CN101966285B - Traditional Chinese medicine and traditional Chinese medicine aerosol for preventing and curing chicken respiratory diseases and preparation method thereof - Google Patents
Traditional Chinese medicine and traditional Chinese medicine aerosol for preventing and curing chicken respiratory diseases and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a traditional Chinese medicine for preventing and curing chicken respiratory diseases, which is prepared from the following raw materials: 2-10g of ephedra, 3-10g of semen armeniacae amarae, 3-10g of liquorice, 15-60g of gypsum, 10-15g of chrysanthemum, 5-10g of folium mori, 6-15g of fructus forsythiae, 3-6g of mint, 3-10g of Platycodon grandiflorum, 10-15g of honeysuckle,10-30g of dandelion, 15-30g of Houttuynia cordata, 10-15g of folium isatidis, 3-10g of Scutellaria baicalensis, 10-15g of Radix Ophiopogonis and 0.015-0.03g of venenum bufonis. The invention adopts pure traditional Chinese medicine preparation, has the functions of diminishing inflammation, sterilizing and relieving cough and asthma, and has good prevention and cure effect on chicken respiratory diseases.
Description
Technical field
The present invention relates to a kind of Chinese medicine of preventing and treating respiratory disease of chicken, relate to a kind of medicinal herbs aerosol inhalation agent that prevents and treats respiratory disease of chicken simultaneously, and the method for preparing of this aerosol.
Background technology
Autumn and winter season, the bad control of environment in the hen house, pathogen is very easily bred, and respiratory disease of chicken increases immediately.This type of respiratory tract disease is the disease that is caused concurrent or mixed infection by multiple reasons such as virus, antibacterial, mycoplasma, immunosuppressant disease pathogen and adverse environments.Modal bacterial disease such as colibacillosis, white diarrhea disease, infectious coryza of chicken all can show respiratory symptom.Many viral diseases such as newcastle, bird flu, infectious bronchitis, infectious laryngotracheitis etc. also can show respiratory symptom, and viral disease sometimes with bacterial disease mixed infection.Respiratory disease of chicken can infect the chicken of various ages in days, and causes the chick growth hypoevolutism, the chicken egg drop reduction of growing up even cause death.The chicken crowd is in case morbidity, will influence culture benefit to raiser's huge economic loss of taking.Yet present many efficacious therapy medicines all are Western medicine, and it is oversize to be used for laying hen or off-drug period, and has certain toxic and side effects, is easy to generate drug resistance, also is easy to generate drug residue.
Summary of the invention
The Chinese medicine that the purpose of this invention is to provide a kind of control respiratory disease of chicken that adopts pure Chinese medicinal preparation, has no side effect, has no drug resistance is to improve the prevention effect to respiratory disease of chicken.
In order to realize above purpose; The technical scheme that Chinese medicine adopted that the present invention prevents and treats respiratory disease of chicken is: a kind of Chinese medicine of preventing and treating respiratory disease of chicken is that the feedstock production by following weight in grams forms: Herba Ephedrae 2～10 grams, Semen Armeniacae Amarum 3～10 grams, Radix Glycyrrhizae 3～10 grams, Gypsum Fibrosum 15～60 grams, Flos Chrysanthemi 10～15 grams, Folium Mori 5～10 grams, Fructus Forsythiae 6～15 grams, Herba Menthae 3～6 grams, Radix Platycodonis 3～10 grams, Flos Lonicerae 10～15 grams, Herba Taraxaci 10～30 grams, Herba Houttuyniae 15～30 grams, Folium Isatidis 10～15 grams, Radix Scutellariae 3～10 grams, Radix Ophiopogonis 10～15 gram, Venenum Bufonis 0.015～0.03 gram.
The present invention also aims to provide a kind of medicinal herbs aerosol inhalation agent that prevents and treats respiratory disease of chicken, to realize colony's administration to chicken.
The present invention prevents and treats the technical scheme that the medicinal herbs aerosol inhalation agent of respiratory disease of chicken adopted: a kind of medicinal herbs aerosol inhalation agent that prevents and treats respiratory disease of chicken, mainly contain the extract and the reagent of following components by weight percent in the said aerosol: Herba Ephedrae 2～10 grams, Semen Armeniacae Amarum 3～10 grams, Radix Glycyrrhizae 3～10 grams, Gypsum Fibrosum 15～60 grams, Flos Chrysanthemi 10～15 grams, Folium Mori 5～10 grams, Fructus Forsythiae 6～15 grams, Herba Menthae 3～6 grams, Radix Platycodonis 3～10 grams, Flos Lonicerae 10～15 grams, Herba Taraxaci 10～30 grams, Herba Houttuyniae 15～30 grams, Folium Isatidis 10～15 grams, Radix Scutellariae 3～10 grams, Radix Ophiopogonis 10～15 gram, Venenum Bufonis 0.015～0.03 gram; Also contain 10% ~ 30% propellant, the propylene glycol of 50～100ml, the ethanol of 100～300ml and the water of surplus.
The present invention also aims to provide a kind of method for preparing of preventing and treating the medicinal herbs aerosol inhalation agent of respiratory disease of chicken.
The technical scheme that method for preparing adopted of medicinal herbs aerosol inhalation agent of the present invention is: a kind of method for preparing of preventing and treating the medicinal herbs aerosol inhalation agent of respiratory disease of chicken, this aerosol adopt following steps to be prepared from:
1) takes by weighing Herba Ephedrae 2～10 gram, Flos Chrysanthemi 10～15 grams, Fructus Forsythiae 6～15 grams, Herba Menthae 3～6 grams, Flos Lonicerae 10～15 grams, Herba Houttuyniae 15～30 grams respectively and extract volatile oil with steam distillation; Extract each time of merging extracting solution more than 3 times, it is subsequent use to obtain extract 1;
2) take by weighing Gypsum Fibrosum 15～60 grams and added decocting earlier 20～40 hours; Taking by weighing Semen Armeniacae Amarum 3～10 gram, Radix Glycyrrhizae 3～10 grams, Folium Mori 5～10 grams, Radix Platycodonis 3～10 grams, Herba Taraxaci 10～30 grams, Folium Isatidis 10～15 grams, Radix Scutellariae 3～10 grams, Radix Ophiopogonis 10～15 grams then respectively adds wherein; The water steaming and decocting that adds 8～10 times of amounts that are equivalent to the medical herbs amount merges each time decocting liquid more than 3 times, filters; Concentrated filtrate, it is subsequent use to obtain extract 2;
3) with step 1) and 2) extract that obtains 1 mixes with extract 2, is concentrated into 80 ℃ of relative densities and is 1.1 ~ 1.3 clear paste, and adjust pH to 5 ~ 6 add Venenum Bufonis 0.015～0.03 gram mixing again;
4) add propylene glycol 50～100ml, ethanol 100～300ml and water in the mixing liquid that step 3) is obtained to 900ml, in filtrating, add activated carbon decolorizing, left standstill after stirring 30 minutes, back sucking filtration obtains the liquid of 900ml.
The effect of each component raw material is following in the Chinese medicine of the present invention: Herba Ephedrae: main component is an ephedrine, and contains a small amount of pseudoephedrine, volatilization wet goods, and ephedrine and pseudoephedrine have the effect of alleviating bronchial muscular spasm.Volatile oil has inhibitory action to influenza virus.Semen Armeniacae Amarum: contain amygdaloside, it can decompose the back and produce little hydrogen cyanic acid, can suppress coughing centre and plays the antitussive and antiasthmatic effect.Radix Glycyrrhizae: ability expelling phlegm for arresting cough, the throat of protection inflammation and the mucosa of trachea.Gypsum Fibrosum: merit with relieving cough and asthma.Flos Chrysanthemi: contain volatile oil, main component all has certain antibacterial action to staphylococcus aureus, multiple pathogenic bacillus and dermatophytes, during high concentration influenza virus is also had inhibitory action.Folium Mori: various pathogens such as staphylococcus aureus, beta hemolytic streptococcus are had inhibitory action.Fructus Forsythiae: Fructus Forsythiae has broad-spectrum antibacterial action, and staphylococcus aureus, Hayes dysentery bacterium are had very strong inhibitory action, and other pathogenic bacterium, influenza virus, fungus are all had certain inhibitory action.Herba Menthae: the master contains volatile oil, and Oleum menthae can promote the respiratory tract glandular secretion and respiratory inflammation is had therapeutical effect, coccus and bacillus etc. is had bacteriostasis.Radix Platycodonis: can reflexive increase the trachea secretion, dilution sputum and stronger phlegm-dispelling functions is arranged, and antitussive effect is arranged.Flos Lonicerae: these article have broad-spectrum antibacterial action, and sick have inhibitory action to influenza virus.Herba Taraxaci: staphylococcus aureus, Hemolytic streptococcus are had stronger inhibitory action, bacillus is also had certain inhibitory action.Herba Houttuyniae:, inhibitory action is in various degree arranged all to multiple Gram-negative and positive bacteria.Folium Isatidis: " effect such as that these article have is antibiotic, antiviral, analgesic, antiinflammatory.Radix Scutellariae: have wider antimicrobial spectrum, influenza virus is also had inhibitory action.Radix Ophiopogonis: can yin nourishing, heat clearing away, moisturize.Venenum Bufonis: the treatment chronic tracheitis has certain curative effect.All medicines merge, anti-inflammation and sterilization, and relieving cough and asthma has the prevention and the therapeutic effect of becoming reconciled to respiratory disease of chicken.
In addition, the Chinese medicine of control respiratory disease of chicken of the present invention adopts aerosol dosage forms, and colony's administration has improved the efficient of administration in can trooping to chickens a large amount of in the hen house.Do not have expensive and rare part in addition in the Chinese medicinal components, the manufacturing of medicinal herbs aerosol inhalation agent is very convenient, is easy to apply.
The specific embodiment
Embodiment 1
The Chinese medicine of the control respiratory disease of chicken of present embodiment is that the feedstock production by following weight in grams forms: Herba Ephedrae 2 grams, Semen Armeniacae Amarum 6 grams, Radix Glycyrrhizae 3 grams, Gypsum Fibrosum 40 grams, Flos Chrysanthemi 13 grams, Folium Mori 5 grams, Fructus Forsythiae 15 grams, Herba Menthae 6 grams, Radix Platycodonis 6 grams, Flos Lonicerae 15 grams, Herba Taraxaci 30 grams, Herba Houttuyniae 30 grams, Folium Isatidis 12 grams, Radix Scutellariae 10 grams, Radix Ophiopogonis 15 gram, Venenum Bufonis 0.03 gram.
The medicinal herbs aerosol inhalation agent of the control respiratory disease of chicken of present embodiment, mainly contain the extract and the reagent of following components by weight percent in said this aerosol: Herba Ephedrae 2 grams, Semen Armeniacae Amarum 6 grams, Radix Glycyrrhizae 3 grams, Gypsum Fibrosum 40 grams, Flos Chrysanthemi 13 grams, Folium Mori 5 grams, Fructus Forsythiae 15 grams, Herba Menthae 6 grams, Radix Platycodonis 6 grams, Flos Lonicerae 15 grams, Herba Taraxaci 30 grams, Herba Houttuyniae 30 grams, Folium Isatidis 12 grams, Radix Scutellariae 10 grams, Radix Ophiopogonis 15 gram, Venenum Bufonis 0.03 gram; The propellant, the propylene glycol of 50ml, the ethanol of 200ml and the water of surplus that also contain 20% gram.
The medicinal herbs aerosol inhalation agent of the control respiratory disease of chicken of present embodiment, method for preparing is following:
1) take by weighing Herba Ephedrae 2 gram, Flos Chrysanthemi 13 grams, Fructus Forsythiae 15 grams, Herba Menthae 6 grams, Flos Lonicerae 15 grams, Herba Houttuyniae 30 grams respectively and extract volatile oil with steam distillation, extract and merge each time extracting solution more than 3 times, it is subsequent use to obtain extract 1;
2) take by weighing Gypsum Fibrosum 40 grams and added decocting earlier 20 hours; Taking by weighing Semen Armeniacae Amarum 6 gram, Radix Glycyrrhizae 3 grams, Folium Mori 5 grams, Radix Platycodonis 6 grams, Herba Taraxaci 30 grams, Folium Isatidis 12 grams, Radix Scutellariae 10 grams, Radix Ophiopogonis 15 grams then respectively adds wherein; The water steaming and decocting that adds 8～10 times of amounts that are equivalent to the medical herbs amount merges each time decocting liquid more than 3 times, filters; Concentrated filtrate, it is subsequent use to obtain extract 2;
3) with step 1) and 2) extract that obtains 1 mixes with extract 2, is concentrated into 80 ℃ of relative densities and is 1.1 ~ 1.3 clear paste, and adjust pH to 5 ~ 6 add Venenum Bufonis 0.03 gram mixing again;
4) add propylene glycol 50ml, ethanol 200ml and water in the mixing liquid that step 3) is obtained to 900ml, in filtrating, add activated carbon decolorizing, left standstill after stirring 30 minutes, back sucking filtration obtains the liquid of 900ml.
5) aerosol is tested, charge into the 20g propellant according to every 90ml after being packed as 10 bottles, embedding, gland after qualified.
Embodiment 2
The Chinese medicine of the control respiratory disease of chicken of present embodiment is that the feedstock production by following weight in grams forms: Herba Ephedrae 10 grams, Semen Armeniacae Amarum 10 grams, Radix Glycyrrhizae 6 grams, Gypsum Fibrosum 60 grams, Flos Chrysanthemi 15 grams, Folium Mori 7 grams, Fructus Forsythiae 10 grams, Herba Menthae 4.5 grams, Radix Platycodonis 10 grams, Flos Lonicerae 12 grams, Herba Taraxaci 20 grams, Herba Houttuyniae 15 grams, Folium Isatidis 10 grams, Radix Scutellariae 7 grams, Radix Ophiopogonis 10 gram, Venenum Bufonis 0.022 gram.
The medicinal herbs aerosol inhalation agent of the control respiratory disease of chicken of present embodiment, mainly contain the extract and the reagent of following components by weight percent in said every 1000ml aerosol: Herba Ephedrae 10 grams, Semen Armeniacae Amarum 10 grams, Radix Glycyrrhizae 6 grams, Gypsum Fibrosum 60 grams, Flos Chrysanthemi 15 grams, Folium Mori 7 grams, Fructus Forsythiae 10 grams, Herba Menthae 4.5 grams, Radix Platycodonis 10 grams, Flos Lonicerae 12 grams, Herba Taraxaci 20 grams, Herba Houttuyniae 15 grams, Folium Isatidis 10 grams, Radix Scutellariae 7 grams, Radix Ophiopogonis 10 gram, Venenum Bufonis 0.022 gram; The propylene glycol that also contains 75ml, the ethanol of 300ml and the water of surplus.Every 1000ml aerosol need charge into the propellant of 110ml.
The medicinal herbs aerosol inhalation agent of the control respiratory disease of chicken of present embodiment, method for preparing is following:
1) take by weighing Herba Ephedrae 10 gram, Flos Chrysanthemi 15 grams, Fructus Forsythiae 10 grams, Herba Menthae 4.5 grams, Flos Lonicerae 12 grams, Herba Houttuyniae 15 grams respectively and extract volatile oil with steam distillation, extract and merge each time extracting solution more than 3 times, it is subsequent use to obtain extract 1;
2) take by weighing Gypsum Fibrosum 60 grams and added decocting earlier 40 hours; Taking by weighing Semen Armeniacae Amarum 10 gram, Radix Glycyrrhizae 6 grams, Folium Mori 7 grams, Radix Platycodonis 10 grams, Herba Taraxaci 20 grams, Folium Isatidis 10 grams, Radix Scutellariae 7 grams, Radix Ophiopogonis 10 grams then respectively adds wherein; The water steaming and decocting that adds 8～10 times of amounts that are equivalent to the medical herbs amount merges each time decocting liquid more than 3 times, filters; Concentrated filtrate, it is subsequent use to obtain extract 2;
3) with step 1) and 2) extract that obtains 1 mixes with extract 2, is concentrated into 80 ℃ of relative densities and is 1.1 ~ 1.3 clear paste, and adjust pH to 5 ~ 6 add Venenum Bufonis 0.022 gram mixing again;
4) add propylene glycol 75ml, ethanol 300ml and water in the mixing liquid that step 3) is obtained to 900ml, in filtrating, add activated carbon decolorizing, left standstill after stirring 30 minutes, back sucking filtration obtains the liquid of 900ml.
5) aerosol is tested, be packed as behind 10 bottles, embedding, gland the propellant that charges into 10g according to every 90ml after qualified.
Embodiment 3
The Chinese medicine of the control respiratory disease of chicken of present embodiment is that the feedstock production by following weight in grams forms: Herba Ephedrae 6 grams, Semen Armeniacae Amarum 3 grams, Radix Glycyrrhizae 10 grams, Gypsum Fibrosum 15 grams, Flos Chrysanthemi 10 grams, Folium Mori 10 grams, Fructus Forsythiae 6 grams, Herba Menthae 6 grams, Radix Platycodonis 3 grams, Flos Lonicerae 15 grams, Herba Taraxaci 10 grams, Herba Houttuyniae 23 grams, Folium Isatidis 15 grams, Radix Scutellariae 3 grams, Radix Ophiopogonis 13 gram, Venenum Bufonis 0.015 gram.
The medicinal herbs aerosol inhalation agent of the control respiratory disease of chicken of present embodiment, mainly contain the extract and the reagent of following components by weight percent in said every 1000ml aerosol: Herba Ephedrae 6 grams, Semen Armeniacae Amarum 3 grams, Radix Glycyrrhizae 10 grams, Gypsum Fibrosum 15 grams, Flos Chrysanthemi 10 grams, Folium Mori 10 grams, Fructus Forsythiae 6 grams, Herba Menthae 6 grams, Radix Platycodonis 3 grams, Flos Lonicerae 15 grams, Herba Taraxaci 10 grams, Herba Houttuyniae 23 grams, Folium Isatidis 15 grams, Radix Scutellariae 3 grams, Radix Ophiopogonis 13 gram, Venenum Bufonis 0.015 gram; Also contain 20% propellant, the propylene glycol of 100ml, the ethanol of 100ml and the water of surplus.The medicinal herbs aerosol inhalation agent of the control respiratory disease of chicken of present embodiment, method for preparing is following:
1) take by weighing Herba Ephedrae 6 gram, Flos Chrysanthemi 10 grams, Fructus Forsythiae 6 grams, Herba Menthae 6 grams, Flos Lonicerae 15 grams, Herba Houttuyniae 23 grams respectively and extract volatile oil with steam distillation, extract and merge each time extracting solution more than 3 times, it is subsequent use to obtain extract 1;
2) take by weighing Gypsum Fibrosum 15 grams and added decocting earlier 30 hours; Taking by weighing Semen Armeniacae Amarum 3 gram, Radix Glycyrrhizae 10 grams, Folium Mori 10 grams, Radix Platycodonis 10 grams, Herba Taraxaci 10 grams, Folium Isatidis 15 grams, Radix Scutellariae 3 grams, Radix Ophiopogonis 13 grams then respectively adds wherein; The water steaming and decocting that adds 8～10 times of amounts that are equivalent to the medical herbs amount merges each time decocting liquid more than 3 times, filters; Concentrated filtrate, it is subsequent use to obtain extract 2;
3) with step 1) and 2) extract that obtains 1 mixes with extract 2, is concentrated into 80 ℃ of relative densities and is 1.1 ~ 1.3 clear paste, and adjust pH to 5 ~ 6 add Venenum Bufonis 0.015 gram mixing again;
4) add propylene glycol 100ml, ethanol 100ml and water in the mixing liquid that step 3) is obtained to 900ml, in filtrating, add activated carbon decolorizing, left standstill after stirring 30 minutes, back sucking filtration obtains the liquid of 900ml.
5) aerosol is tested, charge into 20 gram propellant according to every 90ml after being packed as 10 bottles, embedding, gland after qualified.
Experimental example
Below with concrete description of test drug effect of the present invention:
1 material and method:
1.1 test drug: aerosol of the present invention.Control drug: ofloxacin soluble powder.
1.2 experimental animal: the chicken of 40 ages in days.Sick chicken stream nose liquid, cough; Cockscomb is light white, and the feather pine disorderly; The clavus eyelid swelling that the course of disease is long is shed tears, and sick chicken often is put on tear on the feather of wing.Cut open the sick chicken of inspection and be verified as the slow breathing tract disease.
1.3 EXPERIMENTAL DESIGN
Select 500 chickens at random, be equally divided into 5 groups.Test the product of one group of spray delivery instance 1 of the present invention; Test the product of two groups of spray delivery instances 2 of the present invention; Test the product of three groups of spray delivery instances 3 of the present invention, test four groups of drinking-water administration ofloxacin soluble powder reference substances, test five groups and drink normal saline.
1.4 test method
First three groups is sprayed administration, successive administration 6 days according to the 100ml/ group to each group chicken.The 4th group of consumption 100g that at every turn drinks water mixes water 200Kg, the administration 6 days of drinking water continuously.The 5th group, freely drink water.Situation such as the clinical change of observation every day trouble chicken, healing, improvement, death, the line item of going forward side by side.
1.5 curative effect judging standard
Cure rate: after the off-test, sick chicken transference cure, spirit takes a turn for the better, and appetite increases, and recovers the ratio that normal chicken accounts for this group chicken fully.
Effective percentage: after the off-test, the chicken that fowl disease symptom, spirit, appetite are clearly better accounts for the ratio of this group chicken.
Inefficiency: symptom appears in duration of test, and the chicken that still loses transference cure after the off-test accounts for the ratio of this group chicken.
1.6 clinical test results is seen table 1
Table 1 clinical test results
| Group | Cure rate (%) | Effective percentage (%) | Inefficiency (%) |
| Instance 1 of the present invention | 50 | 82 | 18 |
| Instance 2 of the present invention | 85 | 95 | 5 |
| Instance 3 of the present invention | 53 | 80 | 20 |
| The ofloxacin soluble powder | 79 | 85 | 15 |
| Normal saline | ﹣ | ﹣ | ﹣ |
2 conclusions
Instance 2 made aerosols of the present invention have significant curative effect to the chicken chronic respiratory tract disease, compare cure rate with the ofloxacin soluble powder and improve 10%.Explain that the present invention has the obvious treatment effect to respiratory disease of chicken.
Claims (1)
1. Chinese medicine of preventing and treating the chicken chronic respiratory tract disease is characterized in that: be that feedstock production by following weight forms: Herba Ephedrae 2 ~ 10 grams, Semen Armeniacae Amarum 3 ~ 10 grams, Radix Glycyrrhizae 3 ~ 10 grams, Gypsum Fibrosum 15 ~ 60 grams, Flos Chrysanthemi 10 ~ 15 grams, Folium Mori 5 ~ 10 grams, Fructus Forsythiae 6 ~ 15 grams, Herba Menthae 3 ~ 6 grams, Radix Platycodonis 3 ~ 10 grams, Flos Lonicerae 10 ~ 15 grams, Herba Taraxaci 10 ~ 30 grams, Herba Houttuyniae 15 ~ 30 grams, Folium Isatidis 10 ~ 15 grams, Radix Scutellariae 3 ~ 10 grams, Radix Ophiopogonis 10 ~ 15 gram, Venenum Bufonis 0.015 ~ 0.03 gram.
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| CN102133354B (en) * | 2011-03-24 | 2013-03-13 | 山东省健牧生物药业有限公司 | Chinese medicinal composition for treating respiratory diseases of chicken |
| CN102614296B (en) * | 2012-03-27 | 2013-11-06 | 南京日升昌生物技术有限公司 | Traditional Chinese medicine composition for preventing and curing respiratory diseases of livestock and application of traditional Chinese medicine composition |
| CN104162074B (en) * | 2013-08-30 | 2017-10-03 | 河南后羿实业集团有限公司 | A kind of veterinary Chinese medicinal composition and preparation method for being used to treat respiratory infectious disease |
| CN104666685A (en) * | 2013-12-02 | 2015-06-03 | 天津中敖生物科技有限公司 | Traditional Chinese and western medicine compound aerosol for preventing and treating avian cough and preparation method thereof |
| CN117942405A (en) * | 2016-02-25 | 2024-04-30 | 应用生物实验室公司 | Compositions and methods for protecting airborne pathogens and irritants |
| CN105748682A (en) * | 2016-04-07 | 2016-07-13 | 五河县顺鹏农副产品有限公司 | Traditional Chinese medicine solution for preventing and treating respiratory diseases of pheasant |
| CN107998280A (en) * | 2017-12-05 | 2018-05-08 | 广州白云山宝神动物保健品有限公司 | A kind of poultry Substitutes For Antibiotic and preparation method and application |
| CN111249337A (en) * | 2020-01-15 | 2020-06-09 | 河南天昊鸿发生物技术有限公司 | Microecological preparation for treating respiratory diseases of livestock and poultry and preparation method thereof |
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