CN101439090A - Formulation containing honeysuckle flower and scutellaria, and uses thereof - Google Patents
Formulation containing honeysuckle flower and scutellaria, and uses thereof Download PDFInfo
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Abstract
The invention relates to a Yinhuang Preparation and a usage thereof and further relates to a Yinhuang composition which is composed of honeysuckle extract and a Baikal skullcap root extract. The invention also relates to the usage of the composition in the preparation of drugs for curing chronic respiratory disease infection of beasts and birds.
Description
Technical field
The present invention relates to silver yellow compositions and the purposes in the medicine of preparation treatment domestic animal chronic respiratory infectious disease thereof.
Background technology
Chronic respiratory disease (CRD) is that (Mycoplasma gallisepticum MG) infects the chronic respiratory infectious disease of a kind of chicken cause by chicken virus mycoplasma.Because MG is distributed in countries in the world, can propagate by level and vertical mode, therefore, but its long-term existence and spreading in the chicken group.According to serosurvey, chicken group's infection rate is 70%～80%.The mortality rate of primary disease is generally not high, but when secondary or concurrent other diseases, its case fatality rate can be up to more than 40%.According to another statistics, after MG was chicken infected, the weak young rate of chickling increased about 10%, the laying rate of laying hen descends 10%～20%, broiler lose weight 38%, marketing period prolongs, feed conversion rate reduces by 20%, and can cause a large amount of medication expenses indirectly, causes the poultry product Chinese medicine residual simultaneously.Therefore, this disease has caused enormous economic loss to poultry husbandry, and is affecting indirectly human beings'health.
With the vaccine prevention primary disease is more satisfactory means.Therefore, people do a lot of work aspect vaccine development, and used vaccine mainly contains two kinds of inactivated vaccine and attenuated vaccines.Because mycoplasma condition of culture harshness, so it is higher to make the cost of inactivated vaccine and attenuated vaccine.Sometimes because bad some immune protective antigen of chicken virus mycoplasma that also can make of condition of culture can not give full expression to, cause the inactivated vaccine immunity uncertain.Recent study finds, the immunologic mechanism that chicken MG infects is based on cellular immunization, and the inductive immunity of inactivated vaccine is based on humoral immunization, so inactivated vaccine has certain limitation.Though but weak malicious Seedling both can have been induced also inducing cell immunity of humoral immunization, the virulence of existing attenuated vaccine such as F strain, TS211 strain is stable inadequately, can not produce good immunity sometimes, and shortcomings such as severe reaction are arranged sometimes.In addition, strong inadequately though a common weakness of attenuated vaccine and inactivated vaccine is to produce certain immunoprotection, can not prevent infections, vaccinated flock still can large quantities ofly be fallen ill under the feeding and management condition of poor, and immuning failure happens occasionally.
Because MG is to the part antibiotic sensitive, simultaneously because therefore the infection of other antibacterials of primary disease Chang Bingfa treats the normal antibacterials of using to primary disease.But because pathogen antagonism bacteriums medicine very easily produces drug resistance, when therefore treating primary disease in clinical, curative effect is usually unsatisfactory.And more and more pay close attention to today of food safety people, residues of antibiotics is a problem demanding prompt solution, uses a large amount of antibiotic obviously to be unfavorable for human beings'health for the broiler and the laying hen that are about to listing.
In recent years, research with the Chinese herb prevention chronic respiratory disease has obtained gratifying progress, viral I number (water preparation) that Chinese medicines such as usefulness Radix Bupleuri, Radix Scutellariae, Herba Houttuyniae, Herba Andrographis, Folium Isatidis, Radix Puerariae, Flos Chrysanthemi Indici, Herba Ephedrae, Semen Armeniacae Amarum, Radix Glycyrrhizae such as Yang Bin are made, be used for the treatment of natural infection MG miscellaneous chicken, result's virus reaches the therapeutic effect identical with tylosin tartrate I number.Guo little usefulness Radix Isatidis, Folium Isatidis, Radix Glycyrrhizae, Venenum Bufonis, Herba Artemisiae Scopariae, Fructus Forsythiae, Herba Houttuyniae, Rhizoma Belamcandae, Herba Taraxaci, the Radix Aucklandiae, the Radix Pulsatillae, Borneolum Syntheticum etc. such as clear are made granule, treatment artificial challenge's chickling chronic respiratory tract disease, effect and Danofloxacin mesylate commonly used clinically be (p〉0.05) quite, than Roxithromycin, tylosin tartrate good (p＜0.05), and do not influence the normal weightening finish of chicken.Li Zhongwu etc. are with Concha Haliotidis, Rhizoma Dioscoreae Bulbiferae, Radix Scutellariae, Radix Stephaniae Cepharanthae, Pericarpium Citri Reticulatae, Rhizoma Atractylodis, Radix Platycodonis, Fructus Gardeniae, Radix Curcumae, Radix Gentianae, Hydrargyri Oxydum Rubrum, Herba Houttuyniae, Folium Perillae, Radix Asteris, Radix Et Rhizoma Rhei, Radix Sophorae Flavescentis, Radix Glycyrrhizae prescription, grind into powder, in conjunction with kanamycin sulfate treatment laying hen chronic respiratory tract disease, 3～5 days clinical symptoms disappear substantially after the medication as a result, feed intake increases to some extent, the bottom out of laying eggs after 8～10 days, cure rate reaches more than 98%.
The result of study of all Chinese herb prevention chronic respiratory diseases all proves, Chinese medicine can be prevented and treated this disease fully, and therapeutic effect can meet or exceed the level of antimicrobial drug commonly used, and does not develop immunity to drugs, add the hypotoxicity of Chinese medicine, therefore have obvious superiority with the Chinese herb prevention chronic respiratory disease.But at present some prescribed preparations that use also exist deficiency, if any the traditional Chinese medicine processing technology of use make powder, make some effective ingredient not to be utilized well, bioavailability is low.In order to reach effective dosage, the consumption of formulation of drug is increased, so just caused the waste of natural resources of Chinese medicinal materials.In addition, because most of powders are that Chinese herbal medicine mixes through pulverizing, cellulosic content wherein is very high, and according to the gastral physiological characteristics of chicken, cellulosic increase can influence the normal digestive function of chicken.The preparation prescription that has is huge, for the action effect of bringing into play single medicinal material just must increase total dosage, brings above-mentioned drawback equally.The prescription that has is studied without modern pharmacology, and its mechanism of action is not understood, can't be judged whether it has potential untoward reaction, and there is certain degree of difficulty in clinical application yet.
Swine enzootic pneumonia also claims the popular pneumonia of endemicity, swine mycoplasma pneumonia, mycoplasmal pneumonia of swine, is a kind of chronic respiratory infectious disease of pig.Cardinal symptom is cough and asthma, and the pathological changes characteristic is a confluent bronchopneumonia, often betides the preceding veutro of lungs.Primary disease extensively is present in all over the world, and sickness rate is generally 50%～80%.Part province, pig farm, city check that positive rate is 30%～50% at home.Suffer from the dysplasia of pig long term growth, the speed of growth descends 15.9%, and efficiency of feed utilization reduces by 13.8%.In the ordinary course of things, the primary disease mortality rate is not high, but if having other cause of disease mixed infection, feeding and management to be not good at and factor such as environment difference also can cause serious death, development brings serious harm to pig industry.Even more serious is the decline that primary disease causes pig immunity, causes the secondary infection or the accompanying infection of other diseases, and loss is obviously increased the weight of.For a long time, one of swine diseases that this disease is considered to normal and sends out, economic implications is great.
Though the primary disease pathogen is a mycoplasma hyopneumoniae, but because mycoplasma infection is earlier attached to trachea and bronchial ciliated epithelial cell, causing that cilium shortens tails off, cilium is swung function normally and is significantly descended, so that can't get rid of alveolar juice, air source fine dust and various cause a disease former, for secondary infection is opened the floodgates wide, so abiogenous swine enzootic pneumonia mostly is mixed infection.Common Secondary cases pathogen has pasteurellosis bacillus, streptococcus pneumoniae, haemophilus, Salmonella, various suppurative bacterium, mycoplasma hyorhinis etc.
As if the respiratory mucosa of a mycoplasma hyopneumoniae infected pigs, the pig at various ages is identical to mycoplasma pneumoniae sensitivity.The sick pig and the pig that carries disease germs are the sources of infection of primary disease.Because sow is passed to piglet with the Pulmonis Sus domestica mycoplasma, make that a lot of swinery Zhong chuan disorder of QI continue to exist.In case there are several pigs infected in the swinery, will propagate between the circle pig together, it is later on all the more so particularly when wean pig to be mixed the group.In a single day primary disease imports into, is difficult to thoroughly put out as not taking tight measure.
Now, the commercialization bacterination has become the very important means of pig farm anti-system asthma.The swine enzootic pneumonia vaccine has multiple, comprises the weak malicious Seedling of rabbitization, acellular weak malicious freeze-dried vaccine and inactivated bacteria.It is reported that their effect is all more satisfactory.But need to adopt different vaccination regimens according to the infection type in pig farm type, the mode of production and the pig farm, and also immunity inoculation and management factors should be combined when controlling this disease and carry out, in case some link has deviation, will cause immuning failure, so swine enzootic pneumonia remains the key factor of current harm pig industry.
Pulse feature uses antibiotic can slow down the clinical symptoms of disease and avoids the generation of secondary infection.Antibiosis commonly used have tetracycline, tylosin, lincomycin, quinolones.But in a single day generally speaking, use antibiotic generations that can not prevent infections, and discontinue medication, disease will soon recur.Normal preventative use antibiotic in production practices, this easily causes pathogen to produce drug resistance, and rethink and thoroughly kill pathogen, will be very difficult.Equally, owing to continue to use antibiotic to cause drug residue, thus reduce meat, and expense also improves thereupon.
Accepted by people gradually with the Chinese herb prevention swine enzootic pneumonia.Little clear the grade with Bufo siccus, synthetic Calculus Bovis, Rhizoma Coptidis, Rhizoma Cyperi, Realgar, Fructus Gleditsia, Radix Platycodonis, Radix Sophorae Tonkinensis, Radix Glycyrrhizae, Alumen, Rhizoma Zingiberis etc. of Guo formed " antasthmatic powder ", is used for the treatment of swine enzootic pneumonia, and effect and tylosin phosphonate are suitable, and pig is only had no side effect.Kong Xiangchen etc. form prescription with Herba Ephedrae, Herba Lophatheri, Radix Glycyrrhizae, Radix Platycodonis, Radix Polygalae, Folium Eriobotryae, Fructus Trichosanthis etc., admix after decocting or the pulverizing and feed the morbidity pig in the feedstuff, have obtained satisfactory therapeutic effects.Also have some with the bonded method treatment of Chinese and Western veterinary swine enzootic pneumonia, good curative effect is also arranged.
But some prescribed preparations that use exist deficiency too at present, can't promote the use of in modernized pig farm as decoction.Use traditional processing technology to make powder, make some effective ingredient not to be utilized well, bioavailability is low, causes the waste of natural resources of Chinese medicinal materials.The preparation prescription that has is huge, for the action effect of bringing into play single medicinal material just must increase total dosage, brings above-mentioned drawback equally.The prescription that has is studied without modern pharmacology, and its mechanism of action is not understood, can't be judged whether it has potential untoward reaction yet.
The silver yellow preparation is one of time-honored conventional medicament of China, and by Flos Lonicerae, Radix Scutellariae two flavor drug matchings also are processed into through extraction.Its many-sided pharmacological action and clinical function has been found in research at home and abroad and use, and is used for wind heat model lung and causes diseases such as heating, cough, pharyngopathy.Upper respiratory tract infection, acute tonsillitis, pharyngitis are seen that above-mentioned syndrome person all has better curative effect, and clinical effectiveness is better.
Only silver yellow soup is a kind of for dosage form the earliest, continuous development along with dosage form research, the people doctor has now developed tens kinds of dosage forms such as oral liquid, tablet (comprising buccal tablet), injection, electuary, pill, powder, enema, collutory, spray, aerosol, and be widely used in clinically according to the characteristics of different dosage form, be used for the treatment of human diseases.Specifically see also CN1398626A, CN1323322A, CN101007059A, CN1634461A, CN1506078A, CN1616050A, CN1939409A, CN1939408A, CN1768785A, CN1424084A, CN1939407A, CN1100653A, CN1742776A, CN1114891A and CN1575813A etc.
But in veterinary applications, available silver yellow preparation is still few, also the silver yellow preparation is not used for the treatment of the report of mycoplasma gallisepticam infection and swine enzootic pneumonia aspect at present.The inventor has verified the effect of silver yellow preparation aspect treatment chicken, porcine mycoplasmal infection in the present invention.
Summary of the invention
The present invention relates to a kind of silver yellow compositions, said composition is made up of Flos Lonicerae extract, Radix Scutellariae extract, it is characterized in that in the Flos Lonicerae extract that content of baicalin is than being 1:13.5～6:1 in the chlorogenic acid contents and Radix Scutellariae extract.
On the other hand, the invention further relates to the present composition is made soluble powder, oral liquid or injection.
The invention further relates to the purposes of silver yellow compositions in the medicine of preparation treatment poultry chronic respiratory tract infections, described poultry are chicken, duck, goose, pig, cattle, sheep, and Canis familiaris L. or rabbit are preferably chicken or pig.
The present invention relates to a kind of silver yellow soluble powder preparation as herbal medicine, its main component is Flos Lonicerae extract and Radix Scutellariae extract, contain Flos Lonicerae in chlorogenic acid (C16H1809) in wherein every gram silver yellow soluble powder, must not be less than 1.7mg, contain Radix Scutellariae in baicalin (C21H18011), must not be less than 18.0mg.
The main effect of silver yellow soluble powder of the present invention is heat-clearing and toxic substances removing, anti-inflammation, cures mainly chicken colibacillosis.Compare with similar powder, not only flavour of a drug are few, and extracts active ingredients is complete, make effective ingredient comprehensive, and curative effect is more outstanding.Because the intestinal of chicken is short, digestion crude fibre ability, crude drug has obviously reduced crude fiber content after extracting, and is beneficial to the absorption of chicken.Significantly improve bioavailability of medicament after the extraction, strengthened curative effect.This medicine is through the drinking-water administration, and medication is simple and convenient, and dose is controlled easily; Make powder with extract, volume is little, preserves convenient transportation, is easily accepted by numerous raisers and distributor.Simultaneously, widely popularize the silver yellow soluble powder after, can obviously reduce the application of antibiotic and chemicals, improve meat egg product matter, ensure human health, obvious social benefit.
Drug quality of the present invention is stable, drug effect height, good effect.For showing stability of drug of the present invention, good effect, we have done great deal of experimental.
Specific embodiment:
Embodiment 1: silver yellow soluble powder preparation technique of the present invention
Concrete steps:
Take by weighing Flos Lonicerae extract 48.0g, Radix Scutellariae extract 32.0g, sodium bicarbonate 40g adds glucose to 1000g, mix homogeneously, packing had both got medicine 1.
Take by weighing Flos Lonicerae extract 900.0g, Radix Scutellariae extract 10.0g, sodium bicarbonate 20g adds glucose to 1000g, mix homogeneously, packing had both got medicine 2.
Take by weighing Flos Lonicerae extract 40g, Radix Scutellariae extract 20g, sodium bicarbonate 20g; Add glucose to 1000g, packing had both got medicine 3.
Embodiment 2: YINHUANG KOUFUYE preparation technology of the present invention
Concrete steps:
Take by weighing Flos Lonicerae extract 48.0g, Radix Scutellariae extract 32.0g adds purified water 600ml, stirs to make it to be suspension, regulates pH value to 5.0 ~ 7.0 with sodium hydroxide solution and makes its dissolving, adds purified water to 1000ml, and packing had both got medicine 4.
Take by weighing Flos Lonicerae extract 900.0g, Radix Scutellariae extract 10.0g adds purified water 600ml, stirs to make it to be suspension, regulates pH value to 5.0 ~ 7.0 with sodium hydroxide solution and makes its dissolving, adds purified water to 1000ml, and packing had both got medicine 5.
Take by weighing Flos Lonicerae extract 40g, Radix Scutellariae extract 20g adds purified water 600ml, stirs to make it to be suspension, regulates pH value to 5.0 ~ 7.0 with sodium hydroxide solution and makes its dissolving, adds purified water to 1000ml, and packing had both got medicine 6.
Embodiment 3: YINHUANG ZHUSHEYE preparation technology of the present invention
Concrete steps:
Take by weighing Flos Lonicerae extract 16.0g, Radix Scutellariae extract 10.0g adds injection water 600ml, stirring makes it to be suspension, regulate pH value to 5.0 ~ 7.0 with sodium hydroxide solution and make its dissolving, benefit adds to the full amount of water for injection, filtering with microporous membrane, be sub-packed in the ampoule bottle, sealing by fusing was sterilized lamp inspection 30 minutes for 115 ℃, packing had both got medicine 7.
Take by weighing Flos Lonicerae extract 300.0g, Radix Scutellariae extract 4.0g adds injection water 600ml, stirring makes it to be suspension, regulate pH value to 5.0 ~ 7.0 with sodium hydroxide solution and make its dissolving, benefit adds to the full amount of water for injection, filtering with microporous membrane, be sub-packed in the ampoule bottle, sealing by fusing was sterilized lamp inspection 30 minutes for 115 ℃, packing had both got medicine 8.
Take by weighing Flos Lonicerae extract 14.5g, Radix Scutellariae extract 6.7g adds the injection water to 600ml, stirring makes it to be suspension, regulate pH value to 5.0 ~ 7.0 with sodium hydroxide solution and make its dissolving, benefit adds to the full amount of water for injection, filtering with microporous membrane, be sub-packed in the ampoule bottle, sealing by fusing was sterilized lamp inspection 30 minutes for 115 ℃, packing had both got medicine 9.
Experimental example
Experimental example 1: the silver yellow soluble powder is to the prevention effect of mycoplasma gallisepticam infection
Mycoplasma gallisepticam infection (Mycoplasma gallisepticum infection) is with cough, sneeze, trachea hello sound and full respiratory inflammation are the infectious disease of feature, once are called as chronic respiratory disease (Chronic respiratory disease) clinically.At present also there are some problems in this sick vaccine research of control, and the cause of disease antimicrobial resistance is on the rise, so be still one of disease of serious harm aviculture so far.Therefore, development effectively preventing medicine is with significant.The silver yellow soluble powder is that the soluble powder that processes from strand has heat-clearing and toxic substances removing by Flos Lonicerae extract and Radix Scutellariae extract prescription, and dampness lung qi dispersing effect is effective to the various bacteria infectious disease.In order to show the effectiveness of the present invention to mycoplasma gallisepticam infection, the clinical effectiveness observation has been carried out in this test.
Trial drug: silver yellow soluble powder medicine 1,2,3 of the present invention, tylosin tartrate, penicillin, thaliium acetate, standard chicken virus mycoplasma antigen and serum.
Laboratory animal: 7542 AA broiler, 16～35 ages in days are identified and Serological testing through Epidemiological study, clinical symptoms discriminating, pathology variation, pathogen separation, are diagnosed as mycoplasma gallisepticam infection; 64.56% concurrent infection escherichia coli wherein.From the blood serum sample that sick chicken is mined massively and collects, 4 ℃ of preservations can not be freezed.
Test method:
1 sets up diagnosis
According to Epidemiological study, clinical symptoms differentiates that pathology change, pathogen separation is identified and Serological testing is comprehensively judged.
1.1 Epidemiological study is carried out according to a conventional method.
1.2 cuing open the inspection variation, pathology carries out according to a conventional method.
1.3 lab testing
1.3.1 the antibacterial separation and Culture is carried out according to a conventional method.
1.3.2 mycoplasma is separated, is identified
Live chickens are taken a sample from nasal cavity, trachea, esophagus, cloaca.Dead chicken is hole, trachea sampling under nasal cavity, the socket of the eye, other draws hole and joint exudate or the interior flushing of conjunctival sac thing under the socket of the eye, makes the sample suspension, and every 1mL adds penicillin 2000IU, 1:4000 thaliium acetate, 1:8 is inoculated in the mycoplasma fluid medium with this sample suspension, and 10 times are diluted to 10 successively
-3, sealing, 37 ℃ of cultivations.Sample is inoculated in the mycoplasma solid medium simultaneously, in containing 5% CO2 gas incubator, cultivates.Check the fluid medium acid-base value every day, find to be inoculated in the solid medium immediately after pH value changes and cultivate.Bacterium colony on the solid medium is observed with low power microscope, and visible central protrusion is poached egg sample (not being true to type sometimes), further does serological Identification again.
Quick plate agglutination test (RSA)
Add (0.02mL) serum in the room temperature on white plaque or glass plate, add equivalent violet staining antigen again, it is mixed into the drop that diameter is 2cm, shake the glass plate gently and make it mix homogeneously with cotton swab or match stick.Positive serum and negative serum contrast are established in test.
Under the criterion room temperature condition, complete coagulation takes place in the 2min, be judged to suspiciously, after this serum was handled through 56 ℃ of 30min, repeated trials if strong agglutination still occurs, or the 24h after dilution or highlyer coagulation person occurs, judged that then serum to be checked is positive.
Microhemagglutination inhibition test (HI)
At first do the antigenic hemagglutinative titer of Microhemagglutination test determination standard chicken virus mycoplasma.
HA: in the micro-reaction of " V " shape is pulled, carry out.Every hole adds 25 μ l normal saline.Dip in the dilution rod and to get 1 of chicken virus mycoplasma standard antigen (doing 5 times of dilutions in advance) (25 μ l),, make doubling dilution successively,, abandon or adopt in the dilution rod to last hole from first hole.Every hole adds 0.5% chicken erythrocyte suspension, 25 μ l, and establishes the erythrocyte control wells that does not add the chicken virus mycoplasma standard antigen, shakes up on agitator immediately, puts 37 ℃ of sense works in the wet box, observed result about 30 minutes.With the high dilution of the complete agglutinative viral liquid of erythrocyte is the terminal point of judging the blood clotting valency.
HI: with the blood clotting valency titre in the above-mentioned HA test divided by 4 value as the antigen hemagglutinating antigen (i.e. 4 HAUs) of this test for example the blood clotting titre be 320 times, 4 HAUs are 80 times so, with 80 times of above-mentioned chicken virus mycoplasma standard antigen dilutions, use after the different dilution sick chicken serum neutralizations, do hemagglutination test again, can reach the highly diluted multiple of the serum that suppresses hemagglutinative function fully and be blood clotting and suppress valency.
When the sick chicken chicken virus mycoplasma of criterion antibody titer surpasses 1:40, positive.
Grouping of 2 experimental animals and processing
The sick chicken of the mycoplasma gallisepticam infection of 7542 natural occurrences is divided into 5 groups at random, and testing 1 group is matched group, does not process; Testing 2 groups is silver yellow soluble powder medicine 1 treatment group, and testing 3 groups is silver yellow soluble powder medicine 2 treatment groups, and testing 4 groups is silver yellow soluble powder medicine 3 treatment groups, and the consumption of silver yellow soluble powder is to add silver yellow soluble powder 1g in 1L drinking-water; Testing 5 groups is tylosin tartrate treatment group, converts water 2.0kg with the 1g tylosin, and the free drink-service of chicken, successive administration 5d are suffered from order.
3 observation index
According to Epidemiological study, clinical symptoms differentiates that pathology change, pathogen separation is identified and Serological testing is determined diagnosis.Through Drug therapy, observe die of illness number, rehabilitation number, healing time and the growth promoter situation etc. of each test group; Calculate case fatality rate, cure rate etc., the difference between relatively each is organized with statistical method.
4 results
4.1 sick chicken diagnostic result
Identify that according to Epidemiological study, clinical symptoms discriminating, pathology variation and pathogen separation the disease that 7542 16～35 aa broiler chicken take a disease of making a definite diagnosis 3 chicken farms, 2 small towns, Dehui City, Jilin Province is mycoplasma gallisepticam infection; 64.56% concurrent infection escherichia coli wherein.
4.1.1 main clinic symptoms
Morbidity successively among the chicken group.Sick chicken essence god is depressed, and feed intake reduces, by Mao Songluan; Occur respiratory symptom thereupon, can know the hiss of hearing that disease chicken trachea sends at nightfall.Heavier with the sick chicken symptom of escherichia coli mixed infection, chicken dehisces to pant, and usually peak with heading is straight, and the hat whisker is livid purple; From the nostril, have a running nose, begin to be the clear water sample, can from the nostril, squeeze out heavy-gravity pus sample secretions to the later stage, seriously block the nostril, one or both sides.The also phenomenon that drops tears of commonly encountered diseases chicken, normal branch hole danger swelling, swelling appears in both sides simultaneously sometimes, and by being pressed with slight fluctuation, the eye danger is bonding up and down because of thick secretions makes for the chicken that has, the clavus ball atrophy that has even blind.
4.1.2 cuing open inspection, pathology changes
More mucus or Catarrhal secretions, the slight edema of tracheal wall appear in nostril, nasal sinuses, trachea and the lungs.Air bag muddiness along with having obviously thickens, and follows the string, and occurs lark on the airbag wall to xanchromatic cheesy exudate, is pearl, lamellar or bulk.Most of mixed infection cases when typical airsacculitis takes place, are seen fiber disposition or fibroid pyoperihepatitis and pericarditis.Arthritis appears in indivedual sick chickens, joint surrounding tissue edema, and joint fluid increases, and what have is limpid, the muddiness that has, what have is butyrous.
4.1.3 laboratory diagnosis
4.1.3.1 mycoplasma is separated, qualification result shows that separator is a chicken virus mycoplasma.
4.1.3.2 antibacterial separation and Culture and qualification result, quite a few chicken (64.56%) has escherichia coli mixed infection.
4.2 the silver yellow soluble powder is to the therapeutic effect of mycoplasma gallisepticam infection
Make the state of an illness very fast controlled after taking the silver yellow soluble powder, untreated matched group spontaneous recovery rate is 89.22%, though fatality rate is not high, glucose recovery is slow; Silver yellow soluble powder treatment group cure rate reaches 99.09%, is significantly higher than tylosin tartrate treatment group (P＜0.01), sees Table 1.
Table 1 silver yellow soluble powder is to the therapeutic effect of mycoplasma gallisepticam infection
4.3 the silver yellow soluble powder merges the therapeutic effect of coli-infection to mycoplasma gallisepticam infection
According to Epidemiological study, clinical symptoms differentiate, pathology change and pathogen separation identifies that 7542 virus mycoplasmas being made a definite diagnosis infect in the disease chicken, 4869 concurrent infections have been arranged escherichia coli account for 64.56%.Concurrent infection person not only symptom is heavier, and case fatality rate (16.23%) also is higher than simple virus mycoplasma infection (10.78%, P＜0.01).Yet the silver yellow soluble powder is still effective to this type mycoplasma gallisepticam infection case, and cure rate reaches 98.18%, and do not have significant difference between the cure rate to simple virus mycoplasma infection (P〉0.05), see Table 2.
Table 2 silver yellow soluble powder merges the therapeutic effect of coli-infection to mycoplasma gallisepticam infection
Experimental example 2: YINHUANG KOUFUYE is to the prevention effect of mycoplasma gallisepticam infection
Trial drug: YINHUANG KOUFUYE medicine 4,5,6 of the present invention, tylosin tartrate, penicillin, thaliium acetate, standard chicken virus mycoplasma antigen and serum.
Laboratory animal: 2295 AA broiler, 16～35 ages in days are identified and Serological testing through Epidemiological study, clinical symptoms discriminating, pathology variation, pathogen separation, are diagnosed as mycoplasma gallisepticam infection; 56.73% concurrent infection escherichia coli wherein.From the blood serum sample that sick chicken is mined massively and collects, 4 ℃ of preservations can not be freezed.
Test method
1 sets up diagnosis
According to Epidemiological study, clinical symptoms differentiates that pathology change, pathogen separation is identified and Serological testing is comprehensively judged.
1.1 Epidemiological study is carried out according to a conventional method.
1.2 cuing open the inspection variation, pathology carries out according to a conventional method.
1.3 lab testing
1.3.1 antibacterial separation and Culture conventional method.
1.3.2 mycoplasma is separated, is identified
Live chickens are taken a sample from nasal cavity, trachea, esophagus, cloaca.Dead chicken is hole, trachea sampling under nasal cavity, the socket of the eye, other draws hole and joint exudate or the interior flushing of conjunctival sac thing under the socket of the eye, makes the sample suspension, and every mL adds penicillin 2000IU, 1:4000 thaliium acetate, 1:8 is inoculated in the mycoplasma fluid medium with this sample suspension, and 10 times are diluted to 10 successively
-3, sealing, 37 ℃ of cultivations.Sample is inoculated in the mycoplasma solid medium simultaneously, in containing 5% CO2 gas incubator, cultivates.Check the fluid medium acid-base value every day, find to be inoculated in the solid medium immediately after pH value changes and cultivate.Bacterium colony on the solid medium is observed with low power microscope, and visible central protrusion is poached egg sample (not being true to type sometimes), further does serological Identification again.
Quick plate agglutination test (RSA)
Add (0.02mL) serum in the room temperature on white plaque or glass plate, add equivalent violet staining antigen again, it is mixed into the drop that diameter is 2cm, shake the glass plate gently and make it mix homogeneously with cotton swab or match stick.Positive serum and negative serum contrast are established in test.
Under the criterion room temperature condition, complete coagulation takes place in the 2min, be judged to suspiciously, after this serum was handled through 56 ℃ of 30min, repeated trials if strong agglutination still occurs, or the 24h after dilution or highlyer coagulation person occurs, judged that then serum to be checked is positive.
Microhemagglutination inhibition test (HI)
At first do the antigenic hemagglutinative titer of Microhemagglutination test determination standard chicken virus mycoplasma.
HA: in the micro-reaction of " V " shape is pulled, carry out.Every hole adds 25 μ l normal saline.Dip in the dilution rod and to get 1 of chicken virus mycoplasma standard antigen (doing 5 times of dilutions in advance) (25 μ l),, make doubling dilution successively,, abandon or adopt in the dilution rod to last hole from first hole.Every hole adds 0.5% chicken erythrocyte suspension, 25 μ l, and establishes the erythrocyte control wells that does not add the chicken virus mycoplasma standard antigen, shakes up on agitator immediately, puts 37 ℃ of sense works in the wet box, observed result about 30 minutes.With the high dilution of the complete agglutinative viral liquid of erythrocyte is the terminal point of judging the blood clotting valency.
HI: with the blood clotting valency titre in the above-mentioned HA test divided by 4 value as the antigen hemagglutinating antigen (i.e. 4 HAUs) of this test for example the blood clotting titre be 320 times, 4 HAUs are 80 times so, with 80 times of above-mentioned chicken virus mycoplasma standard antigen dilutions, use after the different dilution sick chicken serum neutralizations, do hemagglutination test again, can reach the highly diluted multiple of the serum that suppresses hemagglutinative function fully and be blood clotting and suppress valency.
When the sick chicken chicken virus mycoplasma of criterion antibody titer surpasses 1:40, positive.
Grouping of 2 experimental animals and processing
993 sick chickens of simple mycoplasma gallisepticam infection and 1302 colibacillary sick chickens of concurrent infection are divided into 5 groups respectively at random, and testing 1 group is matched group, does not process; Testing 2 groups is YINHUANG KOUFUYE medicine 4 treatment groups, and testing 3 groups is YINHUANG KOUFUYE medicine 5 treatment groups, and testing 4 groups is YINHUANG KOUFUYE medicine 6 treatment groups, and the consumption of YINHUANG KOUFUYE is converted water 1.0kg (1.0 ‰) for the 1mL YINHUANG KOUFUYE; Testing 5 groups is tylosin treatment group, converts water 2.0kg with the 1g tylosin, and the free drink-service of chicken, successive administration 5d are suffered from order.
3 observation index
According to Epidemiological study, clinical symptoms differentiates that pathology change, pathogen separation is identified and Serological testing is determined diagnosis.Through Drug therapy, observe die of illness number, rehabilitation number, healing time and the growth promoter situation etc. of each test group; Calculate case fatality rate, cure rate etc., the difference between relatively each is organized with statistical method.
4 results
4.1 sick chicken diagnostic result
Identify according to Epidemiological study, clinical symptoms discriminating, pathology variation and pathogen separation, make a definite diagnosis 2295 AA broiler that Dehui City, Jilin Province reaches tame ditch chicken farm, be mycoplasma gallisepticam infection.Wherein 993 is simple mycoplasma gallisepticam infection, accounts for 43.27%; 1302 concurrent infection escherichia coli account for 56.73%.
4.1.1 main clinic symptoms
Morbidity successively among the chicken group.Sick chicken essence god is depressed, and feed intake reduces, by Mao Songluan; Occur respiratory symptom thereupon, can know the hiss of hearing that disease chicken trachea sends at nightfall.Heavier with the sick chicken symptom of escherichia coli mixed infection, chicken dehisces to pant, and usually peak with heading is straight, and the hat whisker is livid purple; From the nostril, have a running nose, begin to be the clear water sample, can from the nostril, squeeze out heavy-gravity pus sample secretions to the later stage, seriously block the nostril, one or both sides.The also phenomenon that drops tears of commonly encountered diseases chicken, a normal branch hole eyelid swelling, swelling appears in both sides simultaneously sometimes, and by being pressed with slight fluctuation, the chicken that has is because of sticky secretions makes palpebra inferior bonding, the clavus ball atrophy that has even blind.
4.1.2 cuing open inspection, pathology changes
More mucus or Catarrhal secretions, the slight edema of tracheal wall appear in nostril, stuffy nose, trachea and the lungs.Air bag muddiness along with having obviously thickens, and follows the string, and occurs lark on the airbag wall to xanchromatic cheesy exudate, is pearl, lamellar or bulk.Most of mixed infection cases when typical airsacculitis takes place, are seen fiber disposition or fibroid pyoperihepatitis and pericarditis.Arthritis appears in indivedual sick chickens, joint surrounding tissue edema, and joint fluid increases, and what have is limpid, the muddiness that has, what have is butyrous.
4.1.3 laboratory diagnosis
4.1.3.1 mycoplasma is separated, qualification result shows that separator is a chicken virus mycoplasma.
4.1.3.2 antibacterial separation and Culture and qualification result, quite a few chicken (56.73%) has escherichia coli mixed infection.
4.2 YINHUANG KOUFUYE is to the therapeutic effect of simple mycoplasma gallisepticam infection
Make the state of an illness very fast controlled after taking YINHUANG KOUFUYE, untreated matched group spontaneous recovery rate is 85.85%, though fatality rate is not high, glucose recovery is slow; YINHUANG KOUFUYE group cure rate reaches 98.79%, is significantly higher than tylosin treatment group (P＜0.05).
Table 1 YINHUANG KOUFUYE is to the therapeutic effect of mycoplasma gallisepticam infection
4.3 YINHUANG KOUFUYE merges the therapeutic effect of coli-infection to mycoplasma gallisepticam infection
The Epidemiological study of this experimental evidence, clinical symptoms are differentiated, pathology change and pathogen separation identifies that 2295 virus mycoplasmas being made a definite diagnosis infect in the disease chicken, and 1302 concurrent infections have been arranged escherichia coli account for 56.73%.YINHUANG KOUFUYE is still effective to this type mycoplasma gallisepticam infection case, and YINHUANG KOUFUYE treatment group cure rate reaches 98.17%, and do not have significant difference between the cure rate to simple virus mycoplasma infection (P〉0.05, table 2).
Table 2 YINHUANG KOUFUYE merges the therapeutic effect of coli-infection to chicken virus mycoplasma
Experimental example 3: YINHUANG ZHUSHEYE is to the therapeutic effect of swine enzootic pneumonia
Swine enzootic pneumonia be by mycoplasma hyopneumoniae infection cause a kind of be the infectious disease of primary symptom with dyspnea and cough, still widely take place so far because of the vaccine protective rate is high.Mycoplasma hyopneumoniae has produced strong drug resistance to multiple antibiotic such as sulfonamides and penicillins, thereby it is very necessary to develop more efficient, safe medicine.YINHUANG ZHUSHEYE is the injection that extract polish such as Flos Lonicerae, Radix Scutellariae are made, and has the effect of heat-clearing and toxic substances removing, anti-inflammation, the mixed infection that is mainly used in poultry fever caused by exogenous pathogens, pneumonia asthma and is caused by various bacteria.In order to observe the effect of YINHUANG ZHUSHEYE treatment swine enzootic pneumonia, the sick pig of asthma of 315 natural infections is chosen in this test from different pig farms, and the grouping isolated rearing uses YINHUANG ZHUSHEYE to treat, and has observed the effect of this medicine treatment swine enzootic pneumonia.
Trial drug: YINHUANG ZHUSHEYE medicine 7,8,9 of the present invention, the hydroformylation erythrocyte diagnosticum of tylosin tartrate injection, antigen sensibilization, standard positive serum.
Laboratory animal: the swine enzootic pneumonia of natural infection is suffered from 315 of pigs, makes a definite diagnosis through Epidemiological study, clinical symptoms, pathological change, indirect hemagglutination test; Be acute, chronic type does not select; Long white hybridization commodity generations, 3～10 ages in week.
Test method:
1 sets up diagnosis
According to Epidemiological study, clinical symptoms differentiates that pathology change, pathogen separation is identified and Serological testing is comprehensively judged.
Grouping of 2 experimental animals and processing
The sick pig of the swine enzootic pneumonia of 315 natural occurrences is divided into 5 groups separately at random by feed lot, and the grouping situation sees Table 1.Testing 1 group is matched group, does not process, and quarantines; Testing 2 groups is YINHUANG ZHUSHEYE medicine 7 treatment groups, and testing 3 groups is YINHUANG ZHUSHEYE medicine 8 treatment groups, and testing 4 groups is YINHUANG ZHUSHEYE medicine 9 treatment groups, and the consumption of silver yellow soluble powder is by each every intramuscular injection YINHUANG ZHUSHEYE 0.1ml/kg; Testing 5 groups is tylosin tartrate treatment group, by each every intramuscular injection tylosin tartrate injection 10mg/kg.More than each the group, medication every day 2 times, successive administration 5d.
3 observation index
Collect and analytic epidemiology investigation, main clinic symptoms and pathological change, judgement pathogen separation qualification result writes down dying of illness of each test group and counts, cures number, calculates case fatality rate and cure rate, the difference between relatively each is organized with statistical method.
4 results
4.1 diagnosis is set up
4.1.1 incidence and main clinic symptoms
Beginning at the beginning of 2004 12 months because changeable when ground temperature, cause a plurality of 1～2 monthly age of pig farm piglet to take place to breath with cough in a large number be the respiratory tract disease of cardinal symptom.Brood pig almost breaks out simultaneously, and dyspnea, frequency of respiration per minute can reach 60～120 times, are tangible ventral breathing, and severe patient is dog and sits gesture, and straight of peak with heading dehisces to breathe, mouth and nose stream foam, and the cough number of times is few and overcast.Body temperature is generally not high during no secondary infection, indivedual fervescence to 40 ℃, and when sick pig dyspnea, spirit is depressed, and appetite subtracts greatly, even death by suffocation.The slow slightly person of the state of an illness increases of coughing plants oneself during cough, and hogback is stretched neck, and be sagging, firmly cough repeatedly, cases are continuous spasmodic cough individually, and are moving then gasp for breath.
4.1.2 main pathological change
Major lesions sees lung, hilar lymph node and mediastinal lymph nodes.Lungs have edema and edema due to disorder of QI in various degree, every leaf confluent bronchopneumonia appears at lobus cardiacus, sharp leaf, middle leaf and some cases, lateral symmetry, bright with the normal lung tissue boundary; that pathological changes portion is bois de rose or pale red, and is translucent, the matter degree increases; be carnification, its pathological changes is the most remarkable with lobus cardiacus and sharp leaf.The lobe of the lung pathological changes that has is canescence or lark, becomes " pancreas change " or " shrimp carnification ".Cut focus, flow out muddiness, canescence in the bronchia, be with foamy serosity or mucous liquid, the remarkable enlargement of hilar lymph node and mediastinal lymph nodes is canescence, the swelling of marrow sample, and tangent plane turns up slightly, and is moistening, sometimes the edge hyporrhea.
4.1.3 Serological testing is indirect hemagglutination test all positive (12 negative pigs are not elected experimental animal as) as a result.
By above comprehensive analysis and judgement, be diagnosed as swine enzootic pneumonia (mycoplasmal pneumonia).
4.2 the therapeutic effect of YINHUANG ZHUSHEYE
2nd, make the state of an illness very fast controlled after 3 groups of sick pigs are treated with YINHUANG ZHUSHEYE and tylosin injection.Behind the 12h, the state of an illness is tending towards alleviating gradually, breathes and fades in steadily, and spirit takes a turn for the better, existing appetite behind the 24h; The every symptom of sick pig greatly alleviates behind the 48h; Body temperature, breathing, spirit and diet desire recover normal behind the 72h.YINHUANG ZHUSHEYE group and tylosin group cure rate are significantly higher than matched group (P＜0.01); YINHUANG ZHUSHEYE treatment group cure rate and tylosin treatment group therapeutic equivalence (P〉0.05).
Table 1 YINHUANG ZHUSHEYE is to the therapeutic effect of swine enzootic pneumonia
YINHUANG ZHUSHEYE is processed by Chinese medicine Flos Lonicerae and Radix Scutellariae extract, has heat-clearing and toxic substances removing, depressed lung-energy dispersing effect, for animal various bacteria, mycoplasma infection disease good therapeutic effect is arranged.This evidence, YINHUANG ZHUSHEYE also has very high curative effect to the swine enzootic pneumonia of natural infection morbidity.After sick pig was treated with YINHUANG ZHUSHEYE, the very fast alleviation of the state of an illness was also finally controlled.YINHUANG ZHUSHEYE treatment group cure rate and tylosin treatment group do not have significant difference.
Claims (5)
1, a kind of silver yellow compositions, said composition is made up of Flos Lonicerae extract, Radix Scutellariae extract, it is characterized in that in the Flos Lonicerae extract that content of baicalin is than being 1:13.5～6:1 in the chlorogenic acid contents and Radix Scutellariae extract.
2, silver yellow compositions according to claim 1 is characterized in that said composition is made soluble powder, oral liquid or injection.
3, the purposes of silver yellow compositions according to claim 1 in the medicine of preparation treatment poultry chronic respiratory tract infections.
4, purposes according to claim 3 is characterized in that described poultry are chicken, duck, goose, pig, cattle, sheep, Canis familiaris L. or rabbit.
5, purposes according to claim 4 is characterized in that described poultry are chicken or pig.
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