TRADITIONAL CHINESE MEDICINE OINTMENT FOR PREVENTING AND TREATING MASTITIS IN DAIRY COWS, AND PREPARATION METHOD THEREOF

Abstract

Provided are a traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows and a preparation method thereof, belonging to the technical field of veterinary traditional Chinese medicines. The traditional Chinese medicine ointment includes raw materials in parts by mass: 20-30 parts of dandelion, 20-30 parts of herba patriniae, 10-16 parts of Caulis Lonicerae Japonicae, 0.5-1 part of borneol, 0.03-0.05 part of polypeptide, 40-50 parts of auxiliary emulsifier, 40-50 parts of liquid paraffin and 100-130 parts of white vaseline. An amino acid sequence of the polypeptide is: Phe Phe Arg Lys Val Leu Lys Leu Ile Arg Lys Ile Trp Arg (SEQ ID NO. 1). In the application, dandelion, herba patriniae, Caulis Lonicerae Japonicae and borneol 1 are mixed, and polypeptide is added at the same time to prepare ointment.

Claims

1. A traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows, comprising following raw materials in parts by weight: 20-30 parts of dandelion, 20-30 parts of herba patriniae, 10-16 parts of Caulis Lonicerae Japonicae, 0.5-1 part of borneol, 0.03-0.05 part of polypeptide, 40-50 parts of auxiliary emulsifier, 40-50 parts of liquid paraffin and 100-130 parts of white vaseline, 0.5-1 part of antioxidant and 0.5-1 part of preservative; wherein the amino acid sequence of the polypeptide is shown as SEQ ID NO: 1.

2. The traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows according to claim 1, wherein the auxiliary emulsifier is glyceryl monostearate.

3. The traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows according to claim 1, wherein the antioxidant is alkyl gallate or butylated hydroxytoluene; the preservative is ethylparaben.

4. A preparation method of the traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows according to claim 1, comprising following steps: (1), adding the dandelion, the herba patriniae and the Caulis Lonicerae Japonicae into ethanol aqueous solution, extracting, and concentrating to obtain traditional Chinese medicine concentrated solution; and (2), grinding the borneol and part of the liquid paraffin to eutectic; adding remaining liquid paraffin and molten-state white vaseline, keeping a temperature, adding the auxiliary emulsifier, stirring well, adding the traditional Chinese medicine concentrated solution and the polypeptide, homogenizing, and cooling to obtain the traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows.

5. The preparation method according to claim 4, wherein a volume fraction of ethanol in the ethanol aqueous solution in the step (1) is 70-75%, a temperature for the extracting is 80-90? C.

6. The preparation method according to claim 4, wherein a crude drug content in the traditional Chinese medicine concentrated solution in the step (1) is 1-1.5 g/ml.

7. The preparation method according to claim 4, wherein a temperature of the molten-state white vaseline in the step (2) is 50-55? C., keeping the temperature is to keep the temperature at 50-55? C.

8. The preparation method according to claim 4, wherein a speed of the homogenizing in the step (2) is 3500-4000 rpm and time of the homogenizing is 10-15 min.

Description

BRIEF DESCRIPTION OF THE DRAWING

[0037] The figure is a process of a preparation method of the traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0038] A number of exemplary embodiments of the present application will now be described in detail, and this detailed description should not be considered as a limitation of the present application, but should be understood as a more detailed description of certain aspects, characteristics and embodiments of the present application. It should be understood that the terminology described in the present application is only for describing specific embodiments and is not used to limit the present application.

[0039] In addition, for the numerical range in the present application, it should be understood that each intermediate value between the upper limit and the lower limit of the range is also specifically disclosed. Intermediate values within any stated value or stated range, as well as each smaller range between any other stated value or intermediate values within the stated range are also included in the present application. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.

[0040] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application relates. Although the present application only describes the preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present application.

[0041] The terms comprising, including, having and containing used in this article are all open terms, which means including but not limited to.

[0042] Unless otherwise specified, the polypeptide mentioned in the embodiments and comparative examples of the present application all refer to polypeptide with the amino acid sequence of Phe Phe Arg Lys Val Leu Lys Leu Ile Arg Lys Ile Trp Arg (SEQ ID NO. 1), which originates from Hunan University of Science and Engineering, and the preparation technical reference publication number is CN111529683A.

[0043] As shown in the figure, a process of a preparation method of the traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows includes following steps: [0044] S1, adding the dandelion, the herba patriniae and the Caulis Lonicerae Japonicae into ethanol aqueous solution, extracting, and concentrating to obtain traditional Chinese medicine concentrated solution; and [0045] S2, grinding the borneol and part of the liquid paraffin to eutectic; adding remaining liquid paraffin and molten-state white vaseline, keeping a temperature, adding the auxiliary emulsifier, stirring well, adding the traditional Chinese medicine concentrated solution and the polypeptide, homogenizing, and cooling to obtain the traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows. The process will be described with reference to following embodiments.

Embodiment 1

[0046] Preparation of traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows includes following steps: [0047] S1, taking 30 parts of dandelion, 30 parts of herba patriniae and 10 parts of Caulis Lonicerae Japonicae, adding ethanol aqueous solution with volume fraction of 75% according to the material-liquid ratio of 1:10, reflux extracting at 80? C. for 1 hour, filtering, continuing to add ethanol aqueous solution with volume fraction of 75% to filter residue according to the material-liquid ratio of 1:5, reflux extracting at 80? C. for 1 hour, filtering, combining the filtrates extracted twice, and boiling and concentrating until the crude drug content is 1.5 g/mL to obtain traditional Chinese medicine concentrated solution for later use; [0048] S2, heating the white vaseline to 80? C., and cooling to 50? C. after the white vaseline is completely melted to obtain the molten-state white vaseline for later use; and [0049] S3, taking 0.5 part of borneol, adding 10 parts of liquid paraffin, and grinding to eutectic mixture; adding the remaining 30 parts of liquid paraffin and 100 parts of molten-state white vaseline obtained in step (2), keeping the temperature of the mixture at 50-55? C., adding 40 parts of glyceryl monostearate, stifling well, then adding the traditional Chinese medicine concentrated solution obtained in step (1), 0.05 part of polypeptide, 0.5 part of alkyl gallate and 0.5 part of ethylparaben, homogenizing at 3500 revolutions per minute for 15 minutes, cooling to room temperature to obtain the traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows.

Embodiment 2

[0050] Preparation of traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows includes following steps: [0051] S1, taking 25 parts of dandelion, 25 parts of herba patriniae and 16 parts of Caulis Lonicerae Japonicae, adding ethanol aqueous solution with volume fraction of 75% according to the material-liquid ratio of 1:10, reflux extracting at 80? C. for 1 hour, filtering, continuing to add ethanol aqueous solution with volume fraction of 75% to filter residue according to the material-liquid ratio of 1:5, reflux extracting at 80? C. for 1 hour, filtering, combining the filtrates extracted twice, and boiling and concentrating until the crude drug content is 1.0 g/mL to obtain traditional Chinese medicine concentrated solution for later use; [0052] S2, heating the white vaseline to 80? C., and cooling to 50? C. after the white vaseline is completely melted to obtain the molten-state white vaseline for later use; and [0053] S3, taking 0.7 part of borneol, adding 10 parts of liquid paraffin, and grinding to eutectic mixture; adding the remaining 35 parts of liquid paraffin and 120 parts of molten-state white vaseline obtained in step (2), keeping the temperature of the mixture at 50-55? C., adding 50 parts of glyceryl monostearate, stifling well, then adding the traditional Chinese medicine concentrated solution obtained in step (1), 0.03 part of polypeptide, 0.5 part of alkyl gallate and 1 part of ethylparaben, homogenizing at 3500 revolutions per minute for 15 minutes, cooling to room temperature to obtain the traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows.

Embodiment 3

[0054] Preparation of traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows includes following steps: [0055] S1, taking 25 parts of dandelion, 20 parts of herba patriniae and 14 parts of Caulis Lonicerae Japonicae, adding ethanol aqueous solution with volume fraction of 70% according to the material-liquid ratio of 1:10, reflux extracting at 90? C. for 1 hour, filtering, continuing to add ethanol aqueous solution with volume fraction of 70% to filter residue according to the material-liquid ratio of 1:5, reflux extracting at 90? C. for 1 hour, filtering, combining the filtrates extracted twice, and boiling and concentrating until the crude drug content is 1.2 g/mL to obtain traditional Chinese medicine concentrated solution for later use; [0056] S2, heating the white vaseline to 90? C., and cooling to 50? C. after the white vaseline is completely melted to obtain the molten-state white vaseline for later use; [0057] S3, taking 1 part of borneol, adding 15 parts of liquid paraffin, and grinding to eutectic mixture; adding the remaining 35 parts of liquid paraffin and 130 parts of molten-state white vaseline obtained in step (2), keeping the temperature of the mixture at 50-55? C., adding 45 parts of glyceryl monostearate, stirring well, then adding the traditional Chinese medicine concentrated solution obtained in step (1), 0.05 part of polypeptide, 1 part of alkyl gallate and 0.5 part of ethylparaben, homogenizing at 4000 revolutions per minute for 10 minutes, cooling to room temperature to obtain the traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows.

COMPARATIVE EXAMPLE 1

[0058] Preparation of traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows:

[0059] Compared with embodiment 1, the difference is that the addition of polypeptide is omitted, and other steps are the same as embodiment 1.

Effect Verification:

[0060] Fifty dairy cows with mastitis detected by Shanghai Mastitis Test (SMT) are selected from a dairy farm in Hunan Province, of which 20 cows are externally coated with the traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows prepared in embodiment 1 and 20 cows are externally applied with the traditional Chinese medicine ointment for preventing and treating mastitis in dairy cows prepared in comparative example 1, with the dosage of 2 g each time, twice a day for 5 days. The remaining 10 cows are perfused with penicillin 100,000 IU after all the milk is discharged from their breasts, as a control group.

[0061] Cure standard: one day after stopping the ointments, if curdled milk is negative by the sodium hydroxide test, the cows are judged to be cured.

[0062] The therapeutic effects of each experimental group are shown in Table 1.

TABLE-US-00002 TABLE 1 The therapeutic effects of each experimental group Dairy Cows (head) with cows (head) curdled milk being Cure Experimental with negative by sodium rate group mastitis hydroxide test (%) Embodiment 1 20 18 90 group Comparative 20 14 70 example 1 group Control group 10 8 80

[0063] As can be seen from Table 1, the added polypeptide is able to obviously improve the therapeutic effect of traditional Chinese medicine composition, which may be because the added polypeptide is able to improve the sensitivity of pathogenic microorganisms to traditional Chinese medicine extracts, so the therapeutic effect of the prepared traditional Chinese medicine ointment is able to be obviously improved after the added polypeptide is added.

[0064] In order to confirm the above conjecture, the application verifies the antibacterial effect of the traditional Chinese medicine concentrated solution prepared in each embodiment, and the method is as follows: [0065] (1) taking Staphylococcus aureus and Escherichia coli grown on ordinary nutrient agar plates, dipping a small number of colonies with an inoculating loop, inoculating colonies into 5 mL of Mueller-Hinton (MH) broth, and performing shake culture at 37? C. for 6 hours at 200 r/min; taking bacterial liquid in logarithmic growth period and diluting the bacterial liquid with sterilized physiological saline until the bacterial content is 1-2?108 CFU/ml; diluting with sterilized physiological saline for 1000 times for later use; [0066] (2) making a round filter paper with a diameter of 6.5 mm with a puncher, and dry-heat sterilizing it; first, evenly inoculating the bacterial liquid obtained in step (1) into a nutrient agar culture dish, then attaching the sterilized round filter paper to the surface of agar, transferring 20 ?L of liquid medicine (diluted by 10 times) with a micropipette gun, dropping the liquid medicine in the center of the round filter paper, culturing it at 37? C. for 24 hours, and measuring the diameter of the bacteriostatic circle in which different liquid medicines are dropped. The results are shown in Table 2.

TABLE-US-00003 TABLE 2 Diameter of bacteriostatic circle in different liquid medicine groups Diameter of Diameter of Liquid medicine bacteriostatic circle bacteriostatic circle (transferring 20 ?L after of Staphylococcus of Escherichia coli diluting by 10 times) aureus (mm) (mm) Traditional Chinese medicine 8.3 7.8 concentrated solution in embodiment 1 Group Traditional Chinese medicine 23.9 21.5 concentrated solution in embodiment 1 Group + 0.05 g of polypeptide Traditional Chinese medicine 7.6 7.9 concentrated solution in embodiment 2 Group Traditional Chinese medicine 21.1 21.6 concentrated solution in embodiment 2 Group + 0.03 g of polypeptide Traditional Chinese medicine 8.4 8.2 concentrated solution in embodiment 3 Group Traditional Chinese medicine 22.4 21.9 concentrated solution in embodiment 3 Group + 0.05 g of polypeptide Solution with polypeptide 15.3 14.7 content of 1 mg/mL

[0067] As can be seen from Table 2, the simple polypeptide solution has a strong antibacterial effect, but it is able to obviously improve the antibacterial effect of the traditional Chinese medicine concentrated solution for treating mastitis of dairy cows after being added to the traditional Chinese medicine concentrated solution for treating mastitis, indicating that the polypeptide added in the application is able to synergize with the traditional Chinese medicine concentrated solution disclosed by the application and has a stronger antibacterial effect.

[0068] The above-mentioned embodiments only describe the preferred mode of the application, and do not limit the scope of the application. Under the premise of not departing from the design spirit of the application, various modifications and improvements made by ordinary technicians in the field to the technical scheme of the application shall fall within the protection scope determined by the claims of the application.