CN101664421A - Traditional Chinese biological preparation for preventing and treating livestock diarrhea and preparation method thereof - Google Patents
Traditional Chinese biological preparation for preventing and treating livestock diarrhea and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a traditional Chinese biological preparation for preventing and treating livestock diarrhea and a preparation method thereof. The traditional Chinese biological preparation isprepared by combining a traditional prescription and a modern microbiological transformation technology, utilizing the strong transformation capacity of microorganism and fermenting and metabolizing the traditional Chinese pulsatilla root. The preparation method comprises the following steps: crushing the Chinese pulsatilla root; adding bran, glucose, urea, inorganic salt and water as a fermentation medium; adjusting the pH value; adding a strain culture liquid with a certain proportion for fermenting after sterilization; adding ethanol into matured solid leavening for lixiviating, separating,filtering, concentrating and preparing various preparations. The traditional Chinese biological preparation is prepared by the Chinese pulsatilla root after being fermented and metabolized by yeast,and has greatly superior treatment effect to Chinese pulsatilla root medicinal powder and decoctum thereof for the livestock diarrhea.
Description
Technical field
The present invention relates to a kind of traditional Chinese biological preparation of preventing and treating livestock and poultry diarrhea and preparation method thereof, belong to field of veterinary.
Background technology
Dysentery and diarrhoea are digestive tract disease common on the veterinary clinic, and majority causes that by escherichia coli, Salmonella, coccus etc. sickness rate is very high in the poultry various diseases, all can fall ill throughout the year.Antibiotic or chemicals are mainly adopted in treatment to livestock and poultry diarrhea on the veterinary clinic, and therapeutic effect is not good, and the drug resistance of pathogenic bacterium is more and more serious.Treat that this sick Chinese medicine has dysentery relieving to loose, young dysentery is clean, the Radix Pulsatillae looses, Fructus Mume looses etc., these pharmaceutical dosage forms mainly are traditional powder, with low content of technology, quality is coarse, and consumption is big, and palatability is poor, it is slow to take effect, therefore bioavailability is low, and it is undesirable to prevent and treat the livestock and poultry diarrhea disease effects, has caused the significant wastage of natural resources of Chinese medicinal materials simultaneously.
Modern Chinese medicine fermentation pharmacy technology is on the basis of inheriting tradition science of Chinese drug processing fermentation method, drawn the microecology achievement in research, the high-tech herbal pharmaceutical new technique that forms in conjunction with the modern biological project fermentation technique, be the new curative effect of seeking medicine from Chinese medicine (natural drug) pharmacy aspect, have very big researching value and market development meaning.Studies show that, can produce abundant and powerful enzyme system in the growth of microorganism metabolic process, can be with the ingredient decomposition and inversion of Chinese medicine, can not be degraded into small-molecule active substance by the active component of human body or animal body absorption, and directly absorbed by body, or produce new active component, change the property of medicine, reduce toxic and side effects, thereby improve curative effect of medication and therapeutic domain greatly.
The Chinese medicinal formulae of forming by the Radix Pulsatillae, Cortex Phellodendri, Rhizoma Coptidis, Cortex Fraxini, for controlling dysentery name side in the ancient times pharmacy masterpiece treatise on Febrile Diseases, prescription is rigorous, is called as " Radix Pulsatillae Decoction ", and major function is a heat-clearing and toxic substances removing, eliminating pathogenic heat from blood to cure dysentery.Later age, normal alternative use was in multiple disease.Include the Radix Pulsatillae in " Chinese veterinary drug allusion quotation " and loose, be used for treating the livestock and poultry diarrhea disease.At present, do not see both at home and abroad the diffusing complete square research report that carries out microbial transformation and make corresponding preparations of the Radix Pulsatillae.
Summary of the invention
The traditional Chinese biological preparation that the purpose of this invention is to provide a kind of instant effect, control livestock and poultry diarrhea that effect is clear and definite.
Another object of the present invention provides the preparation method of the traditional Chinese biological preparation of this control livestock and poultry diarrhea.
The present invention also provides the feedstuff of the traditional Chinese biological preparation that comprises this control livestock and poultry diarrhea.
Traditional Chinese biological preparation of the present invention is to utilize microorganism that the Radix Pulsatillae is loose to carry out being prepared from behind the fermentating metabolism, has realized the combination of Chinese medicine prescription and modern biotechnology.
The objective of the invention is to be achieved through the following technical solutions:
The invention provides a kind of traditional Chinese biological preparation of preventing and treating livestock and poultry diarrhea, this preparation is that microbial inoculant is carried out fermenting process of preparing in the culture medium that the Radix Pulsatillae looses and forms to comprising.
Following component and parts by weight are drawn together in Radix Pulsatillae Bales Off of the present invention:
20～80 parts of the Radix Pulsatillaes, 30～60 parts of Cortex Phellodendris, 20～80 parts of Cortex Fraxinis, 10～50 parts of Rhizoma Coptidis.
Wherein, described microorganism is a yeast.
Described yeast is Candida utilis and/or saccharomyces cerevisiae.
The traditional Chinese biological preparation of control livestock and poultry diarrhea of the present invention can be acceptable forms on oral liquid, powder, tablet, granule, slow releasing preparation, pill, microcapsule, injection or other pharmaceuticss.
The present invention also provides the preparation method of the traditional Chinese biological preparation of this control livestock and poultry diarrhea, it is characterized in that may further comprise the steps:
(1) preparation bacteria culture fluid
In the YPD culture medium, 28～32 ℃, 150～200rpm shaking table is cultivated the production primary seed solution, when viable bacteria content reaches 1.0 * 10 with the yeast bacterial classification inoculation
8～9Be transferred to secondary seed solution fermentation tank amplification culture during cfu/ml, inoculum concentration is 3～8%, 28～32 ℃ of cultivation temperature, and speed of agitator 180～220rpm, the secondary seed solution viable bacteria content is 1.0 * 10
8～9During cfu/ml, stop to cultivate, standby;
(2) preparation of culture medium: get the Chinese crude drug of formula ratio, be ground into 80～120 purpose medical material fine powders, in the fermentation tank of packing into, add the wheat bran, 1～5% glucose, 0.1～1.0% carbamide, the 0.5～2.0%K that account for Chinese medicine weight 5～10%
2HPO
4, add water to water content 40～60%, regulate pH to 4～5, sterilization 20～25min;
(3) after the cooling, by 5～10% of fermented product gross weight the strain fermentating liquid in (1) is linked in the culture medium of (2), in 28～37 ℃ of constant temperature culture 24～72h, speed of agitator 150～250rpm is when viable bacteria content reaches 1.0 * 10
8～9Stop fermentation during cfu/ml;
(4) sophisticated solid fermentation thing is carried out lixiviate with ethanol, separate, filter, filtrate is refining, and it is 1.0～1.2 that fermentation liquid is evaporated to relative density, adds adjuvant pharmaceutically commonly used and makes corresponding dosage form.
Further, the preparation method of the traditional Chinese biological preparation of control livestock and poultry diarrhea of the present invention adds adjuvant and makes pre-mixing agent after can comprising the steps: that also concentrated broth that step (4) obtains is through spray drying or lyophilization.
The traditional Chinese biological preparation of control livestock and poultry diarrhea of the present invention can also directly be added in the feedstuff, makes the feedstuff that comprises this traditional Chinese biological preparation.
Raw material of the present invention contains the Radix Pulsatillae, Cortex Phellodendri, Cortex Fraxini, Rhizoma Coptidis four Chinese medicine, and modern study shows that the Radix Pulsatillae contains protoanemonin, pulchinenoside; Rhizoma Coptidis, Cortex Phellodendri contain berberine, jateorhizine; Cortex Fraxini contains Esculin, aesculetin isoreactivity composition, and by making the easier absorption of medicine behind the microbial fermentation, drug effect is more abundant.
Traditional Chinese biological preparation of the present invention and traditional herbal medicine relatively have following advantage:
1, poor, for oral administration back of traditional herbal medicine powder palatability effective ingredient is difficult for dissociating out from plant cell fully being absorbed by body, and bioavailability is low, and decoct centering pharmaceutically active ingredient in preparation process is destructive big, has reduced the due curative effect of medicine.Microbial fermentation transforms Chinese medicine, the reaction condition gentleness, pharmaceutically active ingredient exempts from destruction in can protecting to greatest extent, simultaneously, can make some active constituent content increase and produce new ingredient, thereby improve curative effect of medication, expansion indication, minimizing taking dose;
2, the Radix Pulsatillae looses through the traditional Chinese biological preparation for preparing after the microbial metabolism, is better than traditional Radix Pulsatillae greatly and looses and decoct on livestock and poultry diarrhea treatment of diseases effect.
, it should be understood that described embodiment only is for the present invention is described, rather than limit the scope of the invention by any way more specific description the present invention by following examples.
The specific embodiment
The preparation of embodiment 1 traditional Chinese biological preparation of the present invention
The Radix Pulsatillae looses and forms: 60 parts of the Radix Pulsatillaes, 50 parts of Cortex Phellodendris, 60 parts of Cortex Fraxinis, 30 parts of Rhizoma Coptidis
Preparation method:
(1) with the yeast bacterial classification inoculation in the YPD culture medium, 30 ℃, the 180rpm shaking table is cultivated the production primary seed solution, when viable bacteria content reaches 1.0 * 10
8～9Be transferred to secondary seed solution fermentation tank amplification culture during cfu/ml, inoculum concentration is 6%, 30 ℃ of cultivation temperature, and speed of agitator 220rpm, the secondary seed solution viable bacteria content reaches 1.0 * 10
8～9During cfu/ml, stop to cultivate, standby;
(2) get the Chinese crude drug of above-mentioned formula ratio, be ground into 120 purpose medical material fine powders, add the wheat bran, 2% glucose, 0.5% carbamide, the 1.0%K that account for Chinese medicine weight 10%
2HPO
4, adding water to water content is 50%, regulates pH to 4, sterilization 20min;
(3) after the cooling, by 10% of fermented product gross weight the bacteria culture fluid in (1) is linked in the culture medium of (2), in 30 ℃ of constant temperature culture 72h, speed of agitator 250rpm is when viable bacteria content reaches 1.0 * 10
9Stop fermentation during cfu/ml;
(4) sophisticated solid fermentation thing is carried out lixiviate with ethanol, separate, filter, filtrate is refining, and fermentation liquid concentrating under reduced pressure relative density is 1.0;
(5) fermentation liquid after concentrating adds water to 200 parts, filter, and packing, oral liquid is promptly made in sterilization, and 1ml solution is equivalent to crude drug in whole 1g.
The preparation of embodiment 2 traditional Chinese biological preparation of the present invention
The Radix Pulsatillae looses and forms: 80 parts of the Radix Pulsatillaes, 60 parts of Cortex Phellodendris, 80 parts of Cortex Fraxinis, 50 parts of Rhizoma Coptidis
Preparation method:
(1) with the yeast bacterial classification inoculation in the YPD culture medium, 30 ℃, the 180rpm shaking table is cultivated the production primary seed solution, when viable bacteria content reaches 1.0 * 10
8～9Be transferred to secondary seed solution fermentation tank amplification culture during cfu/ml, inoculum concentration is 8%, 30 ℃ of cultivation temperature, and speed of agitator 220rpm, the secondary seed solution viable bacteria content reaches 1.0 * 10
8～9During cfu/ml, stop to cultivate, standby;
(2) get the Chinese crude drug of above-mentioned formula ratio, be ground into 100 purpose medical material fine powders, add the wheat bran, 3% glucose, 0.7% carbamide, the 1.5%K that account for Chinese medicine weight 8%
2HPO
4Adding water to water content is 60%, regulates pH to 4.5, sterilization 25min;
(3) after the cooling, by 8% of fermented product gross weight the bacteria culture fluid in (1) is linked in the culture medium of (2), in 32 ℃ of constant temperature culture 48h, speed of agitator 180rpm is when viable bacteria content reaches 1.0 * 10
9Stop fermentation during cfu/ml;
(4) sophisticated solid fermentation thing is carried out lixiviate with ethanol, separate, filter, filtrate is refining, and it is 1.1 that fermentation liquid is evaporated to relative density, adds an amount of sucrose, dextrin again, the system soft material and the granulation of sieving; Wet grain drying, granulate detect, and packing promptly is prepared into granule, and every 1g is equivalent to crude drug in whole 1g.
The preparation of embodiment 3 traditional Chinese biological preparation of the present invention
The Radix Pulsatillae looses and forms: 50 parts of the Radix Pulsatillaes, 45 parts of Cortex Phellodendris, 50 parts of Cortex Fraxinis, 45 parts of Rhizoma Coptidis
Preparation method:
(1) with the yeast bacterial classification inoculation in the YPD culture medium, 29 ℃, the 190rpm shaking table is cultivated the production primary seed solution, when viable bacteria content reaches 1.0 * 10
8～9Be transferred to secondary seed solution fermentation tank amplification culture during cfu/ml, inoculum concentration is 5%, 29 ℃ of cultivation temperature, and speed of agitator 200rpm, the secondary seed solution viable bacteria content reaches 1.0 * 10
8～9During cfu/ml, stop to cultivate, standby;
(2) get the Chinese crude drug of above-mentioned formula ratio, be ground into 90 purpose medical material fine powders, add the wheat bran, 4% glucose, 0.8% carbamide, the 0.7%K that account for Chinese medicine weight 9%
2HPO
4, adding water to water content is 55%, regulates pH to 5, sterilization 25min;
(3) after the cooling, by 9% of fermented product gross mass the bacteria culture fluid in (1) is linked in the culture medium of (2), in 29 ℃ of constant temperature culture 60h, speed of agitator 200rpm is when viable bacteria content reaches 1.0 * 10
9Stop fermentation during cfu/ml;
(4) sophisticated solid fermentation thing is carried out lixiviate with ethanol, separate, filter, filtrate is refining, it is 1.2 that fermentation liquid is evaporated to relative density, and spray drying obtains pressed powder again, adds an amount of ethanol and makes wetting agent system soft material, granulate, drying adds the magnesium stearate mix homogeneously, tabletting again, promptly make tablet, every is equivalent to crude drug in whole 0.5g.
The preparation of embodiment 4 traditional Chinese biological preparation of the present invention
The Radix Pulsatillae looses and forms: 70 parts of the Radix Pulsatillaes, 40 parts of Cortex Phellodendris, 40 parts of Cortex Fraxinis, 40 parts of Rhizoma Coptidis
Preparation method:
(1) with the yeast bacterial classification inoculation in the YPD culture medium, 30 ℃, the 200rpm shaking table is cultivated the production primary seed solution, when viable bacteria content reaches 1.0 * 10
8～9Be transferred to secondary seed solution fermentation tank amplification culture during cfu/ml, inoculum concentration is 4%, 30 ℃ of cultivation temperature, and speed of agitator 220rpm, the secondary seed solution viable bacteria content reaches 1.0 * 10
8～9During cfu/ml, stop to cultivate, standby;
(2) get the Chinese crude drug of above-mentioned formula ratio, be ground into 100 purpose medical material fine powders, add the wheat bran, 5% glucose, 0.6% carbamide, the 0.5%K that account for Chinese medicine weight 7%
2HPO
4, adding water to water content is 45%, regulates pH to 4, sterilization 20min;
(3) after the cooling, by 7% of fermented product gross weight the bacteria culture fluid in (1) is linked in the culture medium of (2), in 28 ℃ of constant temperature culture 36h, speed of agitator 150rpm is when viable bacteria content reaches 1.0 * 10
9Stop fermentation during cfu/ml;
(4) sophisticated solid fermentation thing is carried out lixiviate with ethanol, separate, filter, filtrate is refining, and it is 1.2 that fermentation liquid is evaporated to relative density, and spray drying adds mix homogeneously behind the appropriate amount of auxiliary materials maize cob meal again, promptly is prepared into pre-mixing agent.
The preparation of embodiment 5 traditional Chinese biological preparation of the present invention
The Radix Pulsatillae looses and forms: 20 parts of the Radix Pulsatillaes, 30 parts of Cortex Phellodendris, 20 parts of Cortex Fraxinis, 10 parts of Rhizoma Coptidis
Preparation method:
(1) with the yeast bacterial classification inoculation in the YPD culture medium, 29 ℃, the 200rpm shaking table is cultivated the production primary seed solution, when viable bacteria content reaches 1.0 * 10
8～9Be transferred to secondary seed solution fermentation tank amplification culture during cfu/ml, inoculum concentration is 7%, 29 ℃ of cultivation temperature, and speed of agitator 200rpm, the secondary seed solution viable bacteria content reaches 1.0 * 10
8～9During cfu/ml, stop to cultivate, standby;
(2) get the Chinese crude drug of above-mentioned formula ratio, be ground into 100 purpose medical material fine powders, add the wheat bran, 1% glucose, 0.4% carbamide, the 1.2%K that account for Chinese medicine weight 5%
2HPO
4, adding water to water content is 40%, regulates pH to 4, sterilization 20min;
(3) after the cooling, by 5% of fermented product gross weight the bacteria culture fluid in (1) is linked in the culture medium of (2), in 31 ℃ of constant temperature culture 24h, speed of agitator 220rpm is when viable bacteria content reaches 1.0 * 10
9Stop fermentation during cfu/ml;
(4) sophisticated solid fermentation thing is carried out lixiviate with ethanol, separate, filter, filtrate is refining, it is 1.2 that fermentation liquid is evaporated to relative density, adds mix homogeneously behind an amount of soluble starch, the dextrin again, and oven dry is pulverized, sieve, promptly make powder, every 1g is equivalent to crude drug in whole 5g.
Below by concrete test example the beneficial effect of traditional Chinese biological preparation of the present invention is described.
Test example 1
This test example is by the oral liquid of the method preparation of embodiment 1 clinical treatment of artificial onset's piglet pujos blancos to be tested.
1, materials and methods
1.1 for reagent thing Radix Pulsatillae decocted medicine (crude drug content is 1g/ml), oral liquid of the present invention.
1.2 strain escherichia coli O
139, China Veterinary Drugs Supervisory Inst. provides.
1.3 200 body weight of animal subject are about the healthy ablactational baby pig about 22 ± 1kg.
1.4 test method
1.4.1 the animal grouping is divided into 5 groups at random with 200 piglets, 40 every group, wherein 1,2,3,4 groups is medication therapy groups, 5 groups of positive matched groups.
1.4.2 artificial onset
To preserve bacterial classification inoculation earlier in common nutrition flat board, 37 ℃ of constant temperature culture 18 hours, the picking colonies typical is inoculated in the ordinary broth then, and 37 ℃ of constant temperature culture were taken out in 18 hours, and being diluted to concentration with normal saline is 5 * 10
8Cfu/ml, after trial test, except that the blank group, other respectively organizes the counteracting toxic substances amount is every pig intramuscular injection 5ml bacterium liquid.Treat that piglet begins medication when classical symptom such as diarrhoea, the white medicated porridge sample feces of row, loss of appetite occurring.
1.4.3 administrated method
1 group is Radix Pulsatillae decocted medicine (1g/ml) treatment group, gavages administration, every pig 3～5ml/ time, one day 2 times, logotype 3 days; 2 groups for oral liquid high dose group of the present invention (1g/ml), gavages administration, every pig 3～5ml/ time, one day 2 times, logotype 3 days; 3 groups for dosage group (0.5g/ml) in the oral liquid of the present invention, gavages administration, every pig 3～5ml/ time, one day 2 times, logotype 3 days; 4 groups for oral liquid low dose group of the present invention (0.25g/ml), gavages administration, every pig 3～5ml/ time, one day 2 times, logotype 3 days; 5 groups of positive matched groups, not administration behind the infection morbidity.Each is organized test pig and all feeds and do not contain the complete feed of medicine.
1.5 therapeutic evaluation index
Cure rate: clinical symptoms complete obiteration after the medication treatment, spirit and diet desire recover the normal healing that is, statistics is cured number, calculates cure rate.
Effective percentage: clinical symptoms obviously alleviates after the medication treatment, spirit and diet desire improvement person or healing, all being considered as effectively, the statistics significant figure calculates effective percentage.
Mortality rate: because of medication is invalid and dead, the statistics death toll is calculated mortality rate during the treatment.
1.6 data analysis and processing
Biometrics is carried out data analysis with t check and X 2 test.Result of the test sees Table 1.
2, result
The therapeutic effect of table 1 pair artificial onset's piglet pujos blancos
| Group | Treatment number/head | Cure number/head | Cure rate (%) | Effective percentage (%) | Mortality rate (%) |
| ??1 | ??40 | ??35 | ??87.5 | ??95.0 | ??0 |
| ??2 | ??40 | ??38 | ??95.0 | ??100 | ??0 |
| ??3 | ??40 | ??34 | ??85.0 | ??94.5 | ??0 |
| ??4 | ??40 | ??30 | ??75.0 | ??87.5 | ??2.5 |
| ??5 | ??40 | ??0 | ??0 | ??0 | ??15 |
High, medium and low dosage group of Chinese medicine oral liquid of the present invention and Radix Pulsatillae decocted medicine group are to the effective percentage and the poor heteropole of positive controls remarkable (P＜0.01) of artificial onset's piglet pujos blancos; The effective percentage of the dosage group difference not significantly (P＞0.05) of comparing with Radix Pulsatillae decocted medicine group in the Chinese medicine oral liquid of the present invention; Illustrating only needs not by fermentation preparation of the present invention the dosage of fermentation preparation 50% just can reach the not therapeutic effect of fermentation preparation.
Test example 2
This test example is with the test of the pre-mixing agent prevention yellow and white dysentery of piglet of the embodiment of the invention 4 preparations, has obtained good test effect.
Select the pig that farrows that 31 sows of HUANGBAI(sic) dysentery medical history are arranged, be divided into 3 groups at random.1 group is pre-mixing agent group of the present invention, and beginning medication in antenatal 5 days is added pre-mixing agent of the present invention, continuous use 5 days by 0.5% in feedstuff; 2 groups is the diffusing group of the Radix Pulsatillae, and beginning medication in same antenatal 5 days is loose continuous use 5 days by the 1.0% interpolation Radix Pulsatillae in feedstuff; 3 groups is matched group, does not add any medicine in the feedstuff; The raising condition of 3 groups is identical.A record birth piglet morbidity number and a dead number calculate and respectively organize sickness rate and mortality rate.Result of the test sees Table 2.
The result of the test of table 2 prevention yellow and white dysentery of piglet
| Group | A sow number | A farrowing number | A piglet morbidity number | A dead number | Sickness rate (%) | Mortality rate (%) |
| ??1 | ??11 | ??122 | ??6 | ??0 | ??4.92 | ??0 |
| ??2 | ??10 | ??113 | ??13 | ??4 | ??11.50 | ??3.54 |
| ??3 | ??10 | ??111 | ??62 | ??27 | ??55.86 | ??24.32 |
Result of the test shows, after the antenatal use of the farrowing sow traditional Chinese biological preparation of the present invention, the sickness rate of the yellow and white dysentery of piglet that produces, mortality rate are compared with ordinary powder group and matched group and are obviously descended, illustrate traditional Chinese biological preparation of the present invention can by sow effectively prevent the birth yellow and white dysentery of piglet, reduce its mortality rate.
Test example 3
This test example is by the Chinese medicine powder of the embodiment 5 preparation viewing test to broiler chicken Hakuri therapeutic effect.
Test method: certain chicken farm provides 20 age in days broiler chicken, through the sick chicken performance of clinical examination lethargy, is afraid of cold, and spirit is depressed, and appetite reduces or do not eat, and wing is sagging, draws white paste sample feces, tangible Pullorum Disease symptom such as adhesion Calx sample feces around the anus.Cut open 10 sick chickens extremely, detecting through laboratory is Salmonella infection.With 750 of the male broiler chicken of the dull and stereotyped agglutination (detecting unknown antigen) of Pullorum Disease whole blood, be divided into 5 groups at random, 150 every group with standard serum.
Wherein 1 group is powder group of the present invention; 2 groups is the gentamycin group; 3 groups is the florfenicol group; 4 groups is Radix Pulsatillae decocted medicine group (crude drug content 1g/ml), and 5 groups is matched group, takes drugless medication, and the dosage and the using method of 4 medication treatment groups see Table 3.Each raising condition of organizing chicken is identical, does not add any medicine in the feedstuff.Continuous use 5 days is observed therapeutic effect.Result of the test sees Table 4.
Table 3 dosage and using method
| Group | Using method | Using dosage | Frequency of usage |
| ??1 | Mix drink | ??0.5g/kg.BW | Every day 1 time, logotype 5 days |
| ??2 | Mix drink | ??30mg/kg.BW | Every day 2 times, logotype 5 days |
| ??3 | Mix drink | ??20mg/kg.BW | Every day 2 times, logotype 5 days |
| ??4 | Mix drink | ??1ml/kg.BW | Every day 2 times, logotype 5 days |
The therapeutic test result of the table 4 pair white scour of chicken
Above-mentioned result of the test shows that traditional Chinese biological preparation of the present invention has excellent curative to Pullorum Disease, can effectively reduce mortality rate, and therapeutic effect is better than Western medicine group and Radix Pulsatillae decocted medicine group.
Test example 4
This test example is by the Chinese medicine granules of the method for the embodiment 2 preparation viewing test to birth lamb diarrheal prevention effect.
1, materials and methods
1.1 320 lambs that ewe produced in 4 months are tested in experimental animal selection pasture, Gansu.
1.2 trial drug Chinese medicine granules of the present invention, Radix Pulsatillae decocted medicine (crude drug content 1g/ml), oxytetracycline.
1.3 the test grouping is divided into 3 groups by the date of birth random packet of lamb.Test 1 group and be preparation group of the present invention; Testing 2 groups is Radix Pulsatillae decocted medicine group; Testing 3 groups is the oxytetracycline matched group.
1.4 administrated method birth lamb eats administration at once behind the colostrum, Chinese medicine granules of the present invention is by per kilogram of body weight 0.5～1.0g administration; Radix Pulsatillae decocted medicine group is by per kilogram of body weight 0.5～1ml administration; Matched group is by the oral oxytetracycline of per kilogram of body weight 0.02～0.05g.The equal day medication of each group 2 times, continuous use 3～5 days.
Observe, write down clinical symptoms such as respectively organizing lamb diarrhoea and death condition every day, add up, analyze.
2, result
Result of the test sees Table 5.
Table 5 pair birth lamb diarrheal preventive effect
| Group | Lamb number (only) | Diarrhoea number (only) | Diarrhea rate (%) | Survive number (only) | Survival rate (only) | Death toll (only) | Mortality rate (%) |
| ??1 | ??110 | ??9 | ??8.2 | ??107 | ??97.3 | ??3 | ??2.7 |
| ??2 | ??110 | ??12 | ??10.9 | ??101 | ??91.8 | ??9 | ??8.2 |
| ??3 | ??100 | ??15 | ??15.0 | ??88 | ??88.0 | ??12 | ??12.0 |
Annotate: survive number, the statistical result when death toll is 45 ages in days.
As can be seen from the test results, preparation of the present invention and Radix Pulsatillae decocted medicine can play the excellent prevention effect to the diarrhoea of birth lamb, compare diarrhea rate with matched group and have reduced by 6.8% and 4.1% respectively, and survival rate improves 9.3% and 3.8% respectively.Difference extremely significantly (P＜0.01) between preparation group of the present invention and the matched group.
Test example 5
This test example is that tablet by the preparation of the method for embodiment 3 is to the various diarrhea treatment observation on effect of dog.
1, experimental animal from several foster dogs field, select to suffer from acute enteritis, mucus enteritis, hemorrhagic enteritis sick dog totally 112 be for experiment.
2,112 dogs that animal divides into groups and medication will be selected are divided into 2 groups, and 1 group of 100 dog is preparation for treating group of the present invention, 72 of dogs that wherein suffer from acute enteritis, 8 of mucus enteritis dogs, 20 of hemorrhagic enteritis dogs, dosage is 1 of a per kilogram of body weight, every day 1 time, logotype 3～5 days; 2 groups of 12 dogs are organized in contrast, do not give any medicine.
3, criterion
Invalid: medication treatment 5 days, symptom do not see change or the state of an illness become serious be considered as invalid.
Effectively: medication treatment 5 days, all being considered as of clinical symptom relief or recovery from illness is effective.
Cure: medication treatment 5 days, clinical symptom disappearance, the dog spirit of suffering from acute enteritis is normal, and vomiting stops fully, and defecation is normal; Dog spirit, body temperature, appetite, the defecation of suffering from mucus enteritis all recover normal; Suffer from the dog stopped bleeding of hemorrhagic enteritis, defecation is normal, and spirit, appetite, temperature recovery are normal, the asoscope moistening.
4, result
Result of the test sees Table 6.
Table 6 pair dog diarrheal therapeutic effect
From result of the test as can be known, the various diarrheal average effectiveness levels of Drug therapy dog of the present invention are more than 90%, and average cure rate is more than 85%, and effect is better.
Claims (8)
1, a kind of traditional Chinese biological preparation of preventing and treating livestock and poultry diarrhea is characterized in that it being that microbial inoculant is carried out fermenting process of preparing in the culture medium that the Radix Pulsatillae looses and forms to comprising.
2, the traditional Chinese biological preparation of control livestock and poultry diarrhea according to claim 1 is characterized in that described Radix Pulsatillae Bales Off draws together following component and parts by weight:
20～80 parts of the Radix Pulsatillaes, 30～60 parts of Cortex Phellodendris, 20～80 parts of Cortex Fraxinis, 10～50 parts of Rhizoma Coptidis.
3, the traditional Chinese biological preparation of control livestock and poultry diarrhea according to claim 1 is characterized in that described microorganism is a yeast.
4, the traditional Chinese biological preparation of control livestock and poultry diarrhea according to claim 3 is characterized in that described yeast is Candida utilis and/or saccharomyces cerevisiae.
5, the traditional Chinese biological preparation of control livestock and poultry diarrhea according to claim 1 is characterized in that described preparation is an acceptable forms on oral liquid, powder, tablet, granule, slow releasing preparation, pill, microcapsule, injection or other pharmaceuticss.
6, according to the preparation method of the traditional Chinese biological preparation of each described control livestock and poultry diarrhea of claim 1-5, it is characterized in that may further comprise the steps:
(1) preparation bacteria culture fluid
In the YPD culture medium, 28～32 ℃, 150～200rpm shaking table is cultivated the production primary seed solution, when viable bacteria content reaches 1.0 * 10 with the yeast bacterial classification inoculation
8～9Be transferred to secondary seed solution fermentation tank amplification culture during cfu/ml, inoculum concentration is 3～8%, 28～32 ℃ of cultivation temperature, and speed of agitator 180～220rpm, the secondary seed solution viable bacteria content is 1.0 * 10
8～9During cfu/ml, stop to cultivate, standby;
(2) preparation of culture medium: get the Chinese crude drug of formula ratio, be ground into 80～120 purpose medical material fine powders, in the fermentation tank of packing into, add the wheat bran, 1～5% glucose that account for Chinese medicine weight 5～10%, 0.1～1.0% carbamide, 0.5～2.0%K
2HPO
4, add water to water content 40～60%, regulate pH to 4～5, sterilization 20～25min;
(3) after the cooling, by 5～10% of fermented product gross weight the bacteria culture fluid in (1) is linked in (2) culture medium, in 28～37 ℃ of constant temperature culture 24～72h, speed of agitator 150～250rpm is when viable bacteria content reaches 1.0 * 10
8～9Stop fermentation during cfu/ml;
(4) sophisticated solid fermentation thing is carried out lixiviate with ethanol, separate, filter, filtrate is refining, and it is 1.0～1.2 that fermentation liquid is evaporated to relative density, adds adjuvant pharmaceutically commonly used and makes corresponding dosage form.
7, the preparation method of the traditional Chinese biological preparation of control livestock and poultry diarrhea according to claim 6, after it is characterized in that also can further comprising the steps: that concentrated broth that step (4) obtains is through spray drying or lyophilization, add adjuvant and make pre-mixing agent.
8, a kind of feedstuff that comprises the traditional Chinese biological preparation of each described control livestock and poultry diarrhea of claim 1-4.
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