Preparation method of special post-medicated bath disinfectant for dairy farm

Abstract

The invention provides a preparation method of a special post-medicated bath disinfectant for dairy farms, which comprises the following steps: S01, adding a micro-nano zinc solution into zinc sulfate hydrate, heating to 40-70 DEG C, and stirring for 1 hour to obtain a solution A; s02, respectively grinding sodium chloride and boric acid into 600-800-mesh powder, then adding a surfactant, continuously grinding for 5 hours, then adding deionized water, stirring for 1 hour, filtering, and reserving a solution as a solution B; s03, stirring and mixing the solution A and the solution B, adding povidone iodine, sodium bicarbonate and water, stirring for 2 hours at the temperature of 65 DEG C, then adding polyvinylpyrrolidone, continuously stirring for 2 hours, and cooling to obtain a post-medicated bath disinfectant; the micro-nano zinc post-medicated bath disinfectant for dairy cows is actually non-toxic, durable in antibiosis and non-irritant to mucous membranes, a breathable film is formed during post-milking medicated bath, nipples of the milked dairy cows are protected against infection for a long time, the micro-nano zinc post-medicated bath disinfectant is easy to clean during pre-medicated bath in next milking, and preparation is made for cleaning treatment before next milking.

Description

Technical Field

[0001] The invention belongs to the technical field of dairy farm animal husbandry, and in particular relates to a method for preparing a special post-medicated bath disinfectant for dairy farms. Background Art

[0002] Mastitis is a common multiple disease in dairy cows. The main cause of the disease is the invasion of pathogens and various causes of infection. The pathogenic microorganisms of mastitis are relatively complex, including bacteria, fungi, mycoplasma; yeast, viruses, etc. can reach more than 80 kinds. Under normal circumstances, Staphylococcus, Streptococcus and Escherichia coli account for more than 70% of clinical mastitis, followed by pyogenic rod bacteria, Pseudomonas aeruginosa, necrotic bacillus, Nocardia, Klebsiella, etc. Asymptomatic latent mastitis is higher than clinical mastitis, and its incidence rate accounts for about 50% of the entire herd. About 90% of latent mastitis is caused by streptococci and staphylococci.

[0003] Post-dip disinfection is to disinfect and protect the cow's nipples from milking to the next milking. After milking, the cow's nipples are easily invaded by pathogens due to the expansion, and the micro-injury during milking will make it easier to cause infection. Cows with udder inflammation will have an increase in somatic cells during milking. The somatic cell count is an important condition for dairy farms to judge the quality of milk. If the somatic cell count in milk exceeds the limit, the entire tank of milk will be unusable. Therefore, preventing cow udder inflammation is an indispensable measure in the milk production process.

[0004] At present, dairy farms usually use 5%-10% povidone-iodine solution for post-dip disinfection. Since iodine evaporates quickly and has low toxicity, it irritates the mucous membrane of the cow's nipples, thus often causing udder inflammation in the group of dairy cows, causing losses to the dairy farm. Summary of the invention

[0005] In order to solve the above problems, the present invention discloses a micro-nano zinc post-milk cow medicated bath disinfectant.

[0006] To achieve the above object, the technical solution of the present invention is as follows:

[0007] The present invention provides a method for preparing a post-medicated dipping disinfectant solution for dairy farms, comprising the following steps:

[0008] Add S01 micro-nano zinc solution to hydrated zinc sulfate, heat to 40-70°C, and stir for 1h-3h to obtain solution A;

[0009] S02: Grind sodium chloride and boric acid into 600-800 mesh powders respectively, then add surfactant, continue grinding for 5h-7h, then add deionized water, stir for 1h-3h and filter, and retain the solution as solution B;

[0010] S03 After stirring and mixing solution A and solution B, add povidone iodine, sodium bicarbonate and water, stir at a temperature of 65-85°C for 2h-4h, then add polyvinyl pyrrolidone, continue stirring for 2h-4h, and obtain the post-medication bath disinfectant after cooling.

[0011] Furthermore, in step S01, the mass ratio of micro-nano zinc to hydrated zinc sulfate is 99-95:1-5; and the concentration of the sodium chloride solution is 10000 ppm.

[0012] Furthermore, in step S02, the mass ratio of sodium chloride, boric acid, surfactant and water is 1-5:1-5:5-10:93-80.

[0013] Furthermore, the surfactant is one or more of sodium stearate and silane.

[0014] Further, in step S03, the volume ratio of solution A to solution B is 1-5:9-5; the mass ratio of povidone iodine, sodium bicarbonate, and water is 1-5:10-15:89-85.

[0015] Furthermore, in step S03, the mass ratio of polyvinylpyrrolidone to the AB mixed solution is 1-5:99-95.

[0016] The beneficial effects of the present invention are:

[0017] The micro-nano zinc disinfectant for post-milk cow medicated bathing of the present invention is practically non-toxic, durable and antibacterial, has no irritation to the mucous membrane, forms a breathable film during post-milk medicated bathing, protects the nipples of the milking cows from infection for a long time, and is easy to clean during the pre-medicated bathing of the next milking, thereby preparing for the cleaning treatment before the next milking.

[0018] The invention can form a layer of antibacterial film which is breathable for a long time, and can protect the nipples of dairy cows from infection for a longer period of time after milking. DETAILED DESCRIPTION

[0019] The present invention will be further explained below in conjunction with specific implementation modes. It should be understood that the following specific implementation modes are only used to illustrate the present invention and are not used to limit the scope of the present invention.

[0020] Example 1

[0021] S01: Add 10 g of hydrated zinc sulfate ZnSO4·nH2O (n=6-7) to 990 ml of 10000 ppm micro-nano zinc, heat to 40°C in a beaker, and stir for 1 hour to obtain solution A;

[0022] S02 Grind 10g of sodium chloride and 10g of boric acid H3BO3 into 600-800 mesh powder respectively, add 50g of sodium stearate in total, and continue grinding for 5h. Then, add 930ml of deionized water, stir for 1h, filter, and retain the solution as solution B.

[0023] S03 Take 100ml of solution A and 900ml of solution B, stir and mix, take 800ml of the mixed solution, add 2g of povidone iodine, add 20g of sodium bicarbonate, add 178ml of deionized water, stir at 65°C for 2h, take 990ml of the mixed liquid after stirring and adding povidone iodine and sodium bicarbonate, add 10g of polyvinyl pyrrolidone, continue stirring for 2h, cool, and obtain the post-medicated bath disinfectant.

[0024] The preparation method of micro-nano zinc is the same as patent 2021107358455.

[0025] 1. Equipment

[0026] 1. Sample: post-dip disinfectant (1:2 dilution);

[0027] 2. Test bacteria: Escherichia coli, Staphylococcus aureus, Candida albicans;

[0028] 3. Neutralizer: 1% lecithin, 1% Tween-80TSB;

[0029] 4. Organic interferences: 3% bovine serum albumin;

[0030] 5. Constant temperature incubator (model: 404L);

[0031] 6. Culture medium: ordinary nutrient agar;

[0032] 7. Adjustable constant temperature water bath (model: SD28R-30-A12Y);

[0033] 8. Standard hard water: hardness 342 mg / L.

[0034] 2. Methods

[0035] 1. Inspection basis: Technical Specifications for Disinfection, 2002 edition, 2.1.1.5.5, 2.1.1.7.4;

[0036] 2. Neutralizer identification test: Test bacteria: Escherichia coli, Staphylococcus aureus, Candida albicans;

[0037] Disinfectant concentration: original solution action time: 2min;

[0038] The test temperature was 20°C and the experiment was repeated 3 times.

[0039] 3. Experimental Results

[0040] The experimental results are shown in Table 1 below.

[0041] Table 1

[0042]

[0043] The rest is the same as Example 1, except that hydrated zinc sulfate ZnSO4·nH2O (n=6-7) is not added. The test results of the post-medicated bath disinfectant prepared without changing other components and processes are shown in Table 2.

[0044] Table 2

[0045]

[0046] Example 2

[0047] S01: Add 20 g of hydrated zinc sulfate ZnSO4·nH2O (n=6-7) to 980 ml of 10000 ppm micro-nano zinc, heat to 45°C in a beaker, and stir for 1.5 h to obtain solution A;

[0048] S02 Grind 20g of sodium chloride and 20g of boric acid H3BO3 into 600-800 mesh powder respectively, add 60g of LS30 in total, and continue grinding for 5.5h. Then, add 900ml of deionized water, stir for 1.5h, filter, and retain the solution as solution B.

[0049] S03 Take 200ml of solution A and 800ml of solution B, stir and mix, take 800ml of the mixed solution, add 4g of povidone iodine, add 22g of sodium bicarbonate, add 174ml of deionized water, stir at 70°C for 2.5h, take 980ml of the mixed liquid after stirring and adding povidone iodine and sodium bicarbonate, add 20g of polyvinyl pyrrolidone, continue stirring for 2.5h, cool, and obtain the post-medication bath disinfectant.

[0050] The preparation method of micro-nano zinc is the same as patent 2021107358455.

[0051] 1. Equipment

[0052] 1. Sample: post-dip disinfectant (1:4 dilution);

[0053] 2. Test bacteria: Escherichia coli, Staphylococcus aureus, Candida albicans;

[0054] 3. Neutralizer: 1% lecithin, 1% Tween-80TSB;

[0055] 4. Organic interferences: 3% bovine serum albumin;

[0056] 5. Constant temperature incubator (model: 404L);

[0057] 6. Culture medium: ordinary nutrient agar;

[0058] 7. Adjustable constant temperature water bath (model: SD28R-30-A12Y);

[0059] 8. Standard hard water: hardness 342 mg / L.

[0060] 2. Methods

[0061] 1. Inspection basis: Technical Specifications for Disinfection, 2002 edition, 2.1.1.5.5, 2.1.1.7.4;

[0062] 2. Neutralizer identification test: Test bacteria: Escherichia coli, Staphylococcus aureus, Candida albicans;

[0063] Disinfectant concentration: original solution action time: 2min;

[0064] The test temperature was 20°C and the experiment was repeated 3 times.

[0065] 3. Experimental Results

[0066] The experimental results are shown in Table 3 below.

[0067] Table 3

[0068]

[0069]

[0070] The rest is the same as in Example 2, except that boric acid is not added. The test results of the post-medicated bath disinfectant prepared without changing other components and processes are shown in Table 4.

[0071] Table 4

[0072]

[0073] Example 3

[0074] S01: Add 30 g of hydrated zinc sulfate ZnSO4·nH2O (n=6-7) to 970 ml of 10000 ppm micro-nano zinc, heat to 50°C in a beaker, and stir for 2 h to obtain solution A;

[0075] S02 Grind 30g of sodium chloride and 30g of boric acid H3BO3 into 600-800 mesh powder respectively, add 70g of diphenyl sulfonyl chloride in total, and continue grinding for 6h. Then, add 870ml of deionized water, stir for 2h, filter, and retain the solution as solution B.

[0076] S03 Take 300ml of solution A and 700ml of solution B, stir and mix, take 800ml of the mixed solution, add 6g of povidone iodine, add 24g of sodium bicarbonate, add 170ml of deionized water, stir at 75°C for 3h, take 970ml of the mixed liquid after stirring and adding povidone iodine and sodium bicarbonate, add 30g of polyvinyl pyrrolidone, continue stirring for 3h, cool, and obtain the post-medicated bath disinfectant.

[0077] The preparation method of micro-nano zinc is the same as patent 2021107358455.

[0078] 1. Equipment

[0079] 1. Sample: post-dip disinfectant (1:6 dilution);

[0080] 2. Test bacteria: Escherichia coli, Staphylococcus aureus, Candida albicans;

[0081] 3. Neutralizer: 1% lecithin, 1% Tween-80TSB;

[0082] 4. Organic interferences: 3% bovine serum albumin;

[0083] 5. Constant temperature incubator (model: 404L);

[0084] 6. Culture medium: ordinary nutrient agar;

[0085] 7. Adjustable constant temperature water bath (model: SD28R-30-A12Y);

[0086] 8. Standard hard water: hardness 342 mg / L.

[0087] 2. Methods

[0088] 1. Inspection basis: Technical Specifications for Disinfection, 2002 edition, 2.1.1.5.5, 2.1.1.7.4;

[0089] 2. Neutralizer identification test: Test bacteria: Escherichia coli, Staphylococcus aureus, Candida albicans;

[0090] Disinfectant concentration: original solution action time: 2min;

[0091] The test temperature was 20°C and the experiment was repeated 3 times.

[0092] 3. Experimental Results

[0093] The experimental results are shown in Table 5 below.

[0094] Table 5

[0095]

[0096] The rest is the same as in Example 3, except that chlorinated diphenyl sulfonate is not added, and the test results of the post-medicated bath disinfectant prepared without changing other components and processes are shown in Table 6.

[0097] Table 6

[0098]

[0099]

[0100] Example 4

[0101] S01: Add 40 g of hydrated zinc sulfate ZnSO4·nH2O (n=6-7) to 960 ml of 10000 ppm micro-nano zinc, heat to 55°C in a beaker, and stir for 3 h to obtain solution A;

[0102] S02 Grind 40g of sodium chloride and 40g of boric acid H3BO3 into 600-800 mesh powder respectively, add 80g of potassium dihydrogen phosphate in total, and continue grinding for 6.5h. Then, add 840ml of deionized water, stir for 2.5h, filter, and retain the solution as solution B.

[0103] S03 Take 400ml of solution A and 600ml of solution B, stir and mix, take 800ml of the mixed solution, add 8g of povidone iodine, add 26g of sodium bicarbonate, add 166ml of deionized water, stir at 80°C for 3.5h, take 960ml of the mixed liquid after adding povidone iodine and sodium bicarbonate after stirring, add 40g of polyvinyl pyrrolidone, continue stirring for 3.5h, cool, and obtain the post-medication bath disinfectant.

[0104] The preparation method of micro-nano zinc is the same as patent 2021107358455.

[0105] 1. Equipment

[0106] 1. Sample: post-dip disinfectant (1:8 dilution);

[0107] 2. Test bacteria: Escherichia coli, Staphylococcus aureus, Candida albicans;

[0108] 3. Neutralizer: 1% lecithin, 1% Tween-80TSB;

[0109] 4. Organic interferences: 3% bovine serum albumin;

[0110] 5. Constant temperature incubator (model: 404L);

[0111] 6. Culture medium: ordinary nutrient agar;

[0112] 7. Adjustable constant temperature water bath (model: SD28R-30-A12Y);

[0113] 8. Standard hard water: hardness 342 mg / L.

[0114] 2. Methods

[0115] 1. Inspection basis: Technical Specifications for Disinfection, 2002 edition, 2.1.1.5.5, 2.1.1.7.4;

[0116] 2. Neutralizer identification test: Test bacteria: Escherichia coli, Staphylococcus aureus, Candida albicans;

[0117] Disinfectant concentration: original solution action time: 2min;

[0118] The test temperature was 20°C and the experiment was repeated 3 times.

[0119] 3. Experimental Results

[0120] The experimental results are shown in Table 7 below.

[0121] Table 7

[0122]

[0123] The rest is the same as in Example 4, except that povidone-iodine is not added, and the test results of the post-medicated bath disinfectant prepared by the other components and process remain unchanged are shown in Table 8.

[0124] Table 8

[0125]

[0126] Example 5

[0127] S01: Add 50 g of hydrated zinc sulfate ZnSO4·nH2O (n=6-7) to 950 ml of 10000 ppm micro-nano zinc, heat to 60°C in a beaker, and stir for 3 h to obtain solution A;

[0128] S02 Grind 50g of sodium chloride and 50g of boric acid H3BO3 into 600-800 mesh powder respectively, add 90g of potassium sorbate in total, and continue grinding for 7h. Then, add 810ml of deionized water, stir for 3h, filter, and retain the solution as solution B.

[0129] S03 Take 500ml of solution A and 500ml of solution B, stir and mix, take 800ml of the mixed solution, add 10g of povidone iodine, add 28g of sodium bicarbonate, add 162ml of deionized water, stir at 85°C for 4h, take 950ml of the mixed liquid after stirring and adding povidone iodine and sodium bicarbonate, add 50g of polyvinyl pyrrolidone, continue stirring for 4h, cool, and obtain the post-medication bath disinfectant.

[0130] The preparation method of micro-nano zinc is the same as patent 2021107358455.

[0131] 1. Equipment

[0132] 1. Sample: post-dip disinfectant (1:4 dilution);

[0133] 2. Test bacteria: Escherichia coli, Staphylococcus aureus, Candida albicans;

[0134] 3. Neutralizer: 1% lecithin, 1% Tween-80TSB;

[0135] 4. Organic interferences: 3% bovine serum albumin;

[0136] 5. Constant temperature incubator (model: 404L);

[0137] 6. Culture medium: ordinary nutrient agar;

[0138] 7. Adjustable constant temperature water bath (model: SD28R-30-A12Y);

[0139] 8. Standard hard water: hardness 342 mg / L.

[0140] 2. Methods

[0141] 1. Inspection basis: Technical Specifications for Disinfection, 2002 edition, 2.1.1.5.5, 2.1.1.7.4;

[0142] 2. Neutralizer identification test: Test bacteria: Escherichia coli, Staphylococcus aureus, Candida albicans;

[0143] Disinfectant concentration: original solution action time: 2min;

[0144] The test temperature was 20°C and the experiment was repeated 3 times.

[0145] 3. Experimental Results

[0146] The experimental results are shown in Table 9 below.

[0147] Table 9

[0148]

[0149] The rest is the same as Example 5, except that sodium bicarbonate is not added. The test results of the post-medicated bath disinfectant prepared without changing other components and processes are shown in Table 10.

[0150] Table 10

[0151]

[0152] Performance test items:

[0153] 1. Antibacterial durability test (before durability treatment)

[0154] Experimental strains: Staphylococcus aureus AS1.89, Escherichia coli AS1.90

[0155] The test process method is as follows:

[0156] Inspection Procedure

[0157] 1.1 Strain preservation

[0158] The strain was inoculated on a nutrient agar (NA) slant, cultured at (37±1)°C for 24 hours, and then stored at (0-5)°C (not more than 1 month) as a slant-preserved strain.

[0159] 1.2 Bacteria Activation

[0160] Use strains that have been stored for no more than 2 weeks, transfer the slant-stored strains to plate nutrient agar medium, and culture at (37±1℃ for (18-20)h. The test should use fresh bacterial culture that has been transferred twice in a row (transferred within 24h).

[0161] 1.3 Preparation of bacterial suspension

[0162] Use an inoculation loop to take a small amount (scrape 1-2 loops) of fresh bacteria from the 1.2 medium, add it to the culture medium, and make 10-fold incremental dilutions in sequence, selecting a concentration of (5.0-10.0)×10 5 cfu / mL of bacterial solution is used as the inoculation solution and the method of GB / T 4789.2 is used for operation.

[0163] 1.4 Sample test

[0164] Take 0.4mL-0.5mL of the test bacterial solution (1.3) and drop it on the negative control sample (A), the blank control sample (B) and the antibacterial coating sample (C).

[0165] Sample preparation: Select the base material, usually the film-coated cardboard is selected as the base material, and the size of the sample is 50mmx50mm.

[0166] Preparation method of antibacterial coating sample:

[0167] (1) Application: Apply a layer of paint on the substrate ( After the surface is dried, apply a second coat. The coating thickness should be less than 100 μm and dry under natural conditions for 7 days to ensure complete drying.

[0168] (2) Preparation of test plate: The dried coating film was made into a 50 mm x 50 mm antibacterial coating sample.

[0169] Use sterile tweezers to pick up the sterilized covering film (the preparation method of the covering film: first, add the antibacterial agent (Example 4) to the resin, melt it at high temperature in a twin-screw device and mix it evenly with the base resin, and then prepare the antibacterial masterbatch through a pelletizer. Then, the antibacterial masterbatch is melted at high temperature, cooled and rolled up through a single-screw extruder to finally obtain a double-layer barrel-shaped film) and cover it on sample (A), sample (B) and sample (C) respectively. Make sure to lay it flat and free of bubbles so that the bacteria can evenly contact the sample, place it in a sterilized plate, and culture it at (37±1)℃ and relative humidity RH>90% for 21 hours. Perform 3 parallel tests on each sample.

[0170] Take out the samples cultured for 24 hours, add 20 mL of washing solution respectively, wash sample (A), sample (B), sample (C) and the covering film repeatedly (it is best to use tweezers to pick up the film and rinse), shake well, take the washing solution and inoculate it on nutrient agar medium (NA) A, culture at (37±1)℃ for (24-48) hours, and count the viable bacteria. Determine the number of viable bacteria in the washing solution according to the method of GB / T 4789.2.

[0171] 2. Calculation of test results

[0172] Multiply the above measured viable count results by 1000 to obtain the actual viable counts of sample A, sample B, and sample C after 24 hours of culture. The values ​​are A, B, and C respectively. Ensure that the test results meet the following requirements, otherwise the test is invalid:

[0173] The actual recovered viable bacteria value A of sample A should not be less than 1.0x10 5 cfu / piece, and the actual recovered viable bacteria value B of sample B should not be less than 1.0x10 4 cfu / piece;

[0174] The three parallel viable bacteria values ​​of the same blank control sample B must meet the requirement of (highest logarithm value - lowest logarithm value) / average viable bacteria value logarithm value less than or equal to 0.3.

[0175] The calculation formula of antibacterial rate is:

[0176] R=(BC) / B×100

[0177] Where:

[0178] R—antibacterial rate, expressed as (%), with four significant figures, in accordance with the provisions of GB / T 1250;

[0179] B—Average number of recovered bacteria in the blank control plate after 24 hours (cfu / plate);

[0180] C—Average number of recovered bacteria (cfu / piece) on the antibacterial coating sample after 24 hours.

[0181] 3. Antibacterial durability test

[0182] A 30W UV lamp with a wavelength of 253.7nm was used, and the UV lamp complied with GB19258. The antibacterial sample test plate was 0.8m-1.0m away from the UV lamp and irradiated for 100h. The antibacterial durability of the treated test plate was calculated according to the above 1 test procedure and 2 test results. The test results are shown in Table 11.

[0183] Table 11

[0184]

[0185] 4. Antibacterial performance test (after durability treatment)

[0186] Experimental bacteria: Staphylococcus aureus AS1.89, Escherichia coli AS1.90.

[0187] The testing process is the same as above.

[0188] The test results are shown in Table 12.

[0189] Table 12

[0190]

[0191] 5 The results of antibacterial performance and antibacterial durability are shown in Table 13;

[0192] Experimental bacteria: Staphylococcus aureus AS1.89, Escherichia coli AS1.90.

[0193] The specific testing process is the same as above.

[0194] Table 13

[0195]

[0196] The antibacterial performance test results are shown in Table 14

[0197] Experimental bacteria: Staphylococcus aureus AS1.89, Escherichia coli AS1.90, Candida albicans; specific test method is the same as above (the post-medicated bath disinfectant prepared in Example 1 was diluted 1:2 for use).

[0198] Table 14

[0199]

[0200] Toxicology Testing

[0201] 1. Acute oral toxicity test (mice);

[0202] The detection method is as follows:

[0203] 1. Test basis: Refer to the Technical Specifications for Disinfection (2002 Edition), Part 2, Technical Specifications for Inspection of Disinfection Products, 2.3.1 Acute Toxicity Test.

[0204] 2. Test method: A maximum test was conducted with a designed dose of 10000 mg / kg b.wt (post-medicated bath disinfectant prepared in Example 4). After the mice were fasted overnight, they were given oral gavage once with a gavage volume of 20 mL / kg b.wt. Food was given 4 hours after gavage and the mice were observed for 14 days.

[0205] 3. Test results: The acute oral LD50 values ​​of the samples for both female and male mice are greater than 10000 mg / kgb.wt, which is practically non-toxic.

[0206] 2. Acute inhalation toxicity test (rat)

[0207] The detection method is as follows:

[0208] 1. Test basis: Refer to "Technical Specifications for Disinfection" (2002 edition), Part 2, 2.3.2 Acute inhalation toxicity test.

[0209] 2. Test method: one maximum test method, 1m 3 The poison cabinet is used for static poisoning. Males and females are poisoned separately. The poisoning space is 50L / h / rat. The sample stock solution is placed in a sprayer and sprayed directly into the poison cabinet for poisoning. At the same time, the stirring system of the poison cabinet is turned on to evenly spread the test substance. The concentration of each poisoning is obtained by the reduction method. The animals are taken out after 2 hours and observed for 2 weeks.

[0210] 3. Test results: The acute oral LC50 (2h) values ​​of the samples for both female and male rats are greater than 10000mg / m3, which is practically non-toxic.

[0211] 3. One complete skin irritation test (maximum application concentration: 33.3%);

[0212] The detection method is as follows:

[0213] 1. Test basis: Refer to "Technical Specifications for Disinfection" (2002 edition), Part 2, 2.3.3 Skin Irritation Test.

[0214] 2. Test method: 24 hours before the animal test, the hair on both sides of the spine were cut and the skin was prepared to an area of ​​about 3cm×3cm each. The next day, 0.5ml of the test substance was applied to the intact skin on one side with an area of ​​2.5cm×2.5cm. It was covered with 2 layers of gauze and 1 layer of cellophane, and then fixed with irritating tape. The other side was left untreated as a blank control. After 4 hours of application, the residual test substance was washed off with warm water. The skin reaction was observed and recorded after 1h, 24h, and 48h.

[0215] 3. Test results: The maximum mean score of the irritation reaction of the sample solution at 3 times the highest application concentration on the complete skin of rabbits is 0, and the irritation intensity is non-irritating.

[0216] 4. Mouse bone marrow polychromatic erythrocyte micronucleus test.

[0217] The detection method is as follows:

[0218] 1. Test basis: Refer to the Technical Specifications for Disinfection (2002 Edition), Part 2, 2.3.8.4 Mouse Bone Marrow Polychromatic Erythrocyte Micronucleus Test.

[0219] 2. Experimental method: The positive control group, the negative control group (purified water) and each dosage group were gavaged twice every 30 hours, each gavage volume was 20 mg / kg (1:2 dilution of the post-medicated bath disinfectant prepared in Example 1), the mice were killed 6 hours after the second gavage, the femoral bone marrow was suspended in calf serum and directly smeared, fixed, stained, 1000 polychromatic erythrocytes were examined under a microscope for each mouse, the cells with micronuclei were counted, 200 polychromatic erythrocytes were observed and the orthochromatic erythrocytes were counted at the same time, the ratio of polychromatic erythrocytes to orthochromatic erythrocytes (PCE / NCE) was calculated, and the chi-square test was used for statistical analysis.

[0220] 3. Test results: The sample was tested at a dose of 5000 mg / kgb.wt and had no micronucleus-inducing effect on the polychromatic erythrocytes in the mouse bone marrow.

[0221] Example 6

[0222] Verification experiment: experimental equipment and materials

[0223] 1. Experimental strains: Commonly used strains include Staphylococcus aureus (ATCC 6538), Escherichia coli (8099), and Candida albicans (ATCC 10231). Other strains can also be selected according to the specific use of the antibacterial agent.

[0224] 2. Phosphate buffer: used to dilute bacterial suspension.

[0225] 3. Culture medium: including tryptone soy agar (TSA), tryptone soy broth (TSB) and sandcastle agar.

[0226] 4. Film: Use materials that do not affect bacterial growth and do not absorb water. The thickness is not specified. One side should have good adhesion and the area should be 40mmx2mm square.

[0227] 5. Sterile plastic bag: used to wash away bacteria.

[0228] 6. Constant temperature water bath: used to culture bacteria.

[0229] 7.37℃ constant temperature incubator: used to culture bacteria.

[0230] 8. Samples: Prepare the sample (prepared in the same way as above) and the control sample (same as the sample but without antibacterial components) into squares with a side length of 50 mm2mm. Before the test, gently wipe the sample 2 to 3 times with cotton wool dipped in alcohol and dry it thoroughly.

[0231] Experimental procedures

[0232] 1. Pour agar medium plates.

[0233] 2. Dilute the bacterial suspension with 1 / 500 nutrient broth to make an experimental bacterial suspension of 2.5×10^5cfu / mL~1.0x10^6cfu / mL.

[0234] 3. Place the sample with the test surface facing up in a sterile dish, take 0.4 mL of bacterial suspension and drop it on the sample, spread it evenly, cover it with a film, carefully press the film to spread the bacterial solution evenly, and cover it with the dish lid.

[0235] 4. Place the plates containing the test samples inoculated with the bacterial solution (3 antibacterial samples and 3 control samples) in an incubator at 35°C ± 1°C and a relative humidity of not less than 90% for 24 hours.

[0236] 5. Aseptically place the cover film and test sample into a plastic bag using tweezers, then add 10 mL of broth culture solution, and rub the test sample and cover film in the bag thoroughly by hand to wash off the bacteria.

[0237] 6. Pipette 1 mL of washed bacterial solution into a test tube containing 9 mL of PBS, make a series of 10-fold dilutions, take appropriate dilutions to inoculate 2 plates, add 15 mL to 20 mL of TSA by pouring method, and culture in a constant temperature incubator at 35°C ± 1°C for 40 h to 48 h.

[0238] 7. Immediately after inoculating the other three control samples, use tweezers to place the cover film and test samples into a plastic bag. The elution and inoculation methods are the same as those for the test samples.

[0239] 8. Set up negative controls using the same batch of diluents, culture media, etc. as used in the experiment.

[0240] 9. The experiment was repeated 3 times.

[0241] Experimental results calculation method

[0242] The formula for calculating the number of viable bacteria is: N=C×D×V, where N is the number of viable bacteria, C is the average number of colonies on two plates, D is the dilution factor, and V is the volume of broth culture medium used for elution (mL).

[0243] The specific experimental data are as follows:

[0244]

[0245] The post-medication bath toxicology test prepared by the present invention is practically non-toxic and non-irritating, and the durability test reaches the Class I standard.

[0246] It should be noted that the above content only illustrates the technical idea of ​​the present invention and cannot be used to limit the protection scope of the present invention. For ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications all fall within the protection scope of the claims of the present invention.

Claims

1. A method for preparing a post-dip disinfectant solution for dairy farms, characterized in that: The following steps are involved: Add S01 micro-nano zinc solution to hydrated zinc sulfate, heat to 40-70°C, and stir for 1h-3h to obtain solution A; S02: Grind sodium chloride and boric acid into 600-800 mesh powders respectively, then add surfactant, continue grinding for 5h-7h, then add deionized water, stir for 1h-3h and filter, and retain the solution as solution B; S03 After stirring and mixing solution A and solution B, add povidone iodine, sodium bicarbonate and water, stir at a temperature of 65-85°C for 2h-4h, then add polyvinyl pyrrolidone, continue stirring for 2h-4h, and obtain the post-medication bath disinfectant after cooling.

2. The method for preparing a post-medicated disinfectant solution for dairy farms according to claim 1, characterized in that: In step S01, the mass ratio of the micro-nano zinc solution to the hydrated zinc sulfate is 99-95:1-5; the concentration of the sodium chloride solution is 10000 ppm.

3. The method for preparing a post-medicated disinfectant solution for dairy farms according to claim 1, characterized in that: In step S02, the mass ratio of sodium chloride, boric acid, surfactant and water is 1-5:1-5:5-10:93-80.

4. The method for preparing a post-medicated disinfectant solution for dairy farms according to claim 3, characterized in that: The surfactant is one or more of sodium stearate and silane.

5. The method for preparing a post-medicated disinfectant solution for dairy farms according to claim 1, characterized in that: In step S03, the volume ratio of solution A to solution B is 1-5:9-5; the mass ratio of povidone iodine, sodium bicarbonate, and water is 1-5:10-15:89-85.

6. The method for preparing a post-medicated disinfectant solution for dairy farms according to claim 1, characterized in that: In step S03, the mass ratio of polyvinylpyrrolidone to the AB mixed solution is 1-5:99-95.

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