly improve the nipple skin condition of dairy cows and can significantly reduce the incidence of subclinical mastitis in dairy cows, which is worthy of popularization in production practice.

[0160] Comparative test: The breast protection film preparations prepared in each example and comparative example were used as test samples respectively. The same amount of the test samples was applied to the nipples of the corresponding dairy cows, and the evaluation was carried out with reference to the 1.3 dairy cow nipple skin condition scoring standard.

[0161] Table 7 Data table of the comparative test

[0162]

[0163]

[0164] Combined with the above experimental results, the breast care film preparation obtained in this application has excellent antibacterial and bactericidal effects, is mild in nature, has no irritation to the skin, and at the same time has a good repair and care effect. At the same time, combined with the test results of Example 1 and Comparative Examples 1-2, there is an obvious synergistic effect between lysostaphin and polyhexamethylene biguanide, and the compound use effect is significantly better than the single use.

[0165] Combined with the test results of Example 1 and Comparative Examples 3-7, the addition of the film-forming agent can effectively improve the comprehensive breast care effect of the breast care film preparation, and there is an obvious compound synergistic relationship between sodium carboxymethylcellulose and plant polysaccharides, and the compound use effect is significantly better when the two are compounded in a certain proportion.

[0166] Combined with the test results of Example 1 and Comparative Examples 8-9, the addition of the moisturizing agent and vitamin B group has obvious positive significance for the moisturizing, moisturizing and breathable properties of the breast care film layer, and has obvious positive significance for improving the breast care effect.

[0167] This specific embodiment is only an interpretation of this application, and it does not limit this application. After reading this specification, those skilled in the art can make modifications without creative contributions to this embodiment as needed, but as long as they are within the scope of the claims of this application, they are protected by the patent law.

Claims

1. A lysostaphin milk film preparation, characterized in that, It comprises raw materials with the following mass percentages: lysostaphin 0.0001 - 2%, polyhexamethylene biguanide 0.0001 - 1%, lubricant 5 - 20%, film-forming agent 0.5 - 2%, thickening agent 1 - 3%, humectant 1 - 3%, vitamin B group 0.1 - 0.5%, and the balance being purified water; the film-forming agent is a mixture composed of sodium carboxymethyl cellulose and plant polysaccharide in a mass ratio of 1:(2.3 - 4.1); the humectant is a mixture composed of hyaluronic acid and vegetable oil in a mass ratio of 1:(1.4 - 1.9); the vegetable oil is a mixture composed of coconut oil, grape seed oil and wheat germ oil in a mass ratio of 1:(2.6 - 3.2):(1.5 - 2.0); the plant polysaccharide is aloe polysaccharide; the vitamin B group is a mixture composed of vitamin B5 and vitamin B12 in a mass ratio of (3.5 - 4.2):

1.

2. The lysostaphin milk film preparation according to claim 1, wherein It comprises raw materials with the following mass percentages: lysostaphin 0.4 - 1%, polyhexamethylene biguanide 0.04 - 0.2%, lubricant 10 - 20%, film-forming agent 1 - 2%, thickening agent 1 - 2%, humectant 1.5 - 2.4%, vitamin B group 0.2 - 0.4%, and the balance being purified water; the film-forming agent is a mixture composed of sodium carboxymethyl cellulose and plant polysaccharide in a mass ratio of 1:(2.8 - 3.5).

3. The lysostaphin milk film preparation according to claim 1, characterized in that, The lubricant includes at least one of glycerol and lanolin.

4. A lysostaphin milk film preparation according to claim 1, characterized in that, The thickening agent includes at least one of sodium alginate, carrageenan and gum arabic.

5. A preparation process of the lysostaphin-containing milk film preparation according to any one of claims 1 to 4, characterized in that, It includes the following steps: First mixing: Mix the film-forming agent, thickening agent, humectant, vitamin B group and part of the purified water, and stir evenly to obtain solution A; Second mixing: Mix lysostaphin and polyhexamethylene biguanide and dissolve them in part of the purified water, stir evenly and then let it stand, and then add the lubricant and stir evenly to obtain solution B; Third mixing: Mix solution A and solution B, stir evenly to obtain the finished product of the nursing film preparation, and store it in the dark.

6. The preparation process of a lysostaphin milk film preparation according to claim 5, characterized in that, In the first mixing and second mixing steps, the purified water is obtained by heating and boiling and then keeping it at 30 - 45°C for insulation.

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