CN101690743A - Traditional Chinese medicine biological agent for improving immunity of livestock and preparation method thereof - Google Patents
Traditional Chinese medicine biological agent for improving immunity of livestock and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a traditional Chinese medicine biological agent for improving immunity of livestock and a preparation method thereof; the traditional Chinese medicine biological agent is prepared by combining traditional Chinese medicine formulas and modern microbiological technology; the biological agent adopts folium cortex eucommiae, grifola frondosa, astragalus root and herba epimedii, which belong to traditional Chinese medicines, as raw materials; bran, bean pulp, ammonium sulfate, magnesium sulfate, calcium carbonate and water are added in the raw materials after the raw materials are crushed, so as to form fermentation medium, strain nutrient solution with a certain proportion is added in the fermentation medium for fermentation, the mature solid fermentation broth is added in alcohol for extraction, separation, filtration, concentration, so as to prepare various dosage forms. The traditional Chinese medicine biological agent prepared by microbial fermentation metabolism can remarkably improve the immunity of livestock organisms, enhance the immunity of the organisms, improves the production performance of the livestock and effectively prevent immunity inhibitive diseases.
Description
Technical field
The present invention relates to a kind of traditional Chinese biological preparation that improves immunity of livestock and preparation method thereof, belong to field of veterinary.
Background technology
In recent years, ubiquity vaccine virus immunization preventive effect is bad in Production of Livestock and Poultry, and dosage strengthens gradually and does not still reach ideal treatment and preventive effect, this is to descend owing to a large amount of for a long time Drug abuse and irrational vaccination cause the poultry body's immunity, particularly at a low ebb the or inhibitory state of immunocyte such as lymphocyte, macrophage; The new eqpidemic disease of animal occurs and the pathogen variation and the virulence enhancing of old eqpidemic disease in recent years in addition.Address this problem, just must take the effective means activating immune system, improve activity of immune cells, so that obtain the immunne response of higher level, the resistance against diseases of enhancing body after the animal body vaccination.The research in this field at present is very active, and particularly the research of chemical medicine immunostimulant has obtained bigger progress, and the sound development of poultry industry is played a positive role.But the chemicals immunostimulant is existing and reduce the body fertility performance in various degree all, causes toxic and side effects and drug residue, and drawback such as be detrimental to health.
The Chinese herbal medicine immunostimulant has multi-functionals such as active stimulation, nutrition, adjusting and antibacterial, antiviral, when it acts on each system of body, can make the equilibrated effect of body recovery.Along with the continuous acceleration of China's aquaculture intensification degree, particularly under the tide of pursuing green, environmental protection, ecology promoted, Chinese herbal medicine in the unique advantage aspect the raising immunologic function, had obtained more and more general applying with it in aquaculture.Research Chinese herbal medicine immunostimulant for the control livestock and poultry, improves Production of Livestock and Poultry power to the influence of animal immune effect, increases the animal husbandry economy benefit and has important practical significance.But used Chinese herbal medicine immunostimulant generally is the lower powder of technology content at present, directly adds in the feedstuff and uses, and bioavailability is low and palatability is poor, and is extremely unbecoming with Developing of Animal Industry.
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.Therefore, modern microbial technique and Chinese medicine prescription are combined, the development of new traditional Chinese biological preparation is a breakthrough to Chinese medicine, has higher technical merit, helps improving China's modernization of Chinese medicine level.
Summary of the invention
The purpose of this invention is to provide a kind of traditional Chinese biological preparation that improves immunity of livestock, said preparation improves young stock poult survival rate in the effectively generation of control fowl poultry immune inhibition disease, when promoting growth, and good palatability, bioavailability height.Traditional Chinese biological preparation of the present invention is to utilize microorganism that Chinese medicine is carried out being prepared from behind the fermentating metabolism, has realized the combination of Chinese medicine prescription and modern biotechnology.
Another object of the present invention provides the preparation method of the traditional Chinese biological preparation of this raising immunity of livestock.
The present invention also provides the feedstuff of the traditional Chinese biological preparation that comprises this raising immunity of livestock.
The objective of the invention is to be achieved through the following technical solutions:
The invention provides a kind of traditional Chinese biological preparation that improves immunity of livestock, this preparation is microbial inoculant to be carried out fermenting process of preparing in the culture medium that comprises Chinese medicine form.
Chinese medicine of the present invention comprises following component and parts by weight:
50～150 parts of Folium Eucommiaes, 50～150 parts of Grifola frondosa, 25～100 parts of the Radixs Astragali, 25～100 parts of Herba Epimedii.
Wherein, described microorganism is a bacillus cereus.
Bacillus cereus of the present invention is bacillus subtilis and/or Bacillus licheniformis.
The traditional Chinese biological preparation of raising immunity of livestock 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 raising immunity of livestock, it is characterized in that may further comprise the steps:
(1) preparation bacteria culture fluid
Bacillus cereus is inoculated in the broth medium, and 35～37 ℃, 200～250rpm 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 8～10%, 25～30 ℃ of cultivation temperature, and speed of agitator 200～250rpm, the secondary seed solution viable bacteria content reaches 1.0 * 10
8～9During cfu/ml, stop to cultivate, standby;
(2) preparation of culture medium: the Chinese crude drug of getting formula ratio, be ground into 80～120 purpose medical material fine powders, pack in the fermentation tank, add 10～30% wheat brans, 5～20% bean cake, 0.1～1.0% ammonium sulfate, 0.01～0.10% magnesium sulfate, 0.01～0.05% calcium carbonate that account for Chinese medicine weight, add water to water content 40～60%, regulate pH to 6.5～7.5, sterilization 20～30min;
(3) after the cooling, by 5～10% of fermented product gross weight the bacteria culture fluid in (1) is linked in the culture medium of (2), in 25～30 ℃ 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 raising immunity of livestock 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 raising immunity of livestock of the present invention can also directly be added in the feedstuff, makes the feedstuff that comprises this traditional Chinese biological preparation.
Material choice Folium Eucommiae of the present invention, Grifola frondosa, the Radix Astragali, Herba Epimedii four Chinese medicine make up, make each component effect produce synergism, by making the easier absorption of medicine behind the microbial fermentation, drug effect is more abundant, thereby can further improve the immunity of poultry body, strengthen resistance against diseases, promote growth.
Raising immunity of livestock traditional Chinese biological preparation of the present invention relatively has following advantage with traditional similar herbal medicine:
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 traditional Chinese biological preparation of Chinese medicine of the present invention through preparing after the microbial metabolism can further improve immunity of livestock and fertility performance, effectively prevents and treats the generation of fowl poultry immune inhibition disease, improves young stock poult survival rate, promotes growth.
, 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
Chinese medicine is formed: 100 parts of Folium Eucommiaes, 100 parts of Grifola frondosa, 50 parts of the Radixs Astragali, 50 parts of preparation methoies of Herba Epimedii:
(1) bacillus cereus is inoculated in the broth medium, 37 ℃, the 220rpm 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 10%, 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 formula ratio, be ground into 120 purpose medical material fine powders, in the fermentation tank of packing into, add 20% wheat bran, 10% bean cake, 0.6% ammonium sulfate, 0.05% magnesium sulfate, 0.02% calcium carbonate that account for Chinese medicine weight, add water to water content 50%, regulate pH to 7.0, sterilization 20min;
(3) after the cooling, by 10% of fermented product gross weight the strain liquid in (1) is linked in the culture medium of (2), in 30 ℃ of constant temperature culture 48h, speed of agitator 220rpm 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 that fermentation liquid is evaporated to relative density;
(5) fermentation liquid after concentrating adds water to 300 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
Chinese medicine is formed: 70 parts of Folium Eucommiaes, 60 parts of Grifola frondosa, 40 parts of the Radixs Astragali, 30 parts of preparation methoies of Herba Epimedii:
(1) bacillus cereus is inoculated in the broth medium, 36 ℃, the 230rpm 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%, 29 ℃ of cultivation temperature, and speed of agitator 230rpm, 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 formula ratio, be ground into 100 purpose medical material fine powders, in the fermentation tank of packing into, add 15% wheat bran, 8% bean cake, 0.4% ammonium sulfate, 0.03% magnesium sulfate, 0.03% calcium carbonate that account for Chinese medicine weight, add water to water content 45%, regulate pH to 7.1, sterilization 25min;
(3) after the cooling, by 9% 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 230rpm 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, 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.
The preparation of embodiment 3 traditional Chinese biological preparation of the present invention
Chinese medicine is formed: 125 parts of Folium Eucommiaes, 125 parts of Grifola frondosa, 80 parts of the Radixs Astragali, 70 parts of preparation methoies of Herba Epimedii:
(1) bacillus cereus is inoculated in the broth medium, 35 ℃, 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 9%, 30 ℃ 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 formula ratio, be ground into 90 purpose medical material fine powders, in the fermentation tank of packing into, add 25% wheat bran, 5% bean cake, 0.8% ammonium sulfate, 0.08% magnesium sulfate, 0.04% calcium carbonate that account for Chinese medicine weight, add water to water content 40%, regulate pH to 6.9, 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 30 ℃ of constant temperature culture 24h, speed of agitator 180rpm 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, it is 1.1 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
Chinese medicine is formed: 50 parts of Folium Eucommiaes, 50 parts of Grifola frondosa, 25 parts of the Radixs Astragali, 25 parts of preparation methoies of Herba Epimedii:
(1) bacillus cereus is inoculated in the broth medium, 37 ℃, the 250rpm 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 9%, 28 ℃ of cultivation temperature, and speed of agitator 250rpm, 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 formula ratio, be ground into 80 purpose medical material fine powders, in the fermentation tank of packing into, add 30% wheat bran, 12% bean cake, 1.0% ammonium sulfate, 0.01% magnesium sulfate, 0.01% calcium carbonate that account for Chinese medicine weight, add water to water content 55%, regulate pH to 7.2, 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 48h, speed of agitator 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 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
Chinese medicine is formed: 150 parts of Folium Eucommiaes, 150 parts of Grifola frondosa, 100 parts of the Radixs Astragali, 100 parts of preparation methoies of Herba Epimedii:
(1) bacillus cereus is inoculated in the broth medium, 36 ℃, the 210rpm 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 10%, 27 ℃ of cultivation temperature, and speed of agitator 210rpm, the secondary seed solution viable bacteria content is 1.0 * 10
8～9During cfu/ml, stop to cultivate, standby;
(2) get the Chinese crude drug of formula ratio, be ground into 100 purpose medical material fine powders, in the fermentation tank of packing into, add 12% wheat bran, 6% bean cake, 0.5% ammonium sulfate, 0.07% magnesium sulfate, 0.05% calcium carbonate that account for Chinese medicine weight, add water to water content 60%, regulate pH to 6.7, sterilization 30min;
(3) after the cooling, by 6% 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 150rpm 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 1.2 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 2g.
Below by concrete test example the beneficial effect of pharmaceutical composition of the present invention is described.
Test example 1
The pre-mixing agent of embodiment 4 preparations is to the fertility performance and the exponential influence of immune organ of broiler.
1, test material
Trial drug: pre-mixing agent of the present invention, lincomycin.
Experimental animal: 300 of the healthy commercial chickens of 1 age in days are provided by chicken farm, Beijing.
2, test method
300 chickens are divided into 5 groups at random, and every group each 60, each organizes 2 repetitions, and each repeats 30 chickens.Raise respectively in 5 chicken hurdles.Wherein 1,2,3 group is pre-mixing agent group of the present invention, adds pre-mixing agent by 0.5%, 0.3%, 0.1% respectively in basal diet; 4 groups is the lincomycin group, adds the 2.5mg lincomycin in the per kilogram basal diet; 5 groups is the basal diet matched group.Experimental period is 40d.
It is consistent with feeding and management method that each organizes feeding environment, and routine immunization is raised before the test routinely, free choice feeding, and temperature, humidity, feedstuff and drinking-water are identical.Each test group disease of duration of test is treated according to a conventional method.
3, index detects
3.1 fertility performance detects the broiler survival rate and adds up survival rate, weightening finish, the feed conversion amount of respectively organizing test chicken respectively at 40d, calculates feedstuff-meat ratio.
3.2 the exponential detection of broiler immune organ selects spleen, fabricius bursa and thymus as immune organ, after the off-test, from each test group, randomly draw 5 chickens, weigh on an empty stomach, butcher back separating spleen, fabricius bursa and thymus, measure its weight, calculate immune organ index (the heavy kg of the heavy g/ live body of organ).
4, result
4.1 fertility performance testing result (seeing Table 1).
4.2 immune organ index testing result (seeing Table 2).
Table 1 traditional Chinese biological preparation of the present invention is to the influence of meat chicken production performance
| Group | Test chicken (only) | Survive number (only) | Survival rate (%) | Feedstuff-meat ratio |
| 1 (high dose group) | ??60 | ??60 | ??100 | ??1.65±0.040 |
| 2 (middle dosage groups) | ??60 | ??60 | ??100 | ??1.68±0.038 |
| 3 (low dose group) | ??60 | ??59 | ??98.3 | ??1.74±0.052 |
| 4 (lincomycin groups) | ??60 | ??55 | ??91.7 | ??1.76±0.035 |
| Group | Test chicken (only) | Survive number (only) | Survival rate (%) | Feedstuff-meat ratio |
| 5 (matched groups) | ??60 | ??54 | ??90.0 | ??1.88±0.021 |
As can be seen from Table 1, biological preparation of the present invention can significantly improve the survival rate of broiler, reduces feedstuff-meat ratio.Hence one can see that, and traditional Chinese biological preparation of the present invention can significantly promote growth of meat chicken, improves speed of weight increment, reduces sickness rate, saves feedstuff, improves the price of deed.
Table 2 traditional Chinese biological preparation of the present invention is to the exponential influence of broiler immune organ
| Group | Index and spleen index | The fabricius bursa index | Thymus index |
| 1 (high dose group) | ??2.13±0.47 | ??2.35±0.35 | ??4.43±0.12 |
| 2 (middle dosage groups) | ??2.01±0.16 | ??1.71±0.21 | ??3.49±0.44 |
| 3 (low dose group) | ??1.72±0.35 | ??1.64±0.12 | ??3.21±1.22 |
| 4 (lincomycin groups) | ??1.69±0.18 | ??1.69±0.10 | ??3.49±0.45 |
| 5 (matched groups) | ??1.64±0.36 | ??1.25±0.17 | ??3.01±0.35 |
As known from Table 2, fabricius bursa index, Chinese medicine microorganism formulation high dose group of the present invention and matched group comparing difference be (P＜0.5) significantly, in, low dose group and matched group comparing difference not remarkable (P＞0.5); Index and spleen index height, middle dosage group and matched group comparing difference be (P＜0.5) significantly, and thymus index high dose group and matched group compare, significant difference (P＜0.5).In sum, traditional Chinese biological preparation of the present invention has the exponential effect of immune organ that increases broiler, promptly can promote immune organ to grow human body immunity improving power.
Test example 2
Granule of the present invention is to according to Sha laying hen egg laying performance and Immune Effects.
1,2400 health of 200 ages in days of experimental animal selection Hebei chicken house is subjects according to Sha laying hen.
2, test method
2400 chickens are divided into 3 groups at random, 2 test group and 1 matched group, 800 every group, every group of 4 repetitions, each repeats 200.1 group is matched group, the basal diet of feeding; 2 groups in basal diet the dosage by every chicken 0.5g/ time add granule of the present invention, one day 2 times; 3 groups in basal diet the dosage by every chicken 0.25g/ time add granule of the present invention.Test group and matched group adopt full staged to raise in cages in same building hen house, freely drink water, and three groups of layer breeding management are in full accord.Experimental period is 60 days.
3, egg-laying test
Omnidistance every day is write down egg number, egg size in test, the diet of viewing test chicken, drinking-water, feces, mental status, record death toll.
1. the egg of average egg weight (g)=randomly draw every day 5% is weighed; 2. in the total egg number/feeding day in laying rate (%)=statistics phase the hen number add up * 100%; 3. mortality rate (%)=dead chicken number/test chicken number * 100%
4, amynologic index is measured and method
During off-test, every group of 10 laying hens at random, blood sample collection requires before the blood sampling the laying hen 12h that stops eating, cuts off the water.Centrifugalize serum, sterilization-20 ℃ of preservations in back detect amynologic index and The Total albumen content.
1. total serum protein (TP) content uses test kit application biuret method to measure; 2. serum immune globulin (IgG) is measured with immunoturbidimetry; 3. the periphery blood T lymphocyte conversion ratio detects with the MTT colorimetric analysis; 4. the newcastle epidemic disease antibody titre is measured with hemagglutination inhibition test (HI).
5, date processing
Data are represented with average ± standard deviation, determine the significance of difference with the t check.
6, result of the test
Table 3 traditional Chinese biological preparation of the present invention is to the influence of laying hen egg laying performance
| Group | Average egg weight (g) | Laying rate (%) | Mortality rate (%) |
| ??1 | ??32.71±1.45 | ??49.45±4.03 | ??3 |
| ??2 | ??38.77±3.12 | ??53.53±1.68 | ??0 |
| ??3 | ??37.01±2.28 | ??51.15±2.54 | ??0 |
As shown in Table 3: testing 2 groups and 3 groups of egg sizes increases by 15.63% and 11.62% respectively than matched group average egg weight, laying rate has improved 7.62% and 3.32% respectively, mortality rate descends 100%, illustrate that traditional Chinese biological preparation of the present invention has the effect that improves egg laying performance and reduce mortality rate to laying hen, thereby increase economic efficiency.
Table 4 traditional Chinese biological preparation of the present invention is to the influence of laying hen total serum protein and amynologic index
| Group | Total serum protein (g.L -1) | T lymhocyte transformation rate (OD 570) | Serum immune globulin (g.L -1) | Newcastle HI (log2) titre |
| ??1 | ??39.21±0.45 | ??0.320±0.001 | ??3.43±0.04 | ??2.70±0.23 |
| ??2 | ??45.29±0.56 | ??0.394±0.003 | ??3.67±0.12 | ??4.10±0.21 |
| ??3 | ??40.53±0.72 | ??0.321±0.001 | ??3.55±0.03 | ??3.56±0.19 |
As shown in Table 4: 1) content of 2 groups of total serum proteins of test is the highest, with matched group relatively, significant difference (P＜0.05), test 3 groups with matched group relatively, difference is significantly (P＞0.05) not; 2) OD
570Value shows that the T lymhocyte transformation rate of 2 groups of tests is significantly higher than test 1 group and matched group (P＜0.05); 3) 2,3 groups IgG level of test all increases than matched group, but group difference not significantly (P＞0.05); 4) 2,3 groups newcastle HI titre of test all is higher than matched group, 2 groups with matched group than difference remarkable (P＜0.05).
This shows in the daily ration of laying hen and adds the content that 0.5g traditional Chinese biological preparation of the present invention can improve serum immune globulin (IgG) by every chicken, significantly improving The Total albumen content, T lymhocyte transformation rate and newcastle epidemic disease antibody tires, illustrate that traditional Chinese biological preparation of the present invention can improve cellular immunization and the humoral immunity level of laying hen, thereby strengthen the specific immune function of chicken.
Test example 3
This test example is by the influence to ablactational baby pig fertility performance and immunity of the powder of the method for embodiment 2 preparation.
1, materials and methods
1.1 material
Experimental animal: the healthy ablactational baby pig of 30 ages in days.
Trial drug: powder of the present invention, the former powder of chlortetracycline.
1.2 method
1.2.1 animal is divided healthy ablactational baby pig about 45 body weight 6kg of group selection, prerun 5 days is weighed by head, is divided into 3 groups at random by body weight, every group of 3 repetitions.Experimental period is 60 days.
3 test group are respectively: 1 group is added powder of the present invention by 5g/kg in basal diet; In basal diet, press 20mg/kg for 2 groups and add chlortetracycline; 3 groups is matched group, does not add any medicine in the feedstuff in the basal diet.Every day, feeding was 3 times, freely drank water, and other management is carried out according to a conventional method.
Table 5 basal diet is formed and trophic level
| Daily ration is formed | ??(％) | Nutritive index | Trophic level |
| Corn | ??63 | Digestible energy (MJ.kg -1) | ??13.23 |
| Bean cake | ??25 | Crude protein (%) | ??18.0 |
| Fish flour | ??2 | Lysine (%) | ??1.0 |
| Wheat bran | ??6 | Calcium (%) | ??0.86 |
| Premix material | ??4 | Total phosphorus (%) | ??0.72 |
1.2.2 performance test
Write down the situation such as appetite, spirit, feces, morbidity of test pig every day, write down the feed intake of every group of every day.Begin respectively at formal test the 1st day, during off-test, morning, every empty stomach was weighed, and calculated daily gain, daily ingestion amount, material anharmonic ratio, the diarrhoea situation of ablactational baby pig is calculated the diarrhoea incidence rate.
1.2.3 immune indexes is measured and method
1.2.3.1 after the blood specimen collection feeding experiment finishes, organize each from each and select 2 of piglets at random, male and female half and half, totally 18 repeating.Piglet is gone on a hunger strike behind the 24h (freely drinking water), blood sample collection and separation of serum, and ultralow temperature is preserved, and is standby.
1.2.3.2 mensuration project and method measure total protein in the serum, albumin, globulin, immunoglobulin (IgA, IgM, IgG) and complement (C
3, C
4) content.Wherein, total serum protein (TP) adopts biuret method to measure; Serum albumin (ALB) adopts the bromocresol green method; Serum globulin (GLOB) is total protein and albumin difference; Serum immune globulin (IgA, IgM, IgG) and complement (C
3, C
4) content adopt immunoturbidimetry to measure.
1.3 date processing
The data average ± standard deviation is represented, determines the significance of difference with the t check.
2, result and analysis
Table 6 traditional Chinese biological preparation of the present invention is to the influence of ablactational baby pig fertility performance
| Group | Number (head) | Starting weight (kg) | End heavy (kg) | Daily gain (g) | Average daily feed intake (g) | The material anharmonic ratio |
| ??1 | ??15 | ??6.0±0.56 | ??40.78±1.23 | ??579.67±47 | ??1100±25 | ??1.90 |
| ??2 | ??15 | ??5.8±0.70 | ??37.04±2.06 | ??520.67±34 | ??1098±56 | ??2.11 |
| ??3 | ??15 | ??6.1±0.23 | ??32.56±5.20 | ??441.0±50 | ??1012±34 | ??2.29 |
Table 7 traditional Chinese biological preparation of the present invention is to the influence of diarrhea of weaned piglets and survival rate
| Group | Number | Diarrhea rate (%) | Survival rate (%) |
| ??1 | ??15 | ??6.7 | ??100 |
| ??2 | ??15 | ??13.3 | ??100 |
| ??3 | ??15 | ??33.3 | ??86.7 |
By table 6 and table 7 as can be known, the piglet daily gain of preparation group of the present invention improves 10.18% and 23.92% respectively than antibiotic group, matched group, the material anharmonic ratio reduces by 9.95% and 17.03% respectively, and diarrhea rate reduces by 49.62% and 79.88%, and survival rate improves 13.3% than matched group.This proves absolutely that traditional Chinese biological preparation of the present invention can improve the fertility performance of ablactational baby pig, effectively improves the premunition of piglet, reduces the generation of piglet diarrhea, improves efficiency of feed utilization simultaneously, reduces aquaculture cost.
Table 8 traditional Chinese biological preparation of the present invention is to the influence of piglet serum biochemistry index
| Group | ??TP(g/L) | ??ALB(g/L) | ??GLOB(g/L) |
| ??1 | ??64.73±2.47a | ??34.60±3.04a | ??32.33±3.47a |
| ??2 | ??53.87±3.98b | ??27.98±4.12b | ??25.89±4.65b |
| ??3 | ??50.43±2.76b | ??25.23±3.67b | ??24.45±5.01b |
Represent difference not remarkable (P＞0.05) with attached same letter person behind the column of figure, different adjacent letters is represented significant difference (P＜0.05), and different alternate letter representation difference is (P＜0.01) extremely significantly.Down together
Table 9 traditional Chinese biological preparation of the present invention is to the influence of piglet immunological index
| Group | ??IgG(g/L) | ??IgA(g/L) | ??IgM(g/L) | ??C 3(g/L) | ??C 4(g/L) |
| ??1 | ??3.56±0.22a | ??0.28±0.13a | ??0.95±0.07a | ??0.37±0.10a | ??0.18±0.01a |
| ??2 | ??2.79±0.13b | ??0.24±0.05b | ??0.79±0.18b | ??0.28±0.08c | ??0.13±0.02b |
| ??3 | ??2.38±0.40c | ??0.20±0.09b | ??0.71±0.05c | ??0.25±0.04c | ??0.12±0.01b |
From table 8 and 9 as can be seen, compare with antibiotic group, matched group (1), and the total protein content that adds traditional Chinese biological preparation group of the present invention improves 20.16% (P＜0.05), 28.36% (P＜0.05) respectively; Globulin content improves 23.64% (P＜0.05), 37.14% (P＜0.05) respectively; Albumin content improves 24.87% (P＜0.05), 32.23% (P＜0.05) respectively.(2) compare with the antibiotic group, add traditional Chinese biological preparation group immunoglobulin IgG of the present invention, IgA, IgM improve 2.76% (P＜0.05), 16.67% (P＜0.05), 20.25% (P＜0.05) respectively; Compare with matched group, improve 49.56% (P＜0.01), 40.0% (P＜0.05), 33.80% (P＜0.01) respectively.(3) compare with the antibiotic group, add traditional Chinese biological preparation group complement C of the present invention
3, C
4Improve 32.14% (P＜0.01), 50.0% (P＜0.01) respectively; Compare with matched group, improve 48.0% (P＜0.05), 38.46% (P＜0.05) respectively.This result of the test shows that traditional Chinese biological preparation of the present invention can significantly improve non-specific antibody level in the pig body, improves the pig body immunity function, strengthens the resistance of pig, helps the growth promoter of ablactational baby pig, reduces the generation of disease.
Test example 4
This test example is by the test to the meat sheep weightening finish of the oral liquid of the method for embodiment 1 preparation.
Select 60 of the cold sheep of healthy little tail of 4 monthly ages, be divided into 4 groups at random, 15 every group.The I group is matched group, and II is a low dose group, and every day, every sheep gavaged oral liquid 0.5ml of the present invention, every day 1 time; III is middle dosage group, and every day, every sheep gavaged oral liquid 0.8ml of the present invention, every day 1 time; IV is a high dose group, and every day, every sheep gavaged oral liquid 1.0ml of the present invention, every day 1 time; Continuous use 60 days.By feeding experiment, observe body weight gains and the feedstuff-meat ratio of the cold sheep of little tail.
The result shows: do not find untoward reaction in the process of the test, each test group sheep mental status is good.The comparison of each test group average weight you can well imagine high 7.43%, 10.62% and 11.40% according to component during off-test; Each test group average daily gain has improved 22.09%, 41.57% and 48.45% than matched group, and test III, IV group average daily gain and matched group more all reach significant level (P＜0.05).The result shows that traditional Chinese biological preparation of the present invention can promote the weightening finish of meat sheep, and can accelerate the speed of fattening of meat sheep.
Test example 5 traditional Chinese biological preparation of the present invention are to the influence of white mice macrophage phagocytic function
1, test method
40 Kunming white mouses are divided into 4 groups, 10 every group, irritate stomach by 8g/kg, 4g/kg, 2g/kg respectively for 1～3 group and give traditional Chinese biological preparation of the present invention, 4 groups is the blank group, gives the normal saline of Isodose, continuous use 7 days.The every Mus lumbar injection of 1～2h 0.4ml mass fraction is 5% chicken red blood cell after the drug withdrawal, behind 8～12h animal is taken off cervical vertebra and puts to death, and faces upward the position and is fixed in the Mus plate.The sterilization abdominal part, cut off skin, inject normal saline or A Shi liquid 2～2.5ml through the abdominal cavity, gently rub abdominal part 1min, extracting peritoneal fluid 1ml then drips and is applied on the clean slide, every 0.2ml, is placed in the enamel box that is lined with wet gauze totally by 2, place 37 ℃ of incubator incubation 30min, take out slide, drop into rinsing in the normal saline, to remove the not cell of paster.Dry, with the fixing 5min of acetone-methanol (1: 1), the female Sa of reuse 4% Ji-Rui Te Albert'stain Albert liquid dyeing 3～5min uses the distilled water rinsing, dries.200 macrophages of counting calculate phagocytic rate and phagocytic index under oily mirror.Phagocytic rate=(engulfing macrophage number/200 macrophage of chicken red blood cell) * 100%; Chicken red blood cell sum/200 macrophage that phagocytic index=quilt is engulfed.
2, result of the test
Result of the test sees Table 10.From table as can be known: the function that traditional Chinese biological preparation of the present invention can the enhance immunity system, phagocytic index, the phagocytic rate of mouse macrophage significantly improve, high dose group, middle dosage group and matched group comparison, significant difference.
Table 10 traditional Chinese biological preparation of the present invention is to the influence of white mice macrophage phagocytic function
* compare with matched group P＜0.5
Test example 6
This test example is by the tablet of the method preparation of embodiment 3 influence of young rabbit weightening finish and survival rate to be tested.
Select 150 weaned newborn rabbits in certain warren, Chicheng County, Hebei province for use, be divided into 2 groups, 75 every group.1 group is test group, and every day, every young rabbit gavaged 2 in tablet of the present invention; 2 groups is matched group.Continuous use 60 days, two groups of young rabbit feeding manners are identical.
The result shows, compare with matched group, test group daily increases weight and improves 22.41% (P＜0.01), feed conversion rate improves 17.37% (P＜0.05), mortality rate decline 6.66% (P＜0.05), this shows that traditional Chinese biological preparation of the present invention has the resistance that strengthens weaned newborn rabbit, significantly improves the survival rate of weaned newborn rabbit, somatotrophic effect.
Claims (8)
1. traditional Chinese biological preparation that improves immunity of livestock is characterized in that it being microbial inoculant to be carried out fermenting process of preparing in the culture medium that comprises Chinese medicine form.
2. the traditional Chinese biological preparation of raising immunity of livestock according to claim 1 is characterized in that described Chinese medicine comprises following component and parts by weight:
50～150 parts of Folium Eucommiaes, 50～150 parts of Grifola frondosa, 25～100 parts of the Radixs Astragali, 25～100 parts of Herba Epimedii.
3. the traditional Chinese biological preparation of raising immunity of livestock according to claim 1 is characterized in that described microorganism is a bacillus cereus.
4. the traditional Chinese biological preparation of raising immunity of livestock according to claim 3 is characterized in that described bacillus cereus is bacillus subtilis and/or Bacillus licheniformis.
5. the traditional Chinese biological preparation of raising immunity of livestock 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 raising immunity of livestock of claim 1-5, it is characterized in that may further comprise the steps:
(1) preparation bacteria culture fluid
Bacillus cereus is inoculated in the broth medium, and 35～37 ℃, 200～250rpm 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 8～10%, 25～30 ℃ of cultivation temperature, and speed of agitator 200～250rpm, the secondary seed solution viable bacteria content reaches 1.0 * 10
8～9During cfu/ml, stop to cultivate, standby;
(2) preparation of culture medium: the Chinese crude drug of getting formula ratio, be ground into 80～120 purpose medical material fine powders, pack in the fermentation tank, add 10～30% wheat brans, 5～20% bean cake, 0.1～1.0% ammonium sulfate, 0.01～0.10% magnesium sulfate, 0.01～0.05% calcium carbonate that account for Chinese medicine weight, add water to water content 40～60%, regulate pH to 6.5～7.5, sterilization 20～30min;
(3) after the cooling, by 5～10% of fermented product gross weight the bacteria culture fluid in (1) is linked in the culture medium of (2), in 25～30 ℃ 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 raising immunity of livestock 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. feedstuff that comprises the traditional Chinese biological preparation of each described raising immunity of livestock of claim 1-4.
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