CN101703601A - Chinese medicinal composition for preventing and treating Mycoplasma gallisepticum and preparation method thereof - Google Patents
Chinese medicinal composition for preventing and treating Mycoplasma gallisepticum and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a Chinese medicinal composition for preventing and treating Mycoplasma gallisepticum and a preparation method thereof. The Chinese medicinal composition is prepared from the following raw material medicaments in part by weight: 1 to 4 parts of baical skullcap root, 1 to 4 parts of honeysuckle, 1 to 4 parts of tatarian aster root, 1 to 4 parts of ephedra herb, 0.2 to 1.5 parts of tangerine peel and 0.2 to 1.5 parts of liquoric root. The Chinese medicinal composition has the effects of clearing heat and releasing toxin, moistening the lung to stop cough, relieving cough and removing asthma, eliminating inflammation and relieving pain, improving immunity and the like, has simple preparation process, low cost, no toxic or side effect, and is not easy to generate medicament resistance. Various active components in the oral liquid of the Chinese medicinal composition are dispersed in a medium in a molecule or microparticle state, have high dispersity, are quickly absorbed, and can quickly begin to work. Clinical pharmacodynamic experiment results show that the Chinese medicinal composition has good treatment effect on chronic respiratory disease and the like caused by the Mycoplasma gallisepticum infection, and can also improve the weight increment of chickens and fodder returns.
Description
Technical field
This aspect relates to a kind of Chinese medicine composition for animals, relates in particular to the Chinese medicine composition of a kind of prevention and treatment Mycoplasma gallisepticum, belongs to field of traditional Chinese veterinary.
Background technology
(Mycoplasma gallisepticum MG) often causes the chronic respiratory tract disease of chicken to Mycoplasma gallisepticum, and this is a kind of chronic infectious disease of height contact, and clinical is principal character with abnormal breathing sound, cough, stream nose liquid, dyspnea etc.This disease is clinical to be chronic process, and the course of disease is long, and normal again and other cause of disease mixed infections cause the respiratory symptom aggravation, and mortality rate increases; And because of the infected chicken growth promoter is obstructed, the discarded rate of the defect ware of trunk improves, and laying rate reduces, and causes serious threat to poultry husbandry.Primary disease all can take place throughout the year, and is comparatively serious with cold season, the sickness rate height.Spread all over all over the world at present, also generally infect China various places, has become the focus that researcher is paid close attention in recent years.
At present still undesirable to the vaccine development of Mycoplasma gallisepticum, only can provide limited protection, Drug therapy is an important means of control primary disease.Each foster fowl country has all done a large amount of work in the world, but because mycoplasma easily produces problems such as toleration to medicine, and therefore how using efficient, medicine low toxicity is the key of treatment primary disease.The acellular wall of mycoplasma, penicillin, sulfa drugs are failed to respond to any medical treatment to it, and medicines such as fluoroquinolones, Macrolide and Tetracyclines commonly used, but long-term prescription all has the problem of Resistant strain, and can not thoroughly eradicate the mycoplasma that has infected in the body, in case drug withdrawal, disease also can be recurred.The tradition herbal medicine has very big advantage improving clinical symptoms, alleviate bacterial drug resistance and reduce aspects such as toxic and side effects.
It is with the Therapeutic Principle of Chinese medicine strengthening vital QI to eliminate pathogenic factors and the theory of modern medicine overall regulation and control that Chinese veterinarian treats this disease, select the medicine prescription in conjunction with characteristics of incidence and pathogen characteristic, treatment safety, untoward reaction is light, toxic and side effects is low, cause of disease is difficult for producing drug resistance, basic noresidue or seldom residual in meat, egg food.Simultaneously, herbal medicine has aboundresources, cheap advantage.Therefore, develop and develop Chinese herbal medicine and preparation energetically,, ensure that human body health is significant, also have good using value and good market prospect promoting the development of poultry husbandry.Many based on powder at this sick Chinese herbal medicine dosage form in the market, liquid preparation temporarily is blank.The effective ingredient of liquid preparation Chinese medicine is dispersed in the medium with molecule or graininess, and dispersion is big, absorbs soon, can bring into play drug effect rapidly.Therefore, using liquid preparation of Chinese medicine to carry out the sick body conditioning has great importance in the control primary disease.
Summary of the invention
The object of the present invention is to provide a kind of have prevention and treatment Mycoplasma gallisepticum safe and effective, toxic and side effects is low, the pure Chinese medicine composition of noresidue, promotion chicken weightening finish.
The present invention also aims to provide a kind of and produce anti-Mycoplasma gallisepticum method for compositions by Chinese crude drug.
The objective of the invention is to be achieved through the following technical solutions:
The Chinese medicine composition of a kind of prevention and treatment Mycoplasma gallisepticum comprises that following bulk drugs makes:
Radix Scutellariae 1-4 part, Flos Lonicerae 1-4 part, Radix Asteris 1-4 part, Herba Ephedrae 1-4 part, Pericarpium Citri Reticulatae 0.2-1.5 part and Radix Glycyrrhizae 0.2-1.5 part;
Preferably, the weight portion of each crude drug is:
1 part in 2 parts of Radix Scutellariaes, 2 parts of Flos Loniceraes, 2 parts of Radix Asteriss, 2 parts in Herba Ephedrae, 1 part of Pericarpium Citri Reticulatae and Radix Glycyrrhizae;
The pure Chinese medicinal preparation that the Chinese medicine composition of the anti-Mycoplasma gallisepticum of the present invention is made up of kinds of traditional Chinese medicines such as Radix Scutellariae, Flos Lonicerae, Radix Asteriss based on heat-clearing and toxic substances removing, is aided with nourishing the lung to arrest cough.The contrary gas of lung falls in lung heat clearing pyretic toxicity bitter cold, protects the multiple gold of cloudy Tianjin Gan Run respectful clearly of lung, then coughs, disease knowing such as heating remove.
The used crude drug of the present invention all can be bought from common Chinese material shop and obtain, and its specification meets national Chinese crude drug standard.
Chinese medicine composition of the present invention can add various adjuvants required when preparing different dosage form, for example disintegrating agent, lubricant, adhesive etc., herbal medicine formulation method with routine is prepared into any oral formulations commonly used, wherein, described oral formulations can be oral liquid, powder or granule etc., is preferably oral liquid.
Preferably, a kind of method for preparing Chinese medicine composition of the present invention may further comprise the steps:
(1) takes by weighing each crude drug by described weight portion;
(2) Flos Lonicerae, Radix Asteris, Herba Ephedrae, Pericarpium Citri Reticulatae and Radix Glycyrrhizae are decocted with water, decoction liquor is filtered, and concentrates, and adds ethanol, and is static, spends the night;
(3) filter when going precipitation, filtrate to be concentrated into nothing alcohol flavor, cooling adds ethanol, spends the night;
(4) precipitation is gone in filtration, and filtrate concentrates, and obtains medicinal liquid 1, and is standby;
(5) baikal skullcap root decoction pieces is decocted with boiling water and 0.5% tween 80, filter;
(6) filtrate concentrates, and is static with 1～2 back insulation of dilute hydrochloric acid adjust pH, filters taking precipitate 20-95% washing with alcohol; Precipitation adds the water mixing, adjusts pH value and makes the baicalin dissolving, and the centrifugal precipitation of going is got supernatant, obtains medicinal liquid 2, and is standby;
(7) medicinal liquid 1 and medicinal liquid 2 are mixed, concentrate, adjust pH to 5.5～6.5 add antiseptic, and are canned behind the mixing, seal, and sterilization, promptly.
In the above-mentioned preparation method, decocting with water described in the step (2) is preferably and decocts with water 2 times, adds 10 times of weight decoctings for the first time and boils 2h, and wherein Flos Lonicerae is put into after decocting 1h, adds 10 times of weight decoctings for the second time and boils 1.5h;
Preferably, in the step (2) filtrate is concentrated into 4 times of crude drug weight, adds final concentration and be 65% ethanol, static, 4 ℃ are spent the night;
Preferably, the adding final concentration is 75% ethanol in the step (3), and 4 ℃ are spent the night;
In the step (4), preferred, it is 1g/mL that filtrate being concentrated into contained the crude drug amount, obtains medicinal liquid 1, standby.
In the step (5), preferred, baikal skullcap root decoction pieces is decocted 2 times with boiling water and 0.5% tween 80, add 10 times of weight decoctings for the first time and boil 2h, add 8 times of weight decoctings for the second time and boil 1h, gradation filters while hot, merging filtrate;
In the step (6), preferred, filtrate is concentrated into relative density 1.0 backs and transfers pH1～2 back insulation 1.5h with 2mol/L hydrochloric acid under 80 ℃, static 3h filters; With precipitate is 50%, 75% and 90% washing with alcohol with final concentration respectively; Precipitation adds the water mixing of 1 times of weight of Radix Scutellariae crude drug, transfers pH8 to make the baicalin dissolving with 0.1mol/L NaOH solution;
In the step (7), medicinal liquid 1 and medicinal liquid 2 are mixed, be concentrated into 1g/mL.
The experiment proved that the anti-Mycoplasma gallisepticum Chinese medicine composition of the present invention has following advantage or beneficial effect:
1. the anti-Mycoplasma gallisepticum Chinese medicine composition of the present invention is to make Chinese medicine preparation by Chinese medicine extract, has the incomparable advantages of medicine such as antibiotic, chemosynthesis antibacterial, and antibacterial is difficult for producing drug resistance, is difficult for producing drug residue in meat, the egg.
2. the anti-Mycoplasma gallisepticum Chinese medicine composition of the present invention is a liquid preparation, and the effective ingredient of its Chinese medicine is dispersed in the medium with molecule or graininess, and dispersion is big, absorbs soon, can bring into play drug effect rapidly.
3. the anti-Mycoplasma gallisepticum oral liquid of the present invention just can effectively suppress the growth of Mycoplasma gallisepticum when in vitro tests proves its concentration at 15.625mg/mL.
4. the anti-Mycoplasma gallisepticum Chinese medicine composition of the present invention is aided with nourishing the lung to arrest cough based on heat-clearing and toxic substances removing.The contrary gas of lung falls in lung heat clearing pyretic toxicity bitter cold, protects the multiple gold of cloudy Tianjin Gan Run respectful clearly of lung, then coughs, disease knowing such as heating remove.The main pharmacological experiment relevant with Mycoplasma gallisepticum treatment proves that the anti-Mycoplasma gallisepticum Chinese medicine composition of the present invention has good analgesic, analgesia, antiinflammatory, cough-relieving, reduces phlegm and antiasthmatic effect, can alleviate the respiratory symptom that Mycoplasma gallisepticum causes effectively.
5. the anti-Mycoplasma gallisepticum Chinese medicine composition of the present invention is certain to the therapeutic effect of tentative manual-induced Mycoplasma gallisepticum, cure rate and effective percentage significantly are better than infecting matched group, and can obviously reduce the serum antibody response positive rate, reduce the air bag damage ratio and improve rate of body weight gain.The anti-Mycoplasma gallisepticum Chinese medicine composition of the present invention can suppress the mycoplasma growth on the one hand, has effects such as nourishing the lung to arrest cough, heat-clearing and toxic substances removing, antalgic and inflammation relieving on the other hand.All medicines cooperate, the performance heat-clearing and toxic substances removing, and the effect of nourishing the lung to arrest cough, treating both the principal and secondary aspects of a disease reaches the effect of preventing and treating Mycoplasma gallisepticum.
6. the anti-Mycoplasma gallisepticum Chinese medicine composition of the present invention is safe and reliable, judgement foreign compound toxicity grading standard according to the United Nations's health organization recommendation, the anti-Mycoplasma gallisepticum Chinese medicine composition of the present invention is to the true border of chickling non-toxic compound, and the result of long term toxicity test shows that Chinese medicine composition of the present invention do not see tangible toxicity.
The usage of Chinese medicine composition of the present invention and consumption; Individual treatment: the chicken that clinical symptoms is arranged is dripped clothes, every day 2 times, 3～5 droplets/time.Colony's prevention: every 100ml converts water 200kg, concentrates dispensing, divides twice dispensing of upper and lower noon, cuts off the water before the dispensing 1 hour, drinks logotype 3～5 days 2～3 hours at every turn.
The specific embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage of the present invention and characteristics will be more clear along with description.But these embodiment only are exemplary, scope of the present invention are not constituted any restriction.It will be understood by those skilled in the art that and down can make amendment or replace without departing from the spirit and scope of the present invention, but these modifications and replacing all fall within the scope of protection of the present invention the details of technical solution of the present invention and form.
Embodiment 1
(1) takes by weighing each raw material (unit: g): Radix Scutellariae 200, Flos Lonicerae 200, Radix Asteris 200, Herba Ephedrae 200, Pericarpium Citri Reticulatae 100, Radix Glycyrrhizae 100 by following weight;
(2) Flos Lonicerae, Radix Asteris, Herba Ephedrae, Pericarpium Citri Reticulatae and Radix Glycyrrhizae are decocted with water 2 times, add 10 times of weight decoctings for the first time and boil 2h, wherein Flos Lonicerae is put into after decocting 1h, adds 10 times of weight decoctings for the second time and boils 1.5h, filters merging filtrate.Filtrate is concentrated into about 4 times of crude drug weight, adds final concentration and be 65% ethanol, static, 4 ℃ are spent the night.Precipitation is gone in filtration, and filtrate is concentrated into when not having the alcohol flavor, and cooling adds final concentration and be 75% ethanol, and 4 ℃ are spent the night.Precipitation is gone in filtration, and filtrate being concentrated into contains the crude drug amount and be 1g/mL, and be standby.Baikal skullcap root decoction pieces is decocted 2 times with boiling water and 0.5% tween 80 (adding tween 80 to its final concentration in boiling water is 0.5%), for the first time add 10 times of weight decoctings and boil 2h, for the second time add 8 times of weight decoctings and boil 1h, gradation filters while hot, merging filtrate, under 80 ℃ filtrate is concentrated into relative density 1.0, and with being incubated 1.5h behind 2mol/L hydrochloric acid accent pH1～2, static 3h, filter, precipitate is 50% with final concentration respectively, 75% and 90% ethanol is washed, and precipitation adds the water mixing of 1 times of weight of Radix Scutellariae crude drug, with making the baicalin dissolving about 0.1mol/L NaOH solution accent pH8, the centrifugal precipitation of going.2 kinds of medicinal liquids of above preparation are mixed, be concentrated into 1g/mL, transfer pH5.5～6.5, add 0.15% sodium benzoate and make antiseptic, canned behind the mixing, seal flowing steam sterilization 30min.
Embodiment 2
(1) takes by weighing each raw material (unit: g): Radix Scutellariae 100, Flos Lonicerae 100, Radix Asteris 100, Herba Ephedrae 100, Pericarpium Citri Reticulatae 20, Radix Glycyrrhizae 20 by following weight;
(2) Flos Lonicerae, Radix Asteris, Herba Ephedrae, Pericarpium Citri Reticulatae and Radix Glycyrrhizae are decocted with water 2 times, add 10 times of weight decoctings for the first time and boil 2h, wherein Flos Lonicerae is put into after decocting 1h, adds 10 times of weight decoctings for the second time and boils 1.5h, filters merging filtrate.Filtrate is concentrated into about 4 times of crude drug weight, adds final concentration and be 65% ethanol, static, 4 ℃ are spent the night.Precipitation is gone in filtration, and filtrate is concentrated into when not having the alcohol flavor, and cooling adds final concentration and be 75% ethanol, and 4 ℃ are spent the night.Precipitation is gone in filtration, and filtrate being concentrated into contains the crude drug amount and be 1g/mL, and be standby.Baikal skullcap root decoction pieces is decocted 2 times with boiling water and 0.5% tween 80 (adding tween 80 to its final concentration in boiling water is 0.5%), for the first time add 10 times of weight decoctings and boil 2h, for the second time add 8 times of weight decoctings and boil 1h, gradation filters while hot, merging filtrate, under 80 ℃ filtrate is concentrated into relative density 1.0, and with being incubated 1.5h behind 2mol/L hydrochloric acid accent pH1～2, static 3h, filter, precipitate is 50% with final concentration respectively, 75% and 90% ethanol is washed, and precipitation adds the water mixing of 1 times of weight of Radix Scutellariae crude drug, with making the baicalin dissolving about 0.1mol/L NaOH solution accent pH8, the centrifugal precipitation of going.2 kinds of medicinal liquids of above preparation are mixed, be concentrated into 1g/mL, transfer pH5.5～6.5, add 0.15% sodium benzoate and make antiseptic, canned behind the mixing, seal flowing steam sterilization 30min.
Embodiment 3
(1) takes by weighing each raw material (unit: g): Radix Scutellariae 400, Flos Lonicerae 400, Radix Asteris 400, Herba Ephedrae 400, Pericarpium Citri Reticulatae 150, Radix Glycyrrhizae 150 by following weight;
(2) Flos Lonicerae, Radix Asteris, Herba Ephedrae, Pericarpium Citri Reticulatae and Radix Glycyrrhizae are decocted with water 2 times, add 10 times of weight decoctings for the first time and boil 2h, wherein Flos Lonicerae is put into after decocting 1h, adds 10 times of weight decoctings for the second time and boils 1.5h, filters merging filtrate.Filtrate is concentrated into about 4 times of crude drug weight, adds final concentration and be 65% ethanol, static, 4 ℃ are spent the night.Precipitation is gone in filtration, and filtrate is concentrated into when not having the alcohol flavor, and cooling adds final concentration and be 75% ethanol, and 4 ℃ are spent the night.Precipitation is gone in filtration, and filtrate being concentrated into contains the crude drug amount and be 1g/mL, and be standby.Baikal skullcap root decoction pieces is decocted 2 times with boiling water and 0.5% tween 80 (adding tween 80 to its final concentration in boiling water is 0.5%), for the first time add 10 times of weight decoctings and boil 2h, for the second time add 8 times of weight decoctings and boil 1h, gradation filters while hot, merging filtrate, under 80 ℃ filtrate is concentrated into relative density 1.0, and with being incubated 1.5h behind 2mol/L hydrochloric acid accent pH1～2, static 3h, filter, precipitate is 50% with final concentration respectively, 75% and 90% ethanol is washed, and precipitation adds the water mixing of 1 times of weight of Radix Scutellariae crude drug, with making the baicalin dissolving about 0.1mol/L NaOH solution accent pH8, the centrifugal precipitation of going.2 kinds of medicinal liquids of above preparation are mixed, be concentrated into 1g/mL, transfer pH5.5～6.5, add 0.15% sodium benzoate and make antiseptic, canned behind the mixing, seal flowing steam sterilization 30min.
Embodiment 4
(1) takes by weighing each raw material (unit: g): Radix Scutellariae 100, Flos Lonicerae 100, Radix Asteris 100, Herba Ephedrae 100, Pericarpium Citri Reticulatae 150, Radix Glycyrrhizae 150 by following weight;
(2) Flos Lonicerae, Radix Asteris, Herba Ephedrae, Pericarpium Citri Reticulatae and Radix Glycyrrhizae are decocted with water 2 times, add 10 times of weight decoctings for the first time and boil 2h, wherein Flos Lonicerae is put into after decocting 1h, adds 10 times of weight decoctings for the second time and boils 1.5h, filters merging filtrate.Filtrate is concentrated into about 4 times of crude drug weight, adds final concentration and be 65% ethanol, static, 4 ℃ are spent the night.Precipitation is gone in filtration, and filtrate is concentrated into when not having the alcohol flavor, and cooling adds final concentration and be 75% ethanol, and 4 ℃ are spent the night.Precipitation is gone in filtration, and filtrate being concentrated into contains the crude drug amount and be 1g/mL, and be standby.Baikal skullcap root decoction pieces is decocted 2 times with boiling water and 0.5% tween 80 (adding tween 80 to its final concentration in boiling water is 0.5%), for the first time add 10 times of weight decoctings and boil 2h, for the second time add 8 times of weight decoctings and boil 1h, gradation filters while hot, merging filtrate, under 80 ℃ filtrate is concentrated into relative density 1.0, and with being incubated 1.5h behind 2mol/L hydrochloric acid accent pH1～2, static 3h, filter, precipitate is 50% with final concentration respectively, 75% and 90% ethanol is washed, and precipitation adds the water mixing of 1 times of weight of Radix Scutellariae crude drug, with making the baicalin dissolving about 0.1mol/L NaOH solution accent pH8, the centrifugal precipitation of going.2 kinds of medicinal liquids of above preparation are mixed, be concentrated into 1g/mL, transfer pH5.5～6.5, add 0.15% sodium benzoate and make antiseptic, canned behind the mixing, seal flowing steam sterilization 30min.
Embodiment 5
(1) takes by weighing each raw material (unit: g): Radix Scutellariae 400, Flos Lonicerae 300, Radix Asteris 300, Herba Ephedrae 200, Pericarpium Citri Reticulatae 20, Radix Glycyrrhizae 20 by following weight;
(2) Flos Lonicerae, Radix Asteris, Herba Ephedrae, Pericarpium Citri Reticulatae and Radix Glycyrrhizae are decocted with water 2 times, add 10 times of weight decoctings for the first time and boil 2h, wherein Flos Lonicerae is put into after decocting 1h, adds 10 times of weight decoctings for the second time and boils 1.5h, filters merging filtrate.Filtrate is concentrated into about 4 times of crude drug weight, adds final concentration and be 65% ethanol, static, 4 ℃ are spent the night.Precipitation is gone in filtration, and filtrate is concentrated into when not having the alcohol flavor, and cooling adds final concentration and be 75% ethanol, and 4 ℃ are spent the night.Precipitation is gone in filtration, and filtrate being concentrated into contains the crude drug amount and be 1g/mL, and be standby.Baikal skullcap root decoction pieces is decocted 2 times with boiling water and 0.5% tween 80 (adding tween 80 to its final concentration in boiling water is 0.5%), for the first time add 10 times of weight decoctings and boil 2h, for the second time add 8 times of weight decoctings and boil 1h, gradation filters while hot, merging filtrate, under 80 ℃ filtrate is concentrated into relative density 1.0, and with being incubated 1.5h behind 2mol/L hydrochloric acid accent pH1～2, static 3h, filter, precipitate is 50% with final concentration respectively, 75% and 90% ethanol is washed, and precipitation adds the water mixing of 1 times of weight of Radix Scutellariae crude drug, with making the baicalin dissolving about 0.1mol/L NaOH solution accent pH8, the centrifugal precipitation of going.2 kinds of medicinal liquids of above preparation are mixed, be concentrated into 1g/mL, transfer pH5.5～6.5, add 0.15% sodium benzoate and make antiseptic, canned behind the mixing, seal flowing steam sterilization 30min.
The outer anti-Mycoplasma gallisepticum activity test of test example 1 antibody
Adopt the anti-Mycoplasma gallisepticum activity of test tube doubling dilution research Chinese medicine compound oral liquid of the present invention.Before the test oral liquid of the present invention (embodiment 1 is prepared) is diluted to 500mg/mL concentration, standby after the filtration sterilization, get 24h and can in the FM-4 culture medium, reach the Mycoplasma gallisepticum culture on logarithmic growth peak through 10
4Doubly the dilution back is as the bacterium liquid of this test usefulness.250.000,125.000,62.500,31.250,15.625,7.812,3.906,1.953,0.977,0.489mg/mL get 10 test tubes, final liquor strength is followed successively by in each pipe:, add the bacterium liquid 0.5mL after the dilution successively in each pipe.Each test repeats 3 times, and establishes blank, culture contrast and the culture medium contrast of medicinal liquid.Test tube adds 37 ℃ of cultivations of plug, treat control tube bacterium liquid color from red stain to yellow (pH7.7 changes to 6.8 below), when control medium change color does not take place, write down the pipe number of generation change color.The result shows, when the drug level of compound preparation in the culture fluid reaches 15.625mg/mL, just can effectively suppress the growth of Mycoplasma gallisepticum.
Analgesic, analgesia, antiinflammatory, the cough-relieving of test example 2 oral liquids of the present invention, reduce phlegm and the antiasthmatic effect test
In order to verify that better oral liquid of the present invention has the effect of anti-Mycoplasma gallisepticum, the present invention has carried out and the relevant main pharmacological experiment of this disease treatment.
(1) to the refrigeration function of heating rat due to the yeast
Adopt yeast pyrogenicity method to make rat fever, select for use healthy and 48 of rat that temperature taking is qualified (body weight 180 ± 10g) is divided into the high, medium and low dosage group of oral liquid of the present invention (embodiment 2 is prepared) (be respectively 2,1,0.5mL/100g body weight), aspirin matched group (20mg/100g body weight), normal control group and model control group at random.Press 1mL/100g subcutaneous injection 20% yeast suspension, normal control group injection equivalent normal saline.Body temperature begins to rise behind the injection yeast 3h, and the animal that behind the 4h body temperature rise is lower than 0.8 ℃ is rejected.4h and 5h intragastrically respectively are administered once by above-mentioned dosage behind the injection yeast.2h begins surveying record body temperature value behind the injection yeast, surveys once until 8h every 1h, and calculates the body temperature changing value.The result shows, test rat 2h body temperature behind the injection yeast slightly descends, and begins to rise to 3h body temperature, and 6h peaks, and shows that inhibitory action that oral liquid height of the present invention, middle dosage group raise to the heating rat temperature all clearly.Results of statistical analysis shows, oral liquid height, middle dosage group to the refrigeration function of heating rat due to the yeast and model control group relatively, difference is extremely significantly (p＜0.01) all, low dose group and model control group comparison, significant difference (p＜0.05).Illustrate that product of the present invention has refrigeration function.
(2) Dichlorodiphenyl Acetate causes the analgesic activity of mouse writhing reaction
Adopt the acetic acid twisting method that male white mouse is carried out the analgesic activity experimentation.Test is divided into the high, medium and low dosage group of oral liquid of the present invention (embodiment 3 is prepared) (be respectively 0.2,0.1,0.05mL/10g body weight), aspirin matched group (2mg/10g body weight), normal control group and model control group.All administration treated animals are by the continuous gastric infusion 3d of above-mentioned dosage, once a day, after the last administration, all test group animals except that the normal control group, inject 1% acetum 0.3mL at the 45min pneumoretroperitoneum, the writhing response of mice (hind leg stretches, and abdominal part shrinks, indent, and buttocks is raised) number of times in the opening entry 10min behind the 5min.Calculate this medicine analgesic percentage rate.
The result shows: inject 1% acetic acid and can make mice at short notice owing to writhing response appears in pain, positive control medicine aspirin can be alleviated the mouse writhing reaction that acetic acid causes.By irritating after stomach gives oral liquid, the body number of times of turning round that each dosage group mice occurs in 10min obviously reduces, and this analgesic activity is strengthened along with the increase of drug dose, and the analgesia rate of high, medium and low three dosage groups is respectively 72.2%, 67.1% and 34.6%.Results of statistical analysis shows, high, medium and low three the dosage groups of oral liquid of the present invention are compared with model control group to the inhibitory action of mouse writhing reaction, difference is extremely significantly (p＜0.01) all, and wherein high, middle dosage group is compared with the aspirin matched group, and difference is remarkable (p＞0.05) not.Show that oral liquid of the present invention has analgesic activity.
(3) to the antiinflammatory action of mice caused by dimethylbenzene xylene dropsy of ear
Adopt caused by dimethylbenzene xylene dropsy of ear method to carry out the antiinflammatory action experiment of mice.Get 60 of mices, body weight 20 ± 2g is divided into the high, medium and low dosage group of oral liquid (embodiment 4 is prepared) (be respectively 0.2,0.1,0.05mL/10g body weight), aspirin matched group (2mg/10g body weight), normal control group and model control group.All administration treated animals are by the continuous gastric infusion 3d of above-mentioned dosage, once a day, behind the last gastric infusion 1h, with microsyringe dimethylbenzene 0.05mL/ only evenly is applied to the mouse right ear two sides, left side ear is not coated with as blank, behind the 4h dislocation of mice cervical vertebra is caused death, and cuts ears along the auricle baseline, lay circular auricle in left and right ear same area respectively with the 6mm card punch, on ten thousand/analytical balance, weigh.Difference with left and right ear is the swelling degree.
The result shows: after smearing the mouse ear face with dimethylbenzene, can cause the obvious swelling of Mus ear.Chmice acute auricle edema due to the high, medium and low dosage group of the oral liquid xylol all has certain inhibitory action, and suppression ratio is respectively 51.4%, 37.6% and 24.8%.Results of statistical analysis shows, oral liquid high dose group of the present invention is compared with model control group to the inhibitory action of mice ear, difference is (p＜0.01) extremely significantly, in, low dose group compares with model control group, significant difference (p＜0.05), high dose group is compared with the aspirin matched group, significant difference (p＜0.05).Oral liquid height, middle dosage group can significantly suppress by the mice auricle swelling due to the dimethylbenzene, and inhibitory rate of intumesce is respectively 51.4%, 37.6%, with the poor heteropole of model control group remarkable (p＜0.01).Show that oral liquid of the present invention has antiinflammatory action.
(4) to mice SO
2Draw the antitussive action of coughing
Adopt SO
2Draw the method for coughing and observe the antitussive action of oral liquid of the present invention the cough mice.Test is divided into the high, medium and low dosage group of oral liquid (embodiment 1 is prepared) (be respectively 0.2,0.1,0.05mL/10g body weight), carbetapentane citrate matched group (0.05mg/10g body weight), normal control group and model control group.1h puts into mice and is placed with SO after the administration
2In the exsiccator of gas, behind the stimulation 15s, taking out immediately, observe mice and draw the cough number of times of coughing in incubation period and the 2min. the result shows: pass through SO
2After mice stimulated, the model group mice can cause cough in 20s, but and the high, medium and low dosage group of oral liquid significant prolongation SO
2Draw and cough incubation period, and can significantly reduce the cough number of times of mice.To SO
2Draw to cough and carry out statistical analysis incubation period, the result shows that the high, medium and low dosage of oral liquid of the present invention all has prolongation SO
2Draw the preclinical effect of coughing, wherein high, middle dosage group is compared with model group, and difference is (p＜0.01) extremely significantly; Low dose group and carbetapentane citrate matched group are compared with model group, significant difference (p＜0.05); High dose group is compared with the carbetapentane citrate matched group, significant difference (p＜0.05).The number of times that cough takes place is carried out statistical analysis, show that the oral liquid high dose group compares with model control group, difference is remarkable (p＜0.01) extremely; Middle dosage group and carbetapentane citrate matched group are compared significant difference (p＜0.0) with model group; High dose group is compared with the carbetapentane citrate matched group, significant difference (p＜0.05).Show that oral liquid of the present invention has antitussive action.
(5) to the resolve phlegm effect of mice
Adopt the phenol red method of trachea section, with Kunming kind white mice, be divided into the high, medium and low dosage group of oral liquid (embodiment 2 is prepared) (be respectively 0.2,0.1,0.05mL/10g body weight), JIZHI TANGJIANG matched group (25mL/kg body weight), normal control group at random by the sex body weight.Every group 10, the blank group gives isometric normal saline.Irritate stomach every day 1 time, successive administration 10d, 30min after the last administration, the phenol red 0.1mL/10g of lumbar injection, inject phenol red back 30min, peel off animal trachea surrounding tissue, cut one section trachea down to the trachea bifurcation, put into the test tube that fills the 2mL normal saline from thyroid cartilage, add the 0.1mL sodium hydroxide solution again, with 721 type spectrophotometers, wavelength 546nm place surveys the OD value, with the phenol red content of OD value representative.The result shows that anti-Mycoplasma gallisepticum oral liquid high dose group can significantly increase the phenol red excretion amount of trachea, with the poor heteropole of matched group remarkable (P＜0.01), shows that product of the present invention has antitussive action.
(6) to the antiasthmatic effect of Cavia porcellus
Adopting spraying to cause the method for breathing heavily tests.Some of test preliminary election the previous day Cavia porcelluss childhood are put respectively in the spray bottle, and constant voltage sprays into the histamine phosphate 1mL of 1mg/mL in bottle, and animal through certain latent time, can produce asthma reaction after sucking medicinal liquid, with " falling " as the asthma reaction index.The time of falling surpasses 150s person will not select for use, and animal one " falling " occur and takes out immediately." asthma " Cavia porcellus that preliminary election is qualified, be divided into 5 groups at random by the sex body weight, every group 8, i.e. normal saline group, the high, medium and low dosage group of anti-Mycoplasma gallisepticum oral liquid (embodiment 3 is prepared) (be respectively 20,10,5mL/kg body weight) and JIZHI TANGJIANG matched group (12mL/kg body weight).Every day, gastric infusion was 1 time, and blank group is given isometric normal saline, behind the 2h by above-mentioned dosage gastric infusion 1 time once more.1h after the administration puts into sprayer unit respectively, the similarity condition when the preliminary election, and the histamine phosphate (concentration 1mg/mL) of spraying respectively spraying beginning symptom (falling) occur as latent time, record asthma reaction incubation period, prolongs one times with latent time and thinks effective.The result show anti-Mycoplasma gallisepticum oral liquid obviously prolonged guinea pig draw and breathe heavily incubation period, improve the effective percentage of relievining asthma, through X
2Check high agent group and blank group comparing difference remarkable (P＜0.01), effective percentage is higher than the JIZHI TANGJIANG matched group.
4. to the therapeutic effect of Mycoplasma gallisepticum infection
By collunarium, eye dripping and air bag injection system 180 22 age in days healthy chicks are carried out artificial challenge's Mycoplasma gallisepticum (MG), be about to Mycoplasma gallisepticum MG S6 freeze-dried vaccine and go down to posterity through 3 times, the growth turbidity reaches 10
7CCU/mL inoculation Frustrate blood and mycoplasma liquid.Except that the normal healthy controls group, all the other are respectively organized the every plumage of chicken left and right sides torso bag (the 2nd intercostal reciprocal) and inject Mycoplasma gallisepticum culture fluid 0.5mL (cooperating collunarium eye dripping 0.5mL).Hold test chicken back and cervical region with left hand during operation, the right hand is held syringe and is drawn MG S6 culture fluid, lentamente it is splashed in the nostril of chicken, keep vertical position a few minutes, utilize the action of chicken naturally aspirated, bacterium liquid is sucked air flue to pulmonary, carry out eye dripping with quadrat method. test is divided into oral liquid (embodiment 1-4 is prepared) height, in, low dose group, the tylosin matched group, infect matched group (infecting not administration) and normal healthy controls group (not infecting not administration), every group 30, male and female have both. adopt MG separation and Culture and mycoplasma preliminary determining method (L-type bacteriologic test, thalli morphology is observed, bacterium colony is painted, cholesterol needs test, the tetrazole reduction test, decomposition glucose and arginine test, growth inhibition test, the chicken red blood cell adsorption test) the positive cases of infection of conclusive evidence Mycoplasma gallisepticum. adopt mixed drink administering mode that the morbidity chicken is carried out therapeutic test. except that normal control group and model group, all the other respectively organize the drug treatment of all drinking water. the oral liquid height, in, the concentration of low dose group drinking-water administration is respectively 1.2%, 0.8% and 0.5%, every group of chicken amount of drinking water every day is about 1500mL, be that every chicken average every day of amount of gathering medicinal herbs is respectively 0.6,0.4 and 0.25mL, be equivalent to crude drug 1.2,0.8 and 0.5g. control drug tylosin group dosage is to contain medicine 0.5g. in every 1000mL water to organize chicken every day early, respectively change water once evening, prohibit water 2h before changing water at every turn, successive administration 5d, observe clinical symptoms and dead chicken is cutd open inspection before observing the each medication of 15d. after the drug withdrawal, observe air bag, the variation of trachea and lung, and make cause of disease and separate, be strictly with proof and die from Mycoplasma gallisepticum infection, from each group, get 1～2 live chickens metainfective the 8th day (after the drug withdrawal the 3rd day) and cut open and kill, observe the air bag pathological changes; All chickens that survive were weighed in metainfective the 20th day average weight, and weightening finish situation; Randomly draw 5 blood sampling separation of serum for every group and make the dull and stereotyped agglutination of serum, detect the intravital Mycoplasma gallisepticum antibody of chicken; At last all chickens are cutd open and are killed, observe the air bag pathological changes, mark to air bag, respectively organize chicken air bag degree of injury with 1～4 fen four grade of every side.
The result shows: when drug level of the present invention is 1.2%, 0.8% in the drinking-water, logotype 5 days, infected chicken there is tangible curative effect, can significantly reduce the serum antibody response positive rate, air bag damage slip is respectively 87.3%, 83.1%, cure rate to infected chicken reaches 71.4%, 67.9%, and effective percentage reaches 92.9%, and rate of body weight gain reaches 107.1%, 105.4%.Illustrate that product of the present invention has the effect of obvious control Mycoplasma gallisepticum, can obtain high economic benefit and social benefit, have wide market application prospect.
5. the clinical efficacy of product of the present invention test
In Harbin, 12 chicken farms, area such as Qiqihar, Jiamusi have carried out clinical expansion group therapeutic test and have applied, the result shows, product of the present invention has notable therapeutic effect to the acute and chronic respiratory tract disease, high economic benefit and social benefit have been obtained, be subjected to raiser's favorable comment and welcome, had wide market application prospect.
The safety testing of test example 3 Chinese medicine oral liquid of the present invention
(1) acute toxicity test.
Adopt and simplify bandit's formula method survey LD
50(median lethal dose(LD 50)) is with its index as evaluation Chinese medicine oral liquid acute toxicity.After trial test, fail to find out the dosage of 100% dead mouse or the dosage range of 0% and 100% dead mouse.Because of administration concentration and administration volume (0.8mL/20g) to maximum, fail to measure LD
50, therefore to observe after carrying out in one day in the maximum tolerance determination 24h irritating stomaches 3 times so that the tolerant Cmax of animal is continuous, the animal spontaneous activity has minimizing slightly, and reaction is seen indifferently slightly to external world, recovers successively in the 30min, and loose stool is arranged around the individual animal anus.Animal does not all observe ANOMALOUS VARIATIONS at aspects such as spontaneous activity, outward appearance, reaction to external world, the colour of skin, secretions, eye, breathing, appetite, muscular movements in the 7d later on.As calculated, this medicine accumulative total dosage in 24h reaches 60g/kg, and the maximum tolerance multiple of extrapolating this medicine is 100 times of chicken therapeutic dose.
(2) long term toxicity test.
With 80 SD is that rat is divided into 4 groups at random, three groups of Chinese medicine oral liquid (40 of irritating the stomach various dose wherein, 20, the 10g/kg body weight) successive administration is 3 months, another group is for the normal control group. be subjected to 24h behind the reagent thing the last time, 3/5 animal is carried out once the complete detection of every index, plucking eyeball gets blood and carries out hematology and blood biochemical and learn index determining, dissect animal main organs is carried out comprehensive and systematic perusal, important organ is weighed, calculate organ coefficient, internal organs are drawn materials, conventional fixing with 10% formalin, dehydration, the paraffin embedding film-making, HE dyeing carrying out histopathologic examination under the light microscopic. remaining 2/5 animal drug withdrawal, after continuing to observe for 2 weeks, reexamine, test item and method are the same, with the degree of reversibility of understanding toxic reaction and the retardance toxic reaction that may occur. the inspection item index comprises: 1. general situation is observed: the outward appearance of observing animal every day, behavioral activity, secretions, the feces character, poisoning symptom etc. administration before measurement normal type, survey body weight after the administration weekly 1 time, observe body weight change. 2. hematological indices: comprise hemoglobin (Hb), erythrocyte (RBC) counting, leukocyte (WBC) counting, platelet (PLT) counting, the coagulant blood time (CT), Fibrinogen (FIB), leukocyte differential count lymphocyte (LY), mononuclear cell (MO), neutrophilic granulocyte, acid granulocyte, alkalescence granulocyte etc. 3. blood biochemical is learned index: detect alanine aminotransferase (ALT), aspartic acid conversion enzyme (AST), alkali phosphatase (ALP), carbamide (BUN), creatinine (Grea), T-CHOL (T-CHO), total bilirubin (T-BIL), total protein (TP), albumin (ALB), blood glucose (GLU), measure with automatic clinical chemistry analyzer. 4. system's postmortem perusal and histopathologic examination: in laboratory animal dissection process, the form of comprehensive careful observation animal main organs and body of gland, size, change in color. win the heart of animal, liver, spleen, lung, kidney, stomach, trachea, small intestinal, large intestine, pancreas, brain, the adrenal gland, prostate, thymus, testis, the uterus, main organs and bodies of gland such as ovary, weigh respectively, calculate the organ coefficient (ratio of organ weights and the weight of animals, with g/100g is unit). histopathologic examination: after the main organs of animal drawn materials, 10% formalin fixed, paraffin embedding, film-making, the histopathologic examination under the light microscopic is carried out in HE dyeing.
The result shows that 1. general situation: in entire test, each is organized, and the rat outward appearance is normal, and behavioral activity is normal, and diet is normal; Except that having the last week, heavy dose group small part animal (recovers normal) the loose stool all the other no abnormal findings after the week. and 2. growing state: heavy dose of the first two all body weight of group small part animal has slightly and alleviates, but relatively there is not significant difference with matched group, recover after one month. by statistics, each administration group rat body weight of whole experimental session increases weight and matched group compares there was no significant difference (P is all＞0.05). and 3. hematological indices detects: each administration group blood routine and leukocyte differential count are all in normal range. compare with matched group, hemoglobin, the RBC counting, WBC counting and classification thereof, the PLT counting, FIB value and coagulation time test etc., difference that there are no significant (P＞0.05). 4. blood biochemical is learned the index detection: learn by statistics and handle, the every blood parameters of each administration group does not all change, compare there was no significant difference (P＞0.05) with the blank group. 5. animal viscera perusal: the form of each main organs of anatomic observation experimental animal and body of gland, size, colors etc. show no obvious abnormalities variation. 6. rat important organ coefficient influenced: learn by statistics and handle, each administration group main organs coefficient compares with matched group respectively, difference that there are no significant (P＞0.05). 7. histopathologic examination: the main organs and the body of gland of each administration group and blank treated animal, as heart, kidney, stomach, large intestine, thymus, prostate, the adrenal gland, pancreas, testis, the uterus, ovary, brain, cerebellum there is no tangible degeneration, pathological changes such as necrosis; The slight variation appears in other internal organs in the individual animal of administration group, but similar pathological change also appears in the individual animal in the blank group. as liver: high dose group has 1 example to occur having lymphoid cell to soak into around the bile duct, the hole expansion, in be dispersed in lymphocyte, 1 example is a vasculitis, and middle dosage group has 1 example the congestion variation to occur. but in the normal control group, also have 2 examples to occur similarly changing; Lungs: the variation of alveolar hyperemia, edema and inflammatory cell infiltration all appears in each test group, and wherein the normal control group has 1 example to see an alveolar wall and a light weight degree hyperemia, 1 routine edema, and visible a little cell infiltration that is dispersed in of 1 example, bronchioles is not seen obvious change; Logical each the dosage group of lung all has above variation to produce, being indicated as due to the spontaneous pathological changes of animal. spleen: blank group and low agent group respectively have a routine red pulp hypertrophy, each dosage group part animal folliculus occurs and boundary is unclear on every side. and small intestinal: the variation of small intestinal all has generation in each test group, mainly show as the fine hair lamina propria lymphocytic infiltration is arranged, edema, epithelial goblet cell increases, the epithelium necrosis of severe patient top, come off. there are 3 examples mucous hyperemia to occur as the normal control group, downright bad and the 3 routine mucosa inflammatory cell infiltrations of 1 routine mucomembranous epithelial cell. the visible mucous hyperemia of high dose group 2 examples, 1 routine mucous epithelium degeneration, 1 routine epithelium necrosis, 2 routine mucosa inflammatory cell infiltrations. middle dosage and low dose group also have individual animal similar variation to occur. this shows, tangible pathological change does not appear in the most of internal organs and the body of gland of each administration group, liver, spleen, lungs, the variation that small intestinal occurs performance to some extent in the normal control group equally, so think that the pathological change of these internal organs may be the spontaneous pathological changes of animal, so do not belong to pathological changes and the damage relevant with drug toxicity., 50 multiple doses that the maximum dose level of this test promptly is equivalent to the clinical usual amounts of people are safety non-toxics. 8. reversibility and delayed toxicity observed result: after remaining 2/5 animal drug withdrawal observed for 2 weeks, carry out convalescent every inspection, result and blank group are relatively, every observation index no abnormality seen, hematology and blood biochemical learn index and matched group compares there was no significant difference, each test group animal main organs and body of gland are not seen obvious damage, each administration group of organ coefficient and matched group be there was no significant difference more also, histopathologic examination, each administration group and matched group more all do not have pathology and change.
Claims (10)
1. the Chinese medicine composition of prevention and treatment Mycoplasma gallisepticum comprises that following bulk drugs makes: Radix Scutellariae 1-4 part, Flos Lonicerae 1-4 part, Radix Asteris 1-4 part, Herba Ephedrae 1-4 part, Pericarpium Citri Reticulatae 0.2-1.5 part and Radix Glycyrrhizae 0.2-1.5 part.
2. according to the described Chinese medicine composition of claim 1, it is characterized in that the weight portion of each crude drug is:
2 parts of Radix Scutellariaes, 2 parts of Flos Loniceraes, 2 parts of Radix Asteriss, 2 parts in Herba Ephedrae, 1 part of Pericarpium Citri Reticulatae, 1 part in Radix Glycyrrhizae.
3. according to claim 1 or 2 described Chinese medicine compositions, it is characterized in that: it is prepared into the oral formulations that poultry is used.
4. according to the described Chinese medicine composition of claim 3, it is characterized in that described oral formulations comprises oral liquid, powder or granule; Be preferably oral liquid.
5. method for preparing claim 1 or 2 described Chinese medicine compositions may further comprise the steps:
(1) takes by weighing each crude drug by described weight portion;
(2) Flos Lonicerae, Radix Asteris, Herba Ephedrae, Pericarpium Citri Reticulatae and Radix Glycyrrhizae are decocted with water, decoction liquor is filtered, and concentrates, and adds ethanol, and is static, spends the night;
(3) filter when going precipitation, filtrate to be concentrated into nothing alcohol flavor, cooling adds ethanol, spends the night;
(4) precipitation is gone in filtration, and filtrate concentrates, and obtains medicinal liquid 1, and is standby;
(5) baikal skullcap root decoction pieces is decocted with boiling water and 0.5% tween 80, filter;
(6) filtrate concentrates, and is static with 1～2 back insulation of dilute hydrochloric acid adjust pH, filters taking precipitate 20-95% washing with alcohol; Precipitation adds the water mixing, adjusts pH value and makes the baicalin dissolving, and the centrifugal precipitation of going is got supernatant, obtains medicinal liquid 2, and is standby;
(7) medicinal liquid 1 and medicinal liquid 2 are mixed, concentrate, adjust pH to 5.5～6.5 add antiseptic, and are canned behind the mixing, seal, and sterilization, promptly.
6. in accordance with the method for claim 5, it is characterized in that: the number of times that decocts with water described in the step (2) is 2 times, adds 10 times of weight decoctings for the first time and boils 2h, and wherein Flos Lonicerae is put into after decocting 1h, adds 10 times of weight decoctings for the second time and boils 1.5h; In the step (2) filtrate is concentrated into 4 times of crude drug weight, adds final concentration and be 65% ethanol, static, 4 ℃ are spent the night.
7. in accordance with the method for claim 5, it is characterized in that: in the step (4) filtrate being concentrated into being contained the crude drug amount is 1g/mL, obtains medicinal liquid 1, standby.
8. in accordance with the method for claim 5, it is characterized in that: in the step (5) baikal skullcap root decoction pieces is decocted 2 times with boiling water and 0.5% tween 80, add 10 times of weight decoctings for the first time and boil 2h, add 8 times of weight decoctings for the second time and boil 1h, gradation filters while hot, merging filtrate.
9. in accordance with the method for claim 5, it is characterized in that: under 80 ℃ filtrate is concentrated into relative density 1.0 backs in the step (6) and also transfers pH1～2 back insulation 1.5h, static 3h, filtration with 2mol/L hydrochloric acid; With precipitate is 50%, 75% and 90% washing with alcohol with final concentration respectively; Precipitation adds the water mixing of 1 times of weight of Radix Scutellariae crude drug, transfers pH8 to make the baicalin dissolving with 0.1mol/L NaOH solution.
10. it is characterized in that in accordance with the method for claim 5: in the step (7) medicinal liquid 1 and medicinal liquid 2 mixing are concentrated into 1g/mL.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103599413A (en) * | 2013-11-12 | 2014-02-26 | 青岛绿曼生物工程有限公司 | Traditional Chinese medicine composition for treating mycoplasmosis of chicken and preparation method thereof |
| CN105641138A (en) * | 2014-11-13 | 2016-06-08 | 天津市中敖饲料有限公司 | Chicken mycoplasmosis treating traditional Chinese medicine composition and preparation method thereof |
| CN106511550A (en) * | 2015-09-09 | 2017-03-22 | 韦爽姬 | Drug for treatment of chicken mycoplasmosis |
| CN106578639A (en) * | 2016-12-20 | 2017-04-26 | 新昌县钧国生物技术有限公司 | Feed additive for treating avian mycoplasmosis |
| CN111388527A (en) * | 2020-04-10 | 2020-07-10 | 聊城创新畜禽养殖有限公司 | Preparation for preventing and treating mycoplasma of laying hens and preparation method thereof |
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2009
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103599413A (en) * | 2013-11-12 | 2014-02-26 | 青岛绿曼生物工程有限公司 | Traditional Chinese medicine composition for treating mycoplasmosis of chicken and preparation method thereof |
| CN103599413B (en) * | 2013-11-12 | 2017-01-11 | 青岛众志合才智能科技有限公司 | Traditional Chinese medicine composition for treating mycoplasmosis of chicken and preparation method thereof |
| CN105641138A (en) * | 2014-11-13 | 2016-06-08 | 天津市中敖饲料有限公司 | Chicken mycoplasmosis treating traditional Chinese medicine composition and preparation method thereof |
| CN106511550A (en) * | 2015-09-09 | 2017-03-22 | 韦爽姬 | Drug for treatment of chicken mycoplasmosis |
| CN106578639A (en) * | 2016-12-20 | 2017-04-26 | 新昌县钧国生物技术有限公司 | Feed additive for treating avian mycoplasmosis |
| CN111388527A (en) * | 2020-04-10 | 2020-07-10 | 聊城创新畜禽养殖有限公司 | Preparation for preventing and treating mycoplasma of laying hens and preparation method thereof |
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