A medicated bath liquid for preventing and treating cow mastitis and a preparation method thereof

Abstract

The application discloses a medicated bath liquid for preventing and treating cow mastitis and a preparation method thereof, and belongs to the technical field of veterinary drug preparations. Each component in the medicated bath liquid is bactericide 1-20 parts, emollient 5-50 parts, auxiliary agent 1-20 parts, toning agent 0.1-4 parts, stabilizer 0.1-2 parts, thickening agent 0.1-2 parts, skin softener 0.1-5 parts, and purified water 20-90 parts in terms of weight ratio. The medicated bath liquid has the advantages of low residue, fast sterilization, persistent sterilization, nipple skin protection and the like, avoids the disadvantages of high residue, drug resistance and nipple skin dryness and the like that may be caused by using conventional iodine preparations, meanwhile, raw materials of the product are easy to obtain, the preparation process is simple, and the product can be mass-produced.

Description

Technical Field

[0001] This invention belongs to the field of veterinary drug preparation technology, specifically relating to a medicated bath solution for preventing and treating mastitis in dairy cows and its preparation method. Background Technology

[0002] Mastitis is the leading cause of death among dairy cows, with clinical mastitis incidence rates ranging from 1% to 10% and subclinical mastitis rates ranging from 7% to 35% in China. Disinfecting teats with disinfectants before and after milking can effectively reduce the incidence of mastitis. Most dairy farms in China use iodine-based teat disinfectants to prevent mastitis.

[0003] Dairy teat disinfectants, used before and after milking, come into direct contact with the cow's teat skin. Therefore, the safety and residue levels of the bactericides used in their formulation are crucial. Iodine in iodine preparations has strong iodination and oxidation capabilities, allowing it to penetrate the skin and enter the bloodstream, potentially increasing the iodine content in milk. The higher the concentration of iodine-containing bath products used on dairy farms, the greater the risk of iodine being introduced into the milk. Furthermore, long-term use of iodine preparations may lead to the development of microbial resistance.

[0004] Patent CN 105596235A uses guanidine disinfectants and alcohols to replace iodine bactericides in the preparation of dairy cow medicated bath solutions. The use of alcohols in this patent leads to peeling and dryness of the cow's skin, making it more susceptible to microbial colonization on the teats. Furthermore, guanidine disinfectants, as surfactants, are significantly affected by organic matter during sterilization. CN106344491 B uses benzalkonium chloride and hydrogen peroxide as bactericides to prepare dairy cow teat medicated bath solutions. Benzalkonium chloride is a cationic surfactant, which is highly effective against Gram-positive bacteria but ineffective against Pseudomonas aeruginosa, acid-fast bacilli, and spores. Its bactericidal effect is significantly reduced in the presence of organic matter. When cow dung or bedding from the barn adheres to the teats, the aforementioned active ingredients cannot effectively kill microorganisms on the teats. The uncertain effective ingredients and content, and insignificant efficacy of traditional Chinese medicine (TCM) medicated bath products hinder their widespread application.

[0005] Finding bactericides that are low in residue, safe, more effective, and gentle on the skin is a major problem that urgently needs to be solved in this field. Summary of the Invention

[0006] The present invention aims to solve the technical problems in the prior art and provide a medicated bath solution for preventing and treating mastitis in dairy cows and its preparation method.

[0007] To achieve the above objectives, the specific technical solution of the present invention is as follows:

[0008] In a first aspect, the present invention provides a medicated bath solution for preventing and treating mastitis in dairy cows, the medicated bath solution comprising the following components: bactericide, emollient, auxiliary agent, thickener, stabilizer, colorant, skin softener, and purified water.

[0009] The medicated bath solution for preventing and treating mastitis in dairy cows comprises, by weight, 1-20 parts bactericide, 5-50 parts emollient, 1-20 parts auxiliary agent, 0.1-2 parts stabilizer, 0.1-4 parts colorant, 0.1-2 parts thickener, 0.1-5 parts skin softener, and 20-90 parts purified water.

[0010] Preferably, the medicated bath solution for preventing and treating mastitis in dairy cows comprises, by weight, 5-15 parts bactericide, 20-50 parts emollient, 1-15 parts auxiliary agent, 0.1-3 parts colorant, 0.1-1.5 parts stabilizer, 0.1-2 parts thickener, 0.2-3 parts skin softener, and 30-70 parts purified water.

[0011] More preferably, the medicated bath solution for preventing and treating mastitis in dairy cows comprises, by weight, 5-13 parts bactericide, 35-50 parts emollient, 3-8 parts auxiliary agent, 0.5-1.5 parts colorant, 0.2-0.8 parts stabilizer, 0.5-1.5 parts thickener, 0.5-2 parts skin softener, and 35-60 parts purified water.

[0012] The bactericide is one or more of the following: peroxides, guanidines, organic acids, quaternary ammonium salts, and alcohols.

[0013] Preferably, the bactericide is one or more of hydrogen peroxide, chlorhexidine gluconate, chlorhexidine acetate, polyhexamethylene biguanide hydrochloride (PHMB), polyhexamethylene monoguanide hydrochloride (PHMG), peracetic acid, lactic acid, heptanoic acid, benzalkonium chloride, and 1,3-propanediol.

[0014] More preferably, the bactericide is a combination of chlorhexidine gluconate, lactic acid, and heptanoic acid.

[0015] The mass ratio of chlorhexidine gluconate, lactic acid, and heptanoic acid in the bactericide is 1:(10-20):(1.25-10).

[0016] More preferably, the mass ratio of chlorhexidine gluconate, lactic acid, and heptanoic acid in the bactericide is 1:10:2.5.

[0017] The selected emollient is one or more of the following: glycerin, urea, sorbitol, water-soluble lanolin, glycerol, and sodium lactate.

[0018] Preferably, the emollient is selected from glycerin and water-soluble lanolin.

[0019] The adjuvant is one or more of polysorbate-60, sodium dodecyl sulfonate, dodecylbenzene sulfonic acid, polyoxyethylene phosphate, PEG400, fatty alcohol polyoxyethylene ether, alkyl glycoside, and polyethylene glycol stearate.

[0020] Preferably, the adjuvant is selected from sodium dodecyl sulfonate and polysorbate-60.

[0021] The colorant includes one or more of brilliant blue, acid blue, lemon yellow, gardenia yellow, and carmine, preferably carmine.

[0022] The thickener includes one or more of hydroxyethyl cellulose, hydroxymethyl cellulose, sodium alginate, guar gum, locust bean gum, and hydroxypropyl guar gum, preferably hydroxypropyl guar gum.

[0023] The stabilizer includes one or more of phosphates, EDTA, lactates, acetates, and sodium metabisulfite, preferably sodium metabisulfite.

[0024] The skin softening agent is one or more of salicylic acid, urea, lactic acid, and allantoin, preferably allantoin.

[0025] A preferred formulation of a medicated bath solution for preventing and treating mastitis in dairy cows is as follows: 8 parts lactic acid, 1-2 parts heptanoic acid, 0.8 parts chlorhexidine gluconate, 41 parts emollient, 5 parts auxiliary agent, 1 part thickener, 1 part colorant, 0.5 parts stabilizer, 1 part skin softener, and 39.7-40.7 parts purified water.

[0026] A more preferred formulation of a medicated bath for preventing and treating mastitis in dairy cows is as follows: 8 parts lactic acid, 2 parts heptanoic acid, 0.8 parts chlorhexidine gluconate, 39 parts glycerin, 2 parts water-soluble lanolin, 3 parts polysorbate-60, 2 parts sodium dodecyl sulfate, 1 part hydroxypropyl guar gum, 1 part carmine, 0.5 parts sodium metabisulfite, 1 part allantoin, and 39.7 parts purified water.

[0027] In a second aspect, the present invention provides a method for preparing a medicated bath solution for preventing and treating mastitis in dairy cows, the preparation steps of which are as follows:

[0028] (1) Dissolve the prescribed amount of thickener in purified water to prepare solution one;

[0029] (2) Dissolve the prescribed amount of bactericide in purified water to prepare solution two;

[0030] (3) Mix the prescribed amounts of emollient, skin softener and colorant to make solution three;

[0031] (4) Mix solution one and solution two and stir well, then add to solution three to obtain solution four with a pH value of 2-6;

[0032] (5) Add the auxiliary agent and stabilizer to solution four, stir evenly and then make up the volume to obtain the medicated bath solution.

[0033] Preferably, the pH value in step (4) is 3-5.

[0034] The chlorhexidine gluconate component in the above-mentioned medicated bath solution for the prevention and treatment of mastitis in dairy cows kills microorganisms by altering the permeability of bacterial cell membranes, destroying their structure and function, and denaturing intracellular macromolecules.

[0035] Both lactic acid and heptanoic acid are food additives. Lactic acid is a colorless, transparent liquid at room temperature and is readily soluble in water. It is weakly acidic and works by creating an acidic environment that inhibits bacterial growth, altering the permeability and stability of bacterial cell membranes, thereby disrupting bacterial structure and function and inhibiting bacterial reproduction. Heptanoic acid, at certain concentrations, can alter the permeability of microbial cell membranes, thereby disrupting cell metabolism and achieving a bactericidal effect. It has a certain bactericidal effect against various microorganisms, spores, and viruses.

[0036] GB 2760-2014, the National Food Safety Standard for the Use of Food Additives, indicates that lactic acid can be used in appropriate amounts as needed in other special dietary foods. Heptanoic acid is naturally found in many plants, hops, dairy products, and fermented products. Chlorhexidine gluconate is widely used for preoperative skin disinfection and wound disinfection. Studies on its skin irritation, acute toxicity, and subacute toxicity show that this disinfectant component is non-irritating to the skin, its acute oral toxicity is practically non-toxic, and no toxic effects were observed in subacute toxicity. It can be used for sterilization and disinfection.

[0037] Beneficial effects:

[0038] (1) This invention provides a novel compound bactericidal medicated bath solution for the prevention and treatment of mastitis in dairy cows. It uses chlorhexidine gluconate, lactic acid and heptanoic acid to enhance the effect. It can achieve effective bactericidal effect in 15 seconds, which is faster and more efficient than the currently used medicated bath solution (30 seconds of bactericidal effect).

[0039] (2) The medicated bath product of the present invention has added stabilizers, which makes the product more stable. The medicated bath liquid is made of food additive raw materials and low-residue bactericidal ingredients. There is no bactericidal residue after sterilization. It can be used as a disinfectant for dairy cow teats, which improves the quality and safety of milk.

[0040] (3) This invention does not contain iodine-based raw materials or alcohols, and is non-irritating to the skin of cow teats, keeping the teats smooth and moist, and effectively preventing the occurrence of mastitis in cows.

[0041] (4) The preparation method of the present invention is simple, the raw materials are readily available, and it is easy to industrialize. Detailed Implementation

[0042] The above content will be further described in detail below through specific embodiments. However, this should not be construed as limiting the scope of the above subject matter to the following embodiments. All technologies implemented based on the content of this invention fall within the scope.

[0043] Examples 1-11

[0044] A medicated bath solution for preventing and treating mastitis in dairy cows:

[0045] Table 1. Formulation and dosage of medicated bath solutions for the prevention and treatment of mastitis in dairy cows.

[0046]

[0047]

[0048] Experimental Example 1: High Temperature Accelerated Stability

[0049] The products of Examples 1-11 were placed into sample bottles and placed in drying ovens at 50 and 60°C respectively. After 10 days, the samples were taken out of the drying ovens and the pH value changes of different samples were detected using a pH meter. The results are shown in Table 2.

[0050] Table 2 pH stability of samples after high-temperature acceleration in different embodiments

[0051]

[0052] Based on the pH stability comparison of Examples 1 to 11 in Table 1, it can be seen that adding the stabilizer sodium metabisulfite to the bath solution can improve the pH stability under high temperature conditions of 50℃ and 60℃. The formulations of Examples 3 to 10 all showed good pH stability, especially the formulation of Example 6.

[0053] Experimental Example 2: Laboratory Sterilization Efficacy Verification

[0054] The laboratory bactericidal effects of the above examples were verified using Staphylococcus aureus (ATCC25923), Streptococcus agalactiae (ATCC13813), Streptococcus lactis (ATCC9927), Escherichia coli (ATCC44102), and Bacillus subtilis (ATCC6633).

[0055] 1. Rapid sterilization test

[0056] Staphylococcus aureus, Streptococcus agalactiae, Streptococcus lactis, Escherichia coli, and Bacillus subtilis were inoculated onto nutrient agar medium and cultured for 24 hours. Then, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus lactis, Escherichia coli, and Bacillus subtilis were inoculated onto nutrient broth medium, blood agar medium, LB medium, and beef extract peptone medium and cultured at 37°C for 18-24 hours for later use.

[0057] For each example and control, povidone-iodine solution was used to prepare one reaction tube and four neutralization tubes for each pathogenic bacterium. 1 mL of bacterial suspension was added to the reaction tube, followed by 1 mL of the corresponding example sample. After mixing, timing was started. 1 mL of the reaction solution was taken out at 15 s, 30 s, 60 s, and 1 min, and 9 mL of neutralizing agent was added (preliminary tests confirmed that the neutralizing agent could completely inactivate the corresponding bactericide in the example). After shaking, the mixture was allowed to stand for 10 min. Then, 1 mL of the solution was taken and 9 mL of the corresponding liquid culture medium was added. After shaking, the mixture was placed in a 37℃ incubator for 24 hours. The bacterial growth in each tube was observed, and the results are shown in Tables 3-7.

[0058] Table 3. Rapid bactericidal ability of different embodiments against Staphylococcus aureus

[0059]

[0060] Note: "-" indicates no bacterial growth, "+" indicates bacterial growth. 5% povidone-iodine is used with commercially available 5% povidone-iodine dairy cow bath solution.

[0061] Table 4. Rapid bactericidal ability of different embodiments against Streptococcus agalactiae

[0062]

[0063] Note: "-" indicates no bacterial growth, "+" indicates bacterial growth. 5% povidone-iodine is used with commercially available 5% povidone-iodine dairy cow bath solution.

[0064] Table 5. Rapid bactericidal ability of different embodiments against Streptococcus mammaria.

[0065]

[0066]

[0067] Note: "-" indicates no bacterial growth, "+" indicates bacterial growth. 5% povidone-iodine is used with commercially available 5% povidone-iodine dairy cow bath solution.

[0068] Table 6. Rapid bactericidal ability of different embodiments against Escherichia coli

[0069]

[0070] Note: "-" indicates no bacterial growth, "+" indicates bacterial growth. 5% povidone-iodine is used with commercially available 5% povidone-iodine dairy cow bath solution.

[0071] The experimental results in Tables 3 to 6 show that Examples 2 and 3 were not effective in killing Staphylococcus aureus, Escherichia coli, Streptococcus agalactiae, and Streptococcus dysgalactiae within 15 and 30 seconds, respectively. Examples 1 with added benzalkonium chloride and Examples 4-10 with added organic acids, along with 5% povidone-iodine, were all effective in killing Staphylococcus aureus, Escherichia coli, Streptococcus agalactiae, and Streptococcus dysgalactiae within 30 seconds. In particular, Examples 5 and 6 could kill these common mastitis pathogens—Staphylococcus aureus, Escherichia coli, Streptococcus agalactiae, and Streptococcus dysgalactiae—within 15 seconds, demonstrating the strongest rapid bactericidal ability.

[0072] Table 7. Rapid bactericidal ability of different embodiments against Bacillus subtilis

[0073]

[0074] Note: "-" indicates no bacterial growth, "+" indicates bacterial growth. 5% povidone-iodine is used with commercially available 5% povidone-iodine dairy cow bath solution.

[0075] The experimental results in Table 7 show that Examples 1, 2, and 3 can effectively kill Bacillus subtilis within 2 minutes. Example 4, with the addition of organic acid, can effectively kill Bacillus subtilis within 60 seconds. Examples 5, 6, 9, and 10, along with 5% povidone-iodine, can all effectively kill Bacillus subtilis within 30 seconds. Examples 7 and 8 require more than 30 seconds, and Example 11 requires at least 60 seconds to kill Bacillus subtilis.

[0076] 2. Antibacterial test

[0077] To further illustrate the effect of adding different organic acids on the antibacterial ability of the products, an antibacterial test was conducted using the Oxford cup double-layer agar diffusion method. The size of the inhibition zone of the products in Examples 2 to 11 was observed and measured, and compared with commercially available 5% povidone-iodine dairy cow teat bath products. The results are shown in Table 8. In Example 4, compared to Example 3, the addition of lactic acid increased the diameter of the inhibition zone against Staphylococcus aureus, Streptococcus agalactiae, Streptococcus lactis, and Escherichia coli compared to using chlorhexidine gluconate alone. In Examples 5 and 6, compared to Example 3, the addition of heptanoic acid further increased the diameter of the inhibition zone against Staphylococcus aureus, Streptococcus agalactiae, Streptococcus lactis, and Escherichia coli. Meanwhile, in Examples 7 and 8, acetic acid and peracetic acid were used instead of heptanoic acid, resulting in a decrease in antibacterial ability. This indicates that the combination of chlorhexidine gluconate, lactic acid, and heptanoic acid can achieve a better antibacterial effect. Examples 5 and 6, and Examples 9 and 10 all showed better antibacterial ability than the 5% povidone-iodine group. The diameter of the inhibition zone in Example 11 was comparable to that of the 5% povidone-iodine group.

[0078] Table 8. Diameter of inhibition zones (mm) for different mastitis pathogens in different embodiments.

[0079]

[0080] Trial Case 3: Clinical Efficacy Trial

[0081] To further verify the effectiveness of this invention in preventing clinical mastitis in dairy cows, experiments were conducted using Examples 1, 6, 9, 10, and 11 with 5% povidone-iodine solution to test the protective effect against mastitis in dairy cows. 480 healthy dairy cows with an average lactation period of 140 days (somatic cell count below 200,000 CFU / mL) were selected and divided into 6 groups of 80 cows each. The experimental period was 30 days. The condition of the teat skin and the incidence of clinical mastitis were observed at the beginning and end of the experiment. Somatic cell counts for each cow were measured using a somatic cell analyzer at the beginning and end of the experiment. The results are shown in Tables 9 and 10.

[0082] Table 9. Clinical mastitis incidence in dairy cows in different embodiments.

[0083]

[0084]

[0085] As shown in Table 9, compared with the commonly used 5% povidone-iodine teat disinfectant, Examples 6, 9, and 10 all reduced the incidence of clinical mastitis in dairy cows and also reduced somatic cell counts. Examples 1 and 11 showed poorer effects in reducing clinical mastitis, and the somatic cell counts were slightly higher at the end of the experiment.

[0086] Table 10. Average teat scores of cows at the start and end of the experiment.

[0087] Example 1 1.344 1.4375 Example 6 1.375 1.125 Example 9 1.406 1.3125 Example 10 1.422 1.2813 Example 11 1.344 1.531 5% povidone-iodine 1.3125 1.35

[0088] Note: Scoring criteria: 1 point - smooth, moisturized skin; 2 points - slightly dry skin; 3 points - dry, flaky skin.

[0089] As shown in Table 10, the teat skin score of the cows in Example 6 decreased from 1.375 to 1.125, while the teat skin score of the cows in the 5% povidone-iodine group increased from 1.3125 to 1.35. Example 6 can better protect the teat skin. The teat skin in Examples 1 and 11 became slightly drier, and the skin scores in Examples 9 and 10 decreased.

[0090] In summary, compared with conventional iodine-based dairy cow teat disinfectants, this invention reduces the residue of active ingredients in milk, has a faster and more sustained bactericidal ability and a preventive effect against clinical mastitis, while also resulting in better dairy cow teat skin and increased farming profits.

[0091] The above embodiments are merely illustrative of the technical methods and bactericidal active ingredients used in this invention. It should be noted that those skilled in the art can make various improvements and modifications to this invention without departing from its principles, and these improvements and modifications also fall within the scope of protection of the claims of this invention.

Claims

1. A medicated bath solution for preventing and treating mastitis in dairy cows, characterized in that, The medicated bath solution comprises the following components: bactericide, emollient, auxiliary agent, thickener, colorant, stabilizer, skin softener, and purified water; the bactericide is a combination of chlorhexidine gluconate, lactic acid, and heptanoic acid; the emollient is selected from glycerin and water-soluble lanolin; the auxiliary agent is selected from sodium dodecyl sulfate and polysorbate-60; the colorant is carmine; the stabilizer is sodium metabisulfite; the thickener is hydroxypropyl guar gum; the skin softener is allantoin; the medicated bath solution for preventing and treating mastitis in dairy cows, by weight, comprises 8 parts lactic acid, 2 parts heptanoic acid, 0.8 parts chlorhexidine gluconate, 39 parts glycerin, 2 parts water-soluble lanolin, 3 parts polysorbate-60, 2 parts sodium dodecyl sulfate, 1 part hydroxypropyl guar gum, 1 part carmine, 0.5 parts sodium metabisulfite, 1 part allantoin, and 39.7 parts purified water.

2. A method for preparing a medicated bath solution for preventing and treating mastitis in dairy cows as described in claim 1, comprising the following steps: (1) Dissolve the prescribed amount of thickener in purified water to prepare solution one; (2) Dissolve the prescribed amount of bactericide in purified water to prepare solution two; (3) Mix the prescribed amounts of emollient, skin softener, and colorant to prepare solution three; (4) Mix solution one and solution two and stir well, then add to solution three to obtain solution four with a pH value of 2-6; (5) Add the auxiliary agent and stabilizer to solution four, stir evenly and make up the volume to obtain the medicated bath solution for preventing and treating mastitis in dairy cows.

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