CN1325077C - Animal immunopotentiator using membranous milk vetch root as basic remedy - Google Patents
Animal immunopotentiator using membranous milk vetch root as basic remedy Download PDFInfo
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- CN1325077C CN1325077C CNB2005100501612A CN200510050161A CN1325077C CN 1325077 C CN1325077 C CN 1325077C CN B2005100501612 A CNB2005100501612 A CN B2005100501612A CN 200510050161 A CN200510050161 A CN 200510050161A CN 1325077 C CN1325077 C CN 1325077C
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Abstract
The present invention discloses an animal immunopotentiator which uses milkvetch roots as a main formula. The potentiator has formula components of shares by weight: 10 to 15 shares of milkvetch root, 5 to 10 shares of golden thread, 10 to 15 shares of baical skullcap root, 20 to 25 shares of weeping forsythia capsule and 15 shares of dyers woad leaf. The animal immunopotentiator which uses milkvetch roots as a main formula in the present invention has the advantages of low cost and capability of enhancing the animal immunizing powder and improving the growth performance of animals.
Description
Technical field
The present invention relates to a kind of prescription of animal immunity potentiator, particularly a kind of is the prescription of the animal immunity potentiator of main formula with the Radix Astragali.
Background technology
Domesticated animal is sick in order to prevent, improves disease resistance of animals, and the poultry raiser generally can add a certain proportion of antibiotic in feedstuff.But so not only can cause animal to develop immunity to drugs, but also can make antibiotic remains in animal body,, can cause great hidden danger healthy if directly or indirectly eaten this kind animal as food chain people topmost.So people urgently wish that a kind of health of pure natural is arranged, the animal immunity potentiator that has no side effect, to improve disease resistance of animals.
Medical herbs in the natural drug is the human material of using the earliest, also is the feed additive that is employed the earliest.There are 12807 kinds of natural goods Chinese herbal medicine resources in China, and wherein has only 10% for commonly used, still need further investigate utilization.The Radix Astragali is " strengthening the body resistance " class tonification medicine in the Chinese herbal medicine, can grow void and help the weak, and conditioning intestinal microecology balance improves the body non-specific immunity.But the Radix Astragali be main formula the animal feed immunostimulant do not see relevant report.
Summary of the invention
At the deficiencies in the prior art part, the invention provides a kind of cost low, can improve the animal immune ability, what improve its growth performance is the animal immunity potentiator of main formula with the Radix Astragali.
The present invention is for reaching above purpose, be to realize by such technical scheme: providing a kind of is the animal immunity potentiator of main formula with the Radix Astragali, and the recipe ingredient of this reinforcing agent (by weight) is: 15 parts of 10～15 parts of the Radixs Astragali, 5～10 parts of Rhizoma Coptidis, 10～15 parts of Radix Scutellariaes, 20～25 parts of Fructus Forsythiaes and Folium Isatidiss.
Of the present invention is the animal immunity potentiator of main formula with the Radix Astragali, and what select for use all is common raw material, therefore can control production cost to greatest extent.Particularly the Radix Astragali of one of raw material has characteristics abundant, cheap in china natural resources, safety non-toxic; Use as a kind of feed additive with its animal immunity potentiator that is raw material is made, can improve the animal immune function, but therefore also supplement feed albumen effect when improving the animal intestinal microecological balance can improve the growth of animal performance.Of the present invention is the animal immunity potentiator of main formula with the Radix Astragali, and suitable animal is taken for a long time, and animal is developed immunity to drugs, can be all or most of antibiotic that substitutes in the feedstuff.
Of the present invention is that the animal immunity potentiator of main formula can prove by following experimental technique to the enhancing of animal immune ability with to the function of improving of animal intestinal microecological balance with the Radix Astragali.
1, choose 60 store pigs that body weight is close, be divided into 2 groups (matched group and test group) at random, every group of 3 repetitions, each repeats 10.Having added the of the present invention of 1% weight ratio in the feedstuff that test group ate is the animal immunity potentiator of main formula with the Radix Astragali, eats the matched group that is called of normal diet.Test was raised 7 days in advance, was just trying 30 days phases, write down feed intake every day.
2, butcher sampling during off-test, weigh respectively internal organs and immune organ calculate organ coefficient; Blood sample collection prepares serum, adopts immune turbidimetry kit measurement serum antibody IgA, IgG, IgM, complement C3, C4, and ELISA method kit measurement Cytokine of Serum IL-1, IL-2 content are measured on the CHEM-5 semi-automatic biochemical analyzer; Get test pig venous blood 10ml, adding is added with in the centrifuge tube of 1% aseptic heparin in advance, takes back laboratory and carries out cell culture and NBT detection.
3, gather fresh blood and spleen, adopt mtt assay to carry out peripheral blood lymphocyte and cultivate and the spleen lymphocyte proliferation test, measure T/B lymphopoiesis effect:
1) gets the aseptic anticoagulation of 2.0ml and mix the dilute suspension cell with equivalent Hanks liquid.Cell suspension carefully slowly is added on the lymphocyte separation medium with blood aliquot (2.0ml), blood is overlapped on the layering liquid, the centrifugal 2000rpm20min of room temperature level.Remove uppermost blood plasma, with capillary tube will be between blood plasma and the layering liquid thin but more clearly milky buffy coat be drawn in the test tube that contains 5ml Hanks liquid, the centrifugal 10min of 1000r/min behind the mixing cleans twice, the lymphocyte separation medium of flush away remnants.Detect cell viability＞95% and counting with trypan blue dyeing.With the cell culture fluid that contains 20% hyclone cell is adjusted into 2 * 10
6Individual cell/ml.
2) the every hole of 96 porocyte culture plates adds single cell suspension 100ul and 100ul PHA, and every group is provided with 5 repeating holes, and the culture fluid blank is set simultaneously, puts 5%CO
2, 37 ℃ of cell culture incubators leave standstill and cultivate 48h, stop cultivating every hole adding 5mg/ml tetramethyl azo azoles salt (MTT) solution 20ul continuation cultivation in preceding 4 hours and add DMSO solution 100ul again in every hole after 4 hours, dissolve black-and-blue Jia Za precipitation, use enzyme-linked immunosorbent assay instrument (BIO-RAD) to detect every hole OD570nm afterwards.Result of the test is represented with stimulation index SI:
The blank OD570 of SI=test group OD570/
4, spleen cell culture experiment ConA (Sigma):
1) the aseptic pig spleen of getting, place serum-free RPMI-1640 culture fluid (containing penicillin and streptomycin) plate, flushing repeatedly, carefully spleen is crushed with pin bucket bolt, filter through 400 order stainless steel filtering nets, clean twice (1000rpm/min, 5 minutes) with culture fluid, make cell suspension with the RPMI-1640 culture fluid that contains 20% hyclone.Adjusting the splenocyte number with the RPMI-1640 culture fluid that contains 20% hyclone is 6 * 10
6Individual cell/ml.
2) cell inoculation: cultivate with 96 holes are dull and stereotyped, every hole adds above-mentioned cell suspension 100ul, adds ConA or PHA in addition, and every group is provided with 5 repeating holes, and the culture fluid blank is set simultaneously, puts 5%CO
2, 37 ℃ of cell culture incubators leave standstill and cultivate 48h.
3) mtt assay is surveyed cell proliferation: add 5mg/mlMTT solution 20ul in stopping cultivating preceding 4 hours every holes, continue to put the incubator cultivation and add DMSO solution 100ul again in every hole after 4 hours, concussion is 10 minutes on miniature oscillator, uses afterwards and detects every hole OD570 value under enzyme-linked immunosorbent assay instrument (BIO-RAD) the 570nm wavelength.Result of the test is represented with the cell proliferation rate percentage rate:
The rate of increase (%)=(the blank OD570 of test group OD570-)/blank OD570 * 100%
5, select fresh blood to adopt the NBT reducing process to measure the neutrophil reducing power: get above-mentioned anticoagulation 0.1ml, the NBT that adds equivalent uses liquid, puts into 12 * 75mm plastic tube mixing, and on mouthpiece cover, test tube can be put 37 degree incubators, and jolting therebetween once.The aforesaid liquid taking-up is shaken up, draw 1 end in slide with capillary burette and do push jack, require to release afterbody, push jack wants thickness suitable, dry up with hair-dryer immediately, methanol is 1-2min fixedly, dries up, with 1% husky yellow fluid solution-dyed 5min, the tap water flushing, oily sem observation, counting.
Result of the test is as shown in the table:
Table 1 is the influence of the present invention to DLY ablactational baby pig daily gain, material anharmonic ratio and diarrhea rate;
Table 2 is the influence of the present invention to DLY ablactational baby pig peripheral blood lymphocyte conversion ratio;
Table 3 is the influence of the present invention to DLY ablactational baby pig spleen lymphocyte propagation;
Table 4 for the present invention to DLY ablactational baby pig serum IgG, IgA and IgM and C3, C4 level influence;
Table 5 is the influence of the present invention to IL-1 and IL-2 level in the DLY ablactational baby pig serum;
Table 1
| Matched group | Test group | |
| Eventually heavy (kg) daily gain of starting weight (kg) (g) material anharmonic ratio diarrhea rate (%) | 21.94±1.97 45.13±0.38 773.1±57.0 2.03±0.29 6.37±0.31 | 21.82±0.51 47.93±1.15 870.3±28.6 1.90±0.06 2.73±3.23 |
Compare with matched group, test group daily gain 12.5% (P＜0.05), material anharmonic ratio 6.4% (p＞0.05), diarrhea rate reduce by 133% (P＜0.05)
Table 2
| Matched group | Test group | |
| Con induces (SI) LPS to induce (SI) | 1.06±0.08 1.11±0.17 | 1.27±0.08 1.26±0.06 |
Compare with matched group, test group peripheral blood T-lymhocyte transformation rate under Con induces has improved 19.8% (P＜0.05) respectively.Induce the B-lymhocyte transformation rate to improve 13.5% but difference is not remarkable at Lps.
Table 3
| Matched group | Test group | |
| Con induces (%) LPS to induce (%) | 84.37±28.28 49.82±13.62 | 153.8±40.02 101.9±22.09 |
Compare with matched group, test group spleen cell conversion ratio under Con induces has improved 82.3% (P＜0.05) respectively.Inducing down at LPS, the spleen cell conversion ratio has improved 105% (P＜0.05).
Table 4
| Matched group | Test group | |
| IgG(g/l) IgA(g/l) IgM(g/l) C3(g/l) C4(g/l) | 4.41±0.168 0.529±0.007 0.63±0.09 0.173±0.018 0.021±0.006 | 4.81±0.310 0.658±0.017 0.68±0.06 0.201±0.017 0.024±0.004 |
Compare IgG, C in the test group piglet serum with matched group
3And C
1Level has improved 9.1%, 16.2% and 14.2% but difference is all not remarkable respectively, makes that the IgA level has improved 24.4% (P＜0.05) in the serum, makes in the serum IgM level reduce by 8.6% but difference is not remarkable.
Table 5
| Matched group | Test group | |
| IL-1(pg/ml) IL-2(pg/ml) | 8.48±1.39 13.95±1.18 | 19.33±1.89 27.28±1.18 |
Compare with matched group, the IL-1 alpha levels has improved 128% (P＜0.05) in the test group serum, makes that the IL-2 level has improved 95.6% (P＜0.05) in the serum.
The specific embodiment
Embodiment 1, a kind of be the animal immunity potentiator of main formula with the Radix Astragali, take by weighing raw material (kg) by following proportioning:
The Radix Astragali 12, Rhizoma Coptidis 8, Radix Scutellariae 13, Fructus Forsythiae 22 and Folium Isatidis 15.
Above-mentioned raw materials is simply mixed.
Embodiment 2, a kind of be the animal immunity potentiator of main formula with the Radix Astragali, take by weighing raw material (kg) by following proportioning:
The Radix Astragali 10, Rhizoma Coptidis 10, Radix Scutellariae 10, Fructus Forsythiae 25 and Folium Isatidis 15.
Above-mentioned raw materials is simply mixed.
Embodiment 3, a kind of be the animal immunity potentiator of main formula with the Radix Astragali, take by weighing raw material (kg) by following proportioning:
The Radix Astragali 15, Rhizoma Coptidis 5, Radix Scutellariae 15, Fructus Forsythiae 20 and Folium Isatidis 15.
Above-mentioned raw materials is simply mixed.
At last, it is also to be noted that what more than enumerate only is several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.
Claims (1)
1, is the animal immunity potentiator of main component with the Radix Astragali, it is characterized in that the recipe ingredient (by weight) of this reinforcing agent is: 15 parts of 10～15 parts of the Radixs Astragali, 5～10 parts of Rhizoma Coptidis, 10～15 parts of Radix Scutellariaes, 20～25 parts of Fructus Forsythiaes and Folium Isatidiss.
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|---|---|---|---|
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| CNB2005100501612A CN1325077C (en) | 2005-06-20 | 2005-06-20 | Animal immunopotentiator using membranous milk vetch root as basic remedy |
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| CN1325077C true CN1325077C (en) | 2007-07-11 |
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| CN102940717A (en) * | 2012-12-05 | 2013-02-27 | 邓凯伟 | Chinese medicament for treating cattle foot-and-mouth diseases |
| CN108272910A (en) * | 2018-03-20 | 2018-07-13 | 长沙小新新能源科技有限公司 | It is a kind of to antiviral veterinary drug and preparation method thereof |
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|---|---|---|---|---|
| CN1471836A (en) * | 2003-07-03 | 2004-02-04 | 中国农业科学院饲料研究所 | Birds feed additive |
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| CN1471836A (en) * | 2003-07-03 | 2004-02-04 | 中国农业科学院饲料研究所 | Birds feed additive |
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