Composition for preventing and treating mastitis and application thereof

Abstract

The present invention discloses a composition for preventing and treating mastitis and its application, relating to the technical field of traditional Chinese medicine preparation. The composition for preventing and treating mastitis comprises a sunflower extract, a ginkgo leaf extract, and aloe vera gel. The aloe vera gel, sunflower extract, and ginkgo leaf extract are compounded and used to obtain a composition having the effects of clearing heat and detoxifying, antibacterial and anti-inflammatory, reducing swelling and relieving pain, and promoting menstruation and lactation. Furthermore, hydrogen has antioxidant properties, high permeability, and biosafety, and can enhance the effect of the composition on the site of inflammation.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine preparation, and in particular to a composition for preventing and treating mastitis and application thereof. Background Art

[0002] Mastitis in dairy cows is a defensive response to pathological changes in the mammary glands of dairy cows, adversely affecting milk production, milk physicochemical properties, and reproductive function, making it a crucial component of dairy cow disease prevention and control. Traditional Chinese veterinary medicine believes that mastitis is caused by the stagnation of phlegm, dampness, qi, and blood, which then transforms into inflammation. This is often caused by poor husbandry and management, prolonged exposure to hot, damp soil, which allows toxic dampness and heat to invade the udders, nipple bites that allow pathogenic toxins to enter, or excessive stomach fire, liver qi stagnation, and blocked milk channels, which lead to condensation of the mammary meridians and the development of carbuncles. Essentially, mastitis in dairy cows is an inflammation of the udder caused by microorganisms invading the mammary glands. Escherichia coli, Staphylococcus aureus, and Streptococcus uberis are the main pathogens of udder infections in dairy cows.

[0003] Currently, cow mastitis is primarily treated with antibiotics to kill pathogens and eliminate inflammatory responses. However, Escherichia coli, Staphylococcus aureus, and Streptococcus are highly resilient and prone to mutation under natural conditions, leading to antibiotic resistance. Furthermore, the use of antibiotics can lead to excessive levels of antibiotics in milk and the increasing number of drug-resistant strains. Traditional Chinese medicine, as a purely natural substance, contains a variety of biologically active ingredients, each with its own distinct efficacy and effects. Traditional Chinese medicine possesses antitoxin and antibacterial properties, enhancing animal immunity and disease resistance. Furthermore, bacteria are less likely to develop drug resistance, making it a promising alternative to antibiotics.

[0004] The paper "Analysis of the Antibacterial Effect of Saltwort Compound Extract on Cow Milk-derived Staphylococcus aureus" (Dong Gui, Tarim University) disclosed that Saltwort and its extract can effectively inhibit Staphylococcus aureus. "Effect of Artemisinin on Mastitis Induced by Escherichia coli" (Li Zhaoming, Shandong Agricultural University) disclosed that artemisinin can effectively alleviate Escherichia coli-induced mammary tissue damage and reduce inflammatory factors in serum. It can be seen that traditional Chinese medicine and its extracts in the existing technology mostly show good antibacterial effects on one of the pathogens of cow mastitis. At the same time, traditional Chinese medicine emphasizes reasonable compatibility and uses different drugs in combination to improve the efficacy of the drugs and reduce or eliminate the toxic side effects of the drugs to achieve the best therapeutic effect.

[0005] Therefore, it is very necessary to rationally select a combination of Chinese medicinal herbs to prepare a composition with antibacterial and anti-inflammatory effects, which can be used to prevent and treat cow mastitis caused by Escherichia coli, Staphylococcus aureus and Streptococcus uberis. Summary of the Invention

[0006] In response to the above-mentioned prior art, the present invention aims to provide a composition for preventing and treating mastitis, as well as its preparation method and application. The composition, which is prepared by combining aloe vera gel, sunflower extract, and ginkgo biloba extract, has the effects of clearing heat and detoxifying, providing antibacterial and anti-inflammatory effects, relieving swelling and pain, and promoting menstruation and lactation. It also exhibits excellent antibacterial effects against Escherichia coli, Staphylococcus aureus, and Streptococcus uberis. Furthermore, hydrogen gas has antioxidant properties, high permeability, and safety, which can enhance the effectiveness of the composition at the site of inflammation.

[0007] To achieve the above object, the present invention adopts the following technical solutions:

[0008] The first aspect of the present invention provides a composition for preventing and treating mastitis, characterized in that it comprises the following raw materials in parts by weight:

[0009] 20-25 parts of aloe vera gel, 6-9 parts of amomum villosum extract, and 8-10 parts of ginkgo biloba extract.

[0010] Astragalus membranaceus, as recorded in the National Compendium of Chinese Herbal Medicine, clears heat and dampness, draws out toxins and discharges pus. It cures amoebic dysentery and urinary stones. The entire plant can be used as medicine. It has a slightly sweet taste and a cold nature. It enters the heart and lung meridians and has the effects of clearing heat and detoxifying, promoting lactation and relieving constipation. It is mainly used to treat persistent high fever, lung heat cough, dysentery, constipation, postpartum milk stagnation, fractures, carbuncles, abscesses, unidentified swellings and burns.

[0011] Ginkgo leaves have the effects of promoting blood circulation, removing blood stasis and unblocking meridians. They are used to treat chest pain, heart pain, stroke, hemiplegia, tongue stiffness, dysphasia, speech impairment, coronary heart disease, stable angina pectoris and cerebral infarction caused by blood stasis. Pharmacological studies on ginkgo leaves have shown that this drug has the effects of dilating blood vessels, resisting myocardial ischemia, resisting cerebral ischemia and resisting thrombosis.

[0012] Preferably, the sunflower extract is prepared by the following method:

[0013] Add ethanol to the sunflower sunflower powder and ultrasonically extract it 2-3 times to obtain an extract; the extract is concentrated and dried to obtain the sunflower sunflower extract.

[0014] Further preferably, the mass ratio of the sunflower oil powder to ethanol is 1:(20-40); the volume fraction of the ethanol is 50-70%; the ultrasonic extraction time is 1-3 hours, and the ultrasonic power is 120-180W.

[0015] Preferably, the preparation method of the ginkgo leaf extract is prepared by the following method:

[0016] Add ethanol to ginkgo leaf powder, heat and reflux for 1-2 times to obtain an extract; concentrate and dry the extract to obtain ginkgo leaf extract.

[0017] Further preferably, the mass ratio of the ginkgo leaf powder to ethanol is 1:(5-10), the volume fraction of the ethanol is 50-70%; the heating reflux temperature is 75-85° C., and the extraction time is 3-5 hours.

[0018] The second aspect of the present invention provides the use of the above composition in the following (1)-(2):

[0019] (1) Preparation of products for inhibiting Escherichia coli, Staphylococcus aureus and Streptococcus uberis;

[0020] (2) Preparation of medicines for preventing and treating mastitis.

[0021] The third aspect of the present invention provides a drug for preventing and treating mastitis, which comprises the above composition and hydrogen-rich water.

[0022] A fourth aspect of the present invention provides a method for preparing a drug for preventing and treating mastitis, comprising the following steps:

[0023] Add hydrogen-rich water to the composition and mix them evenly to obtain a medicine for preventing and treating mastitis.

[0024] Preferably, the mass ratio of the composition to hydrogen-rich water is 1:(5-10), and the hydrogen content in the hydrogen-rich water is 8ppm-10ppm.

[0025] Beneficial effects of the present invention:

[0026] 1. The present invention adopts aloe vera gel, sunflower extract and ginkgo leaf extract to prevent and treat mastitis of dairy cows caused by Escherichia coli, Staphylococcus aureus and Streptococcus uberis. Ginkgo leaf extract contains multiple active substances such as flavonoids, lactones, phenols and polysaccharides, and has multiple biological activities such as antioxidant, antibacterial, anti-inflammatory and antiviral. Sunflower extract contains active substances such as flavonoids, polysaccharides, alkaloids and multiple amino acids, and has the effects of antioxidant, improving the body's immunity and scavenging free radicals. Aloe vera gel also has anti-inflammatory and bactericidal effects. Therefore, the present invention uses aloe vera gel, sunflower extract and ginkgo leaf extract in combination, and the resulting composition has the effects of clearing away heat and detoxification, antibacterial and anti-inflammatory, reducing swelling and relieving pain, and promoting menstruation and promoting lactation.

[0027] 2. Hydrogen, as a reducing diatomic gas molecule, has anti-inflammatory, antioxidant, anti-apoptotic, and autophagy-regulating effects. Its selective antioxidant properties only scavenge free radicals and do not affect normal cellular metabolism. Furthermore, its small molecular mass allows it to penetrate lipid membranes and reach organelles such as mitochondria and the nucleus to exert its biological effects. Its high permeability allows it to quickly reach sites of inflammation and exert its effects. Furthermore, hydrogen is highly safe, and no negative effects have been observed in either laboratory experiments or clinical applications. DETAILED DESCRIPTION

[0028] It should be noted that the following detailed descriptions are illustrative and intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present application belongs.

[0029] As described in the background technology, Escherichia coli, Staphylococcus aureus and Streptococcus uberis are among the main pathogens that cause mastitis in dairy cows. In the existing technology, antibiotics are mostly used to treat mastitis in dairy cows. However, the use of antibiotics will cause the antibiotic content in milk to exceed the standard, and the number of drug-resistant strains caused by strain mutations will continue to increase.

[0030] Based on this, in view of the fact that Chinese veterinary medicine theory attaches importance to holistic concept, when Chinese veterinary medicine is a compound preparation, its chemical composition has diversity, and content changes greatly, and can influence each other, generate some stable or metastable complex compounds. The present invention is according to Chinese veterinary medicine theory, in conjunction with the pathogenic mechanism of cow mastitis, selects multi-flavor Chinese herbal medicine to be applied to the in vitro antibacterial test of cow mastitis Staphylococcus aureus, and by checking the antibacterial concentration index (FICI) of different Chinese medicine extracts to characterize its combined inhibitory effect on Staphylococcus aureus, obtains aloe vera gel, sunflower extract and ginkgo biloba extract being used in collaboration to prepare the composition for preventing and treating cow mastitis, the composition has heat-clearing and detoxicating, antibacterial and anti-inflammatory, detumescence and analgesia and the effect of promoting menstruation and promoting lactation. And, in order to further improve the anti-inflammatory effect of composition, composition is used in combination with hydrogen-rich water, the hydrogen in hydrogen-rich water has antioxidant, high permeability and biosafety, can effectively improve the anti-inflammatory and antibacterial effect of composition.

[0031] In order to enable those skilled in the art to more clearly understand the technical solution of the present application, the technical solution of the present application will be described in detail below with reference to specific embodiments.

[0032] The test materials used in the examples of the present invention are all conventional test materials in the art and can be purchased through commercial channels.

[0033] The aloe vera gel was purchased from Wuhan Lvjing Fenghua Biotechnology Co., Ltd., model number CB2311.

[0034] Example 1: Preparation of a composition for preventing and treating mastitis

[0035] 1. Raw material composition:

[0036] 23 parts aloe vera gel, 8 parts ginkgo biloba extract and 9 parts ambrette extract.

[0037] 2. Preparation process:

[0038] (1) The whole plant of sunflower is dried and crushed, and then passed through a 100-mesh sieve to obtain sunflower powder. Then, 60% ethanol by volume is added to the sunflower powder, wherein the mass ratio of sunflower powder to ethanol is 1:30. Ultrasonic extraction is performed three times at 150W, each extraction time is 2 hours, and the solutions are combined to obtain an extract. The extract is concentrated and dried at 140°C to obtain a sunflower extract.

[0039] (2) After drying and crushing the ginkgo leaves, pass them through a 100-mesh sieve to obtain ginkgo leaf powder, add 60% ethanol by volume to the ginkgo leaf powder, wherein the mass ratio of ginkgo leaf powder to ethanol is 1:8, heat and reflux extract at 80°C for 2 times, each extraction time is 4 hours, combine the solutions to obtain an extract, concentrate the extract, and dry it at 120°C to obtain ginkgo leaf extract;

[0040] (3) Add the sunflower extract and the ginkgo leaf extract to the aloe vera gel and mix well to obtain a composition for preventing and treating mastitis.

[0041] Example 2: Preparation of a composition for preventing and treating mastitis

[0042] 1. Raw material composition:

[0043] 20 parts aloe vera gel, 6 parts ginkgo biloba extract and 8 parts ambrette extract.

[0044] 2. Preparation process:

[0045] (1) The whole plant of sunflower is dried and crushed, and then passed through a 100-mesh sieve to obtain sunflower powder. Then, 50% ethanol by volume is added to the sunflower powder, wherein the mass ratio of sunflower powder to ethanol is 1:30. Ultrasonic extraction is performed twice at 120W, each extraction time is 3 hours, and the solutions are combined to obtain an extract. The extract is concentrated and dried at 140°C to obtain a sunflower extract.

[0046] (2) After drying and crushing the ginkgo leaves, pass them through a 100-mesh sieve to obtain ginkgo leaf powder, and then add 50% ethanol by volume to the ginkgo leaf powder, wherein the mass ratio of ginkgo leaf powder to ethanol is 1:5, and heat reflux extraction at 75°C once, each extraction time is 5 hours, combine the solutions to obtain an extract, concentrate the extract, and dry it at 120°C to obtain ginkgo leaf extract;

[0047] (3) Add the sunflower extract and the ginkgo leaf extract to the aloe vera gel and mix well to obtain a composition for preventing and treating mastitis.

[0048] Example 3: Preparation of a composition for preventing and treating mastitis

[0049] 1. Raw material composition:

[0050] 25 parts aloe vera gel, 9 parts ginkgo biloba extract and 10 parts ambrette extract.

[0051] 2. Preparation process:

[0052] (1) The whole plant of sunflower is dried and crushed, and then passed through a 100-mesh sieve to obtain sunflower powder. Then, 70% ethanol by volume is added to the sunflower powder, wherein the mass ratio of sunflower powder to ethanol is 1:40. Ultrasonic extraction is performed three times at 120W, each extraction time is 1 hour, and the solutions are combined to obtain an extract. The extract is concentrated and dried at 140°C to obtain a sunflower extract.

[0053] (2) Drying and crushing the ginkgo leaves, and then passing through a 100-mesh sieve to obtain ginkgo leaf powder, and then adding 70% ethanol by volume to the ginkgo leaf powder, wherein the mass ratio of ginkgo leaf powder to ethanol is 1:10, heating and refluxing at 85°C for 2 times, each extraction time is 3 hours, combining the solutions to obtain an extract, concentrating the extract, and drying at 120°C to obtain the ginkgo leaf extract;

[0054] (3) Add the sunflower extract and the ginkgo leaf extract to the aloe vera gel and mix well to obtain a composition for preventing and treating mastitis.

[0055] Example 4: Preparation of a drug for preventing and treating mastitis

[0056] Hydrogen-rich water was added to the composition prepared in Example 1 and mixed to obtain a drug for preventing and treating mastitis, wherein the mass ratio of the composition to the hydrogen-rich water was 1:8, and the hydrogen content in the hydrogen-rich water was 10 ppm.

[0057] Example 5: Preparation of a drug for preventing and treating mastitis

[0058] Hydrogen-rich water was added to the composition prepared in Example 2 and mixed to obtain a drug for preventing and treating mastitis, wherein the mass ratio of the composition to the hydrogen-rich water was 1:5, and the hydrogen content in the hydrogen-rich water was 8 ppm.

[0059] Example 6: Preparation of a drug for preventing and treating mastitis

[0060] Hydrogen-rich water was added to the composition prepared in Example 3 and mixed to obtain a drug for preventing and treating mastitis, wherein the mass ratio of the composition to the hydrogen-rich water was 1:10, and the hydrogen content in the hydrogen-rich water was 9 ppm.

[0061] Comparative Example 1:

[0062] The difference between this comparative example and Example 4 is that the composition is only the sunflower extract prepared in step (1) of Example 1.

[0063] Comparative Example 2:

[0064] The difference between this comparative example and Example 4 is that the composition is only the Ginkgo biloba extract prepared in step (2) of Example 1.

[0065] Comparative Example 3:

[0066] The difference between this comparative example and Example 4 is that the composition is only aloe vera gel.

[0067] Test Example 1: Antibacterial Test

[0068] The antibacterial test was conducted with reference to the journal "Study on the Antibacterial Effect of Aqueous Extract of Melia azedarach Fruit" (Peng Hong, Zhou Gang et al. Industrial Microbiology Vol. 50 No. 4 Aug. 020). The specific steps are as follows:

[0069] (1) Escherichia coli and Staphylococcus aureus were inoculated on NA (normal nutrient agar) slant culture, and the viable counts were diluted to 5×10 7 CFU / mL of Escherichia coli suspension and Staphylococcus aureus suspension were set aside; Streptococcus uberis was inoculated on THB agar medium and cultured at 37°C for 22-24 hours, and then diluted to obtain a viable cell count of 1×10 5 CFU / mL of Streptococcus uberis suspension for later use;

[0070] Among them, Escherichia coli was purchased from the China Industrial Culture Collection of Microorganisms with a strain collection number of CICC10899, Staphylococcus aureus was purchased from the China Industrial Culture Collection of Microorganisms with a strain collection number of CICC10384, and Streptococcus uberis was purchased from the Shanghai Collection Biotechnology Center with a strain collection number of ATCC 700407; the main components of the NA (normal nutrient agar medium) were: 10.0 g peptone, 3.0 g beef extract powder, 5.0 g sodium chloride, 15.0 g agar, and 1000 mL water; the main components of the THB agar medium were: 10 g / L beef powder, 20 g / L tryptone, 2 g / L glucose, 2 g / L sodium chloride, 2 g / L sodium bicarbonate, and 0.4 g / L disodium hydrogen phosphate.

[0071] (2) Equal amounts of the composition for preventing and treating mastitis prepared in Examples 1-3, the sunflower extract prepared in step (1) of Example 1, the ginkgo leaf extract prepared in step (2) of Example 1, and aloe vera gel were respectively placed in sterile test tubes as the test group, and equal amounts of distilled water were placed in sterile test tubes as the blank control group. Each group was water-bathed at 20°C ± 1°C for 5 minutes, and then 1 mL of bacterial suspension was added to each group. The bacterial suspension was prepared by compounding equal volumes of Escherichia coli suspension, Staphylococcus aureus suspension, and Streptococcus uberis suspension. The mixture was mixed and cultured at 37°C. After 2 hours, the live bacteria in each test tube were counted, and the antibacterial rate was calculated. The results are shown in Table 2.

[0072] Inhibition rate (%) = [(total number of colonies in the blank control group - total number of colonies in the test group) / total number of colonies in the blank control group] × 100%

[0073] Table 2 Inhibition rate against Escherichia coli, Staphylococcus aureus and Streptococcus uberis (%)

[0074]

[0075] As can be seen from Table 1, the composition prepared by compounding aloe vera gel with ambrette extract and ginkgo biloba extract has an antibacterial rate of up to 98.58% against Escherichia coli, up to 96.58% against Staphylococcus aureus, and up to 90.56% against Streptococcus uberis. In contrast, the antibacterial rate of ambrette extract alone against Escherichia coli was only 35.58%, the antibacterial rate against Staphylococcus aureus was only 37.47%, and the antibacterial rate against Streptococcus uberis was up to 90.15%. Therefore, it can be seen that the compounding of aloe vera gel, ambrette extract, and ginkgo biloba extract in the present invention can significantly improve the antibacterial rate against Staphylococcus aureus.

[0076] Test Example 2: Mouse Test

[0077] The mouse experiment was conducted with reference to Chapter 5 of the paper "Screening of Traditional Chinese Medicines Against Major Cow Mammary Pathogens and Study on the Mechanism of Action of Aesculetin in Treating Mouse Mastitis" (Zhang Wuhao, Henan Agricultural University), which studies the therapeutic effect of aesculetin on Streptococcus mastitis in mice. The mice were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., and the Staphylococcus aureus was purchased from the China Industrial Microbiology Culture Collection Center, with the strain collection number being CICC 10384. The specific steps are as follows:

[0078] (1) Staphylococcus aureus was inoculated on BP agar plates and cultured at 37°C for 22-24 hours. After enrichment and dilution, the viable count was 1×10 4 CFU / mL of Staphylococcus aureus bacterial solution for later use.

[0079] (2) Female mice 8 days postpartum were anesthetized with ether, and 50 μL of Staphylococcus aureus solution was injected into the milk duct using a microsyringe to establish the mouse model;

[0080] (3) The mice with successful modeling were randomly divided into 5 groups, with 6 mice in each group, namely experimental group 1, experimental group 2, experimental group 3, experimental group 4 and control group. Each group received the following treatments:

[0081] The mice in test group 1 were smeared with the drug prepared in Example 4 on their nipples every day;

[0082] The mice in the experimental group 2 were smeared with the drug prepared in the comparative example 1 on their nipples every day;

[0083] The mice in the experimental group 3 were smeared with the medicine prepared in the comparative example 2 on their nipples every day;

[0084] The mice in the experimental group 4 were smeared with the drug prepared in the comparative example 3 on their nipples every day;

[0085] The mice in the control group were given hydrogen-rich water applied to their nipples daily;

[0086] Apply twice a day at a fixed time for seven consecutive days, with an application amount of 0.01g each time. The feeding management and feeding conditions of each group of mice are kept consistent.

[0087] (4) Seven days after application, the mice were killed and their left and right mammary gland tissues were collected. The mammary gland tissues were homogenized and centrifuged, and the supernatant was collected. The ELISA detection procedure was used to determine the levels of TNF-α, IL-1β, and IL-6. The results are shown in Table 1.

[0088] Table 1 TNF-α, IL-1β and IL-6 levels in each group of mice

[0089]

[0090]

[0091] As can be seen from Table 1, the compound of aloe vera gel, sunflower extract and ginkgo leaf extract in the present invention can significantly reduce the content of TNF-α, IL-1β and IL-6. Therefore, it can be seen that the medicine prepared by the present invention can significantly reduce the inflammatory response of mastitis in mice.

[0092] The above description is merely a preferred embodiment of the present application and is not intended to limit the present application. Various modifications and variations are possible for those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present application shall be included within the scope of protection of the present application.

Claims

1. A composition for preventing and treating mastitis, characterized in that: It is composed of the following raw materials in parts by weight: 20-25 parts aloe vera gel, 6-9 parts sunflower extract, 8-10 parts ginkgo biloba extract; The sunflower extract is prepared by the following method: sunflower powder and 50-70% ethanol by volume are mixed in a mass ratio of 1:(20-40), and ultrasonically extracted 2-3 times at 120-180W, each ultrasonic extraction time being 1-3 hours, to obtain an extract; the extract is concentrated and dried to obtain the sunflower extract; The ginkgo leaf extract is prepared by the following method: mixing ginkgo leaves and 50-70% ethanol by volume in a mass ratio of 1:(5-10), heating and refluxing at 75-85° C. for 1-2 times, with each extraction time being 3-5 hours, to obtain an extract; and concentrating and drying the extract to obtain the ginkgo leaf extract.

2. Use of the composition according to claim 1 in the following (1)-(2): (1) Preparation of drugs for inhibiting Escherichia coli, Staphylococcus aureus and Streptococcus uberis; (2) Preparation of medicines for preventing and treating mastitis.

3. A drug for preventing and treating mastitis, characterized in that: The drug for preventing and treating mastitis is composed of the composition according to claim 1 and hydrogen-rich water; The drug for preventing and treating mastitis is prepared by the following method: the composition according to claim 1 and hydrogen-rich water are uniformly mixed in a mass ratio of 1:(5-10) to obtain the drug for preventing and treating mastitis; wherein the hydrogen content in the hydrogen-rich water is 8ppm-10ppm.

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