A dairy cow teat disinfectant that promotes the repair of damaged epidermis, its preparation method and application

Abstract

This invention relates to the field of disinfectant technology, and more particularly to a dairy cow teat disinfectant that promotes the repair of damaged epidermis, its preparation method, and its application. By weight, the dairy cow teat disinfectant that promotes the repair of damaged epidermis comprises the following raw materials: 0.2-2 parts iodine, 0.1-5 parts sodium iodide, 0.1-0.5 parts sodium iodate, 0.1-10 parts skin-repairing active ingredients, 1-10 parts surfactant, 0.05-0.5 parts thickener, 2-10 parts lubricant, 0.1-0.5 parts wetting agent, and 0.15-1 part pH adjuster, with the balance being water. A novel surfactant, alkyl glycoside, is selected. In the disinfectant system, alkyl glycoside can efficiently complex with iodine to form highly stable complexed iodine, extending the shelf life of the formulation. In addition, the cow teat disinfectant in this solution also contains skin repair ingredients. While maintaining a highly effective bactericidal effect, it can also promote the repair of skin damaged by bacterial infection, promote the healing of abrasions on the cow teat epidermis, and reduce the risk of infection by pathogens due to exposed teat epidermal wounds.

Description

Technical Field

[0001] This invention relates to the field of disinfectant technology, and in particular to a dairy cow teat disinfectant that can promote the repair of damaged epidermis, its preparation method, and its application. Background Technology

[0002] Currently, dairy cow teat bath agents mainly include iodine preparations, chlorhexidine, chlorine dioxide preparations, dodecylbenzenesulfonic acid preparations, glycolic acid preparations, and hydrogen peroxide preparations. Many new disinfectants have also emerged in domestic and international markets. Among all the above-mentioned dairy cow teat bath agents, iodine preparations are the most mainstream type, especially in the Chinese and Japanese markets, where they account for as much as 85-90%.

[0003] Among them, the most commonly used iodine preparation on the market is povidone-iodine, which is a disinfectant formed by complexing iodine with polyvinylpyrrolidone, also known as povidone-iodine. As a first-generation iodine preparation, povidone-iodine is included in various pharmacopoeias both domestically and internationally. It is a medium-efficiency chemical disinfectant, widely used in human and veterinary surgical disinfection due to its good bactericidal effect and low irritation. Compared to iodine tincture, while povidone-iodine improves the stability of the solution to some extent, it still has shortcomings: because it contains some free iodine, it is prone to sublimation with increasing storage time and can easily undergo complex chemical reactions in aqueous systems, leading to a rapid decrease in the effective iodine content; furthermore, the high povidone-iodine content in povidone-iodine disinfectant solution can easily lead to residues in milk when used for disinfecting dairy cow teats.

[0004] There are many causes of mastitis in dairy cows, including climate change, pasture management, cleanliness and hygiene of the milking parlor, use of disinfectants before and after milking, and physical injuries to the cows themselves. Proper use of disinfectant baths before and after milking can effectively kill pathogens attached to the cow's teats, reducing the chance of infection and lowering the risk of subclinical mastitis. However, most teat disinfectants on the market currently use povidone-iodine, which often has an unsuitable consistency. If the consistency is too thick, more disinfectant is needed, increasing costs; if it is too thin, the disinfectant cannot be evenly distributed on the udder surface, leading to excessive dripping, which not only contaminates the milking parlor and barn but also reduces the effectiveness of disinfection.

[0005] Furthermore, commercially available povidone-iodine disinfectants have limited effectiveness, mostly only providing disinfection and sterilization without promoting the healing of wounds on the cow's teats. In dairy farms, cows frequently experience varying degrees of damage to their teats, such as injuries from rubbing against the ground while lying down, friction with milking equipment, or skin abrasions caused by friction from swollen udders. Damaged udder skin impairs the natural skin barrier, weakening the body's natural protective mechanisms and making it easier for pathogens in the environment to invade the animal's body, increasing the risk of mastitis. Ordinary commercially available udder bath solutions only meet basic disinfection requirements and are insufficient for complex clinical situations, failing to adequately address the current challenges of mastitis in dairy cows. Only by providing continuous and stable disinfection while promoting the healing of the cow's udder skin and accelerating the recovery of its own skin barrier can the risk of pathogens invading the body be reduced at its source, thus lowering the probability of mastitis in dairy cows.

[0006] Therefore, there is a need to develop a dairy cow teat disinfectant with good adhesion, long-term stability, and the ability to promote the repair of damaged epidermis. Summary of the Invention

[0007] The main objective of this invention is to provide a cow teat disinfectant that can promote the repair of damaged epidermis, its preparation method, and its application. This invention aims to improve the technical problems of existing cow teat disinfectants, such as the rapid decline of effective ingredients after long-term storage, unsuitable texture, and the inability to promote the healing of cow teat skin wounds despite only having a disinfecting effect.

[0008] To achieve the above objectives, this invention proposes a dairy cow teat disinfectant that can promote the repair of damaged epidermis. By weight, it comprises the following ingredients: 0.2-2 parts iodine, 0.1-5 parts sodium iodide, 0.1-0.5 parts sodium iodate, 0.1-10 parts skin-repairing active ingredients, 1-10 parts surfactant, 0.05-0.5 parts thickener, 2-10 parts lubricant, 0.1-0.5 parts wetting agent, and 0.15-1 part pH adjuster, with the remainder being water.

[0009] Preferably, the skin-repairing active ingredient is any one of glyceryl glucoside, aloe vera extract, or γ-polyglutamic acid.

[0010] Preferably, the surfactant is an alkyl glycoside surfactant.

[0011] Preferably, the molecular weight of the alkyl glycoside surfactant is 320.42-348.47.

[0012] Preferably, the thickener is at least one of glucomannan, guar gum, tara gum, and locust bean gum; the lubricant is at least one of glycerol, propylene glycol, PEG4000, and PEG6000; and the wetting agent is at least one of sodium docusate, AEROSOL OT-75, and BETTERSOL 607.

[0013] The pH adjuster includes an acidic adjuster and an alkaline adjuster. By weight, the amount of the acidic adjuster added is 0.1-0.5 parts, and the amount of the alkaline adjuster added is 0.05-0.5 parts.

[0014] The acidity regulator is at least one of anhydrous citric acid, malic acid, and mandelic acid, and the alkalinity regulator is at least one of sodium hydroxide, potassium hydroxide, and sodium carbonate.

[0015] Preferably, by weight, it comprises the following raw materials: 1 part iodine, 0.5 parts sodium iodide, 0.1 parts sodium iodate, 6 parts skin repair functional ingredients, 5 parts surfactant, 0.5 parts thickener, 0.2 parts wetting agent, 10 parts lubricant, 0.3 parts acidity regulator, 0.15 parts alkalinity regulator, and 77.45 parts water.

[0016] In addition, the present invention also proposes a method for preparing a bovine teat disinfectant that can promote the repair of damaged epidermis as described in any of the above claims, comprising the following preparation steps:

[0017] S1. Under constant temperature conditions in a water bath, the surfactant, acid regulator, wetting agent and 60%-70% of the total weight of water are mixed according to the formula and stirred until evenly dispersed to obtain mixture A;

[0018] S2. Disperse the thickener evenly in the lubricant to obtain mixture B;

[0019] S3. Under constant temperature conditions in a water bath, add mixture B to mixture A and stir to disperse evenly to obtain mixture C;

[0020] S4. After cooling mixture C to ≤40℃, sodium iodide is added and stirred until evenly dispersed to obtain mixture D;

[0021] S5. Under closed conditions, add 30%-40% of the total weight of water to mixture D, add iodine and stir to disperse evenly, then add an alkaline regulator, and then add sodium iodate and stir to disperse evenly to obtain mixture E;

[0022] S6. Add the skin-repairing active ingredients to mixture E and stir to disperse them evenly to obtain the cow teat disinfectant solution that can promote the repair of damaged epidermis.

[0023] Preferably, the water bath temperature in steps S1 and S3 is 60-70℃, and the water bath time is 15-45 min.

[0024] Preferably, the pH value of the cow teat disinfectant that promotes the repair of damaged epidermis is 4.5-4.8.

[0025] In addition, the present invention also proposes an application of a cow teat disinfectant that can promote the repair of damaged epidermis as described in any of the above claims, wherein the cow teat disinfectant is used by spraying, rinsing, soaking, or applying, whether diluted or undiluted.

[0026] Compared with the prior art, the technical solution of the present invention has the following beneficial effects:

[0027] 1. Addressing the issue of rapid decline in the effective components of existing dairy cow teat disinfectants after long-term storage, this solution differs from traditional povidone-iodine disinfectants by using a novel surfactant, alkyl glycoside (APG2000-decyl glucoside). In the disinfectant system, alkyl glycosides can efficiently complex with iodine to form highly stable complexed iodine, allowing iodine to dissolve in water and release free iodine. The effective iodine content suffers minimal damage over a longer period, extending the shelf life of the commercial formulation and improving its protective efficacy in clinical use. This also prolongs the effective sterilization time and further reduces the possibility of pathogenic microorganisms invading the animal's body.

[0028] 2. Furthermore, the cow teat disinfectant in this formulation also contains skin-repairing ingredients such as glyceryl glucoside, aloe vera extract, and γ-polyglutamic acid. While maintaining highly effective bactericidal action, it also promotes the repair of skin damaged by bacterial infection, accelerates the healing of abrasions on the cow teat epidermis, and reduces the risk of pathogen infection due to exposed teat epidermal wounds. Experimental verification shows that this disinfectant can inhibit inflammatory cell infiltration in wounds, accelerate the resolution of inflammatory responses, promote the healing of damaged skin, restore the body's own skin barrier, and fundamentally improve the body's ability to defend against pathogens.

[0029] 3. Addressing the issue of high povidone-iodine content in povidone-iodine disinfectants potentially leaving residues in milk, this solution avoids the use of traditional povidone-iodine. Instead, it employs iodine, sodium iodide, and sodium iodate in combination with other ingredients, thus preventing residue contamination in milk. Furthermore, the disinfectant formulation of this invention uses completely biodegradable materials. The surfactants, repairing agents, thickeners, and other excipients are all natural extracts or food additives obtained through bacterial fermentation, making it both environmentally friendly and safe, effectively preventing residual substances from affecting human health or contaminating milk sources and the environment.

[0030] 4. In response to the common problem of unsuitable texture in existing povidone-iodine disinfectants, this solution introduces thickeners and lubricants into the raw material system. By utilizing the organic combination of these two excipients, the skin feel and adhesion of the disinfectant are enhanced, resulting in a suitable texture that allows the disinfectant to be evenly distributed on the surface of the cow's udder through a medicated bath. Attached Figure Description

[0031] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0032] Figure 1 Results of wound healing rate measurement for bacterial infections in mice;

[0033] Figure 2 Results of bacterial load measurement on bacterial wounds in mice on day 8 (Note: **p<0.01);

[0034] Figure 3 The mean optical density of iNOS on the wound skin eight days after Staphylococcus aureus ATCC29213 infection (Note: **p<0.01);

[0035] Figure 4 The mean optical density of the skin at F4 / 80 on the wound surface eight days after infection with Staphylococcus aureus ATCC29213 is measured (Note: **p<0.01);

[0036] Figure 5 The mean optical density of Ly6G in the skin wound eight days after Staphylococcus aureus ATCC29213 infection was measured (Note: *p<0.05, **p<0.01).

[0037] Figure 6 The mean optical density of the skin P-P38 on the wound surface eight days after infection with Staphylococcus aureus ATCC29213 is measured (Note: **p<0.01).

[0038] The realization of the purpose, functional features and advantages of this application will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0039] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0040] Furthermore, the technical solutions of the various embodiments can be combined with each other, but only if they are feasible for those skilled in the art. If the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such combination of technical solutions does not exist and is not within the scope of protection claimed by this invention.

[0041] A disinfectant for dairy cow teats that can promote the repair of damaged epidermis, comprising, by weight, the following ingredients: 0.2-2 parts iodine, 0.1-5 parts sodium iodide, 0.1-0.5 parts sodium iodate, 0.1-10 parts skin-repairing active ingredients, 1-10 parts surfactant, 0.05-0.5 parts thickener, 2-10 parts lubricant, 0.1-0.5 parts wetting agent, and 0.15-1 part pH adjuster, with the balance being water.

[0042] In the above disinfectant formula, sodium iodide is used as a co-solvent, according to the chemical equation... Therefore, iodine molecules, as Lewis acids, react with I... －The lone pair electrons combine to form triiodide ions, which are much more soluble in water than iodine molecules. Sodium iodide increases the iodide ion content in the system, significantly increasing the solubility of iodine in water and accelerating its dissolution. A pH adjuster maintains the pH stability of the entire system, thereby enhancing the stability of each active ingredient to cope with the complex and variable environmental climate of pastures in practical applications. It also makes the pH of the disinfectant solution close to the pH of the cow's skin, reducing irritation to the animals during use. The cow teat medicated bath disinfectant solution of this invention also contains excipients such as thickeners and lubricants. The organic combination of these two excipients enhances the skin feel and adhesion of the disinfectant solution, allowing it to be evenly distributed on the surface of the cow's udder during the medicated bath. Commercially available traditional film-forming povidone-iodine disinfectants are often too sticky, leading to increased usage per bath and causing dripping waste that is difficult to clean. This invention, through the combination of thickeners and lubricants, significantly reduces dripping, ensuring long-lasting and effective disinfection and care, while also lubricating the udder skin. This facilitates cleaning after each bath and effectively reduces the risk of friction damage during milking. More importantly, in addition to the aforementioned thickeners and lubricants, the system also contains wetting agents and repair-promoting ingredients, allowing it to penetrate deep into the folds of the cow's udder skin. The solution remains effective for a long time after bathing, killing pathogens hidden in the skin folds, continuously and comprehensively maintaining and repairing the udder epidermis, preventing skin cracking, promoting the healing of bacterial infections, and reducing the risk of mastitis in cows.

[0043] Furthermore, the skin-repairing active ingredient is any one of glyceryl glucoside, aloe vera extract, or γ-polyglutamic acid.

[0044] This formula contains three skin-repairing ingredients: glyceryl glucoside, aloe vera extract, and γ-polyglutamic acid. Glyceryl glucoside promotes the expression of AQP3 mRNA and protein in the stratum corneum of the skin. AQP3 is crucial for maintaining high glycerol levels in the stratum corneum, achieving a highly effective moisturizing effect. It also increases the production of the antioxidant skin-protective enzyme superoxide dismutase 1 (SOD1), enhancing the skin's own immune resistance, improving the skin barrier, and promoting the repair of dry and damaged skin. The main active component of aloe vera extract is aloe polysaccharide. Large-molecule aloe polysaccharides have good film-forming properties, forming a thin film on the epidermis, reducing skin moisture evaporation, and effectively promoting the reverse migration of inflammatory cells. It is highly effective in treating various skin and mucous membrane injuries, rapidly stopping bleeding, promoting rapid wound tissue regeneration, and relieving pain. Furthermore, aloe vera extract also has a certain bactericidal effect, working in conjunction with free iodine in wound healing treatment to effectively inhibit the continuous proliferation of microorganisms. γ-Polyglutamic acid is a natural water-soluble polyglutamic acid synthesized by specific microorganisms. It has good moisturizing and water-locking abilities, biocompatibility and biodegradability, as well as the ability to inhibit hyaluronidase activity. When used in disinfectants, it can reduce skin moisture loss and improve skin elasticity. It can also promote skin wound healing by promoting cell migration and cell proliferation.

[0045] Skin-repairing ingredients such as glyceryl glucoside, aloe vera extract, and γ-polyglutamic acid work together with the aforementioned lubricants, thickeners, and pH adjusters to moisturize and nourish the epidermis, promote the healing of damaged cow teat epidermis, accelerate the repair of the skin's natural barrier, maintain the health of the mammary epidermis, and reduce the risk of mammary infection by pathogenic microorganisms.

[0046] Furthermore, the surfactant is an alkyl glycoside surfactant. The iodine-based cow nipple disinfectant of the present invention contains an alkyl glycoside (APG) surfactant. APG is composed of renewable natural fatty alcohols and glucose, and is widely used in daily personal care products for sensitive groups such as pregnant women and infants, including facial cleansers, shampoos, and laundry detergents.

[0047] However, this solution utilizes alkyl glycosides in iodine-based disinfectants. Alkyl glycosides themselves exhibit high stability, maintaining high solubility even in strong acids, strong alkalis, and high-concentration salt solutions. Furthermore, in the disinfectant system, alkyl glycosides can efficiently complex with iodine to form highly stable iodine complexes, allowing iodine to dissolve in water and release free iodine. This results in a disinfectant with high effective iodine content, high long-term stability, and high safety. The disinfectant also boasts advantages such as high bactericidal efficiency, long-lasting bactericidal effect, and suitability for long-term use, effectively preventing common bovine mastitis pathogens such as Staphylococcus aureus, Escherichia coli, and Streptococcus from infecting animal bodies. Simultaneously, APG is a novel, environmentally friendly, mild, and highly detergency green nonionic surfactant. Its high surface activity reduces the surface tension of the disinfectant and provides strong wetting ability, making it easier to remove dirt from the surface of the cow's teats. This gives the disinfectant both strong cleaning power and gentle, skin-friendly properties, minimizing skin irritation and providing protection.

[0048] Furthermore, the molecular weight of the alkyl glycoside surfactant is 320.42-348.47. Within this molecular weight range, the prepared disinfectant exhibits better effective iodine content characteristics.

[0049] Further, the thickener is at least one of glucomannan, guar gum, tara gum, and locust bean gum; the lubricant is at least one of glycerol, propylene glycol, PEG4000 (polyethylene glycol), and PEG6000 (polyethylene glycol); and the wetting agent is at least one of sodium docusate, AEROSOL OT-75 (sodium dioctyl sulfosuccinate), and BETTERSOL 607 (acetylenic glycol surfactant).

[0050] The pH adjuster includes an acidic adjuster and an alkaline adjuster. By weight, the amount of the acidic adjuster added is 0.1-0.5 parts, and the amount of the alkaline adjuster added is 0.05-0.5 parts.

[0051] The acidity regulator is at least one of anhydrous citric acid, malic acid, and mandelic acid, and the alkalinity regulator is at least one of sodium hydroxide, potassium hydroxide, and sodium carbonate.

[0052] This solution uses glucomannan as a thickener. Taking glucomannan as an example, it has strong hydration capacity, is easily soluble in water, and is one of the polysaccharides with the highest self-adhesion. This increases the viscosity of the disinfectant solution, allowing it to better adhere to the surface of the cow's udder and effectively kill pathogenic microorganisms and repair the udder epidermis. Glucomannan also has excellent film-forming properties; when used at low or normal temperatures, it forms a thin film that prevents contaminants from directly contacting the animal's udder epidermis. The compatibility of glucomannan with glycerol, sodium docusate, and other components in this invention improves the film's softness, making it more adaptable to the wrinkles on the surface of the cow's udder.

[0053] This solution uses glycerol and other raw materials as lubricants. Taking glycerol as an example, glycerol is a small molecule moisturizing ingredient and one of the important skin care ingredients. It has a strong hygroscopic property and can help the skin lock in moisture by absorbing moisture from the air. It can keep moisture in the stratum corneum, making the skin moist and soft, and effectively prevent nipple cracking. Combined with the functional ingredients in the invention, it can effectively promote the repair of the skin barrier.

[0054] This solution uses sodium docusate and other raw materials as wetting agents, which can reduce the surface tension of the disinfectant solution of this invention, allowing it to more completely soak the cow's udder during medicated baths.

[0055] The disinfectant of this invention is stable in nature. Through the combined action of acid and alkaline regulators, the pH value of the solution is stabilized at around 4.5-4.8, which is weakly acidic and close to the pH value of cow skin, thus reducing irritation.

[0056] Further, by weight, it includes the following raw materials: 1 part iodine, 0.5 parts sodium iodide, 0.1 parts sodium iodate, 6 parts skin repair functional ingredients, 5 parts surfactant, 0.5 parts thickener, 10 parts lubricant, 0.3 parts acidity regulator, 0.15 parts alkalinity regulator, and 77.45 parts water.

[0057] The present invention also proposes a method for preparing a bovine teat disinfectant that can promote the repair of damaged epidermis as described in any of the above claims, comprising the following preparation steps:

[0058] S1. Under constant temperature conditions in a water bath, the surfactant, acid regulator, wetting agent and 60%-70% of the total weight of water are mixed according to the formula and stirred until evenly dispersed to obtain mixture A;

[0059] S2. Disperse the thickener evenly in the lubricant to obtain mixture B;

[0060] S3. Under constant temperature conditions in a water bath, add mixture B to mixture A and stir to disperse evenly until the thickener is completely dissolved in the system to obtain mixture C;

[0061] S4. Remove the water bath and wait for mixture C to cool to ≤40℃. Then add sodium iodide and stir for about 1-2 hours to disperse it evenly to obtain mixture D.

[0062] S5. Under closed conditions, add 30%-40% of the total weight of water to mixture D, add iodine, stir vigorously for about 12-24 hours until the iodine is completely dissolved in the system, then add an alkaline regulator to adjust the pH, and then add sodium iodate and stir to disperse evenly to obtain mixture E;

[0063] S6. Add the skin-repairing active ingredients to mixture E while stirring, and continue stirring and dispersing for 1-2 hours to obtain the dairy cow teat disinfectant that can promote the repair of damaged epidermis.

[0064] Furthermore, the water bath temperature in steps S1 and S3 is 60-70℃, and the water bath time is 15-45 minutes.

[0065] The temperature range of 60℃-70℃ is selected in step S3 because this temperature accelerates the dissolution rate of the surfactant (APG2000), ensuring its complete dissolution in water. Simultaneously, the addition of an acidic regulator in step S1 lowers the pH of the system, and an acidic environment is more conducive to the subsequent dissolution of iodine. The mixture in step S3 also requires a water bath condition of 60℃-70℃ to ensure that the glucomannan dispersed in glycerol is completely dispersed in water at this temperature, providing uniform thickening. If the temperature is too low, the thickener (glucomannan) will agglomerate. In step S4, the system temperature is lowered to below 40℃ to prevent the iodine from sublimating due to excessively high temperatures during vigorous stirring of the APG2000, thus preventing a reduction in the effective iodine content in the disinfectant. In step S6, an alkaline regulator is added. Taking anhydrous citric acid and sodium hydroxide as an example, anhydrous citric acid and sodium hydroxide undergo the following neutralization reaction to produce sodium citrate: C6H8O7 + 3NaOH → Na3C6H5O7 + 3H2O. This adjusts the pH of the system to 4.5-4.8 while enhancing the buffering capacity of the system's pH. Sodium iodate is added after sodium hydroxide mainly because of the above reaction principle; if a large amount of H2O is released from citric acid... + Adding sodium iodate under certain conditions will reverse the chemical equilibrium, rapidly consuming iodate ions and preventing sodium iodate from effectively stabilizing elemental iodine in the system over a long period. Finally, in step S6, a skin-repairing active ingredient (glyceryl glucoside) is added to ensure that this active ingredient is not affected by high temperature and low pH, resulting in a dairy cow teat disinfectant with better performance.

[0066] Furthermore, the pH value of the dairy cow teat disinfectant that promotes the repair of damaged epidermis is 4.5-4.8.

[0067] The present invention also proposes an application of a cow teat disinfectant that can promote the repair of damaged epidermis as described in any of the above claims, wherein the cow teat disinfectant is used by spraying, rinsing, soaking, or applying, whether diluted or undiluted.

[0068] The specific usage instructions for this dairy cow teat disinfectant are as follows: Choose either the undiluted or diluted disinfectant solution based on the climate of the dairy farm's location. In cold, dry, and windy climates, the undiluted solution can be used directly. The high concentration of glucomannan, glycerol, and glyceryl glucoside in the disinfectant can prevent frostbite on the cow's teats. In hot and humid climates, dilute the disinfectant solution at a ratio of 1:3 (water to disinfectant). This will shorten the drying time, prevent teat contamination when the cow is lying down, and help the teat skin recover or maintain its health. This disinfectant solution can be used before and after milking. Before milking, apply the disinfectant solution to the cow's teats for a medicated bath for about 30 seconds, then wipe clean with a clean towel. After milking, apply the disinfectant solution to the teats and allow them to air dry naturally.

[0069] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are only used to explain the present invention and are not intended to limit the present invention.

[0070] Example 1

[0071] A method for preparing a bovine teat disinfectant that can promote the repair of damaged epidermis includes the following preparation steps:

[0072] S1. Disperse 4 parts of surfactant (APG2000), 0.3 parts of acid regulator (anhydrous citric acid), and 0.3 parts of wetting agent (sodium docusate) into deionized water (70% of 76.25 parts of deionized water) at 60°C by stirring and heating. Keep the mixture in a water bath for 40 minutes to obtain mixture A.

[0073] S2. Disperse 0.5 parts of thickener (glucomannan) evenly in 10 parts of lubricant (glycerol) to obtain mixture B;

[0074] S3. Under constant temperature conditions in a water bath, add mixture B to mixture A and stir to disperse evenly to obtain mixture C;

[0075] S4. After cooling mixture C to ≤40℃, add 0.5 parts of sodium iodide and stir to disperse evenly to obtain mixture D;

[0076] S5. Under sealed conditions, add the remaining water to mixture D, add 1 part of iodine and stir to disperse evenly, then add 0.15 parts of alkaline regulator (sodium hydroxide), and then add 0.1 parts of sodium iodate and stir to disperse for 1 hour to obtain mixture E;

[0077] S6. Add 8 parts of the skin repair efficacy ingredient (glyceryl glucoside) to mixture E and stir and disperse for 1.5 hours to obtain the dairy cow teat disinfectant that can promote the repair of damaged epidermis.

[0078] The bovine teat disinfectant prepared in Example 1, which promotes the repair of damaged epidermis, was subjected to the following performance tests:

[0079] Stability testing of disinfection products

[0080] According to the "Disinfection Technical Specifications 2002 Edition", different batches of disinfectant solutions prepared according to the preparation steps and parameters of Example 1 were placed in a 54℃ constant temperature incubator for 14 days. The content of the bactericidal active ingredient in the disinfectant was measured before and after placement. Three batches of samples were tested each time, and each batch was tested twice, and the average value was taken. The results of the accelerated test method were evaluated as follows: if the decrease rate of the active ingredient exceeded 10%, it was considered unacceptable; if the decrease rate of the bactericidal active ingredient after 14 days of storage at 54℃ was ≤10%, the shelf life could be set at 1 year.

[0081]

[0082]

[0083] All six batches of the disinfectant solution of this invention were placed in a comprehensive stability test chamber at 54℃ for 14 days. The test results show that the effective iodine content of the bactericidal active ingredient in the disinfectant solution of this invention decreased by 2.53%-4.77%, which meets the requirements stipulated in the "Disinfection Technical Specifications 2002 Edition". Therefore, the disinfectant solution of this invention is judged to have high stability and a shelf life of more than one year.

[0084] Quantitative sterilization test of suspension

[0085] The quantitative bactericidal test of the suspension was conducted in accordance with the "Evaluation Method of Antibacterial and Bacteriostatic Effects" (WS / T 650—2019).

[0086] First, prepare a disinfectant solution by diluting the disinfectant solution obtained in Example 1 with sterile deionized water at a ratio of 1:3, and then place it in a water bath at 20℃±1℃ for later use. Under aseptic conditions, open the lyophilized bacterial culture tube, draw an appropriate amount of nutrient broth, and add it, gently blowing and aspirating several times. Add a small amount of bacterial suspension to a test tube containing 5mL-10mL of nutrient broth and incubate at 37℃ for 4 hours. Take the bacterial suspension from the first generation culture, inoculate it onto an agar plate, and incubate at 37℃ for 18-24 hours. Pick typical colonies from the second generation culture and inoculate them again at 37℃ for 18-24 hours. Repeat the third generation culture. Take the third generation culture, transfer it to another sterile test tube using a pipette, and vortex it for 20 seconds to ensure uniform suspension of bacteria. Dilute the bacterial suspension with PBS buffer to the required concentration and store at 20℃ for later use. The culture medium for *Escherichia coli* ATCC25922 and *Staphylococcus aureus* ATCC29213 was LB medium; the culture medium for *Streptococcus agalactiae* BNCC336970 was soybean casein medium containing 5% virgin bovine serum. A bacterial suspension was prepared for the experiments at a concentration of 1 × 10⁻⁶.8 cfu / ml ~5×10 8 CFU / ml. Take a sterile test tube, add 5.0 mL of the prepared antibacterial agent, place in a 20℃±1℃ water bath for 5 min, then add 0.1 mL of the test bacterial suspension, mix quickly and start timing immediately. After the test bacteria and antibacterial agent have interacted for the predetermined time (15 s, 30 s, 45 s respectively), take 0.5 mL of the test bacteria and antibacterial agent mixture and add it to 4.5 mL of neutralizing agent, mix well. After each tube of test bacteria and antibacterial agent mixture has been reacted with the neutralizing agent (1% sodium thiosulfate solution) for 10 min, take 1.0 mL of the sample solution and determine the number of viable bacteria according to the viable bacteria culture counting method. Inoculate two petri dishes with each sample solution. If the number of colonies growing on the plate is large, it can be serially diluted 10 times with PBS before viable bacteria culture counting. At the same time, use PBS instead of disinfectant to perform parallel tests as a positive control. The number of recovered colonies in the positive control is 1.0 × 10⁻⁶. 4 CFU / mL~9.0×10 4 CFU / mL. Diluent, neutralizing agent, and culture medium from the same batch were used as negative controls. All test and control samples were incubated at 36℃±1℃, and the final results were observed after 48 hours of bacterial vegetative culture. The experiment was repeated three times, and the sterilization rate was calculated. Specific data are shown in the table below:

[0087]

[0088] As shown in the table above, the cow teat disinfectant provided in Example 1 of this invention has a rapid and efficient bactericidal effect against common mastitis pathogens such as Escherichia coli, Staphylococcus aureus, Streptococcus agalactiae, and Klebsiella pneumoniae, and can kill 99.999% of pathogens within 15 seconds.

[0089] Experiment on factors affecting disinfection effectiveness

[0090] Determination of the effect of organic matter on the bactericidal effect: According to the "Disinfection Technical Specifications 2002 Edition", calf serum was used as the representative of organic matter, and three groups were set up: a control group without calf serum; a group containing 25% calf serum; and a group containing 50% calf serum. The concentration of the disinfectant used in each group was the concentration of the disinfectant of this invention diluted with deionized water at a ratio of 1:3, and the action time was 15s, 30s, and 45s. The microbial suspension prepared with the diluted solution was mixed with sterile calf serum at ratios of 1:1, 3:1, and 19:1 to prepare microbial suspensions containing 50%, 25%, and 5% calf serum, respectively. This microbial suspension containing calf serum was used for quantitative bactericidal tests. As needed, quantitative bactericidal tests were selected for determination. Escherichia coli ATCC25922 was selected as the representative of Gram-negative bacteria, Staphylococcus aureus ATCC29213 was selected as the representative of Gram-positive bacteria, and Streptococcus agalactiae BNCC336970 was selected as the representative of streptococci. Each group of tests should be repeated 3 times, and the sterilization rate should be calculated according to the corresponding microbial killing test.

[0091]

[0092] Determination of the effect of temperature on the effectiveness of microbial killing: According to the "Disinfection Technical Specifications 2002 Edition", temperatures were set at 5℃±1℃, 15℃±1℃, 25℃±1℃, 35℃±1℃, and 45℃±1℃, with 10℃ intervals. The concentration of the disinfectant used in each group was the same as that of the present invention diluted with deionized water at a ratio of 1:3, and the contact time was 15s, 30s, and 45s. For tests requiring a temperature higher than room temperature, a constant temperature water bath (electric heating) was used for adjustment; for tests requiring a temperature lower than room temperature, a cold water bath (with an appropriate amount of ice water) was used for adjustment. After the temperature adjustment device reached the required temperature, a test tube containing the test sample solution was placed in the test tube, along with a test tube containing an equal volume of distilled water and an inserted thermometer. When the thermometer in the test tube indicated the required temperature, the subsequent test began. As needed, select the appropriate suspension quantitative bactericidal test. For the microorganisms, *Escherichia coli* ATCC25922 is selected as the representative Gram-negative bacterium, *Staphylococcus aureus* ATCC29213 as the representative Gram-positive bacterium, and *Streptococcus agalactiae* BNCC336970 as the representative streptococcus. Each test should be repeated three times. Calculate the bactericidal rate according to the corresponding microbial bactericidal test, as shown in the table below:

[0093]

[0094] Determination of the effect of pH value on the bactericidal effect: The pH of the disinfectant obtained in Example 1 of this invention was measured to be 4.5. According to the "Disinfection Technical Specifications 2002 Edition," to investigate the effect of acidity and alkalinity on the bactericidal effect of the disinfectant, the disinfectant of this invention was adjusted with hydrochloric acid and sodium hydroxide respectively to prepare disinfectants with pH values ​​of 2.5, 3.5, 4.5, 5.5, and 6.5. Quantitative suspension killing tests were conducted using disinfectants with different pH values. The tests were repeated three times, and the sterilization rate was calculated.

[0095]

[0096]

[0097] Because dairy cows live in complex and diverse environments, such as barns, milking parlors, and pastures, they are subject to interference from organic matter such as mud, sewage, and manure, as well as fluctuations in temperature and pH. Therefore, experiments were conducted on the disinfectant obtained in this invention to test the factors affecting its disinfection effectiveness, simulating various environments in practical applications.

[0098] The above experimental results show that when the organic matter content is added at 5-50%, the disinfectant solution of this invention can kill 99.999% of pathogenic microorganisms within 15 seconds, and the disinfection and sterilization effect is not affected. When the temperature is between 5 and 45°C, the disinfectant solution prepared by this invention can kill 99.999% of pathogenic microorganisms within 15 seconds, and the disinfection and sterilization effect is not affected. When the pH value varies between 2.5 and 6.5, the disinfectant solution prepared by this invention can kill 99.999% of pathogenic microorganisms within 15 seconds, and the disinfection and sterilization effect is not affected in any way, making it applicable to a very wide range of situations.

[0099] Safety test

[0100] According to the "Disinfection Technical Specifications 2002 Edition," skin disinfectants, in addition to undergoing toxicological testing as required for Class I, II, or III disinfectants, must also undergo a complete skin irritation test. For disinfectants used occasionally or with intervals of several days, a single complete skin irritation test is required; for disinfectants used daily or for several consecutive days, multiple complete skin irritation tests are required. For disinfectants that come into contact with skin wounds, an additional broken skin irritation test is required; for disinfectants that come into contact with wounds, an eye irritation test is required. Other disinfectants that must come into contact with the skin during use should also undergo a complete skin irritation test. If, based on the disinfectant's composition, it is estimated that it may cause sensitization, a skin allergy test is also required. See the table below for specific test data:

[0101] Skin Irritation Response Scoring Standard

[0102]

[0103] Skin irritation intensity grading

[0104] Stimulation intensity level Non-irritating Mild irritation moderately irritating Strong irritant Skin irritation index 0～0.5 0.5～2.0 2.0～6.0 6.0～8.0

[0105] Multiple complete skin irritation tests: 24 hours before the test, three New Zealand rabbits weighing 2.0-2.5 kg were selected. Hair was removed from both sides of the spine on the back of the rabbits using a hair removal agent, taking care not to damage the skin. The hair removal area was approximately 3cm × 3cm on each side. The next day, 0.5mL of the undiluted disinfectant solution was applied to 2-4 layers of gauze covering an area of ​​2.5cm × 2.5cm and placed on one side of the hair-removed skin. Then, a layer of non-irritating plastic film was used to cover it, and finally, non-irritating adhesive tape was used to secure it. The other side of the hair-removed skin served as a blank control (using physiological saline). The application time was 4 hours. After the test, the residual test substance was removed with warm water or a non-irritating solvent. Local skin reactions were observed at 1 hour, 24 hours, and 48 hours after removal of the test substance, and the irritation response was scored according to the table. The average score (irritation index) per animal per day was calculated using the following formula, and the skin irritation intensity was determined using Table 2-12. Average score per animal per day = ∑(total score of erythema and edema per animal over 14 days) ÷ (number of test animals × 14).

[0106] Results of multiple complete skin irritation tests

[0107]

[0108]

[0109]

[0110] The test results above show that when the disinfectant of this invention is used on the intact skin of rabbits for 14 consecutive days, the average score per animal per day is 0, that is, the skin irritation index is equal to 0. The range of 0 to 0.5 indicates that the skin irritation intensity of the disinfectant of this invention is non-irritating and can be safely applied to the disinfection of the udder skin of dairy cows.

[0111] A single skin irritation test: Before applying the disinfectant prepared in Example 1 of this invention, clean and disinfect the exposed skin (2.5cm × 2.5cm) with 75% alcohol. After the alcohol evaporates, make a "well"-shaped incision in the skin area using a sterile scalpel or injection needle, and then apply the disinfectant to the incision area. Note that the skin incision should only reach the epidermis, avoiding damage to the dermis. The scoring method for preoperative skin preparation, postoperative application of the test substance, and observation of local skin reactions is the same as in the table. Pay attention to differentiating between infection and primary irritation reactions. If infection is suspected, the test should be repeated.

[0112] Rabbit skin damage rating chart

[0113]

[0114] In practical applications, dairy cows' udder skin may suffer various injuries, such as cuts from gravel during grazing, abrasions from rubbing against the ground while lying down, or burns caused by quicklime on the cowshed floor. Therefore, a skin irritation test was conducted on the disinfectant of this invention to examine its irritant properties. The test results in the table above show that when the disinfectant of this invention was applied to a "well"-shaped wound on the skin of rabbits, no skin irritation reactions such as edema or erythema occurred at 4 hours, 1 day, 2 days, and 3 days later, with a reaction score of 0. This proves that the disinfectant of this invention is safe and non-irritating when applied to broken skin wounds.

[0115] Acute eye irritation test

[0116] To test the acute irritation and corrosive effects of disinfectants on the eyes of experimental animals, three New Zealand rabbits were used. Before the experiment, both eyes of the rabbits were examined; those with abnormalities were not used. 0.1 mL of the test substance (the disinfectant prepared in Example 1 of this invention) was instilled into the conjunctival sac of one eye of the rabbit, with physiological saline used as a normal control in the other eye. After instilling the test substance, the eye was passively closed for 4 seconds, and then rinsed with physiological saline after 30 seconds. The damage and recovery of the conjunctiva, iris, and cornea of ​​the rabbits were observed visually at 1 hour, 24 hours, 48 ​​hours, 72 hours, 7 days, 14 days, and 21 days after instillation. If no irritation reaction occurred within 72 hours, or if the eye irritation reaction completely recovered on day 7 or 14, the experiment could be terminated early. If necessary, changes in the cornea and iris were examined using 2% sodium fluorescein solution, a slit lamp, or a magnifying glass. The acute irritation response of the rabbit's cornea, iris, and conjunctiva was scored according to the table below. The "average score" for each animal at three different observation times (24h, 48h, and 72h) was calculated for corneal damage, iris damage, conjunctival hyperemia, and conjunctival edema (i.e., the sum of the scores for each animal at 24h, 48h, and 72h divided by the number of observations, 3). The average scores and recovery times for corneal, iris, and conjunctival hyperemia and edema were used to determine the intensity of the test substance's irritation to the eye according to the ocular irritation response grading criteria in the table below.

[0117] See the table at the bottom:

[0118]

[0119]

[0120] Grading Standards for Eye Irritation Reactions

[0121]

[0122] Note: Recovery time is the time it takes for the animal's stimulus response score to recover to 0 for corneal damage, 0 for iris damage, 0 or 1 for conjunctival hyperemia, and 0 or 1 for conjunctival edema.

[0123] Rabbit Acute Eye Irritation Response Scoring Sheet

[0124]

[0125] In poorly managed dairy farms, cows may have poor udder health, and their skin may be damaged by external factors, leading to tissue loss and wounds. Since the disinfectant of this invention may come into contact with wounds, according to the "Disinfection Technical Specifications 2002 Edition," an eye irritation test should be performed before any disinfectant comes into contact with wounds. The test data in the table above shows that in the rabbit eye irritation test, the average scores for corneal damage, iris damage, and conjunctival edema were all less than 1, and conjunctival congestion was less than 2, indicating no irritation. In conclusion, the disinfectant of this invention is non-irritating and safe for use on skin wounds.

[0126] Acute oral toxicity test: This test examines the acute toxicity and intensity of the disinfectant on experimental animals. Twenty 3-week-old Kunming mice (half male and half female) were selected and acclimatized for 3 days before the formal test. A single maximum limit test was used, where 20 animals (half male and half female) were administered a single gavage dose of 5000 mg / kg body weight. If no deaths occurred within 14 days, the LD50 was determined to be greater than 5000 mg / kg body weight.

[0127] Toxicity evaluation of disinfectants: LD50 50 LD50+ is practically non-toxic; 50 Those with a concentration of 501 mg / kg to 5000 mg / kg body weight are considered to have low toxicity; LD50 50 A dose of 51 mg / kg to 500 mg / kg body weight is considered moderately toxic; LD50 50 A dose of 1 mg / kg to 50 mg / kg body weight is considered highly toxic; LD50 50 Less than 1 mg / kg of body weight is considered highly toxic.

[0128] Note: To evaluate the safety of disinfectants to humans in actual application, when the original LD50 of the product is ≤5000mg / kg body weight, an acute oral toxicity test of a solution with a concentration 5 times the highest application concentration of the disinfectant must be performed, and its LD50 must be calculated.

[0129] Mouse weight record in acute oral toxicity test

[0130]

[0131] Acute oral toxicity test mortality rate

[0132] gender Dosage (mg / kg bw) Number of animals (individuals) Number of dead animals (individuals) mortality rate(%) female 5000 10 0 0 male 5000 10 0 0

[0133] Since the disinfectant of this invention is used year-round on dairy cow udders, and the collected milk is used to make dairy products for human consumption, an acute oral toxicity test was conducted on the disinfectant according to the "Disinfection Technical Specifications 2002 Edition" to examine its safety to humans in practical application and to avoid any harmful residues. The mouse weight records for the acute oral toxicity test are shown in the table above. The mice's weight steadily increased from day 1 to day 14 without any obvious abnormalities. The mortality rate for the acute oral toxicity test is shown in the table above. Using a single maximum-limit test protocol, the results showed that after 14 days of administration, the number of dead animals was 0, and the mortality rate was 0%. Using mice lethal by cervical dislocation, gross observation revealed no ocular lesions. Therefore, based on the results, the LD50 of the disinfectant of this invention is determined to be greater than 5000 mg / kg body weight, indicating practical non-toxicity and safety for dairy cows and humans.

[0134] On-site dairy cow teat disinfection test

[0135] Twelve Holstein lactating cows without clinical symptoms of mastitis were selected as experimental animals. Two mammary folds on the left side of each cow were designated as the experimental group, and two mammary folds on the right side as the control group. The experimental group was immersed in the cow teat disinfectant solution provided in Example 1, while the control group was immersed in commercially available povidone-iodine disinfectant solution for 15 seconds. Before and at 1 min, 5 min, 60 min, and 120 min after treatment, sterile cotton swabs were used to apply disinfectant to a 2×2 cm area on the outer side of the cow teats. 2 Wipe the area five times, cut off the swab tip and put it into a sterile EP tube containing 3mL of neutralizing agent, vortex to mix, take 20μL and spread it on nutrient agar medium, incubate at 37℃ for 12h, count the colonies, and calculate the sterilization rate and sterilization index.

[0136]

[0137]

[0138] The test data in the table above show that after using the disinfectant provided in Example 1 and the commercially available cow teat disinfectant, the sterilization rate of the teat skin surface in both the experimental and control groups was above 99% after 1 minute, with a sterilization index above 3.0. According to the "Disinfection Technical Specifications" (2002), a sterilization index ≥1 against natural bacteria is considered qualified. Both disinfectants exceeded the qualified standard and effectively killed bacteria on the surface of cow teats. After the cow teat bath, the number of colonies on the surface of the teats in both groups gradually increased over time. At 60 minutes and 120 minutes after disinfection, the sterilization effect of the control group decayed more rapidly, with a sterilization efficiency of only 26.47% at 120 minutes. The experimental group showed a significantly better sustained sterilization effect than the control group, with sterilization rates of over 74.55% and 50.79% at 60 minutes and 120 minutes, respectively, thus prolonging the protection time of the cow teats to a certain extent. After the experimental group was treated with disinfectant, the cows' udders were observed and touched. The udders appeared rosy, the skin was smooth and soft, the sphincter at the end of the teat was smooth and without a protruding ring, and no adverse reactions such as redness, swelling or cracking were observed.

[0139] Comparative Example 1

[0140] Commercially available povidone-iodine disinfectant.

[0141] Example 2

[0142] In this embodiment, all preparation steps and parameters are the same as in Example 1, except that glycerol glucoside is replaced with Aloe vera extract.

[0143] Example 3

[0144] In this embodiment, all preparation steps and parameters are the same as in Example 1, except that glycerol glucoside is replaced with γ-polyglutamic acid.

[0145] Example 4

[0146] A method for preparing a bovine teat disinfectant that can promote the repair of damaged epidermis includes the following preparation steps:

[0147] S1. Stir and disperse 3 parts of surfactant (APG2000), 0.2 parts of acid regulator (anhydrous citric acid), and 0.2 parts of wetting agent (sodium docusate) into deionized water (60% of 81.3 parts of deionized water) at 55°C, and keep the water bath at a constant temperature for 45 min to obtain mixture A;

[0148] S2. Disperse 0.2 parts of thickener (glucomannan) evenly in 8 parts of lubricant (glycerol) to obtain mixture B;

[0149] S3. Under constant temperature conditions in a water bath, add mixture B to mixture A and stir to disperse evenly to obtain mixture C;

[0150] S4. After cooling mixture C to ≤40℃, add 0.5 parts of sodium iodide and stir to disperse evenly to obtain mixture D;

[0151] S5. Under sealed conditions, add the remaining water to mixture D, add 1 part of iodine and stir to disperse evenly, then add 0.5 parts of alkaline regulator (sodium hydroxide), and then add 0.1 parts of sodium iodate and stir to disperse for 2 hours to obtain mixture E;

[0152] S6. Add 5 parts of the skin repair efficacy ingredient (glyceryl glucoside) to mixture E and stir and disperse for 1 hour to obtain the dairy cow teat disinfectant solution that can promote the repair of damaged epidermis.

[0153] Example 5

[0154] A method for preparing a bovine teat disinfectant that can promote the repair of damaged epidermis includes the following preparation steps:

[0155] S1. Mix and disperse 5 parts of surfactant (APG2000), 0.2 parts of acid regulator (anhydrous citric acid), and 0.2 parts of wetting agent (sodium docusate) in deionized water (70% of 81.85 parts of deionized water) at 62°C, and keep the water bath at a constant temperature for 15 minutes to obtain mixture A;

[0156] S2. Disperse 0.2 parts of thickener (glucomannan) evenly in 8 parts of lubricant (glycerol) to obtain mixture B;

[0157] S3. Under constant temperature conditions in a water bath, add mixture B to mixture A and stir to disperse evenly to obtain mixture C;

[0158] S4. After cooling mixture C to ≤40℃, add 0.1 parts of sodium iodide and stir to disperse evenly to obtain mixture D;

[0159] S5. Under sealed conditions, add the remaining water to mixture D, add 1 part of iodine and stir to disperse evenly, then add 0.5 parts of alkaline regulator (sodium hydroxide), and then add 0.5 parts of sodium iodate and stir to disperse for 0.5 hours to obtain mixture E;

[0160] S6. Add 5 parts of the skin repair efficacy ingredient (glyceryl glucoside) to mixture E and stir and disperse for 1 hour to obtain the dairy cow teat disinfectant solution that can promote the repair of damaged epidermis.

[0161] The bovine teat disinfectant solutions prepared in Examples 1, 4, and 5, which can promote the repair of damaged epidermis, were subjected to the following performance tests:

[0162] Test for determination of effective iodine content

[0163] The effective iodine content of the disinfectants prepared in Examples 1, 4, and 5 was determined according to Appendix A of GB / T26368-2020 "Hygienic Requirements for Iodine-Containing Disinfectants" issued by the State Administration for Market Regulation and the Standardization Administration of China. 10.00 g of the iodine-containing disinfectant was accurately weighed and placed in a 100 mL iodine flask. 50 mL of deionized water and 5 drops of 36% acetic acid solution were added. Titration was performed with 0.1 mol / L sodium thiosulfate titrant, shaking constantly. When the solution turned pale yellow, 10 drops of 5 g / L starch solution were added, and the solution immediately turned blue. Titration continued until the blue color disappeared. The total amount of sodium thiosulfate titrant used was recorded, and the titration result was corrected using a blank test. The sample was measured twice, and the average of the two results was used for calculation.

[0164] Since 1 mL of 1 mol / L sodium thiosulfate titrant is equivalent to 0.1269 g of available iodine, the available iodine content is calculated using the following formula:

[0165]

[0166] In the formula: X is the effective iodine content, g / L; c is the concentration of sodium thiosulfate titrant, mol / L; Vst is the volume of sodium thiosulfate titrant used for titration, mL; m is the weight of the disinfectant stock in the iodine flask, g; V is the volume of the liquid disinfectant stock in the iodine flask, mL.

[0167]

[0168] As shown in the table above, the effective iodine content of the disinfectant provided in Example 1 immediately after preparation is higher than that of the disinfectants provided in Examples 4 and 5, exhibiting the highest effective iodine content and best performance. This is mainly due to the difference in the amount of surfactant (APG2000) used in the three examples.

[0169] The bovine teat disinfectant solutions prepared in Examples 1-3 that promote the repair of damaged epidermis and the disinfectant solution of Comparative Example 1 were subjected to the following performance tests:

[0170] Tests to promote healing of skin damaged by bacterial infection

[0171] A mouse skin abrasion model was established using 4-week-old SPF-grade female ICR mice. Mice were divided into 5 groups of 8 mice each: ① model group, ② commercially available povidone-iodine disinfectant treatment group, ③ disinfectant treatment group provided in Example 1, ④ disinfectant treatment group provided in Example 2, and ⑤ disinfectant treatment group provided in Example 3. Mice in each group were anesthetized with propofol (150 mg / kg), and hair was removed from the midline of their backs. For mice in groups ①, ②, ③, ④, and ⑤, their backs were wiped with alcohol swabs, and then the exposed skin was gently rubbed with 1cm*1cm sterile fine sandpaper until the skin became shiny but did not bleed, with a wound area of ​​approximately 60 mm². 2 Staphylococcus aureus ATCC 29213 was cultured overnight at a concentration of 2.5 x 10⁻⁶. 9 CFU was administered to the wound via 5 µL PBS for bacterial infection. Twenty-four hours after bacterial colonization, the model group (Group ①) received no treatment, while the other treatment groups received treatment with the corresponding disinfectant applied morning and evening. Simultaneously, the injured skin area was measured and photographed every morning to record the wound area, and the skin healing rate was calculated using Image Pro Plus 6.0 software.

[0172] Eight days after Staphylococcus aureus ATCC29213 infection of mouse epidermis, mice were euthanized by cervical dislocation. The injured skin was cut off, aseptically minced and ground, and 3 mL of sterile PBS was added. The mixture was vortexed and mixed. 20 μL of the grinding solution was taken and serially diluted for plate colony counting to calculate the bacterial load on the mouse bacterial infection wound.

[0173] After establishing a mouse skin abrasion model, the injured skin was cut off on day 8, fixed with 4% polymethyl methacrylate and embedded in sections. The sections were then incubated with Ly6G, F4 / 80, iNOS and p-p38 antibodies to observe the inflammation of the bacterial infection wounds on the mouse skin.

[0174] Bacterial wound healing rate in mice

[0175]

[0176]

[0177] Bacterial load on bacterial wounds in mice on day 8

[0178]

[0179] By establishing a mouse skin abrasion model, the ability of various medicated bath solutions to promote wound healing was evaluated. For example... Figure 1As shown in the table above, the bacterial wounds treated with the disinfectant solutions prepared in Examples 1, 2, and 3, and the commercially available povidone-iodine disinfectant solution, healed significantly faster than the untreated model group. Specifically, the disinfectant solution treatment groups in Examples 1 and 2 showed the fastest healing rate, with over 93.2% healing by day 8. The disinfectant solution treatment group in Example 3 showed over 75.6% healing by day 8, while the model group only showed 37.5% healing. The healing rate of Staphylococcus aureus infection wounds in mice treated with the disinfectant solutions in Examples 1 and 2 on day 8 was significantly different from that in the commercially available povidone-iodine disinfectant solution treatment group (p<0.01), while there was no significant difference between the disinfectant solution treatment groups in Examples 1 and 2 (p>0.05). Furthermore, as shown in the table above... Figure 2 As shown, the bacterial load on the bacterial infection wounds of mice in each treatment group was significantly reduced on day 8 compared with the model group. The treatment groups of Example 1, Example 3 and commercial povidone-iodine disinfectant were significantly different from the model group (p<0.05). Among them, Example 2 significantly reduced the bacterial load on the bacterial infection wounds compared with the model group (p<0.01), which was more conducive to the repair of bacterial infection damage.

[0180] Immunohistochemical experiments were performed on the skin of the wound eight days after infection with Staphylococcus aureus ATCC29213 to analyze the average optical density of different antibodies and understand the inflammation status of the wound skin.

[0181] Many diseases trigger inflammation in the body. Inducible nitric oxide synthase (iNOS), induced by injury, is a key mediator of immune activation and inflammation, and a hallmark molecule of M1 macrophages, promoting the massive release of inflammatory factors and chemokines. When the body is severely infected, macrophages initially exhibit the M1 phenotype, releasing various inflammatory factors to combat the stimulus. However, since the acute inflammatory phase has largely passed by day 8 after bacterial infection of the wound, continued release of large amounts of inflammatory factors can cause tissue damage and hinder tissue repair and regeneration. Figure 3 The results showed that, compared with the commercially available povidone-iodine disinfectant group, the expression of iNOS in the Example 1 group, Example 2 group, and Example 3 group was significantly reduced (p<0.01), which helped M2 macrophages release cytokines and promote tissue de-inflammation and tissue healing.

[0182] F4 / 80 macrophages are not only widely distributed in the stroma of organs and connective tissues, but also closely associated with endothelial cells and epithelial cells. Macrophages are the most abundant type of immune cells in the skin, composed of a heterogeneous and plastic group of cells, and are crucial for skin homeostasis and host defense. However, dysregulated macrophages can lead to negative effects such as poor infection control, impaired wound healing, and tissue fibrosis. F4 / 80 is mainly expressed on the surface of macrophages and is used as a marker for mature mouse macrophages. Figure 4As shown, compared with the commercially available povidone-iodine disinfectant group, the infiltration of mature macrophages in the Example 1, Example 2 and Example 3 groups was significantly reduced (p<0.01), which made the skin cell condition tend to be stable and more conducive to the healing of skin wounds.

[0183] Neutrophils are the most abundant type of white blood cell in the body's circulation and have been shown to mediate alternative pathways in systemic allergic reactions and participate in allergic skin reactions. Following trauma, neutrophil activation can trigger local inflammation. Acute inflammation can protect the body against pathogen invasion, but if inflammation persists, it can affect the subsequent repair process. Ly6G is a specific marker of neutrophils, and its immunohistochemical staining is used to observe neutrophil infiltration in wounds. Figure 5 As shown, compared with the commercially available povidone-iodine disinfectant group, the neutrophil infiltration in the Example 2 group was significantly improved (p<0.01), which reduced the cumulative infiltration of neutrophils and the release of their cytotoxic products. This can promote the resolution of inflammation to a certain extent, prevent the occurrence of chronic inflammation, and facilitate the healing of damaged skin.

[0184] p38 kinase is a proline-directed serine / threonine kinase in the mitogen-activated protein kinase (MAPK) family, typically activated by environmental stress and inflammatory signals. p38 kinase is activated via double phosphorylation of MAP2K, and the p38α pathway plays a crucial role in innate immune responses and defense against bacterial and viral pathogens. Studies have shown that strong and sustained p38 activation is generally associated with detrimental effects and cell death, while cell survival and homeostasis often depend on milder or transient levels of p38α activation. Figure 6 As shown, on day 8 after bacterial infection of the wound, the skin was nearing the end of the healing process, making a lower p38α activation level more suitable. The p38 activation level in the Example 1 group was significantly lower (p<0.01) than that in the commercially available povidone-iodine disinfectant group, which was more conducive to the restoration of normal homeostasis of the animal's epidermis.

[0185] In summary, the disinfectants in Examples 1, 2, and 3 can all reduce the colonization of pathogens on skin wounds to a certain extent, accelerate the resolution of epidermal inflammation, and promote the healing of skin wounds. Moreover, their effects are significantly better than those of commercially available dairy cow teat disinfectants. They can restore the natural barrier function of animal epidermis in a short time, fundamentally improve the animal's resistance to environmental pathogens, and are suitable for actual application scenarios in farms, thus having significant clinical application value.

[0186] Comparative Example 2

[0187] In this comparative example, all preparation steps and parameters are the same as in Example 1, except that different surfactants OP-10, AEO-9, and AES are used.

[0188] The sterilization stability of the dairy cow teat disinfectant prepared in Example 1 and Comparative Example 2, which can promote the repair of damaged epidermis, was determined. The data are shown in the table below:

[0189]

[0190] The test results above show that, compared with the three disinfectants prepared in Comparative Example 2, the disinfectant prepared in Example 1 has the highest stability. The effective iodine content loss rate after 14 days is 2.70%. That is, after replacing the alkyl glycoside surfactant with other types of surfactants, the stability of the prepared disinfectant decreases significantly, and the effective iodine content damage rate after 14 days is 7.52% to 10.79%.

[0191] Example 6

[0192] In this embodiment, all preparation steps and parameters are the same as in Example 1, except that the APG type and molecular weight used are different. The effective iodine content of the bovine teat disinfectant solutions prepared in Examples 1 and 6 that promote the repair of damaged epidermis was determined (tested immediately after preparation). The data are shown in the table below:

[0193] model APG molecular weight Available iodine content (g / L) Example 1 APG2000 320.42 9.96 Example 6-1 APG0810 320.42 9.87 Example 6-2 APG1214 238.41 9.25 Example 6-3 APG0814 348.47 9.63

[0194] The test results above show that the surfactant APG used in this method has the best effect when its molecular weight is in the range of 320.42 to 348.47, and the disinfectant prepared using APG2000 in Example 1 has the highest effective iodine content.

[0195] Example 7

[0196] A method for preparing a bovine teat disinfectant that can promote the repair of damaged epidermis includes the following preparation steps:

[0197] S1. Disperse 5 parts of surfactant (APG2000) and 0.3 parts of acid regulator (anhydrous citric acid) into deionized water (70% of 77.45 parts of deionized water) at 65°C by stirring and heating in a water bath for 15 minutes to obtain mixture A;

[0198] S2. Disperse 0.5 parts of thickener (glucomannan) evenly in 10 parts of lubricant (glycerol) to obtain mixture B;

[0199] S3. Under constant temperature conditions in a water bath, add mixture B to mixture A and stir to disperse evenly to obtain mixture C;

[0200] S4. After cooling mixture C to ≤40℃, add 0.5 parts of sodium iodide and stir to disperse evenly to obtain mixture D;

[0201] S5. Under sealed conditions, add the remaining water to mixture D, add 1 part of iodine and stir to disperse evenly, then add 0.15 parts of alkaline regulator (sodium hydroxide), and then add 0.1 parts of sodium iodate and stir to disperse for 1 hour to obtain mixture E;

[0202] S6. Add 6 parts of the skin repair efficacy ingredient (Aloe vera extract) to mixture E and stir and disperse for 1 hour to obtain the cow teat disinfectant that can promote the repair of damaged epidermis.

[0203] The sterilization product stability test was conducted on the bovine teat disinfectant prepared in Example 7, which can promote the repair of damaged epidermis. The data are shown in the table below:

[0204] Tests to promote healing of skin damaged by bacterial infection

[0205]

[0206] The test results above show that, through optimization of raw materials, dosage, and various parameters, the performance of the disinfectant in promoting the healing of damaged skin has been further improved.

[0207] The above description is merely a preferred embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structural transformations made using the contents of the present invention under the inventive concept of the present invention, or direct / indirect applications in other related technical fields, are included within the patent protection scope of the present invention.

Claims

1. A disinfectant for cow teats that promotes the repair of damaged epidermis, characterized in that, consists of iodine 1 part, sodium iodide 0.5 part, sodium iodate 0.1 part, skin repair efficacy component 6 parts, surfactant 5 parts, thickening agent 0.5 part, wetting agent 0.2 part, lubricant 10 parts, acid regulator 0.3 part, basic regulator 0.15 part and water 77.45 parts by weight. The skin repair efficacy component is any one of glycerol glucoside, aloe vera extract or gamma-polyglutamic acid; The surfactant is APG2000 with a molecular weight of 320.42; the thickening agent is glucomannan; and the lubricant is glycerol; The wetting agent is at least one of docusate sodium, AEROSOL OT-75 and BETTERSOL 607; The pH regulator comprises an acid regulator and a basic regulator, the acid regulator is added in an amount of 0.1-0.5 parts by weight, and the basic regulator is added in an amount of 0.05-0.5 parts by weight; The acid regulator is at least one of anhydrous citric acid, malic acid and mandelic acid, and the basic regulator is at least one of sodium hydroxide, potassium hydroxide and sodium carbonate.

2. The disinfectant solution for cow teat which can promote the repair of damaged epidermis according to claim 1, characterized in that, consists of iodine 1 part, sodium iodide 0.5 part, sodium iodate 0.1 part, skin repair efficacy component 6 parts, surfactant 5 parts, thickening agent 0.5 part, wetting agent 0.2 part, lubricant 10 parts, acid regulator 0.3 part, basic regulator 0.15 part and water 77.45 parts by weight.

3. A method of preparing a disinfectant solution for promoting the repair of damaged skin of a cow's teat according to any one of claims 1-2, characterized in that, The preparation steps include: S1. Under the condition of water bath constant temperature, the surfactant, the acid regulator, the wetting agent and water with a total weight of 60%-70% are mixed according to the ratio and stirred to disperse uniformly to obtain a mixture A; S2. The thickening agent is dispersed uniformly in the lubricant to obtain a mixture B; S3. Under the condition of water bath constant temperature, the mixture B is added to the mixture A and stirred to disperse uniformly to obtain a mixture C; S4. After the mixture C is cooled to ≤40℃, sodium iodide is added thereto and stirred to disperse uniformly to obtain a mixture D; S5. Under the condition of airtightness, water with a total weight of 30%-40% is added to the mixture D, iodine is added and stirred to disperse uniformly, then the basic regulator is added, and then sodium iodate is added and stirred to disperse uniformly to obtain a mixture E; S6. The skin repair efficacy component is added to the mixture E and stirred to disperse uniformly to obtain the dairy cow teat disinfectant capable of promoting repair of damaged epidermis.

4. The method for preparing the bovine teat disinfectant that promotes the repair of damaged epidermis according to claim 3, characterized in that, The water bath temperature in steps S1 and S3 is 60-70℃, and the water bath time is 15-45 min.

5. The method for preparing a disinfectant solution for cow teat which can promote the repair of damaged epidermis according to claim 3, characterized in that, The pH value of the dairy cow teat disinfectant capable of promoting repair of damaged epidermis is 4.5-4.8.

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