CN107496905B

CN107496905B - Traditional Chinese medicine composition for improving reproductive capacity of female animals and preparation method thereof

Info

Publication number: CN107496905B
Application number: CN201710119666.2A
Authority: CN
Inventors: 张洪涛; 王双
Original assignee: Individual
Current assignee: Individual
Priority date: 2017-03-02
Filing date: 2017-03-02
Publication date: 2021-05-04
Anticipated expiration: 2037-03-02
Also published as: CN107496905A

Abstract

The invention relates to the technical field of veterinary drug preparations, in particular to a traditional Chinese medicine composition for improving reproductive capacity of female animals and a preparation method thereof. The traditional Chinese medicine composition is mainly prepared from the following raw materials in parts by weight: 5-10 parts of cecropin antibacterial peptide, 10-20 parts of epimedium herb, 15-20 parts of motherwort herb, 8-15 parts of safflower, 8-15 parts of salvia miltiorrhiza, 15-20 parts of sheep red meal, 5-15 parts of honeysuckle and 2-5 parts of bacillus subtilis. The cecropin antibacterial peptide has broad-spectrum antibacterial performance, is a natural antibiotic, has the effect of bacillus subtilis, repairs endometrial injury and balances endometrial microbial flora; the cecropin, the bacillus subtilis and the traditional Chinese medicine components have synergistic effect, so that the body is promoted to absorb the traditional Chinese medicine components, the traditional Chinese medicine components are promoted to exert the medicine effect, the treatment effect of the traditional Chinese medicine composition on the infertility of female livestock such as endometritis, static ovary, permanent corpus luteum infertility and retained afterbirth is improved, and the conception rate of the female livestock is improved.

Description

Traditional Chinese medicine composition for improving reproductive capacity of female animals and preparation method thereof

Technical Field

The invention relates to the technical field of veterinary drug preparations, in particular to a traditional Chinese medicine composition for improving reproductive capacity of female animals and a preparation method thereof.

Background

With the improvement of living standard of people, the demand of people for meat products such as pork, beef, mutton and the like is increasing, the supply of the meat products must be increased by increasing the number of female animals, and the supply amount of livestock is larger than the demand amount according to the conjecture of the female animals raised at present, but the situation is not so. In recent years, the meat products often have the current situation of short supply and short demand, and the prices of meat products such as pork, beef, mutton and the like are high. In addition to epidemic and nutritional causes, the method is closely related to the reduction of the production capacity of female animals per se. The low production capacity of female animals mainly shows that the low oestrus rate, the low mating rate and the small litter size cause the reduction of the breeding efficiency. The investigation results show that the infertility caused by endometritis accounts for the highest proportion, and then the ovary is still and the corpus luteum lasts.

The breeding cow oestrus cycle is abnormal and the reproductive performance is low when the ovary is in the stationary infertility, and one of the main reasons for causing the cow infertility comprises cow ovary stationary, persistent corpus luteum, ovary stationary, ovarian cyst, alternate follicle development, ovulation delay and other diseases. The incidence of the diseases is high, so that the influence on the cattle raising industry is serious, and the cattle raising industry is concerned at home and abroad. Hollis regularly counts the morbidity of 3172 cows, and the finding shows that 210 cows with ovarian cyst exist within 150 days after delivery, and the morbidity is 6.26%; ahmadjaz et al report that the probability of ovarian cyst in dairy cows is 6-19%; according to the investigation of 6158 cows by northeast agriculture university, 325 cows with ovarian disease have the incidence rate of 5.29%, wherein 124 cows with ovarian insufficiency account for 38.15% of ovarian disease, 45 cows with lasting corpus luteum account for 13.85%, 123 cows with ovarian cyst account for 37.85%, 25 cows with alternate follicular development account for 7.69%, and 8 cows with delayed ovulation account for 2.46%; the research institute of Lanzhou livestock and veterinary drug of Chinese academy of agricultural sciences has investigated 21596 adult cows of 83 dairy farms in provinces and cities, such as Heilongjiang, Tianjin, Jiangsu, Hunan, Gansu, Sichuan and Hebei, and the number of infertile cows caused by ovarian diseases is 8.45%, wherein 555 of the corpus luteum accounts for 30.32% of ovarian diseases, 443 of the ovaries are static, 24.20% of ovarian cysts, 379 of ovarian cysts, 21.69% of ovarian atrophy, 184 of the ovaries, 10.05% of ovarian atrophy, 64 of ovulation delay, 3.50% of ovaries and salpingitis, 70 of the ovaries and salpingitis, 3.82% of ovaries and 135 of other diseases (ovarian tumors, salpingo-scleroses, adhesion and the like), and 7.38% of ovarian and salpingitis. As a result, the infertility caused by ovarian diseases is a common disease and frequently encountered disease of dairy cows.

At present, female animal infertility research in China has achieved good results, but most of the researched treatment medicines are chemical medicines and are relatively high in price, western veterinarians for treating endometritis mainly use antibiotics and antisepsis disinfection medicines to wash and perfuse the uterus, the method is effective, but needs to be used repeatedly, and frequent use of antibiotics can cause certain bacteria to generate drug resistance, so that the treatment effect is worse and worse, the dosage is continuously increased, and long-term use of antibiotics can increase the residual quantity in animal products, thereby bringing about the problem of food safety. In addition, western veterinarians mainly adopt hormone therapy for treating ovary quiescence and persistent corpus luteum, but are difficult to activate expected effects, and hormone residues in animal products are easy to cause and harm to human health.

The hormone medicines clinically applied at present for treating the ovarian disease infertility have the following side effects: the activity condition of the cow ovary is judged according to rectal examination in clinic, subjective misjudgment is easily caused due to different clinical experiences of technical operators, and the hormone of a treatment medicine is selected improperly. Therefore, the regulation disorder of the reproductive hormone of the organism is easily caused, and the female animals are not easy to conceive; in order to improve economic benefit, farmers use hormone therapy for a large number of times, which is very easy to cause harm to female animal bodies, and use hormone therapy for a large number of times, which is very easy to cause hormone residue in meat and milk, thus threatening human health.

In order to overcome the side effects of hormone medicines, medicines of certain traditional Chinese medicine preparations are gradually developed to treat female animal ovarian disease infertility, but the effective traditional Chinese medicine components for treating female animal ovarian disease infertility are single at present, only 2 varieties are collected in two departments of Chinese veterinary pharmacy (2000 th edition), are aphrodisiac perfusate and ancient Chinese herbal compound, and students carry out pharmacological research at present, although the curative effect is exact, the production process is extensive, and the quality standard needs to be continuously improved so as to carry out large-scale production; the traditional Chinese medicines for treating the female livestock ovarian disease infertility have poor overall treatment effect, and have low cure rate and low conception rate for the female livestock ovarian disease infertility.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention aims to provide the traditional Chinese medicine composition for improving the reproductive capacity of the female animals, which can obviously treat the ovarian disease infertility of the female animals, promote the oestrus of the female animals and improve the reproductive capacity of the female animals.

Meanwhile, the invention also provides a preparation method of the traditional Chinese medicine composition for improving the reproductive capacity of the female animals.

In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:

a traditional Chinese medicine composition for improving reproductive capacity of female animals is mainly prepared from the following raw materials in parts by weight: 5-10 parts of cecropin antibacterial peptide, 10-20 parts of epimedium herb, 15-20 parts of motherwort herb, 8-15 parts of safflower, 8-15 parts of salvia miltiorrhiza, 15-20 parts of sheep red meal, 5-15 parts of honeysuckle and 2-5 parts of bacillus subtilis.

Optionally, the dosage form of the traditional Chinese medicine composition is perfusate; the perfusate preparation process also comprises the following raw materials in parts by weight: 10-20 parts of glycerol, 1-5 parts of polyvinylpyrrolidone, 0.15-0.5 part of preservative and 0.01-0.1 part of antioxidant.

The preparation method of the traditional Chinese medicine composition for improving the reproductive capacity of the female animals comprises the following operation steps:

1) extracting the active ingredient extract of the traditional Chinese medicine: weighing herba Epimedii, herba Leonuri, Carthami flos, Saviae Miltiorrhizae radix, sheep Red meal and flos Lonicerae according to weight parts, mixing, pulverizing, and adding supercritical CO₂Extracting with fluid as extractant and mixed solvent of water, ethanol and acetone as entrainer, decompressing, separating, and discharging to obtain extract of effective components of Chinese medicinal materials;

2) preparing a perfusate: taking the effective components of the traditional Chinese medicine prepared in the step 1), adding cecropin, bacillus subtilis, glycerol, polyvinylpyrrolidone, preservative and antioxidant, mixing uniformly, and adding distilled water to a constant volume to obtain perfusate.

Optionally, the specific method for extracting the effective components of the traditional Chinese medicine in the step 1) comprises the following steps:

a: weighing herba Epimedii, herba Leonuri, Carthami flos, Saviae Miltiorrhizae radix, sheep Red meal and flos Lonicerae according to weight parts, mixing, pulverizing, adding into an extractor, introducing supercritical CO into the extractor₂Extracting the fluid and the entrainer for 3-5 hours under the conditions of 30-40 MPa pressure and 40-75 ℃ temperature;

b: after the extraction in the step A is finished, the effective components of the traditional Chinese medicine and the supercritical CO are sequentially treated by a first-stage separator and a second-stage separator₂The fluid is subjected to separation, wherein supercritical CO is adopted₂The fluid returns to the extractor for recycling, the first-stage separator discharges a small amount of traditional Chinese medicine ointment with high viscosity, no traditional Chinese medicine active ingredient is detected, the second-stage separator discharges brown traditional Chinese medicine active ingredient extract, and the separation column discharges clear entrainer liquid; wherein supercritical CO₂The pressure of the fluid flowing through the primary separator for primary separation is 30MPa, and the temperature is 50-75 ℃; flowing through a secondary separatorThe pressure of the secondary separation is 30MPa, and the temperature is 35 ℃.

Optionally, the entrainer is water, ethanol, acetone in a volume ratio of 1: 4-7: 2-5.

Optionally, the mass amount of the entrainer is 5-20% of the total mass of the epimedium herb, the motherwort herb, the safflower, the red sage root, the sheep red meal and the honeysuckle flower.

Optionally, supercritical CO₂The flow rate of the fluid is 15-30L/h.

The invention relates to a Chinese medicinal composition for improving reproductive capacity of female animals, which is a perfusate in a dosage form, can be used for treating infertility of female animals such as endometritis, ovarian quiescence, permanent corpus luteum infertility, retained afterbirth and the like, and improves reproductive capacity of female animals, and comprises the following specific application methods: sterilizing instruments and pudendum of female livestock conventionally, delivering the vas deferens into uterus by a method similar to that of semen deference of female livestock or a holding method of uterus, injecting 100ml of the perfusate of the invention each time, 1 time every day, continuously using for 3 days, and 3 times as 1 course of treatment;

the invention has the beneficial effects that:

1) the invention relates to a traditional Chinese medicine composition for improving reproductive capacity of female animals, which takes cecropin, bacillus subtilis, epimedium, motherwort, safflower, salvia miltiorrhiza, sheep red meal and honeysuckle as main components in a synergistic way, wherein the cecropin has broad-spectrum antibacterial performance, has strong bactericidal action on 12 pathogenic bacteria such as salmonella, escherichia coli, staphylococcus aureus and the like, is a natural antibiotic, is safe and efficient, does not generate drug resistance and has no residual side effect; the cecropin antibacterial peptide and the bacillus subtilis act to repair endometrial injury and balance endometrial microbial flora; the cecropin, the bacillus subtilis and the traditional Chinese medicine components have synergistic effects, so that the body is promoted to absorb the traditional Chinese medicine components, the traditional Chinese medicine components are promoted to exert the medicine effect, the treatment effect of the traditional Chinese medicine composition on infertility of female animals such as endometritis, static ovary, permanent corpus luteum infertility and retained afterbirth is improved, the conception rate of the female animals is improved, and the conception rate is improved to more than 80 percent and even reaches 100 percent. Meanwhile, a manner of perfusate uterine perfusion is further adopted, so that the medicine can exert the maximum efficacy;

2) when most of the traditional Chinese medicines are extracted, a water boiling decoction method is generally adopted, so that the loss of active ingredients of the traditional Chinese medicines is more, and particularly, the waste of water-insoluble ingredients is caused. In order to overcome the defects in the prior art, the invention adopts CO₂The supercritical fluid extraction technology has the advantages that the solution obtained by the supercritical fluid extraction technology has less residue, low toxicity and no pollution, the CO2 supercritical fluid extraction technology can avoid the defects in the traditional preparation process of decocting the traditional Chinese medicines by adding water, and the oxidation and the dissipation of heat-sensitive substances are effectively prevented, so that the curative effect is improved; the SFE can more effectively extract volatile components in complex traditional Chinese medicines, completely retain the biological activity of medicines, extract substances with high boiling point, low volatility and easy pyrolysis at the boiling point temperature of the substances, combine extraction and separation into a whole, and when CO-SF saturated with dissolved substances flows through a separator, CO2 and the extract are rapidly separated into two phases (gas-liquid separation) due to pressure reduction, so that the extraction efficiency is high, the energy consumption is low, the extraction fluid is economical and pollution-free, and the extraction fluid can be recycled, thereby really realizing the greenization of the production process;

3) furthermore, in the process of extracting the effective components of the traditional Chinese medicine, the invention limits the pressure, the temperature and the time of extraction, limits the adoption of two-stage separation, limits the temperature and the pressure of each separation and limits the supercritical fluid CO₂The flow rate ensures that the effective components in the traditional Chinese medicine are extracted to the maximum limit, improves the purity of the effective components of the traditional Chinese medicine, and finally improves the medicine curative effect of the traditional Chinese medicine composition.

Detailed Description

The technical solution of the present invention will be described in detail by specific examples.

Example 1

A Chinese medicinal composition for improving reproductive capacity of female animals is a perfusate, and is prepared from the following raw materials in parts by weight: 8 parts of cecropin antibacterial peptide, 15 parts of epimedium herb, 18 parts of motherwort herb, 12 parts of safflower, 12 parts of salvia miltiorrhiza, 18 parts of sheep red meal, 10 parts of honeysuckle, 4 parts of bacillus subtilis, 15 parts of glycerol, 4 parts of polyvinylpyrrolidone, 0.25 part of preservative and 0.07 part of antioxidant.

The preparation method of the perfusate for improving the reproductive capacity of the female animals comprises the following operation steps:

1) extracting effective components of the traditional Chinese medicine:

a: weighing herba Epimedii, herba Leonuri, Carthami flos, Saviae Miltiorrhizae radix, sheep Red meal and flos Lonicerae according to weight parts, mixing, pulverizing, adding into an extractor, and introducing supercritical CO into the extractor at flow rate of 20L/h₂Extracting fluid and entrainer for 4 hours under the conditions of 35MPa pressure and 55 ℃; wherein the entrainer is water, ethanol and acetone in a volume ratio of 1: 5: 3, the dosage of the mixed solution is 12 percent of the total mass of the epimedium, the motherwort, the safflower, the salvia miltiorrhiza, the sheep red meal and the honeysuckle;

b: after the extraction in the step A is finished, the effective components of the traditional Chinese medicine and the supercritical CO are sequentially treated by a first-stage separator and a second-stage separator₂The fluid is subjected to separation, wherein supercritical CO is adopted₂The fluid returns to the extractor for recycling, the first-stage separator discharges a small amount of traditional Chinese medicine ointment with high viscosity, no traditional Chinese medicine active ingredient is detected, the second-stage separator discharges brown traditional Chinese medicine active ingredient extract, and the separation column discharges clear entrainer liquid; wherein supercritical CO₂The pressure of the fluid flowing through the first-stage separator for primary separation is 30MPa, and the temperature is 62 ℃; the pressure of the secondary separation carried out by the secondary separator is 30MPa, and the temperature is 35 ℃.

2) Preparing a perfusate: taking the effective components of the traditional Chinese medicine prepared in the step 1), adding cecropin, bacillus subtilis, glycerol, PVP, a preservative and an antioxidant, uniformly mixing, and adding distilled water to a constant volume to obtain a perfusate.

Example 2

A Chinese medicinal composition for improving reproductive capacity of female animals is a perfusate, and is prepared from the following raw materials in parts by weight: 5 parts of cecropin antibacterial peptide, 10 parts of epimedium herb, 15 parts of motherwort herb, 8 parts of safflower, 8 parts of salvia miltiorrhiza, 15 parts of sheep red meal, 5 parts of honeysuckle, 2 parts of bacillus subtilis, 10 parts of glycerol, 1 part of polyvinylpyrrolidone, 0.15 part of preservative and 0.0.1 part of antioxidant.

1) extracting effective components of the traditional Chinese medicine:

a: weighing herba Epimedii, herba Leonuri, Carthami flos, Saviae Miltiorrhizae radix, sheep Red meal and flos Lonicerae according to weight parts, mixing, pulverizing, adding into an extractor, and introducing supercritical CO into the extractor at a flow rate of 15L/h₂Extracting fluid and entrainer for 5h under the conditions of 30MPa pressure and 40 ℃; wherein the entrainer is water, ethanol and acetone in a volume ratio of 1: 4: 2, the dosage of the mixed solution is 5 percent of the total mass of the epimedium, the motherwort, the safflower, the red sage root, the sheep red meal and the honeysuckle;

b: after the extraction in the step A is finished, the effective components of the traditional Chinese medicine and the supercritical CO are sequentially treated by a first-stage separator and a second-stage separator₂The fluid is subjected to separation, wherein supercritical CO is adopted₂The fluid returns to the extractor for recycling, the first-stage separator discharges a small amount of traditional Chinese medicine ointment with high viscosity, no traditional Chinese medicine active ingredient is detected, the second-stage separator discharges brown traditional Chinese medicine active ingredient extract, and the separation column discharges clear entrainer liquid; wherein supercritical CO₂The pressure of the fluid flowing through the first-stage separator for primary separation is 30MPa, and the temperature is 50 ℃; the pressure of the secondary separation carried out by the secondary separator is 30MPa, and the temperature is 35 ℃.

Example 3

A Chinese medicinal composition for improving reproductive capacity of female animals is a perfusate, and is prepared from the following raw materials in parts by weight: 10 parts of cecropin antibacterial peptide, 20 parts of epimedium, 20 parts of motherwort, 15 parts of safflower, 15 parts of salvia miltiorrhiza, 20 parts of sheep red meal, 15 parts of honeysuckle, 5 parts of bacillus subtilis, 20 parts of glycerol, 5 parts of polyvinylpyrrolidone, 0.5 part of preservative and 0.1 part of antioxidant.

1) extracting effective components of the traditional Chinese medicine:

a: weighing epimedium, motherwort, safflower,Mixing Saviae Miltiorrhizae radix, YANGSHONG diet and flos Lonicerae, pulverizing, adding into an extractor, and introducing supercritical CO into the extractor at a flow rate of 30L/h₂Extracting fluid and entrainer for 3 hours under the conditions of 40MPa pressure and 75 ℃; wherein the entrainer is water, ethanol and acetone in a volume ratio of 1: 7: 5, the dosage of the mixed solution is 20 percent of the total mass of the epimedium, the motherwort, the safflower, the salvia miltiorrhiza, the sheep red meal and the honeysuckle;

b: after the extraction in the step A is finished, the effective components of the traditional Chinese medicine and the supercritical CO are sequentially treated by a first-stage separator and a second-stage separator₂The fluid is subjected to separation, wherein supercritical CO is adopted₂The fluid returns to the extractor for recycling, the first-stage separator discharges a small amount of traditional Chinese medicine ointment with high viscosity, no traditional Chinese medicine active ingredient is detected, the second-stage separator discharges brown traditional Chinese medicine active ingredient extract, and the separation column discharges clear entrainer liquid; wherein supercritical CO₂The pressure of the fluid flowing through the first-stage separator for primary separation is 30MPa, and the temperature is 75 ℃; the pressure of the secondary separation carried out by the secondary separator is 30MPa, and the temperature is 35 ℃.

Comparative example 1

This comparative example is different from example 1 in that cecropin antimicrobial peptide in example 1 is replaced with mammalian antimicrobial peptide, and the other example is the same as example 1.

Comparative example 2

This comparative example is different from example 1 in that cecropin in example 1 was replaced with fish antimicrobial peptide, and the same example 1 was repeated.

Comparative example 3

The comparison example is different from the example 1 in that cecropin and bacillus subtilis in the example 1 are omitted, and the dosage of the effective components of the traditional Chinese medicine is adjusted to be equal to the sum of the dosage of the cecropin, the bacillus subtilis and the effective components of the traditional Chinese medicine in the example 1.

Comparative example 4

The comparative example is different from example 1 in that Bacillus subtilis and Chinese medicinal components are omitted, and the dosage of cecropin is adjusted to be equal to the sum of the dosage of cecropin, Bacillus subtilis and Chinese medicinal effective components in example 1.

Comparative example 5

The difference between the comparative example and the example 1 is that cecropin and traditional Chinese medicine components are omitted, and the dosage of the bacillus subtilis is adjusted to be equal to the sum of the dosage of the cecropin, the bacillus subtilis and the traditional Chinese medicine active components in the example 1.

Test example 1

1. Experiment site

Test animals were selected from Rongsen pasture (cattle farm) in the double city of Harbin, and Borui genetics Ltd (cattle farm) in the double city of Harbin.

2. Materials and methods

2.1 test animals

And randomly selecting test cows meeting the test conditions.

2.2 test milk cow inclusion criteria

Bringing the endometritis-affected cows into the standard: through detection and observation, the cow is found to have high body temperature, reduced appetite and listlessness; abnormal purulent gray secretion flows out of the vulva when the waist is bowed and the vulva is blamed, and sometimes blood is mixed and odor is generated; the rectum is used for checking the swelling and the poor sensitivity or the pain of the uterus.

Bringing the cow suffering from infertility into the standard: three continuous oestrus cycles have no or no obvious oestrus, and the oestrus is disordered or the mating of the three oestrus cycles is carried out after the oestrus but the pregnancy is not realized.

Inclusion criteria of cows with retained afterbirth: the placenta of the cow is not completely discharged 12 hours after delivery.

2.3 test drugs

The perfusion solution prepared in the example 1, the perfusion solutions prepared in the comparative examples 1-5 and the Aufulongjimycin injection.

2.4 test grouping and handling

Randomly selecting 70 cows with mild endometritis and 10 cows with severe endometritis, wherein the 70 cows with mild endometritis are randomly divided into 7 groups, each group comprises 10 cows, 6 groups of cows with mild endometritis are respectively perfused with the perfusates prepared in the example 1 and the perfusates prepared in the comparative examples 1-5 in a one-to-one correspondence manner, and the cows with severe endometritis are perfused with the perfusate prepared in the example 1; the using amount of the perfusates prepared in the example 1 and the comparative examples 1 to 5 is 100 ml/time/day, and the perfusates are used for 3 days continuously; the remaining group is perfused with the Aufulongotmycin injection for uterus perfusion, the dosage is 100 ml/time/day, and the interval between two times of medication is one day;

randomly selecting 70 cows with infertility, randomly dividing into 7 groups, and correspondingly perfusing the perfusates prepared in example 1 and comparative examples 1-5 respectively, wherein the dosage of the perfusates is 100 ml/time/day, and the continuous use lasts for 3 days; the remaining group is perfused with the Aufulongotmycin injection for uterus perfusion, the dosage is 100 ml/time/day, and the interval between two times of medication is one day;

randomly selecting 70 cows with postpartum retained afterbirth, and randomly dividing the cows into 7 groups, wherein 6 groups are respectively perfused with the perfusates prepared in the example 1 and the comparative examples 1-5 in a one-to-one correspondence manner, and the using amount is 100 ml/time/day and is continuously used for 3 days; the remaining group was infused with the Aufulongtumycin injection for uterine perfusion at a dose of 100 ml/time/day with a one-day interval between two administrations.

2.5 assessment of therapeutic Effect

The treatment effect is divided into a curative effect and an ineffective effect:

and (3) curing: the recovery standard of the endometritis of the milk cow is as follows: the uterus inflammation disappears after the medicine is used, the uterus of the cattle in palpation has no lump and no pain reaction, abnormal secretion is discharged in the uterus, the estrus is obvious, and the cattle is pregnant in 3 estrus after artificial fertilization.

And the infertility rehabilitation standard of the dairy cattle: after the medicine is taken, the cow is in normal estrus and is fertilized in 3 estrus after artificial fertilization.

The recovery standard of cow placenta is as follows: after the medicine is taken, the placenta of the cow is completely discharged.

And (4) invalidation: after the medicine is taken, the uterus symptoms are not obviously changed, and no oestrus phenomenon occurs in 3 oestrus periods.

3. And (5) testing results.

TABLE 1 statistics of the clinical efficacy of the perfusates and the Maculopsis chinensis prepared in example 1 and comparative examples 1-5 on the treatment of endometritis in cows

TABLE 2 statistics of the clinical efficacy of the perfusates prepared in example 1 and comparative examples 1-5 and the Auofilan for the treatment of cow infertility

Group of

Example 1

Comparative example 1

Comparative example 2

Comparative example 3

Comparative example 4

Comparative example 5

Maculon (Maculon)

Conception rate

76.92％

45％

42％

40％

42％

2％

40％

TABLE 3 statistics of the clinical efficacy of the perfusates and the Syngnathus prepared in example 1 and comparative examples 1-5 for treating retained fetal membranes in dairy cows

Group of

Example 1

Comparative example 1

Comparative example 2

Comparative example 3

Comparative example 4

Comparative example 5

Maculon (Maculon)

High efficiency

100％

40％

45％

25％

52％

2％

50％

4. Analysis of results

As can be seen from the data shown in tables 1, 2 and 3, the perfusate prepared in the embodiment 1 of the present invention has significant therapeutic effects on mild and severe endometritis, infertility and retained afterbirth of dairy cows, and the therapeutic effects are significantly better than those of the comparative examples 1 to 5, and the pharmaceutical efficacy of the perfusate prepared in the embodiment 1 of the present invention is better than that of the comparative examples 1 to 5 by analyzing the clinical aspects such as uterine recovery, ovarian activity, corpus luteum, oestrus and conception, so that the present invention adopts cecropin and bacillus subtilis to combine the traditional Chinese medicine components as the main preparation raw materials of the perfusate, so as to generate synergistic interaction between the cecropin and the bacillus subtilis, and improve the pharmaceutical efficacy of the perfusate. The perfusion liquid level pure traditional Chinese medicine green product prepared by the invention is safe, has obvious treatment effect and does not have the problem of medicine residue.

5. Conclusion

The estrus-promoting pregnancy-promoting perfusion liquor is used for 3 days and 3 times of a treatment course according to 100 ml/time/day, and has obvious treatment effects on the endometritis, the infertility and the retained afterbirth of the dairy cow.

Test example 2

1. Test site

Harbin city Xussen animal husbandry

2. Test materials and methods

2.1 test animals

Test cows were randomly selected according to the test requirements.

2.2 Experimental drugs

The perfusate prepared in example 1; the perfusate prepared in comparative example 3; qianggong (24 g/zhi containing chlorhexidine acetate), supplied by ruicheng weierfu veterinary drug limited, production lot 0812284, gongjing oil (compound traditional Chinese medicine, 200 ml/bottle), supplied by hangzhou xinyuanshan pharmaceutical industry limited, production lot: 0902294.

2.3 test milk cow inclusion standard

3. Test grouping and processing

3.1 clinical trial grouping of cows with light and severe endometritis

Randomly selecting 40 cows with mild endometritis, randomly dividing into 4 groups, and perfusing the cow uterus with the perfusate prepared in example 1, the perfusate prepared in comparative example 3, Qianggong and uterus oil in each group in one-to-one correspondence; wherein the use amount of the perfusates prepared in the example 1 and the comparative example 3 is 100 ml/time/day, and 3 days is a treatment course; the dosage of the strong uterine health-care product is one dose every time, once every two days, and 4 times of perfusion; the usage amount of the cervical oil is 100 ml/time/day, and 3 days is a treatment course;

randomly selecting 40 cows with severe endometritis, randomly dividing into 4 groups, and perfusing the cow uterus with the perfusate prepared in example 1, the perfusate prepared in comparative example 3, Qianggong and uterus oil in each group in one-to-one correspondence; wherein the use amount of the perfusates prepared in the example 1 and the comparative example 3 is 100 ml/time/day, and 3 days is a treatment course; the dosage of the strong uterine health-care product is one dose every time, once every two days, and 3 times of infusion; the usage amount of the cervical oil is 100 ml/time/day, and 3 days is a treatment course;

3.2 clinical trial grouping of infertile cows

Randomly selecting 80 frequently-matched infertile cows, randomly dividing into 4 groups, each group having 20 heads, and carrying out uterus perfusion on the frequently-matched infertile cows before the test cows are matched with the perfusate prepared in the example 1, the perfusate prepared in the comparative example 3, the Qianggong and the uterine cleansing oil one by one; wherein the use amount of the perfusates prepared in the example 1 and the comparative example 3 is 100 ml/time/day, and 3 days is a treatment course; the dosage of the strong uterine health-care product is one dose every time, once every two days, and 4 times of perfusion; the usage amount of the cervical oil is 100 ml/time/day, and 3 days is a treatment course;

3.3 grouping of postpartum retained fetal membranes in clinical trials of cows

Randomly selecting 80 postpartum cows without placenta, and randomly dividing into 4 groups, wherein the uterus of each group is perfused with the perfusate prepared in example 1, the perfusate prepared in comparative example 3, Qianglongkang and Gongjing oil one by one; wherein the use amount of the perfusates prepared in the example 1 and the comparative example 3 is 100 ml/time/day, and 3 days is a treatment course; the dosage of the strong uterine health-care product is one dose every time, once every two days, and 4 times of perfusion; the application amount of cervical oil is 100 ml/time/day, and 3 days is a treatment course.

4. Treatment outcome assessment

The treatment effect is divided into cure and ineffective.

And (3) curing: the recovery standard of the endometritis of the milk cow is as follows: the uterus inflammation disappears after the medicine is used, the palpation cattle uterus has no lump and no pain reaction, abnormal secretion is discharged in the uterus, the estrus is obvious, and the artificial insemination is performed in 3 estrus.

5. Test results

TABLE 4 statistics of clinical effects of different drugs on treatment of endometritis in cows

TABLE 5 statistics of clinical efficacy of different drugs for treating cow infertility

TABLE 6 statistics of clinical effects of different drugs on retained afterbirth of dairy cow

6. Test results

As can be seen from the data shown in tables 4 to 6, the perfusate prepared in example 1 of the present invention has better curative effects on reproductive diseases such as cow endometritis, infertility and retained afterbirth than the pure traditional Chinese medicine perfusate prepared in comparative example 3, and the clinical effects of the perfusate prepared in example 1 are better as the clinical aspects of uterine recovery, ovarian activity, corpus luteum, oestrus, conception and the like are analyzed due to the commercially available strong uterus rehabilitation and uterus oil.

7. Conclusion

The cecropin and the bacillus subtilis are added into the perfusate, so that the damage to the endometrium of a dairy cow can be repaired, the microbial flora of the endometrium is balanced, and the cecropin has a strong sterilization effect on 12 pathogenic bacteria such as salmonella, escherichia coli, staphylococcus aureus and the like.

Test example 3

In order to verify the treatment effect of the perfusate on the endometritis and the infertility of sows, clinical tests are carried out in an ecological breeding park in Qingcheng mountain of Koelken county, Kouyang city, and the pregnancy-promoting perfusate is divided into a high group and a low group according to different medicament dosages to carry out curative effect comparison so as to determine the treatment effect and the curative effect of the perfusate on the endometritis and the infertility of sows.

1. Materials and methods

1.1 test animals

Selecting sow with endometritis and infertility.

1.2 test drugs

Perfusate prepared in example 1.

1.3 inclusion criteria of test sows

The inclusion standard of the sow with the endometritis is as follows: 5-7 days after the sow is born, mucus or purulent secretion with fishy smell flows out of the vulva of the sow, and the mucus or purulent secretion is white, grey red and yellow.

1.4 test grouping and handling

Randomly selecting 40 sows with severe endometritis, randomly dividing into two groups, wherein each group has 20 sows, and carrying out uterine perfusion on one group of sows according to the dose of 100 ml/time/day for 3 days; the other group carries out uterine perfusion on sows according to the dosage of 50 ml/time/day for 3 days;

randomly selecting 40 sows with mild endometritis, randomly dividing into two groups, wherein each group comprises 20 sows, and carrying out uterine perfusion on one group of sows according to the dose of 100 ml/time/day for 3 days; the other group carries out uterine perfusion on sows according to the dosage of 50 ml/time/day for 3 days;

randomly selecting 40 sows suffering from infertility, randomly dividing the sows into two groups, wherein each group comprises 20 sows, and carrying out uterine perfusion on the sows in a dose of 100 ml/time/day for 3 days; the other group was administered to sows at a dose of 50 ml/time/day for 3 consecutive days.

Note: in order to bring the drug into full contact with the endometrium during the test, the low dose group used 50ml of drug in combination with 50ml of 0.9% saline.

1.5 assessment of therapeutic Effect

The treatment effect is divided into healing and ineffectiveness.

And (3) curing: endometritis recovery criteria: the lochia disappears, and the uterus recovers normally; the amount of lochia is reduced, the viscosity is reduced, and the lochia is light white or egg white like lochia appears; the clinical symptoms were not reduced and the amount of lochia was not reduced or increased to be ineffective.

The infertility recovery standard is as follows: the sow has normal oestrus and can be pregnant after artificial insemination.

And (4) invalidation: the clinical symptoms were not alleviated, the amount of lochia was not reduced or increased, and there was no normal oestrus.

2. Test results

TABLE 7 statistics of clinical efficacy of high and low doses of the perfusate prepared in example 1 for treating endometritis of sows

TABLE 8 statistics of clinical efficacy of high and low doses of the perfusate prepared in example 1 for treating endometritis of sows

4. Analysis of results

The data shown in the tables 7 and 8 show that the groups with high and low dosages of the perfusate have remarkable effects of treating the endometritis and the infertility of the sows, and the treatment of the groups with high and low dosages of the estrus-promoting and pregnancy-promoting perfusate can achieve obvious treatment effects by analyzing the clinical aspects of uterine secretion, oestrus, conception and the like.

5. Conclusion

The perfusate is used for 3 days continuously according to 100 ml/time/day, 3 times are a treatment course, 50ml of the aphrodisiac pregnancy-promoting perfusate and 50ml of 0.9 percent physiological saline are used for 3 days continuously; the two dosage have obvious clinical use effect on treating the endometritis and the infertility of the sows.

Test example 4

In order to verify the treatment effect of the perfusate on the endometritis and the infertility of the sows, clinical tests are carried out on Xinye county and Xinye county farms in Nanyang city, and the clinical effects and the treatment effects of the perfusate on the endometritis and the infertility of the sows are determined by comparing the two groups of treatment effects of the perfusate prepared in the embodiment 1 and the perfusate prepared in the comparative example 3 according to high and low doses with the clinical effects of similar products in the market.

1 materials and methods

1.1 test animals

Selecting test sow meeting test conditions

1.2 test drugs

Test drug the perfusate prepared in example 1 was tested; the perfusate prepared in comparative example 3; gongyanjing, provided by the pharmaceutical industry of agricultural animals in Hunan province; the commercial pregnancy-promoting perfusion fluid is provided by Jiangxi Borai pharmaceutical industry.

1.3 inclusion criteria of test sows

1.4 test grouping and handling

1.41 clinical trial group of sows with light and severe endometritis

Randomly selecting 80 sows with mild endometritis, randomly dividing into 4 groups, and correspondingly perfusing the perfusate prepared in example 1, the perfusate prepared in comparative example 3, GONGYANJING and commercial pregnancy-promoting perfusate in every 20 groups; the use amounts of the perfusate prepared in example 1, the perfusate prepared in comparative example 3 and the commercial pregnancy-promoting perfusate are 100 ml/time/day for continuous 3 days for the perfusion of the uterus of the sow; the GONGYANJING is used for uterus perfusion of test sow, 100 ml/time/day, 1 time every two days, and 3 times;

randomly selecting 80 sows with severe endometritis, randomly dividing into 4 groups, and correspondingly perfusing the perfusate prepared in example 1, the perfusate prepared in comparative example 3, GONGYANJING and commercial pregnancy-promoting perfusate in every 20 groups; the use amount of the perfusate prepared in the example 1, the perfusate prepared in the comparative example 3 and the commercial pregnancy-promoting perfusate is that the uterus of the sow is perfused by 50 ml/time/day for 3 days; the GONGYANJING is used for uterus perfusion of test sow, 50 ml/time/day, 1 time every two days, and 3 times;

1.42 clinical trial groups for sows with infertility

Randomly selecting 80 barrenness sows, randomly dividing into 4 groups, and correspondingly perfusing the perfusate prepared in the example 1, the perfusate prepared in the comparative example 3, GONGYANJING and the commercial pregnancy promoting perfusate in every 20 groups; the use amounts of the perfusate prepared in example 1, the perfusate prepared in comparative example 3 and the commercial pregnancy-promoting perfusate are 100 ml/time/day for continuous 3 days for the perfusion of the uterus of the sow; the GONGYANJING is used for uterus perfusion of test sow, 100 ml/time/day, 1 time every two days, and 3 times;

randomly selecting 80 barrenness sows, randomly dividing into 4 groups, and correspondingly perfusing the perfusate prepared in the example 1, the perfusate prepared in the comparative example 3, GONGYANJING and the commercial pregnancy promoting perfusate in every 20 groups; the use amount of the perfusate prepared in the example 1, the perfusate prepared in the comparative example 3 and the commercial pregnancy-promoting perfusate is 50 ml/time/day for continuous use for 3 days; gongyanjing is used for uterine perfusion of test sows, 50 ml/time/day, 1 time every two days and 3 times of medicine application.

Note: in order to achieve adequate contact between the drug and the endometrium during the test, the low dose group used 50ml of drug in combination with 50ml of 0.9% saline

2. Evaluation of test Effect

The treatment effect is divided into healing and ineffectiveness.

3. Test results

TABLE 9 statistics of clinical efficacy of high-dose different drugs for treating endometritis of sows

TABLE 10 statistics of clinical efficacy of low-dose different drugs for treating endometritis of sows

TABLE 11 clinical curative effect statistics of high-dose different drugs for treating sow infertility

TABLE 12 clinical curative effect statistics of low-dose different drugs for treating sow infertility

4. Results

The data shown in the tables 9-12 show that the perfusate prepared in the embodiment 1 has obvious effect on treating the endometritis and the infertility of the sows in the high and low dose groups, the treatment effect of the perfusate prepared in the embodiment 1 is obviously better than that of the pure traditional Chinese medicine perfusate and the commercially available pregnancy promoting perfusate prepared in the comparative example 3 and the commercially available metritis-eliminating medicines by analyzing the clinical aspects of uterine secretion, oestrus, conception and the like and combining the treatment effective rate on diseases,

5. analysis of

The two using doses of the perfusate in the embodiment 1 of the invention are 100 ml/time/day and 50 ml/time/day for treating the endometritis of the sow and the retention of the fetal membranes after delivery, and researches prove that the perfusate has obvious effect of treating the reproductive diseases of the sow, good curative effect, short course of treatment, time and labor saving, and better clinical effect than a control product; the result shows that the cecropin antibacterial peptide and the bacillus subtilis are added into the perfusate, so that the endometrium injury can be repaired, the endometrial microbial flora can be balanced, and the cecropin antibacterial peptide has a strong bactericidal effect on 12 pathogenic bacteria such as salmonella, escherichia coli, staphylococcus aureus and the like; the Chinese herbal medicines, the cecropin and the bacillus subtilis are used for dual treatment, so that the synergistic effect is achieved, and the treatment effect of the medicine is improved.

Note: experiments prove that the treatment effect of the perfusates prepared in the examples 2 and 3 on the diseases such as endometritis, infertility and retained fetal membranes of female animals is basically equal to that of the perfusate prepared in the example 1.

Claims

1. A traditional Chinese medicine composition for improving reproductive capacity of female animals is characterized by being mainly prepared from the following raw materials in parts by weight: 5-10 parts of cecropin antibacterial peptide, 10-20 parts of epimedium herb, 15-20 parts of motherwort herb, 8-15 parts of safflower, 8-15 parts of salvia miltiorrhiza, 15-20 parts of sheep red meal, 5-15 parts of honeysuckle and 2-5 parts of bacillus subtilis; the dosage form of the traditional Chinese medicine composition is perfusate; the perfusate preparation process also comprises the following raw materials in parts by weight: 10-20 parts of glycerol, 1-5 parts of polyvinylpyrrolidone, 0.15-0.5 part of preservative and 0.01-0.1 part of antioxidant; the preparation method of the traditional Chinese medicine composition for improving reproductive capacity of female animals comprises the following operation steps:

1) extracting the active ingredient extract of the traditional Chinese medicine: A. weighing herba Epimedii, herba Leonuri, Carthami flos, Saviae Miltiorrhizae radix, sheep Red meal and flos Lonicerae according to weight parts, mixing, pulverizing, adding into an extractor, and extracting with supercritical CO₂The fluid is an extracting agent, and the volume ratio of water, ethanol and acetone is 1: 4-7: 2-5 mixed solution is used as entrainer, and supercritical CO is introduced into the extractor₂A fluid and an entrainer, wherein the entrainer,extracting for 3-5 h under the conditions of 30-40 MPa pressure and 40-75 ℃ temperature; B. after the extraction in the step A is finished, the effective components of the traditional Chinese medicine and the supercritical CO are sequentially treated by a first-stage separator and a second-stage separator₂The fluid is subjected to separation, wherein supercritical CO is adopted₂The fluid returns to the extractor for recycling, the first-stage separator discharges a small amount of traditional Chinese medicine ointment with high viscosity, no traditional Chinese medicine active ingredient is detected, the second-stage separator discharges brown traditional Chinese medicine active ingredient extract, and the separation column discharges clear entrainer liquid; wherein supercritical CO₂The pressure of the fluid flowing through the primary separator for primary separation is 30MPa, and the temperature is 50-75 ℃; the pressure of the secondary separation of the mixture flowing through the secondary separator is 30MPa, and the temperature is 35 ℃; the entrainer is 5-20% of the total mass of the epimedium herb, the motherwort herb, the safflower, the red sage root, the sheep red meal and the honeysuckle flower; the supercritical CO₂The flow rate of the fluid is 15-30L/h;