CN104971111A - Traditional Chinese medicine bio-preparation of improving immunity of livestock and poultry, preparation method thereof and feed therewith - Google Patents
Traditional Chinese medicine bio-preparation of improving immunity of livestock and poultry, preparation method thereof and feed therewith Download PDFInfo
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Abstract
The invention relates to a traditional Chinese medicine bio-preparation of improving the immunity of livestock and poultry, a preparation method thereof and a feed therewith. According to the invention, the traditional Chinese medicine bio-preparation is prepared by combining a traditional Chinese medicine preparation with modern microorganism technologies, wherein the bio-preparation is prepared through the steps of: (1) employing the traditional Chinese medicines comprising oldenlandia diffusa, scutellaria baicalensis, astragalus membranaceus and liquorice as raw materials; (2) crushing the raw materials and adding bran, soybean meal, magnesium sulfate, manganese sulfate, sodium acetate and water to form a fermentation culture medium; (3) adding a bacterial strain culture solution in a certain ratio and performing fermentation; and (4) adding alcohol to a matured solid fermentation product to perform extraction, filtration and concentration to prepare various preparations. The traditional Chinese medicine bio-preparation, prepared through microorganism fermentation and metabolism, can significantly improve the immunity of livestock and poultry, can enhance body immunity, can increase production performance of the livestock and poultry, and can effectively prevent and treat immunosuppressive diseases.
Description
Technical field
The present invention relates to a kind of traditional Chinese biological preparation improving immunity of livestock and preparation method thereof and feedstuff, belong to veterinary drug and field of fodder.
Background technology
Current animal epidemic epidemic characteristic presents the situation of atypical bacteria viral disease, bacterial disease, mixed infection, secondary infection more, can give full play to the advantage of herbal medicine at the weak area of Western medicine.Especially along with the continuous acceleration of China's aquaculture intensive degree, particularly pursuing under green, environmental protection, the promotion of ecological tide, herbal medicine improving the unique advantage in immunologic function, obtains more and more general applying with it in aquaculture.Research Chinese veterinary drug preparation, on the impact of animal immune effect, for control livestock and poultry, improves Production of Livestock and Poultry power, increases animal husbandry economy benefit and has important practical significance.But Chinese veterinary drug preparation used is at present generally the powder that technology content is lower, directly adds in feedstuff and uses, and bioavailability is low and palatability is poor, unbecoming with the development pole of animal husbandry.
Modern Chinese medicine fermentation pharmacy technology is on the basis of inheriting tradition science of Chinese drug processing fermentation method, draw Microecology achievement, the high-tech herbal pharmaceutical new technique formed in conjunction with modern biological project fermentation technique, be the new curative effect finding medicine from herbal pharmaceutical aspect, there is very large researching value and market development meaning.Research shows, abundant and powerful enzyme system can be produced in growth of microorganism metabolic process, can by the ingredient decomposition and inversion of Chinese medicine, small-molecule active substance can not be degraded into by the active component that human body or animal body absorb, and directly absorbed by body, or produce new active component, change the property of medicine, reduce toxic and side effects, thus greatly improve curative effect of medication and therapeutic domain.Therefore, Modern microbiological technology and Chinese medicine prescription are combined, development of new traditional Chinese biological preparation is a breakthrough to Chinese medicine, has higher technical merit, is conducive to improving China's modernization of Chinese medicine level.
Summary of the invention
The object of this invention is to provide a kind of traditional Chinese biological preparation improving immunity of livestock, said preparation is is effectively preventing and treating the generation of livestock immunosuppressive disease, and improve young stock poult survival rate, while growth promoting effects, good palatability, bioavailability is high.Traditional Chinese biological preparation of the present invention is prepared from after utilizing microorganism to carry out fermentating metabolism to Chinese medicine, achieves the combination of Chinese medicine prescription and modern biotechnology.
Another object of the present invention is to provide the preparation method of the traditional Chinese biological preparation of this raising immunity of livestock.
Present invention also offers the feedstuff of the traditional Chinese biological preparation comprising this raising immunity of livestock.
The object of the invention is to be achieved through the following technical solutions:
The invention provides a kind of traditional Chinese biological preparation improving immunity of livestock, this preparation is prepared from carrying out fermentation in microbial inoculant to the culture medium comprising Chinese medicine composition, described Chinese medicine composition is made up of the component of following parts by weight: Herba Hedyotidis Diffusae 60 ~ 180 parts, Radix Scutellariae 50 ~ 150 parts, the Radix Astragali 20 ~ 100 parts, 30 ~ 120 parts, Radix Glycyrrhizae, described enterococcus is enterococcus.
Chinese medicine composition of the present invention is made up of the component of following parts by weight: Herba Hedyotidis Diffusae 80 ~ 150 parts, Radix Scutellariae 70 ~ 120 parts, the Radix Astragali 30 ~ 80 parts, 40 ~ 100 parts, Radix Glycyrrhizae.
Further, Chinese medicine composition of the present invention is made up of the component of following parts by weight: Herba Hedyotidis Diffusae 100 ~ 130 parts, Radix Scutellariae 80 ~ 110 parts, the Radix Astragali 40 ~ 70 parts, 50 ~ 80 parts, Radix Glycyrrhizae.
Further, Chinese medicine composition of the present invention is made up of the component of following parts by weight: Herba Hedyotidis Diffusae 120 parts, Radix Scutellariae 100 parts, the Radix Astragali 50 parts, 60 parts, Radix Glycyrrhizae.
Enterococcus of the present invention is preferably enterococcus faecalis and/or enterococcus faecalis.
The present invention does not limit the Strain type of enterococcus faecalis and enterococcus faecalis, and any existing bacterial strain may be used to the present invention, and preferred enterococcus faecalis is CGMCC No.5092, and it has significant advantage in the generation of stopping fermentation raw material mycete.
The traditional Chinese biological preparation of raising immunity of livestock of the present invention, can be acceptable dosage form on oral liquid, powder, tablet, granule, slow releasing preparation, pill, microcapsule, injection or other pharmaceuticss.
Present invention also offers the preparation method of the traditional Chinese biological preparation of this raising immunity of livestock, it is characterized in that comprising the following steps:
(1) prepare bacteria culture fluid: be inoculated into by enterococcus in MRS culture medium, 35 ~ 37 DEG C, production primary seed solution cultivated by 200 ~ 250rpm shaking table, 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%, cultivation temperature 25 ~ 30 DEG C, and speed of agitator 200 ~ 250rpm, secondary seed solution viable bacteria content reaches 1.0 × 10
8 ~ 9during cfu/ml, stop cultivating, for subsequent use;
(2) preparation of culture medium: the Chinese crude drug getting formula ratio, be ground into 80 ~ 120 object fine medicinal material powder, load in fermentation tank, add 10 ~ 30% wheat brans, 5 ~ 20% bean cake, 0.01 ~ 0.10% magnesium sulfate, 0.002 ~ 0.05% manganese sulfate, 0.01 ~ 0.10% sodium acetate that account for Chinese medicine weight, add water to water content 30 ~ 50%, regulate pH to 6.5 ~ 7.5, sterilizing 20 ~ 30min;
(3), after cooling, the bacteria culture fluid in (1) is linked in the culture medium of (2), in 25 ~ 30 DEG C of constant temperature culture 24 ~ 72h, speed of agitator 150 ~ 250rpm, when viable bacteria content reaches 1.0 × 10 by 5 ~ 10% of fermented product gross weight
8 ~ 9fermentation is stopped during cfu/ml;
(4) the solid fermentation thing 30-80% ethanol of maturation is carried out reflux, extract, filter, filtrate is refined, and it is 1.0 ~ 1.2 that fermentation liquid is evaporated to relative density, adds pharmaceutically conventional adjuvant and makes corresponding dosage form.
Further, the preparation method of the traditional Chinese biological preparation of raising immunity of livestock of the present invention also can comprise the steps: that concentrated broth that step (4) obtains is after spraying dry or lyophilization, adds adjuvant and makes pre-mixing agent.
The traditional Chinese biological preparation of raising immunity of livestock of the present invention can also directly be added in feedstuff, makes the feedstuff comprising this traditional Chinese biological preparation.
Material choice Herba Hedyotidis Diffusae of the present invention, Radix Scutellariae, the Radix Astragali and Radix Glycyrrhizae four Chinese medicine combine, each component effect is made to produce synergism, more easily absorb by making medicine after fermentable, drug effect is more abundant, thus the immunity of poultry body can be improved further, strengthen resistance against diseases, growth promoting effects.
Raising immunity of livestock traditional Chinese biological preparation of the present invention herbal medicine similar with tradition compares and has the following advantages:
1, poor, for oral administration rear effective ingredient of traditional herbal medicine powder palatability is not easily free from plant cell is out fully absorbed by body, and bioavailability is low, and decoct is destructive large to Effective Component of Chinese Medicine in preparation process, reduces the due curative effect of medicine.Fermentable transforms Chinese medicine, reaction condition is gentle, can Effective Component of Chinese Medicine be protected to greatest extent to exempt from destruction, simultaneously, some active constituent content can be made to increase and produce new ingredient, thus improve curative effect of medication, expansion indication, minimizing taking dose;
2, the traditional Chinese biological preparation prepared after microbial metabolism of Chinese medicine of the present invention, can improve immunity of livestock and fertility performance further, effectively prevent and treat the generation of livestock immunosuppressive disease, improves young stock poult survival rate, growth promoting effects.
Accompanying drawing explanation
Figure 1 shows that the picture of enterococcus faecalis (CGMCC No.5092) cassiri.Wherein, A and B is respectively the picture that enterococcus faecalis (CGMCC No.5092) and blank ferment 3 days; C and D is respectively the picture that enterococcus faecalis (CGMCC No.5092) and blank ferment 7 days.
Detailed description of the invention
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.
Embodiment 1 enterococcus faecalis (CMGCC No.5092) effect in cassiri process
Semen Maydis powder and water are fully mixed with the ratio of 1:0.5, with the inoculum concentration inoculation enterococcus faecalis CGMCC No.5092 mycopowder of 1%, addition is 10
7/ g, fermentation temperature is 37 DEG C, and fermentation time is 3d.Round is the conical flask of 250mL, then inserts in incubator and ferments.Blank is do not inoculate the Semen Maydis powder of enterococcus faecalis mycopowder and the mixture of water (1:0.5), and contrast 2 is the Semen Maydis powder of 1% inoculum concentration inoculation bacillus subtilis mycopowder and the mixture of water (1:0.5).
Result: compared with blank, Semen Maydis powder is after Enterococcus faecalis fermentation, its color, viscosity and local flavor all obviously improve (Fig. 1), wherein inoculate the color of the cassiri of enterococcus faecalis CGMCC No.5092 and the cassiri of inoculation bacillus subtilis, viscosity and Improving flavor more obvious.Continue to place non-cassiri and cassiri after one week in room temperature, observe non-fermented maize pruinescence mould contamination, fermented maize powder is contaminated.After placing two weeks, the Semen Maydis powder of bacillus subtilis and Enterococcus faecalis fermentation is not still by mould contamination, but its local flavor of the Semen Maydis powder of Enterococcus faecalis fermentation is better, illustrate that enterococcus faecalis CGMCC No.5092 has the effect preventing fungal infection, and effect is better than bacillus subtilis effect.
The preparation of embodiment 2 oral liquid of the present invention
Chinese medicine forms: Herba Hedyotidis Diffusae 100 parts, Radix Scutellariae 100 parts, the Radix Astragali 50 parts, 50 parts, Radix Glycyrrhizae
Preparation method:
(1) be inoculated in MRS culture medium by enterococcus faecalis (commercially available, ACCC 00661), 37 DEG C, production primary seed solution cultivated by 250rpm shaking table, 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%, cultivation temperature 30 DEG C, and speed of agitator 220rpm, secondary seed solution viable bacteria content reaches 1.0 × 10
8 ~ 9during cfu/ml, stop cultivating, for subsequent use;
(2) get the Chinese crude drug of formula ratio, be ground into 120 object fine medicinal material powder, load in fermentation tank, add 20% wheat bran, 10% bean cake, 0.06% magnesium sulfate, 0.005% manganese sulfate, 0.02% sodium acetate that account for Chinese medicine weight, add water to water content 50%, regulate pH to 7.0, sterilizing 20min;
(3), after cooling, be linked in the culture medium of (2) by 10% of fermented product gross weight by the strain liquid in (1), in 30 DEG C of constant temperature culture 48h, speed of agitator 220 rpm, when viable bacteria content reaches 1.0 × 10
8 ~ 9fermentation is stopped during cfu/ml;
(4) the solid fermentation thing of maturation is carried out reflux, extract, with 30% ethanol, filter, filtrate is refined, and it is 1.00 that fermentation liquid is evaporated to relative density;
(5) fermentation liquid after concentrating adds water to 300 parts, and filter, subpackage, sterilizing, namely makes oral liquid, and 1ml solution is equivalent to crude drug in whole 1g.
The preparation of embodiment 3 powder of the present invention
Chinese medicine forms: Herba Hedyotidis Diffusae 75 parts, Radix Scutellariae 50 parts, the Radix Astragali 100 parts, 75 parts, Radix Glycyrrhizae
Preparation method:
(1) be inoculated in MRS culture medium by enterococcus faecalis (CGMCC No.5092), 36 DEG C, production primary seed solution cultivated by 230rpm shaking table, 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%, cultivation temperature 29 DEG C, and speed of agitator 230rpm, secondary seed solution viable bacteria content reaches 1.0 × 10
8 ~ 9during cfu/ml, stop cultivating, for subsequent use;
(2) get the Chinese crude drug of formula ratio, be ground into 100 object fine medicinal material powder, load in fermentation tank, add 15% wheat bran, 10% bean cake, 0.08% magnesium sulfate, 0.004% manganese sulfate, 0.04% sodium acetate that account for Chinese medicine weight, add water to water content 45%, regulate pH to 7.2, sterilizing 25min;
(3), after cooling, be linked in the culture medium of (2) by 9% of fermented product gross weight by the bacteria culture fluid in (1), in 28 DEG C of constant temperature culture 36h, speed of agitator 230 rpm, when viable bacteria content reaches 1.0 × 10
8 ~ 9fermentation is stopped during cfu/ml;
(4) the solid fermentation thing of maturation is carried out reflux, extract, with 50% ethanol, filter, filtrate is refined, and it is 1.08 that fermentation liquid is evaporated to relative density, add mix homogeneously after appropriate soluble starch, dextrin again, dry, pulverize, sieve, namely make powder, every 1g is equivalent to crude drug in whole 1g.
The preparation of embodiment 4 Tablets
Chinese medicine forms: Herba Hedyotidis Diffusae 150 parts, Radix Scutellariae 100 parts, the Radix Astragali 100 parts, 50 parts, Radix Glycyrrhizae
Preparation method:
(1) be inoculated in MRS culture medium by enterococcus faecalis (CGMCC No.5092), 35 DEG C, production primary seed solution cultivated by 200rpm shaking table, 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%, cultivation temperature 30 DEG C, and speed of agitator 200rpm, secondary seed solution viable bacteria content reaches 1.0 × 10
8 ~ 9during cfu/ml, stop cultivating, for subsequent use;
(2) get the Chinese crude drug of formula ratio, be ground into 80 object fine medicinal material powder, load in fermentation tank, add 25% wheat bran, 5% bean cake, 0.10% magnesium sulfate, 0.006% manganese sulfate, 0.06% sodium acetate that account for Chinese medicine weight, add water to water content 40%, regulate pH to 6.8, sterilizing 25min;
(3), after cooling, be linked in the culture medium of (2) by 8% of fermented product gross weight by the bacteria culture fluid in (1), in 30 DEG C of constant temperature culture 24h, speed of agitator 180 rpm, when viable bacteria content reaches 1.0 × 10
8 ~ 9fermentation is stopped during cfu/ml;
(4) the solid fermentation thing of maturation is carried out reflux, extract, with 70% ethanol, filter, filtrate is refined, and it is 1.10 that fermentation liquid is evaporated to relative density, spraying dry obtains pressed powder again, add ethanol in proper amount and make wetting agent soft material, granulate, dry, add magnesium stearate mix homogeneously again, tabletting, namely makes tablet, and every sheet is equivalent to crude drug in whole 0.5g.
The preparation of embodiment 5 pre-mixing agent of the present invention
Chinese medicine forms: Herba Hedyotidis Diffusae 120 parts, Radix Scutellariae 100 parts, the Radix Astragali 50 parts, 60 parts, Radix Glycyrrhizae
Preparation method:
(1) be inoculated in MRS culture medium by enterococcus faecalis (commercially available, ACCC 00661), 37 DEG C, production primary seed solution cultivated by 220rpm shaking table, 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%, cultivation temperature 28 DEG C, and speed of agitator 250rpm, secondary seed solution viable bacteria content reaches 1.0 × 10
8 ~ 9during cfu/ml, stop cultivating, for subsequent use;
(2) get the Chinese crude drug of formula ratio, be ground into 80 object fine medicinal material powder, load in fermentation tank, add 30% wheat bran, 12% bean cake, 0.05% magnesium sulfate, 0.005% manganese sulfate, 0.08% sodium acetate that account for Chinese medicine weight, add water to water content 35%, regulate pH to 7.2, sterilizing 20min;
(3), after cooling, be linked in the culture medium of (2) by 7% of fermented product gross weight by the bacteria culture fluid in (1), in 28 DEG C of constant temperature culture 48h, speed of agitator 250 rpm, when viable bacteria content reaches 1.0 × 10
8 ~ 9fermentation is stopped during cfu/ml;
(4) the solid fermentation thing of maturation is carried out reflux, extract, with 75% ethanol, filter, filtrate is refined, and it is 1.04 that fermentation liquid is evaporated to relative density, spraying dry, then adds mix homogeneously after appropriate amount of auxiliary materials maize cob meal, is namely prepared into pre-mixing agent.
The preparation of embodiment 6 granule of the present invention
Chinese medicine forms: Herba Hedyotidis Diffusae 75 parts, Radix Scutellariae 150 parts, the Radix Astragali 50 parts, 100 parts, Radix Glycyrrhizae
Preparation method:
(1) be inoculated in MRS culture medium by enterococcus faecalis (commercially available, ACCC 00661), 36 DEG C, production primary seed solution cultivated by 210rpm shaking table, 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%, cultivation temperature 27 DEG C, and speed of agitator 210rpm, secondary seed solution viable bacteria content is 1.0 × 10
8 ~ 9during cfu/ml, stop cultivating, for subsequent use;
(2) get the Chinese crude drug of formula ratio, be ground into 100 object fine medicinal material powder, load in fermentation tank, add 12% wheat bran, 6% bean cake, 0.08% magnesium sulfate, 0.01% manganese sulfate, 0.04% sodium acetate that account for Chinese medicine weight, add water to water content 30%, regulate pH to 6.7, sterilizing 30min;
(3), after cooling, the bacteria culture fluid in (1) is linked in the culture medium of (2), in 30 DEG C of constant temperature culture 72h, speed of agitator 150rpm, when viable bacteria content reaches 1.0 × 10 by 6% of fermented product gross weight
8 ~ 9fermentation is stopped during cfu/ml;
(4) the solid fermentation thing of maturation is carried out reflux, extract, with 50% ethanol, filter, filtrate is refined, and it is 1.14 that fermentation liquid is evaporated to relative density, then adds suitable amount of sucrose, dextrin, soft material processed granulation of sieving; Wet grain drying, granulate, detect, subpackage, is namely prepared into granule, and every 1g is equivalent to crude drug in whole 2g.
The beneficial effect of traditional Chinese biological preparation of the present invention is described below by way of concrete test example.
Test example 1
This test example has investigated the fertility performance of pre-mixing agent on broiler and the impact of Immune Organs Index of various dose.
1, test material
Trial drug: pre-mixing agent, lincomycin prepared by embodiment 4.
Experimental animal: the healthy commercial chicken 300 of 1 age in days, is provided by chicken farm, Beijing.
2, test method
300 chickens are divided into 5 groups at random, often organize each 60, each group 2 repetitions, each repetition 30 chickens.Wherein 1,2,3 group is pre-mixing agent group of the present invention, in basal diet, add pre-mixing agent respectively by 0.5%, 0.3%, 0.1%; 4 groups is antibiotic group, adds 2.5mg lincomycin in per kilogram basal diet; Daily ration matched group based on 5 groups.Experimental period is 40 days.
Each group of feeding environment is consistent with feeding and management method, and routine immunization, raises before test routinely, free choice feeding, and temperature, humidity, feedstuff are identical with drinking-water.The each test group disease of duration of test is treated according to a conventional method.
3, Indexs measure
3.1 fertility performance detect broiler survival rates and added up the survival rate of each group of test chicken, weightening finish, feed conversion amount at the 40th day respectively, calculate feedstuff-meat ratio.
The detection of 3.2 broiler Immune Organs Indexes selects spleen, fabricius bursa and thymus as immune organ, after off-test, 5 chickens are randomly drawed from each test group, weigh on an empty stomach, butcher rear separating spleen, fabricius bursa and thymus, measure its weight, Computation immunity shoot formation (the heavy kg of Organs Weight g/ live body).
4, result
4.1 fertility performance testing results (see table 1).
4.2 Immune Organs Index testing results (see table 2).
Table 1 pre-mixing agent of the present invention is on the impact of meat chicken production performance
| Group | Test chicken (only) | Survive number (only) | Survival rate (%) | Feedstuff-meat ratio |
| 1(high dose group) | 60 | 60 | 100 | 1.67±0.035 |
| Dosage group in 2() | 60 | 60 | 100 | 1.71±0.042 |
| 3(low dose group) | 60 | 58 | 96.7 | 1.73±0.029 |
| 4(antibiotic group) | 60 | 56 | 93.3 | 1.76±0.046 |
| 5(matched group) | 60 | 54 | 90.0 | 1.87±0.033 |
As can be seen from Table 1, pre-mixing agent of the present invention can significantly improve the survival rate of broiler, reduces feedstuff-meat ratio.It can thus be appreciated that pre-mixing agent of the present invention significantly can promote growth of meat chicken, improve speed of weight increment, reduce sickness rate, save feedstuff, improve the price of deed.
Table 2 pre-mixing agent of the present invention is on the impact of broiler Immune Organs Index
| Group | Index and spleen index | Bursal index | Thymus index |
| 1(high dose group) | 2.27±0.55 | 2.17±0.44 | 4.45±0.32 |
| Dosage group in 2() | 2.07±0.43 | 1.72±0.35 | 3.56±0.53 |
| 3(low dose group) | 1.81±0.44 | 1.65±0.42 | 3.19±0.67 |
| 4(antibiotic group) | 1.70±0.28 | 1.69±0.23 | 3.32±0.45 |
| 5(matched group) | 1.66±0.54 | 1.37±0.26 | 3.03±0.45 |
As known from Table 2, bursal index, pre-mixing agent high dose group of the present invention and matched group comparing difference significantly (P < 0.05), in, low dose group and matched group comparing difference remarkable (P > 0.05); Index and spleen index, significantly (P < 0.05), thymus index, high dose group compares with matched group, significant difference (P < 0.05) for high, middle dosage group and matched group comparing difference.In sum, pre-mixing agent of the present invention has the effect of Immune Organs Index increasing broiler, namely can Promote immunity allelotaxis, improves immunity of organisms.
Test example 2
This test example granule of having investigated different Chinese medicine composition is on the impact of Layer Production Performance and immunologic function.
1, test material
Trial drug: granule prepared by embodiment 5, the granule that different Chinese medicine composition is made.
Spend Radix Astragali group in vain, get Herba Hedyotidis Diffusae 75 parts, Radix Scutellariae 150 parts, be prepared into by the method for embodiment 5 and spend Astragalus granules in vain, every 1g is equivalent to crude drug in whole 2g;
Radix Astragali Radix Glycyrrhizae group, get the Radix Astragali 50 parts, 100 parts, Radix Glycyrrhizae, be prepared into the agent of Radix Astragali licorice root particles by the method for embodiment 5, every 1g is equivalent to crude drug in whole 2g.
Drug control group: disclosed in referenced patent CN201010573231, the one-tenth of Chinese medicine feed additive is grouped into, be prepared into by the method for embodiment 5 and comprise Herba Hedyotidis Diffusae, Herba Violae, Sha Jiang, Herba Patriniae, Spica Prunellae, Herba Houttuyniae, Radix Scutellariae, the granule of Folium Isatidis, Radix Glycyrrhizae, Radix Codonopsis, the Radix Astragali, Radix Angelicae Sinensis, Maifanitum, Fructus Schisandrae Chinensis, Semen Cuscutae, Herba Scutellariae Barbatae and Fructus Forsythiae, every 1g is equivalent to crude drug in whole 2g.
Experimental animal: 200 age in days laying hens 1500, certain chicken house of effluent north provides.
2, test method
1500 chickens are divided into 5 groups at random, 3 test group, 1 drug control group and 1 blank group, often organize 300, often organize 3 repetitions, each repetition 100.1 group of dosage by every chicken 0.5g/ time in basal diet adds granule of the present invention, one day 2 times; 2 groups of granules adding to spend the Radix Astragali in vain by the dosage of every chicken 0.5g/ time in basal diet, one day 2 times; 3 groups in basal diet by the granule of the dosage interpolation Radix Scutellariae Radix Glycyrrhizae of every chicken 0.5g/ time, one day 2 times; 4 groups in basal diet by the granule of the dosage interpolation drug control group of every chicken 0.5g/ time, one day 2 times; 5 groups is blank group, basal diet of feeding.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 completely the same.Experimental period is 60 days.
3, egg-laying test
Omnidistance every day is recorded egg number, egg size, the diet of viewing test chicken, drinking-water, feces, mental status in test, record death toll.
1. the egg randomly drawing 5% average egg weight (g)=every day is weighed; Hen number cumulative × 100% in total egg number/feeding day of 2. laying rate (%)=in the statistics phase; 3. mortality rate (%)=dead chicken number/test chicken number × 100%
4, amynologic index measures and method
During off-test, often organize random 10 laying hens, blood sample collection, before blood sampling, requirement is stopped eating to laying hen, cut off the water 12h.Centrifugalize serum ,-20 DEG C of preservations after sterilizing, 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) measures with immunoturbidimetry; 3. periphery blood T lymphocyte conversion ratio MTT colorimetric analysis detects; 4. antibody titer of ND hemagglutination inhibition test (HI) measures.
5, date processing
Data represent with average ± standard deviation, determine the significance of difference with t inspection.
6, result of the test
Table 3 granule of the present invention is on the impact of Layer Production Performance
As shown in Table 3: test group average egg weight ratio spends Radix Astragali group, Radix Scutellariae Radix Glycyrrhizae group, drug control group and blank group average egg weight in vain increases by 5.70%, 7.65%, 10.54% and 17.76% respectively, laying rate improves 4.56%, 7.49%, 7.75% and 11.68% respectively, the mortality rate of 3 medicine groups is 0, illustrate that granule of the present invention is improved egg laying performance to laying hen and reduces the effect of mortality rate, thus increase economic efficiency.
Table 4 granule of the present invention is on the impact of Laying-hen Serum total protein and amynologic index
As shown in Table 4: 1) the Total albumen content of test group is the highest, with spend Radix Astragali group, Radix Scutellariae Radix Glycyrrhizae group and matched group in vain and compare, significant difference (P < 0.05); 2) OD
570the T lymhocyte transformation rate of value display test group is significantly higher than spends Radix Astragali group, Radix Scutellariae Radix Glycyrrhizae group and matched group (P < 0.05) in vain; 3) the IgG level of test group is comparatively spent Radix Astragali group, Radix Scutellariae Radix Glycyrrhizae group and matched group in vain and is all increased, but difference is not significantly (P > 0.05); 4) the newcastle HI titre of test group is all higher than spending Radix Astragali group, Radix Scutellariae Radix Glycyrrhizae group and matched group in vain, significant difference (P < 0.05).
This shows to add by every chicken the content that 0.5g granule of the present invention can improve serum immune globulin (IgG) in the daily ration of laying hen, significantly improve the Total albumen content, T lymhocyte transformation rate and antibody titres to newcastle disease virus, illustrate that granule of the present invention can improve cellular immunization and the humoral immunity level of laying hen, thus strengthen the specific immune function of chicken.
Test example 3
This test example powder of having investigated Different Extraction Method is on the impact of Production Performance of Weaning Pigs and immunity.
1, test material
Trial drug: with different fermentations bacterium, powder, the unleavened powder prepared by embodiment 2.Test group: be prepared into powder by the method for embodiment 2, every 1g is equivalent to crude drug in whole 1g.
Bacillus cereus group: take bacillus subtilis as zymocyte, be prepared into powder by the method for embodiment 2, every 1g is equivalent to crude drug in whole 1g.
Saccharomyces cerevisiae group: take saccharomyces cerevisiae as zymocyte, be prepared into powder by the method for embodiment 2, every 1g is equivalent to crude drug in whole 1g.
Not ferment group: get Herba Hedyotidis Diffusae 75 parts, Radix Scutellariae 50 parts, the Radix Astragali 100 parts, 75 parts, Radix Glycyrrhizae, be ground into 100 object fine medicinal material powder, add the alcohol reflux 3 times of 10 times amount 75%, each 90 minutes, merge extractive liquid, being evaporated to relative density is 1.08, then adds mix homogeneously after appropriate soluble starch, dextrin, dries, pulverize, sieve, namely make powder, every 1g is equivalent to crude drug in whole 1g.
Experimental animal: the healthy ablactational baby pig 120 of 28 ages in days, is provided by pig farm, Beijing.
2 test methods
2.1 animal groupings: select 120 healthy ablactational baby pig of about body weight 6kg, prerun 5 days, weighs by head, is divided into 4 groups at random by body weight, often organizes 3 repetitions, each repetition 10.Experimental period is 60 days.
5 test group are respectively: 3 fermentation components do not add through the powder made that ferments by 2g/kg body weight in basal diet; Group of not fermenting is added without the powder made that ferments by 2g/kg body weight in basal diet; Any medicine is not added in basal diet in matched group feedstuff.Every day, feeding 3 times, freely drank water, and other management is carried out according to a conventional method.
Table 5 basal diet composition and trophic level
| Daily ration forms | （％） | Nutritive index | Trophic level |
| Semen Maydis | 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 |
2.2 performance test
Record the situations such as the appetite of test pig, spirit, feces, morbidity every day, the feed intake of every day often organized in record.The 1st day that starts respectively at formal test, off-test time, morning every, empty stomach was weighed, calculate daily gain, daily ingestion amount, feed-weight ratio, the diarrhoea situation of ablactational baby pig, calculate Incidence of Diarrhea.
2.3 immune indexes measure and method
2.3.1 after blood specimen collection feeding experiment terminates, Stochastic choice piglet 2 from each group of each repetition, totally 18.Piglet 24 h(that go on a hunger strike freely drink water) after, blood sample collection separation of serum, Excised Embryos, for subsequent use.
2.3.2 total protein, albumin, globulin, immunoglobulin (IgA, IgM, IgG) and complement (C in project and method mensuration serum is measured
3, C
4) content.Wherein, total serum protein (TP) adopts biuret method to measure; Serum albumin (ALB) adopts Bromocresol green; Serum globulin (GLB) is total protein and albumin difference; Serum immune globulin (IgA, IgM, IgG) and complement (C
3, C
4) content adopt immunoturbidimetry measure.
2.4 date processing
Data acquisition average ± standard deviation represents, determines the significance of difference with t inspection.
3, results and analysis
Table 6 powder of the present invention is on the impact of Production Performance of Weaning Pigs
| Group | Head number (head) | Starting weight (kg) | End heavy (kg) | Daily gain (g) | Average daily feed intake (g) | Feed-weight ratio |
| 1(test group) | 30 | 6.08±0.56 | 41.65±1.23 | 594.17±47 | 1100±25 | 1.85 |
| 2(bacillus cereus group) | 30 | 6.10±0.61 | 39.74±2.33 | 560.67±35 | 1080±45 | 1.93 |
| 3(fermented glutinous rice yeast group) | 30 | 6.12±0.48 | 38.89±3.23 | 546.17±47 | 1090±33 | 2.00 |
| 4(does not ferment group) | 30 | 6.18±0.52 | 38.74±2.06 | 540.83±34 | 1130±56 | 2.09 |
| 5(matched group) | 30 | 6.15±0.23 | 35.22±5.20 | 485.33±50 | 1150±34 | 2.37 |
Table 7 powder of the present invention is on the impact of diarrhea of weaned piglets and survival rate
| Group | Head number | Diarrhea rate (%) | Survival rate (%) |
| 1(enterococcus group) | 30 | 1.2 | 100 |
| 2(bacillus cereus group) | 30 | 5.6 | 100 |
| 3(fermented glutinous rice yeast group) | 30 | 7.8 | 93.33 |
| 4(does not ferment group) | 30 | 8.3 | 96.67 |
| 5(matched group) | 30 | 15.3 | 86.67 |
From table 6 and table 7, the piglet daily gain of test group of the present invention improves 5.97%, 8.79%, 11.44% and 22.43% respectively than bacillus cereus group, fermented glutinous rice yeast group, group of not fermenting, matched group, feed-weight ratio reduces by 4.15%, 7.50%, 11.81% and 21.94% respectively, diarrhea rate reduces by 7.85%, 8.46%, 85.54% and 92.16%, and survival rate improves 6.67%, 3.33% and 13.33% respectively than fermented glutinous rice yeast group, group of not fermenting and matched group.This absolutely proves that powder 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 powder of the present invention is on the impact of Serum Biochemical Index in Piglets
| Group | TP(g/L) | ALB(g/L) | GLB(g/L) |
| 1(test group) | 66.90±2.47a | 34.60±3.04a | 32.33±3.47a |
| 2(bacillus cereus group) | 60.31±3.01b | 30.32±2.08b | 28.64±2.37b |
| 3(fermented glutinous rice yeast group) | 55.37±2.22c | 29.55±1.39b | 26.03±3.06b |
| 4(does not ferment group) | 54.65±3.98c | 28.78±4.12b | 25.89±4.65b |
| 5(matched group) | 50.43±2.76d | 25.60±3.67c | 24.45±5.01b |
Difference is represented not significantly (P > 0.05) with same letter person attached after column of figure, different adjacent letters represents significant difference (P < 0.05), and different alternate letter representation difference extremely significantly (P < 0.01).Lower same
Table 9 powder of the present invention is on the impact of piglet immunological index
| Group | IgG(g/L) | IgA(g/L) | IgM(g/L) | C 3(g/L) | C 4(g/L) |
| 1(test group) | 3.65±0.33a | 0.30±0.18a | 0.96±0.14a | 0.38±0.19a | 0.15±0.08a |
| 2(bacillus cereus group) | 3.27±0.56b | 0.28±0.12a | 0.86±0.18b | 0.35±0.20a | 0.14±0.10a |
| 3(fermented glutinous rice yeast group) | 3.15±0.45b | 0.27±0.14a | 0.85±0.20b | 0.32±0.18a | 0.12±0.11b |
| 4(does not ferment group) | 2.97±0.28b | 0.27±0.13a | 0.81±0.22c | 0.30±0.15a | 0.12±0.13b |
| 5(matched group) | 2.42±0.51c | 0.22±0.07b | 0.73±0.10d | 0.24±0.08b | 0.11±0.04b |
As can be seen from table 8 and 9, (1) compare with matched group with bacillus cereus group, fermented glutinous rice yeast group, group of not fermenting, the total protein content of test group, globulin content and albumin content all have significant difference (P < 0.05), bacillus cereus group, fermented glutinous rice yeast group, do not ferment group difference significantly (P > 0.05).(2) compare with matched group with bacillus cereus group, fermented glutinous rice yeast group, group of not fermenting, test group immunoglobulin IgG and IgM all have significant difference (P < 0.05); Bacillus cereus group, fermented glutinous rice yeast group, IgG and IgA difference remarkable (P > 0.05) between group of not fermenting.(3) compared with matched group, test group complement C
3, C
4improve 25.00%(P < 0.05 respectively), 36.36%(P < 0.05).This result of the test shows, powder of the present invention can significantly improve non-specific antibody level in pig body, improves pig body immunity function, strengthens the resistance of pig, is conducive to the growth promoter of ablactational baby pig, reduces the generation of disease.
Test example 4
This test example has investigated the tablet of different strain fermentation to the impact of white mice macrophage phagocytic function.
1, test material
Trial drug: tablet prepared by embodiment 3, the tablet through fermentation of bacillus.
Bacillus cereus group: get Herba Hedyotidis Diffusae 150 parts, Radix Scutellariae 100 parts, the Radix Astragali 100 parts, 50 parts, Radix Glycyrrhizae, through fermentation of bacillus, carries out lixiviate by the solid fermentation thing ethanol of maturation, be separated, filter, filtrate is refined, it is 1.10 that fermentation liquid is evaporated to relative density, then spraying dry obtains pressed powder, adds ethanol in proper amount and makes wetting agent soft material, granulate, drying, then add magnesium stearate mix homogeneously, tabletting, namely make the tablet through fermentation of bacillus, every sheet is equivalent to crude drug in whole 0.5g.
Experimental animal: Kunming white mouse 60, male and female half and half, are provided by laboratory animal field, Beijing.
2, test method
40 Kunming white mouses are divided into 3 groups, and often organize 20,1 group gives tablet of the present invention by 2g/kg body weight gavage, continuous use 7 days; 2 groups give the tablet through fermentation of bacillus by 2g/kg body weight gavage, continuous use 7 days; 3 groups is blank group.After drug withdrawal, 1 ~ 2h every Mus lumbar injection 0.4ml mass fraction is 5% chicken red blood cell, after 8 ~ 12h, animal is taken off cervical vertebra and puts to death, face upward position and be fixed on Mus plate.Sterilization abdominal part, cut off skin, through Intraperitoneal injection normal saline or A Shi liquid 2 ~ 2.5ml, gently rub abdominal part 1min, then extraction peritoneal fluid 1ml drips and is applied on clean slide, every sheet 0.2ml, is placed on and is lined with in the enamel box of wet gauze by totally 2, be placed in 37 DEG C of incubator incubation 30min, take out slide, drop into rinsing in normal saline, to remove the cell of non-paster.Dry, with the fixing 5min of acetone-methanol (1:1), then with 4% Ji female Sa-Rui Te Albert'stain Albert liquid dyeing, 3 ~ 5min, use distilled water rinsing, dry.Under oily mirror, count 200 macrophages, calculate phagocytic rate and phagocytic index.Phagocytic rate=(engulfing macrophage number/200 macrophage of chicken red blood cell) × 100%; Phagocytic index=by chicken red blood cell sum/200 macrophages engulfed.
3, result of the test
Result of the test is in table 10.As seen from the table: Tablets can strengthen immune function, phagocytic index, the phagocytic rate of mouse macrophage significantly improve, compared with bacillus cereus group, matched group, and significant difference.
Table 10 tablet of the present invention is on the impact of white mice macrophage phagocytic function
* P < 0.05 compares with matched group
Test example 5
This test example has investigated traditional Chinese biological preparation of the present invention prepared by the different embodiment impact on meat sheep weightening finish and immunologic function.
1, test material
Trial drug: pre-mixing agent prepared by embodiment 4, the oral liquid of embodiment 1 preparation.
Experimental animal: healthy 3 monthly age Small-fat-tail sheep 60, plant provides by Beijing.
2, test method:
Be divided into 3 groups at random, often organize 20.1 group is pre-mixing agent group, the pre-mixing agent prepared with embodiment 4, mixed feeding administration, and every 1kg feedstuff adds pre-mixing agent 5g of the present invention, free choice feeding; 2 groups is oral liquid group, the oral liquid prepared with embodiment 1, and oral administration, gavages oral liquid of the present invention by every sheep 0.5ml/kg body weight, every day 1 time; 3 groups is blank group, normally feeds, not administration; 60 days test periods.By feeding experiment, observe the body weight gains of Small-fat-tail sheep.
Before test, after test 30 days and off-test, gather the peripheral blood of sheep respectively, measure erythrocyte, leukocyte and T cell quantity.Erythrocyte and quantity of leucocyte adopt conventional method to measure, and T cell adopts E-rosette formation test to measure.
3, result of the test
Result of the test is in table 11,12.
Table 11 traditional Chinese biological preparation of the present invention is on the impact of Small-fat-tail sheep body weight gains
Table 12 traditional Chinese biological preparation of the present invention is on the impact of erythrocyte, leukocyte, T cell content
Result shows: do not find untoward reaction in process of the test, and each test group sheep mental status is good.During off-test, pre-mixing agent group and the average daily gain of oral liquid group you can well imagine high 35.05% and 29.10% than contrast component, show that traditional Chinese biological preparation of the present invention can promote the weightening finish of meat sheep, and can accelerate meat sheep fatten speed.
Before test, in each group of blood, erythrocyte, leukocyte and T cell content are substantially identical, after test group medication the erythrocyte of 30 days and 60 days, leukocyte and T cell content than before medication and matched group be significantly increased, show that the generation of traditional Chinese biological preparation of the present invention to blood formed element has stimulation, there is obvious incentive action to immunologically competent cell, the immunologic function of body can be improved.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the prerequisite not departing from the technology of the present invention principle; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (9)
1. one kind is improved the traditional Chinese biological preparation of immunity of livestock, it is characterized in that, be prepared from carrying out fermentation in microbial inoculant to the culture medium comprising Chinese medicine composition, described Chinese medicine composition is made up of the component of following parts by weight: Herba Hedyotidis Diffusae 60 ~ 180 parts, Radix Scutellariae 50 ~ 150 parts, the Radix Astragali 20 ~ 100 parts, 30 ~ 120 parts, Radix Glycyrrhizae, described microorganism is enterococcus.
2. the traditional Chinese biological preparation improving immunity of livestock as claimed in claim 1, it is characterized in that, described Chinese medicine composition is made up of the component of following parts by weight: Herba Hedyotidis Diffusae 80 ~ 150 parts, Radix Scutellariae 70 ~ 120 parts, the Radix Astragali 30 ~ 80 parts, 40 ~ 100 parts, Radix Glycyrrhizae.
3. the traditional Chinese biological preparation improving immunity of livestock as claimed in claim 2, it is characterized in that, described Chinese medicine composition is made up of the component of following parts by weight: Herba Hedyotidis Diffusae 100 ~ 130 parts, Radix Scutellariae 80 ~ 110 parts, the Radix Astragali 40 ~ 70 parts, 50 ~ 80 parts, Radix Glycyrrhizae.
4. the traditional Chinese biological preparation improving immunity of livestock as claimed in claim 3, it is characterized in that, described Chinese medicine composition is made up of the component of following parts by weight: Herba Hedyotidis Diffusae 120 parts, Radix Scutellariae 100 parts, the Radix Astragali 50 parts, 60 parts, Radix Glycyrrhizae.
5. the traditional Chinese biological preparation of the raising immunity of livestock according to any one of Claims 1 to 4, is characterized in that, described enterococcus is enterococcus faecalis and/or enterococcus faecalis.
6. the traditional Chinese biological preparation of the raising immunity of livestock according to any one of Claims 1 to 4, it is characterized in that, described preparation is acceptable dosage form on oral liquid, powder, tablet, granule, slow releasing preparation, pill, microcapsule, injection or other pharmaceuticss.
7. the preparation method of the traditional Chinese biological preparation of the raising immunity of livestock according to any one of claim 1 ~ 6, is characterized in that, comprise the following steps:
(1) prepare bacteria culture fluid: be inoculated into by enterococcus in MRS culture medium, 35 ~ 37 DEG C, production primary seed solution cultivated by 200 ~ 250rpm shaking table, 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%, cultivation temperature 25 ~ 30 DEG C, and speed of agitator 200 ~ 250rpm, secondary seed solution viable bacteria content reaches 1.0 × 10
8 ~ 9during cfu/ml, stop cultivating, for subsequent use;
(2) preparation of culture medium: the Chinese crude drug getting formula ratio, be ground into 80 ~ 120 object fine medicinal material powder, load in fermentation tank, add 10 ~ 30% wheat brans, 5 ~ 20% bean cake, 0.01 ~ 0.10% magnesium sulfate, 0.002 ~ 0.05% manganese sulfate, 0.01 ~ 0.10% sodium acetate that account for Chinese medicine weight, add water to water content 30 ~ 50%, regulate pH to 6.5 ~ 7.5, sterilizing 20 ~ 30min;
(3), after cooling, the bacteria culture fluid in (1) is linked in the culture medium of (2), in 25 ~ 30 DEG C of constant temperature culture 24 ~ 72h, speed of agitator 150 ~ 250rpm, when viable bacteria content reaches 1.0 × 10 by 5 ~ 10% of fermented product gross weight
8 ~ 9fermentation is stopped during cfu/ml;
(4) the solid fermentation thing 30-80% ethanol of maturation is carried out reflux, extract, filter, filtrate is refined, and it is 1.0 ~ 1.2 that fermentation liquid is evaporated to relative density, adds pharmaceutically conventional adjuvant and makes corresponding dosage form.
8. the preparation method of the traditional Chinese biological preparation of raising immunity of livestock according to claim 7, it is characterized in that, also comprise the steps: that concentrated broth that step (4) obtains is after spraying dry or lyophilization, adds adjuvant and makes pre-mixing agent further.
9. one kind comprises the feedstuff of the traditional Chinese biological preparation of the raising immunity of livestock described in any one of claim 1 ~ 6.
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| CN105963344A (en) * | 2016-07-13 | 2016-09-28 | 中国科学院海洋研究所 | Fermented traditional Chinese medicine preparation for improving immunity of aquaculture animals and preparation method thereof |
| CN109247445A (en) * | 2018-09-30 | 2019-01-22 | 广西乐业康辉生态养殖专业合作社 | A kind of feed and preparation method thereof improving immunity of livestock |
| CN110692806A (en) * | 2019-10-30 | 2020-01-17 | 石家庄飞龙饲料有限公司 | Chinese herbal medicine fermented biological feed for improving immunity of livestock and poultry and preparation method thereof |
| CN111011597A (en) * | 2019-11-25 | 2020-04-17 | 内蒙古斯隆生物技术有限责任公司 | Functional animal growth and immune regulator and preparation method thereof |
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