KR19990084250A - Composition for preventing and treating cow mastitis - Google Patents

Abstract

The present invention relates to a composition for the prevention and treatment of cow mastitis, the composition is 1 to 5% by weight of chitosan, 0.01 to 0.1% by weight sodium benzoate, 0.1 to 1% by weight allantoin, 0.1 to 0.3% by weight sodium propionate, water-soluble It is characterized by consisting of a base and the remainder of distilled water.

The composition according to the present invention is excellent in the prevention and treatment of mastitis and safe for humans and animals, and does not require a separate period of absence, and shows effects on acute and chronic mastitis and subclinical mastitis as well as latent mastitis. It is also effective in mastitis and fungal mastitis caused by various bacteria that have become resistant to antibiotics and chemotherapeutic agents.

Description

Composition for preventing and treating cow mastitis

The present invention relates to a composition for the prevention and treatment of cow mastitis, in particular chitosan, sodium benzoate, allantoin and the like as an active ingredient, and excellent in the prevention and treatment effect for mastitis while safe to humans and animals separate period of absence The present invention relates to a composition for preventing and treating cow mastitis that does not require milk.

Mastitis that occurs in cows is an inflammation of the mammary gland tissue caused by various microbial infections distributed in nature. It shows symptoms such as decreased flow rate, lowering of oil, fever, and loss of appetite depending on the degree of symptoms. Not only do they lose their function, they are also contagious and are in danger of dying. The decrease in flow and foreclosure due to mastitis is directly related to the economic loss of dairy farmers, and is especially significant under the oil-based differential payment system (June 1, 1993 in Korea).

Mastitis is largely divided into clinical, subclinical, chronic mastitis, and latent mastitis.

In most cases, clinical mastitis develops from subclinical or chronic mastitis, and inflammatory symptoms such as swelling, hot flashes, redness, pain, metaphoric disorder, and breast necrosis are clearly identified with the naked eye. In addition, clinical mastitis can be divided into subacute mastitis, acute mastitis, and acute mastitis. Subacute mastitis has been found to have pus in milk squeezed as part of the preparation for milking. Acute mastitis has edema, fever, redness, pain and Other causes of dysfunction and abnormal squeezing of the breast with wet naked eyes are also confirmed. Acute mastitis also causes systemic fever, chills, loss of appetite, and general weakness in the form of mastitis with rapid and severe clinical symptoms.

Subclinical mastitis is the most common form of mastitis in cows, and pathogenic bacteria invade and proliferate in the mammary gland, but cannot be confirmed by visual milk test. Subclinical mastitis can be diagnosed by physicochemical testing of the California Mastitis Test (CMT) or by laboratory tests by milk culture or by somatic cell counting. Not only will it cause economic losses, but it is also likely to develop into a clinical form.

Chronic mastitis refers to mastitis, a condition in which preclinical infections last for a long time or continue to the next inorganic period, and are characterized by irregular repeated symptoms such as sudden subacute or acute exacerbation and recovery to subclinical conditions. There is this.

Latent mastitis is a condition before pathogenic bacteria invade the mammary gland and cause inflammatory changes in the breast.

The causative agents of mastitis range from Gram-positive bacteria such as Staphylococcus, Streptococcus and Corynebacterium, to Gram-negative bacteria such as Escherichia coli, Pneumococcus pneumoniae, and Pseudomonas aeruginosa, and to bacteria, fungi and yeasts such as Mycobacteria, Mycoplasma and Bacillus.

In the conventional mastitis treatment, various kinds of antibiotics and chemotherapeutic agents have been commonly used, and most of them have been applied for dry and non-organic treatments, and mainly ointment drugs have been injected into the breast through nipples to treat them.

However, these conventional antibiotics or chemotherapeutic agents continue to be used, causing resistance to various mastitis causative organisms, and thus not only exhibiting efficacy but also affecting normal flora in the breast. Therefore, the administration of such antibiotics or chemotherapy agents causes a decrease in breast resistance and changes in the microflora in the breast, in particular in the bacterial flora. Recently, many kinds of fungal infections, including Aspergillus and Candida, are occurring. Duplicate infection with bacteria, such as Streptococcus, causes mastitis to progress to incurable condition. However, existing antibiotics and chemotherapeutic agents are almost insensitive to fungi, so new treatments have been required for fungal mastitis, and there is a need for drugs that are simultaneously sensitive to bacteria and fungi.

In addition, conventional antibiotics and chemotherapeutic agents have been required to set a separate drug holiday period after administration due to drug residues in crude milk and cow's milk when they are administered in the non-organic period. It has been a loss.

In addition, since the conventional ointment mastitis treatment agent mainly used oily substances as a base, oil may float in the milk, and thus there is a problem in that the loss of milk is also large.

The present invention is excellent in the prevention and treatment of mastitis in order to solve the problem of mastitis treatment of cows by conventional antibiotics and chemotherapeutic agents, while safe for human body and animals, it can be used without distinction between inorganic and dry season To provide a mastitis prevention and treatment composition.

In addition, the present invention is to provide a prophylactic and therapeutic composition for mastitis, which has an effect on acute and chronic mastitis and subclinical mastitis as well as latent mastitis.

In addition, the present invention is to provide a mastitis preventive and therapeutic composition that is effective in fungal mastitis as well as bacteria that have developed resistance to various antibiotics and chemotherapeutic agents.

To this end, the present invention is composed of chitosan, which is safe for humans and animals and has antibacterial action, and sodium benzoate, which enhances the action of chitosan, allantoin, which promotes healing of wounds, and the like, as a main component thereof, thereby making up the components in an appropriate ratio. It provides a composition that acts red to synergistically to exhibit an excellent effect in the prevention and treatment of mastitis.

The prophylactic and therapeutic composition for cow mastitis according to the present invention is characterized in that it contains chitosan, sodium benzoate, allantoin, sodium propionate and the like. The preferable composition of this composition is as follows.

1-5% by weight of chitosan,

0.01-0.1 wt% sodium benzoate,

Allantoin 0.1-1% by weight,

0.1-0.3 wt% sodium propionate,

0-0.02% by weight of triprenamine hydrochloride,

Water soluble base,

Balance of distilled water.

(100 ml total)

Chitosan is a natural polymer (β-poly-D-glucosamine) made by deacetylation of chitin (β-poly-N-acetyl-D-glucosamine) extracted from crustaceans such as crabs. . Chitin is a kind of nitrogen-containing polysaccharide obtained by removing protein from the crab shell, and is widely present in nature. However, it is insoluble in aqueous solution, which is a barrier to utilization. On the other hand, chitosan obtained by deacetylation treatment such as heating chitin in sodium hydroxide is widely used because it is easily used because it shows water solubility in weakly acidic liquid. That is, chitosan is safe for human body, so it is used for additives such as foods, surgical sutures, artificial blood vessels, artificial skins, medical materials, cosmetics, etc. Also, when hydrolyzed chitosan with enzymes, it becomes Ν-acetyl oligosaccharide, resulting in an immune responsive effect. It is used as a health food because it shows the effect of lowering cholesterol and the like, in addition, it is usefully used in the field of biotechnology, agricultural, chemical, environmental, such as immobilized carrier of microorganisms.

Chitosan in the composition replaces antibiotics or chemotherapeutic agents to prevent and treat mastitis by antibacterial action. In addition, chitosan has a wound healing promoting action to help the healing of the inflammatory site to speed up the healing, and immune boosting action, such as increased phagocytes and interferon to help the healing and prevention of the disease.

As is known from various studies and literature, chitosan is a safe ingredient for the human body, so when the present composition containing chitosan as an active ingredient is used for treating mastitis of cows, unlike a conventional antibiotic or chemotherapeutic agent, no need.

The content of chitosan for showing the proper antibacterial action is preferably 1 to 5% by weight in the present composition.

Sodium benzoate in the composition is a drug that is commonly used as a preservative of the drug because it has an antiseptic effect, in the composition plays a more important role to increase the action of the chitosan described above. The preferred composition ratio of sodium benzoate for synergy with chitosan is 0.01 to 0.1% by weight in the composition.

Allantoin (all-uratin hydantoin) in the composition is a substance produced by the decomposition of uric acid by uricase, and allantoin is synergistic with the above-mentioned chitosan because it has a function of promoting wound healing to revive cells of wound tissue. It acts as a further promoting the repair of inflammatory tissues. The preferred content of allantoin for synergy with chitosan is 0.1 to 1% by weight in the composition.

Sodium propionate in the composition is an antifungal substance that acts to inhibit fungi that cause mastitis. Therefore, the present composition exhibits an effective effect on mastitis caused by fungi by containing sodium propionate as an active ingredient together with chitosan. The preferred content of sodium propionate for antifungal action is 0.1 to 0.3% by weight in the composition.

In the present composition, an appropriate amount of antihistamine may be added for vasoconstriction, secretion suppression and sedation of the inflammation site. As the antihistamines used in the composition, all of the conventional antihistamines, which have been proved to be effective and safe in humans and animals, may be used, and preferably, 0.005 to 0.02% by weight of triprenamine hydrochloride.

In addition, a small amount of a steroidal anti-inflammatory agent may be added to the present composition to assist the anti-inflammatory action. As a steroidal anti-inflammatory agent, a conventional steroidal anti-inflammatory agent that has been proved to be effective and safe for humans and animals may be used, and preferably, dexamethasone sodium phosphate may be used in a small amount of 0.005 to 0.05% by weight.

As the base of the composition, various bases suitable for infusion administration through nipples, including water-soluble ointment, may be used. However, as mentioned above, oil-based bases such as petroleum jelly, vegetable oil, lard, etc. are not preferable because oil may be oiled. Considering the mode of use of the present composition usually injected and the possibility of remaining in crude oil, propylene glycol can be preferably used as the base of the present composition.

In addition, the present composition may include formulation aids such as dissolution aids, moisturizers, and diluents necessary for formulation into pharmaceuticals, as necessary.

The composition according to the present invention, each of the above-mentioned components complementary or synergistically combined to exhibit an excellent effect on the prevention and treatment of cow mastitis while safe for humans and animals.

The composition is usually prepared by the following method.

(a) 1 to 5 parts by weight of chitosan was dissolved in an acidic aqueous solution and then heat-treated at 80 to 120 ° C. for 20 to 60 minutes;

(b) 0.01 to 0.1 parts by weight of sodium benzoate, 0.1 to 1 part by weight of allantoin and 0.1 to 0.3 parts by weight of sodium propionate are dissolved in an alkaline aqueous solution and then mixed with the heat treated chitosan solution obtained in (a);

(c) A suitable amount of the water-soluble base and distilled water are equally mixed with the mixture obtained in (b), and the liquidity of the mixed solution is adjusted to pH 5.8 to 6.8 to prepare a composition according to the present invention.

In addition, the present composition is preferably filled and packaged in an injector in a single dose unit for ease of use. PH of this composition manufactured by the above method is about 5.8-6.8 normally.

The composition according to the invention can be administered to the cow by a variety of drug administration methods commonly performed on animals, preferably by injection into the quarters of the nipples suffering from mastitis. The dosage is preferably 10ml (100-500mg in chitosan content) twice a day once per minute when injected into the nipple, and the dose may be increased or decreased depending on the symptoms and the cow.

The present invention will be described in more detail with reference to the following Examples. However, the following examples are not intended to limit the scope of the present invention, it is a matter of course that can be changed by those skilled in the art within the scope of the technical idea of the present invention.

Example 1 (Manufacturing Example)

(Iii) 200 ml of distilled water was added to the preparation tank, and 21.3 g of chitosan was added and stirred. Separately, 3.5 ml of HCl and 2.5 ml of acetic acid were added to 71.5 ml of distilled water and added to the chitosan under stirring. It is in a state. The temperature of the preparation tank was raised to 100 degreeC, and 50 minutes of heat processing was made into A liquid.

At this time, chitosan was used by purchasing FL80 of Koyo Chemical Co. Ltd. The product properties of the used chitosan are as follows.

Particle Size: 80 mesh

Loss on drying: 6.5%

Viscosity: 7.5 cps

Deacetylation degree: 86.5%

(Ii) 17.5 ml of 2 Mol NaOH was added to 50 ml of sterile distilled water, and 2 g of allantoin, 0.6 g of sodium benzoate, 0.08 g of trifereneamine hydrochloride, and 3 g of sodium propionate were dissolved to prepare B solution.

(Iii) After cooling A liquid to 30 degreeC, 5 ml of 10% NaOH was added and mixed with stirring, and after standing still for 20 minutes, B liquid was mixed with A liquid, stirring. 50 g of propylene glycol and 600 ml of distilled water were added thereto, followed by mixing well to obtain 1,000 ml of a pale yellow brown liquid which is a composition according to the present invention.

(Iv) 10 ml of the liquid composition obtained in the above (iv) was packed and packed in a disposable injector.

Example 2

Susceptibility testing for mastitis-derived causative organisms

The composition prepared according to Example 1 was used for the susceptibility test against mastitis-derived bacteria of the composition according to the present invention. A total of 253 milk cows with mastitis were tested. Strains were used as a total of 253 strains isolated from milk powder. The test method was disk diffusion. The test results are shown in Table 1 below.

Table 1

Mastitis Number of Tests Susceptibility Results (%) Staphylococcus aureus Coagulase negative Staphylococci Strep. dysgalactiaeMicrococcus sp.Gram positive organism 1033728843313 45 (43.6) 20 (54.0) 13 (41.0) 6 (75.0) 24 (55.8) 12 (38.7) 2 (66.6) Sum 253 122 (48.2)

As a result, it was found that the present composition exhibited high sensitivity of 50% or higher for Gram-positive bacteria, micrococcus and yeast, and about 40% sensitivity to Staphylococcus aureus, Streptococci and Gram-negative bacteria.

Example 3

Minimal Growth Inhibition Test for Mastitis-causing Bacteria

After the composition was prepared in the same manner as in Example 1, the composition was 15 mg / ml, 7.5 mg / ml, 3.75 mg / ml, 1.87 mg / ml, 0.93 mg / ml, 0.46 mg / Diluted to ㎖, 0.23 mg / ㎖ was tested for the minimum growth inhibitory concentration against the causative agent of mastitis. The test results are shown in Table 2.

TABLE 2

Strain Minimum growth inhibitory concentration (mg / ml) 15 7.5 3.75 1.87 0.93 0.46 0.23 Sta. aureus - - - - - + + Sta. epidermis - - - - - + + Strep. agalactiae - - - - + + + Strep. dysgalactiae - - - - + + + E. coli - - - - + + + Proteus sp. - - - - - + + Enterobacter sp. - - - - - + + Bacilli sp. - - - - + + + Yeast - - - - + + +

(-: Growth inhibition, +: growth)

As a result, there was a slight difference depending on the strain, but it was confirmed that most strains were inhibited at a concentration of 1.87 mg / ml.

Example 4

Efficacy test for mastitis of cows

In order to test the efficacy against mastitis, the effect of somatic cell number reduction and somatic cell number distribution was investigated after administration of the composition according to the present invention. When mastitis occurs, a large amount of somatic cells are detected in milk due to inflammation of the mammary gland tissue. The method of diagnosing mastitis by measuring the somatic cell count is called somatic cell counting.

A total of 56 cows of 14 cows from 6 ranches were injected and injected into the composition prepared according to Example 1 through nipples twice a day after milking. Somatic cell counts were measured at 7 days and 14 days after the start of administration.

(1) Study on the effect of reducing somatic cell count

TABLE 3

(Mean ± S.E)

Neck sheet Experiment head Minute Somatic cell count (× 1,000) Before administration 7 days after the start of administration 14 days after start of administration ABCDEF 686866 9999911 2,067 ± 4782,070 ± 4664,383 ± 8713,139 ± 7862,887 ± 7583,519 ± 896 1,024 ± 2911,358 ± 5612,143 ± 6811,869 ± 6271,643 ± 6445,187 ± 997 596 ± 2161,085 ± 1581,393 ± 7311,532 ± 5921,468 ± 7062,644 ± 722 Sum 40 56 3,029 ± 310 2,310 ± 337 ^** (76.2) ^* 1,496 ± 245 ^** (49.3)

^* Somatic cell count after administration / Somatic cell count before administration × 100

^** Significant difference between 7 and 14 days after administration

(P <0.05).

(2) Survey of Somatic Cell Number Distribution

Table 4

Somatic cell count Positive stimulus (%) Before dosing (n = 56) 7 days after the start of administration (n = 56) 14 days after start of administration (n = 56) Less than 500,000 1 (1.7) 7 (12.5) 18 (32.1) 50 to 99 million 8 (14.2) 19 (33.9) 14 (25.0) 1 to 3 million 26 (46.4) 18 (32.1) 16 (28.5) More than 3 million 21 (37.5) 12 (21.4) 8 (14.2)

As a result of the test, when the composition according to the present invention is administered to the mammary glands, significant somatic cell number reduction effect was observed (P <0.05). In other words, there was a slight difference in each ranch, but showed a decrease rate of about 23.7% at 7 days after administration and a high reduction rate of about 50.6% at 14 days after administration.

Example 5

Safety test

In order to test the safety of the composition according to the present invention, the rats were continuously injected with twice the usual dosage and observed clinical symptoms such as papillomatosis. Samples were prepared using the composition prepared according to Example 1, and 10 cows were injected and injected into the teat twice daily with 300 mg of chitosan content. The clinical symptoms of the cow were observed during the administration period and one week after the end of the administration period.

Table 5

division object Dosage Route of administration Duration of administration Experiment head Clinical observation (2 weeks) Test group milk cow 300mg × 2 / day Intramammary injection 2 weeks 10 normal

Observational results showed no clinical changes such as papillomatous swelling in any cow, and clinical signs showed that the composition was safe for cows.

The composition according to the present invention is excellent in the prevention and treatment effect for the mastitis of cows, but has no side effects on humans and animals, and excellent safety does not require a separate drug holiday. In addition, the composition according to the present invention is effective in acute and chronic mastitis and subclinical mastitis as well as latent mastitis, and has a composition that is completely different from conventional therapeutic agents, resulting in resistance to various antibiotics and chemotherapeutic agents. It can also be effective for mastitis caused by and fungal mastitis, which has recently been a problem.

Claims (6)

A composition for the prevention and treatment of cow mastitis consisting of 1 to 5% by weight of chitosan, 0.01 to 0.1% by weight of sodium benzoate, 0.1 to 1% by weight of allantoin, 0.1 to 0.3% by weight of sodium propionate, a water-soluble base and the remainder of distilled water. The composition of claim 1 wherein the water soluble base is propylene glycol. The composition of claim 1, wherein the composition comprises 2 wt% chitosan, 0.06 wt% sodium benzoate, 0.2 wt% allantoin, 0.3 wt% sodium propionate, 5 wt% propylene glycol and the remainder of distilled water. The composition according to claim 1 or 3, wherein the composition further comprises 0.005 to 0.02% by weight of triprenamine hydrochloride. The composition of claim 1 or 3, wherein the composition further comprises 0.005 to 0.05% by weight of dexamethasone sodium phosphate. (a) 1 to 5 parts by weight of chitosan was dissolved in an acidic aqueous solution and then heat-treated at 80 to 120 ° C. for 20 to 60 minutes; (b) 0.01 to 0.1 parts by weight of sodium benzoate, 0.1 to 1 part by weight of allantoin and 0.1 to 0.3 parts by weight of sodium propionate are dissolved in an alkaline aqueous solution and then mixed with the heat treated chitosan solution obtained in (a); (c) A method for producing a prophylactic and therapeutic composition for cow mastitis, comprising mixing the water-soluble base and distilled water in an appropriate amount equally with the mixture obtained in (b) and adjusting the liquidity of the mixed solution to pH 5.8 to 6.8.