CN111494499A - Astragalus membranaceus and acanthopanax senticosus soluble powder for poultry - Google Patents
Astragalus membranaceus and acanthopanax senticosus soluble powder for poultry Download PDFInfo
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- CN111494499A CN111494499A CN202010339743.7A CN202010339743A CN111494499A CN 111494499 A CN111494499 A CN 111494499A CN 202010339743 A CN202010339743 A CN 202010339743A CN 111494499 A CN111494499 A CN 111494499A
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- soluble powder
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Abstract
The invention discloses a soluble powder of radix astragali and acanthopanax for poultry, belonging to the technical field of veterinary medicines. The soluble powder is prepared by decocting radix astragali, radix Acanthopanacis Senticosi, fructus Schisandrae chinensis, radix Sophorae Flavescentis, caulis Kadsurae Longipedunculatae and herb Polygoni chinensis in water, filtering, concentrating to obtain extract, extracting radix Rosae Laevigatae and fructus Toosendan by subcritical extraction to obtain mixed extract, mixing the mixed extract and extract under heating, spray drying, adding glucose, and mixing. The radix astragali and acanthopanax soluble powder prepared by the invention has the effects of resisting infection, enhancing lymphocyte immunocompetence and promoting growth, has high safety and no adverse reaction, and is applied to anti-infection treatment of genitourinary tracts when chicks have low immunity.
Description
Technical Field
The invention belongs to the technical field of veterinary medicines, and particularly relates to radix astragali and acanthopanax soluble powder for poultry.
Background
Because the tail end of the urogenital tract of the poultry is directly communicated with the outside, and the sanitary conditions around the anus are poor, a plurality of pathogenic microorganisms such as bacteria, viruses and the like can be bred at the tail end of the urogenital tract and further enter the genital tract, the ovary and the oviduct are easy to be damaged by the pathogenic microorganisms, and a plurality of lymphatic tissues existing in the mucous membrane of the genital tract form a first immune barrier for resisting the invasion of the external pathogenic microorganisms into the genital tract of the poultry, and the lymphatic tissues are called genital tract-related lymphatic tissues and have important functions on maintaining reproductive physiology such as follicular development, ovulation, fertilization, egg formation and the like of the poultry. If poultry are suffering from genital infections such as salmonellosis, especially if the eggs are infected, the germs will be further transmitted longitudinally through the fertilized eggs to the offspring, and in addition these infected eggs, once taken to the table, will directly endanger human health.
In recent years, various additives are generated for anti-infection treatment of poultry, mainly including chemically synthesized additives, mainly including antibiotics, hormones, chemically synthesized drugs, and the like, and the additives are widely used for prevention and growth promotion and the like, and play a positive role in promoting the development of poultry industry. However, with the continuous and intensive research, people find that huge disadvantages are hidden behind the huge economic benefits brought by the chemical synthetic additives, for example, the excessive use of antibiotics can cause the quality of poultry meat products and products thereof to be reduced and the antibiotic resistance to be enhanced, thereby bringing serious potential safety hazards to human beings. The natural additive is mainly substances separated from plants or microbial metabolites, has the characteristics of high safety, small toxic and side effects, no drug resistance and the like, and is valued for having the functions of nutrition, resisting microorganisms, enhancing immunity and the like.
Therefore, in order to resist the condition that various infectious diseases and the like are easy to spread, and especially when the immunity of the chicken is low, the chicken is easy to be infected with diseases, the application aims to provide a natural additive for treating the chicken against infection, and simultaneously enhancing the immunity, increasing the intake and promoting the growth.
Disclosure of Invention
The invention aims to provide the soluble powder of radix astragali and acanthopanax for poultry, which has the effects of resisting infection, enhancing the immune capacity of lymphocytes and promoting growth, is applied to the anti-infection treatment of genitourinary tracts when the immunity of chickens is low, is safe and effective, and has no adverse reaction. The preparation method is simple, rapid and low in cost.
In order to achieve the purpose, the invention adopts the following technical scheme:
on one hand, the soluble powder of radix astragali and acanthopanax for poultry comprises the following raw materials in parts by weight: 20-30 parts of astragalus membranaceus, 20-30 parts of acanthopanax senticosus, 10-20 parts of schisandra chinensis, 15-25 parts of cherokee rose root, 10-20 parts of radix sophorae flavescentis, 10-20 parts of szechwan chinaberry fruit, 8-15 parts of lygodium japonicum, 8-15 parts of Chinese knotweed herb and 20-30 parts of glucose.
Further, the feed comprises the following raw materials in parts by weight: 23-27 parts of astragalus membranaceus, 23-27 parts of acanthopanax senticosus, 13-17 parts of schisandra chinensis, 18-22 parts of cherokee rose root, 13-17 parts of radix sophorae flavescentis, 13-17 parts of szechwan chinaberry fruit, 10-13 parts of lygodium japonicum, 10-13 parts of Chinese knotweed herb and 23-27 parts of glucose.
Further, the feed comprises the following raw materials in parts by weight: 25 parts of astragalus, 25 parts of acanthopanax, 15 parts of schisandra, 20 parts of cherokee rose root, 15 parts of sophora flavescens, 15 parts of szechwan chinaberry fruit, 12 parts of lygodium japonicum stem, 12 parts of Chinese knotweed herb and 25 parts of glucose.
The traditional Chinese medicine used in the invention
Astragalus root: the product is dried root of Astragalus membranaceus (Fisch.) Bge. of Leguminosae family or Astragalus membranaceus (Fisch.) Bge. Hsiao. Collected in spring and autumn, removed fibrous root and head, and dried in the sun. Sweet in nature and taste, warm. It enters lung and spleen meridians. The functional indications are as follows: tonify qi, strengthen superficies, induce diuresis, expel pus, heal wound and promote tissue regeneration. Can be used for treating deficiency of vital energy, asthenia, anorexia, loose stool, collapse of middle-warmer energy, chronic diarrhea, rectocele, hematochezia, metrorrhagia, superficial deficiency, spontaneous perspiration, qi deficiency, edema, carbuncle, cellulitis, intractable ulcer, blood deficiency, debility with yellowish complexion, internal heat, and diabetes; proteinuria due to chronic nephritis and diabetes.
Acanthopanax root: the product is dried root, rhizome or stem of Harms of Acanthopanax senticosus (Rupr. et Maxim.) Harms of Araliaceae. Collected in spring and autumn, cleaned and dried. Pungent, slightly bitter and warm in nature. It enters spleen, kidney and heart meridians. The functional indications are as follows: to replenish qi, invigorate the spleen, tonify the kidney and induce tranquilization. Can be used for treating spleen and kidney yang deficiency, asthenia, anorexia, soreness of waist and knees, insomnia, and dreaminess.
Schisandra chinensis: the product is dried mature fruit of Schisandra chinensis (Turcz.) Baill of Magnoliaceae or Schisandra sphenanthera Rehd of Wils. The former is called as "Bei Wu Wei Zi", the latter is called as "nan Wu Wei Zi". Picking in autumn when the fruit is ripe, sun-drying or steaming, sun-drying, and removing fruit stalks and impurities. Sour, sweet and warm in nature. It enters lung, heart and kidney meridians. The functional indications are as follows: astringe to arrest discharge, benefit qi and promote the production of body fluid, tonify kidney and calm heart. Can be used for treating chronic cough, asthma, nocturnal emission, enuresis, frequent micturition, chronic diarrhea, spontaneous perspiration, night sweat, thirst due to body fluid consumption, short breath, weak pulse, internal heat, diabetes, palpitation, and insomnia.
Root of cherokee rose: the product is root or root bark of Rosaceae plant fructus Rosae Laevigatae. Digging out from 8 months to 2 months in the next year, cleaning, cutting and drying in the sun. The nature and taste are sour and astringent; sweet; flattening; is nontoxic. It enters kidney and large intestine meridians. The functional indications are as follows: astringe, stop bleeding, heal wound, expel internal blood, activate blood, alleviate pain, kill parasites. Performing primary spermatorrhea; enuresis; dysentery; diarrhea; hemoptysis; hematochezia; (ii) metrorrhagia and metrostaxis; leucorrhea; rectocele; uterine prolapse; rheumatic arthralgia; traumatic injury; sores and ulcers; scalding; toothache; stomach pain; ascariasis; choking on bones; chyluria
Charcoal mother: the product is whole plant of Polygonaceae plant Polygoni chinensis mother grass. Collected in summer and autumn and dried in the sun. Pungent in nature and taste; bitter; cooling; is toxic. The functional indications are as follows: clearing heat and promoting diuresis; cooling blood and removing toxic substance; calming the liver and improving eyesight; promoting blood circulation and relaxing muscles and tendons. Major dysentery; diarrhea; swollen and sore throat; diphtheria; cough due to lung heat; (ii) pertussis; hepatitis; leucorrhea; cancer and swelling; otitis media; eczema; vertigo and tinnitus; corneal clouding; traumatic injury
Toosendan fruit: the product is dried mature fruit of Melia toosendan Sieb. et Zucc. Collected in winter when the fruit is ripe, impurities are removed, and the fruit is bitter, cold and toxic in dryness. The meridians enter liver, stomach and small intestine meridians. The functions of the medicine are mainly used for removing damp-heat, clearing liver fire, relieving pain and killing parasites. It is indicated for heart pain due to heat syncope, hypochondriac pain, hernia pain, abdominal pain due to parasitic infestation.
The lygodium japonicum is prepared from lygodium japonicum L ygodiumjaponicum (Thunb.) Sw., lygodium japonicum L, microphyllum (Cav.) R.Br. or crankshaft lygodium japonicum L, fiexuosum (L.) Sw., wherein the dried aerial parts of lygodium japonicum and lygodium japonicum are 2 medicinal materials with different medicinal parts from the same medicinal plant source, and lygodium japonicum is collected from the second book of the Guangdong Chinese medicinal material standards in 1990 edition and 2010 edition, sweet and cold can clear away heat and toxic materials, promote diuresis and treat stranguria, and can be used for treating heat stranguria, stranguria with sand, stranguria with urinary stone, common cold with blood, stranguria with chyluria, damp-heat jaundice, wind heat, cough, throat, diarrhea and dysentery with swelling and pain.
Glucose: the most widely distributed and important monosaccharide in nature is a polyhydroxyaldehyde. Pure glucose is colorless crystals, has sweetness less than sucrose, is readily soluble in water, is slightly soluble in ethanol, and is insoluble in diethyl ether. Aqueous natural glucose solution rotates to the right and is thus "dextrose". Glucose plays an important role in the field of biology, and is an energy source and a metabolic intermediate product of living cells, namely a main energy supply substance of organisms.
On the other hand, the preparation method of the soluble powder of radix astragali and acanthopanax for poultry comprises the following steps:
s1, weighing astragalus membranaceus, acanthopanax senticosus, schisandra chinensis, sophora flavescens, lygodium japonicum and Chinese knotweed, adding water for decocting twice, adding 8-12 times of water by weight for each time, decocting for 1-2 hours, filtering decoction solutions, combining the filtrate, and concentrating to obtain an extract;
s2, crushing and sieving cherokee rose root and szechwan chinaberry fruit, performing subcritical extraction by using ethyl acetate as a solvent, feeding the pithecellobium clypearia and the ethyl acetate according to a feeding ratio of 1g to 1m L-1 g to 5m L, performing subcritical extraction at a temperature of 35-50 ℃, extracting for 60-80 min at an extraction pressure of 18-22 MPa and a volume flow of 2-4 kg/h, and removing the ethyl acetate by reduced pressure concentration to obtain a mixed extract;
and S3, heating and mixing the extract and the mixed extract, carrying out spray drying at the temperature of 60-70 ℃, adding the glucose, and uniformly mixing to obtain the glucose-.
Further, the relative density of the thick paste in the step S1 is 1.25-1.40.
Further, the process conditions of the spray drying in the step S3 are as follows: the air temperature of the inlet tower is 170 ℃, the flow rate of the peristaltic pump is 400 ml/min-600 ml/min, and the centrifugal speed is 200 r/min-400 r/min.
On the other hand, the application of the soluble powder of radix astragali and acanthopanax for poultry is provided, the soluble powder of radix astragali and acanthopanax has the effects of resisting infection, enhancing lymphocyte immunocompetence and promoting growth, and is applied to anti-infection treatment of genitourinary tracts when the immunity of chickens is low.
The invention has the beneficial effects that:
1. according to the formula, poultry infection is caused by dampness syndrome clinically according to the theory principle of Chinese veterinarian syndrome differentiation, and cherokee rose root, lightyellow sophora root, szechwan chinaberry fruit, lygodium japonicum and Chinese knotweed have the effects of killing insects, clearing heat, drying dampness, cooling blood and stopping bleeding, so that the poultry infection caused by parasites, bacteria and the like can be prevented and treated; radix astragali, radix Acanthopanacis Senticosi, and fructus Schisandrae chinensis have effects of invigorating qi, consolidating superficial resistance, promoting urination, and removing toxic substances, and can be used for enhancing immunity; the glucose powder is easy to absorb, and the prepared soluble powder is quick to absorb, can quickly achieve the purpose of treatment and has long curative effect time. The medicines are matched, so that the formula has the effects of killing insects, clearing heat and drying dampness, cooling blood and stopping bleeding and enhancing immunity, and has high safety and long curative effect.
2. The radix astragali acanthopanacis soluble powder prepared by the invention is high in safety and free of adverse reaction through acute toxicity tests of mice and long-term toxicity tests of rats, female chicks are subjected to resistance treatment after urogenital tract infection ureaplasma urealyticum (mycoplasma) infection, the generation of I L-1 β and I L-6 can be inhibited, pathological damage caused by mycoplasma infection inflammatory reaction is reduced, germ cell apoptosis caused by mycoplasma infection is reduced, the level of antibodies of chickens for resisting newcastle disease viruses can be improved by matching with newcastle disease vaccines, the immunosuppressive action of dexamethasone is resisted, the lymphocyte proliferation of chicks is promoted, the forming rate of red cell C3bR yeast rosette is increased, the development of immune organs such as thymus, spleen, bursa of Fabricius and the like of the chicks is promoted, and the immunity enhancing capability is proved to be very good.
3. According to the preparation method, the ethyl acetate extract prepared in advance by the cherokee rose root and the toosendan fruit through subcritical fluid has stronger anti-infection effect.
Detailed Description
The present invention is further described in the following examples, which should not be construed as limiting the scope of the invention, but rather as providing the following examples which are set forth to illustrate and not limit the scope of the invention.
Example 1
A preparation method of radix astragali and Acanthopanax senticosus soluble powder for poultry comprises:
s1, weighing 25g of astragalus membranaceus, 25g of acanthopanax senticosus, 15g of schisandra chinensis, 15g of sophora flavescens, 12g of lygodium japonicum and 12g of Chinese knotweed, adding water for decocting twice, adding 10 times of water by weight for each time, decocting for 2 hours, filtering decoction solutions, combining filtrate, and concentrating to obtain an extract with the relative density of 1.29;
s2, taking 20g of cherokee rose root and 15g of szechwan chinaberry fruit, crushing, sieving, performing subcritical extraction by using ethyl acetate as a solvent, wherein the feeding ratio of the cherokee rose root to the ethyl acetate is 1g:4m L, the subcritical extraction temperature is 40 ℃, the extraction time is 70min, the extraction pressure is 20MPa, the volume flow is 3kg/h, and performing reduced pressure concentration to remove the ethyl acetate to obtain a mixed extract;
s3, heating and mixing the extract and the mixed extract, spray-drying at 65 ℃ under the process conditions that the air temperature in a tower is 170 ℃, the flow rate of a peristaltic pump is 500ml/min and the centrifugal speed is 300r/min, adding 25g of glucose, and uniformly mixing to obtain the glucose-containing water-soluble granules.
Example 2
A preparation method of radix astragali and Acanthopanax senticosus soluble powder for poultry comprises:
s1, weighing 20g of astragalus membranaceus, 30g of acanthopanax senticosus, 20g of schisandra chinensis, 20g of radix sophorae flavescentis, 12g of lygodium japonicum and 12g of Chinese knotweed herb, adding water for decocting twice, adding 10 times of water by weight for each time, decocting for 2 hours, filtering decoction solutions, combining filtrate, and concentrating to obtain an extract with the relative density of 1.32;
s2, taking 15g of cherokee rose root and 20g of szechwan chinaberry fruit, crushing, sieving, performing subcritical extraction by using ethyl acetate as a solvent, wherein the feeding ratio of the cherokee rose root to the ethyl acetate is 1g:4m L, the subcritical extraction temperature is 40 ℃, the extraction time is 70min, the extraction pressure is 20MPa, the volume flow is 3kg/h, and performing reduced pressure concentration to remove the ethyl acetate to obtain a mixed extract;
s3, heating and mixing the extract and the mixed extract, carrying out spray drying under the process conditions that the air temperature in a tower is 60 ℃, the flow rate of a peristaltic pump is 500ml/min and the centrifugal speed is 300r/min, adding 25g of glucose, and uniformly mixing to obtain the product.
Example 3
A preparation method of radix astragali and Acanthopanax senticosus soluble powder for poultry comprises:
s1, weighing 30g of astragalus membranaceus, 20g of acanthopanax senticosus, 15g of schisandra chinensis, 10g of radix sophorae flavescentis, 12g of lygodium japonicum and 12g of Chinese knotweed herb, adding water for decocting twice, adding 10 times of water by weight for each time, decocting for 2 hours, filtering decoction solutions, combining filtrate, and concentrating to obtain an extract with the relative density of 1.38;
s2, taking 25g of cherokee rose root and 10g of szechwan chinaberry fruit, crushing, sieving, performing subcritical extraction by using ethyl acetate as a solvent, wherein the feeding ratio of the cherokee rose root to the ethyl acetate is 1g:4m L, the subcritical extraction temperature is 40 ℃, the extraction time is 70min, the extraction pressure is 20MPa, the volume flow is 3kg/h, and performing reduced pressure concentration to remove the ethyl acetate to obtain a mixed extract;
s3, heating and mixing the extract and the mixed extract, spray-drying at the temperature of 70 ℃, under the process conditions that the air temperature of an inlet tower is 170 ℃, the flow rate of a peristaltic pump is 500ml/min and the centrifugal speed is 300r/min, adding 25g of glucose, and uniformly mixing to obtain the glucose-glucose mixed material.
Example 4
Decocting radix Rosae Laevigatae, fructus Toosendan and other herbs together in the same manner as in example 1
S1, weighing 25g of astragalus membranaceus, 25g of acanthopanax senticosus, 15g of schisandra chinensis, 15g of sophora flavescens, 12g of kadsura pepper stem, 12g of Chinese knotweed herb, 20g of cherokee rose root and 15g of szechwan chinaberry fruit, adding water for decocting twice, adding 10 times of water for each time, decocting for 2 hours, filtering decoction solutions, combining filtrate, and concentrating to obtain an extract with the relative density of 1.32;
s2, spray-drying the extract under the process conditions that the air temperature of the inlet tower is 170 ℃, the flow rate of a peristaltic pump is 500ml/min and the centrifugal speed is 300r/min, adding 25g of glucose, and uniformly mixing to obtain the glucose-glucose mixed material.
Example 5
Test one safety test-mouse acute toxicity test (L D50 determination)
1. Test drug
The soluble powder of radix astragali and acanthopanax prepared in example 1 is dissolved by ultrasonic with distilled water to prepare a solution with the concentration of 0.48g crude drug/ml, 0.96g crude drug/ml and 1.60g crude drug/ml for later use.
2. Test method
Taking 40 healthy mice, wherein the weight of each mouse is 18-22 g. Fasting for 12 hours without water prohibition, weighing, randomly dividing into 3 groups, respectively intragastrically administering the soluble powder solution of radix astragali and radix Acanthopanacis Senticosi (0.2g crude drug/ml, 0.4g crude drug/ml, 0.8g crude drug/ml, 1.6g crude drug/ml concentration), 0.4ml/10g body weight each time, administering 3 times within 24 hours, feeding and observing for one week. The results are shown in Table 1.
TABLE 1 acute toxicity test (L D)50) Pretest report
The animals are all healthy and active, and none of the animals die, L D50 of gastric lavage administration cannot be detected because the concentration of the drug and the dosage of the drug cannot be increased, and L D50 of gastric lavage administration cannot be detected because the toxicity of the product is low.
And (2) test II: safety test-Long-term toxicity test in rats
1. Test drug
The soluble powder of radix astragali and acanthopanax prepared in example 1 is dissolved by ultrasonic with distilled water to prepare a solution with the concentration of 0.48g crude drug/ml, 0.96g crude drug/ml and 1.60g crude drug/ml for later use.
2. Test method
80 SD rats are taken, are subjected to adaptive observation for 1 week before experiment, and are randomly divided into four groups according to weight balance, namely large, medium and small dose groups of soluble powder of radix astragali and acanthopanax senticosus and a control group, wherein each group comprises 20 rats, and 5 rats are bred in each cage. The large dose group is administered with 38.4g crude drug/kg of radix astragali acanthopanacis soluble powder (corresponding to 80 times of the dosage for clinical poultry), the medium dose group is administered with 19.2g crude drug/kg of radix astragali acanthopanacis soluble powder (corresponding to 40 times of the dosage for clinical poultry), the small dose group is administered with 9.6g crude drug/kg of radix astragali acanthopanacis soluble powder (corresponding to 20 times of the dosage for clinical poultry), the control group is administered with physiological saline with equal volume, the administration volume is 1ml/100g body weight, 1 time per day, 1 month of continuous administration is performed, the general signs of animal mental state, behavior, activity, hair color, stool and the like are observed every day, 1 time of weighing is performed every week, and the dosage is adjusted according to the change of body weight. After the administration, 10 rats were taken from each group, fasted for 12 hours, measured for clotting time by the slide method, hemolyzed by the femoral artery, serum was separated, and blood biochemical examination, systemic autopsy and pathological examination were performed. The remaining animals in each group were discontinued and observed for 2 weeks before being treated as in the previous batch.
3. Observation and detection of indicators
The general behavioral activities, feces, hair, etc., of each group of animals were observed and recorded daily during the test period, weighed 1 time per week, clotting time was measured using a slide method, Hemoglobin (HGB), Red Blood Cells (RBC), platelets (P L T), White Blood Cells (WBC) and classification, etc., were measured using a cytometric analyzer, serum aspartate Aminotransferase (AST), alanine aminotransferase (A L T), alkaline phosphatase (A L P), urea nitrogen (BUN), Total Protein (TP), albumin (Alb), blood glucose (G L U), total bilirubin (T-Bili), creatinine (Crea), and total cholesterol (T-CHO), etc., were measured using a biochemical analyzer.
After dissection, observing whether abnormal organs exist or not by naked eyes, and performing pathological histological examination on the abnormal organs in an important point; organ index: accurately weighing the weights of the centering, liver, spleen, lung and kidney, and calculating the organ coefficient (organ weight g/100g body weight); and (3) pathological examination: taking main organs or tissues such as heart, liver, spleen, lung, kidney, stomach and the like, carrying out gross autopsy observation, fixing with 10% formalin, sending to a pathological tissue morphology subject group, conventionally taking materials, dehydrating, dipping wax, embedding, HE staining, and carrying out pathological examination under a light microscope.
4. Results
In the administration period and within 2 weeks of drug withdrawal, the general signs of the rats, such as spirit, behavior, activity, hair color, stool and the like, are not abnormal, and the death caused by drug toxicity does not occur. The body weight of each administration group after 1 month administration and 2 weeks withdrawal was significantly increased, and the results of t-test showed no significant change (P >0.05) compared with the control group, as shown in tables 2 and 3.
TABLE 2 weight change table of rats administered with soluble powder of radix astragali and Acanthopanax senticosus for 1 month
n＝20)
Compared with the normal control group, the ratio is: p >0.05
TABLE 3 weight change table for rats 2 weeks after drug withdrawal: (
n＝10)
Compared with the normal control group, the ratio is: p >0.05
The test results of Red Blood Cell (RBC) count, White Blood Cell (WBC) count, white blood cell classification (DC) and Hemoglobin (HGB) content and blood coagulation time of each administration group after 1 month administration and 2 weeks after drug withdrawal showed no significant change (P >0.05) compared with the control group by t test, and the results are shown in Table 4.
TABLE 4 Effect of Long-term toxicity test of soluble powder of radix astragali and Acanthopanax gracilistylus on hematological index of rat
Compared with the normal control group, the ratio is: p >0.05
The test results of AST, A L T, A L P, Crea, BUN, TP, Alb, G L U, T-Bili and T-CHO in each administration group after 1 month administration and 2 weeks after drug withdrawal have no obvious change (P >0.05) compared with the normal control group by T test, and the results are shown in Table 5.
TABLE 5 Effect of Long-term toxicity test of soluble powder of radix astragali and Acanthopanax gracilistylus on biochemical index of blood of rat
Compared with the normal control group, the ratio is: p >0.05
After administration for 1 month and 2 weeks after discontinuation, the animals were immediately dissected after sacrifice and the organs were observed with naked eyes one by one without abnormal changes. Accurately weighing the weights of the centering, liver, spleen, lung and kidney, calculating organ coefficients (organ weight g/100g body weight), wherein the organ coefficients among all groups have no obvious change (P is more than 0.05), and the results are shown in Table 6; optical microscopy: after administration for 1 month and 2 weeks after discontinuation, no obvious pathological damage related to the drug in heart, liver, spleen, lung, kidney and other major organs or tissues of each administration group is observed, and the difference is not obviously changed compared with the control group.
TABLE 6 Effect of St.radix Acanthopanacis Senticosi solubility long-term toxicity test on rat organ coefficients
Compared with the normal control group, the ratio is: p >0.05
From the above results, no obvious toxic reaction was observed in the large, medium and small dose groups after 1 month of gavage administration and 2 weeks of withdrawal of the soluble powder of radix astragali and acanthopanax in example 1. General conditions and behavior, feces and the like have no abnormal changes; the weight and growth condition of the rat are good, and no obvious abnormal condition occurs; the indexes of hematology, blood biochemistry, organ coefficients of main organs and the like are measured, and the indexes have no significant change (P is more than 0.05) compared with a normal control group; no apparent pathological lesions were seen at systematic necropsy and histopathological examination. Therefore, the soluble powder of radix astragali and acanthopanax is considered to be safe for long-term administration.
And (3) test III: anti-infection test of genitourinary tract in clinical test
1. Reagent and reagent
Reagent testing: soluble powder of radix astragali and Acanthopanax gracilistylus prepared in example 1 and comparative example 1
Reagent: uu liquid sterile culture solution, Uu solid sterile culture medium (provided by Shanghai Enkang biological science and technology Co., Ltd.), serum type 8 Uu strain (purchased from American strain Collection center, resuscitated passage), azithromycin tablet (specification: 0.25 g/tablet), and estradiol benzoate (specification: 2 mg/ml).
2. Molding die
140 Hailan brown hatchlings of 1 day old purchased from a breeding chicken farm of the agricultural science research institute in Changchun city were raised to 7 days old, and 20 were randomly selected as a blank control group. Then injecting 0.4 mg/female benzoic acid estradiol into the rest 120 female chicks at the neck under the skin, once per week, and totally four times; one week after the injection of estradiol benzoate, ureaplasma urealyticum (Uu) was reinfected. The specific method comprises the following steps: sucking 50 mu l of Uu bacterial liquid in logarithmic phase by using a micropipettor, extending the Uu bacterial liquid into the chicken vagina for about 0.5cm, slowly injecting the Uu bacterial liquid into the chicken vagina, and inverting the chicken for half a minute after the injection is finished so that the Uu bacterial liquid can be fully contacted with the inner wall of the reproductive tract. After one week, the chicken vaginal secretion is taken for culture, and Uu bacterial colony observed under a microscope indicates that the molding is successful.
3. Grouping and administration of drugs
The 120 female chicks successfully molded are randomly divided into 6 groups, each group comprises 20 female chicks, and the female chicks are divided into a model group, a high-dose, a medium-dose and a low-dose radix astragali and acanthopanax soluble powder and a drug control group. The medicines with different dosages in the administration groups are dissolved in warm boiled water and diluted into solution drops with the same volume. Specific grouping and handling is as in table 7.
Table 7 test grouping and handling
4. Observation and detection of indicators
In the experimental process, the diet and drinking conditions, the weight change, the hair color change, the genital secretion and the defecation conditions of the female chick are observed.
4.1 Interleukin detection
A femoral artery blood taking method is adopted before the mother chick is killed, the taken blood is timely centrifuged, then supernatant is taken, and the concentration of interleukin 1 β (I L-1 β) and interleukin 6(I L-6) in the serum of the mother chick is detected by a double-antibody sandwich ABC-E L ISA method.
4.2 apoptosis
After the experiment, each group of female chicks are killed, the abdomen is cut open, the conditions in the abdominal cavity such as vaginal secretion, uterus and the like of the female chicks are observed by naked eyes, partial vaginal and uterine tissues are taken, fixed by 10 percent formaldehyde, and subjected to pathological HE staining and slicing, and the number of apoptotic cells in the vaginal and uterine tissues of the female chicks is detected by a TUNE L method.
4.3 statistical analysis
The method is completed by SPSS18.0 software, and an analysis of variance method is adopted (numerical values with too large deviation are removed, and the detection of variance is uniform).
5. Results
5.1 observations
After the Uu bacterial liquid is inoculated, the water consumption of all the other groups of the female chicks except the normal control group begins to be obviously increased, the periphery of the vagina is red and swollen, secretion is increased, and the phenomena of depilation and liquid exudation are generated around the reproductive tracts of individual female chicks. The urine of the mother and the young baby is smelly and filthy, the urine volume is increased, and the color is yellow. When the female chicks are killed, the fact that the uterus of part of the female chicks is thickened and edema, the wall of the uterus is thinned is found, the uterus tissues are cut, the phenomenon that transparent liquid flows out from the cavity can be seen, and the phenomenon that the uterus of part of the female chicks is hardened can be seen. It is also seen that purulent secretion in the bladder of the mother bird makes the bladder wall thicker and even necrotized.
The phenomenon above the model group is the most serious in the inoculated Uu bacteria liquid group; in the administration group, the phenomena except the low dose group are closer than those in the model group, and the pathological phenomena in the other medium dose group, the high dose group and the drug control group are much better than those in the model group.
5.2 comparison of I L-1 β and I L-6
After the administration is finished, the content changes of interleukin 1 β (I L-1 β) and interleukin 6(I L-6) in the serum of the female chick are detected, the model group has very significant difference compared with a blank control group, the low-dose group and the comparative example 1 have significant difference compared with the model group, and the high-dose group and the medium-dose group have very significant difference compared with the model group, and the result shows that the soluble powder of radix astragali and acanthopanax senticosus prepared in the example 1 can resist the infection of ureaplasma urealyticum and can effectively inhibit the content of interleukin 1 β and interleukin 6, and the example 1 is better than the comparative example 1 when the soluble powder is administered with the medium dose, which shows that the extract obtained after subcritical extraction is carried out by adopting cherokee rose root and szechwan chinaberry fruit has stronger resistance, and the result is shown in a table 8.
TABLE 8 dynamic changes of I L-1 β and I L-6
Note: capital letters differ to indicate a very significant difference (P <0.01), lower case letters differ to indicate a significant difference (P <0.05), the same letters indicate no significant difference, and the following table is the same.
5.3 comparison of apoptosis in uterine tissue and vaginal tissue of each group of female chicks
Compared with the model group, the mean value shows that the uterine tissue apoptosis rate interference has a tendency of reducing in each group compared with the model group, particularly the high-dose group, the medium-dose group and the azithromycin group are most obvious, and the lower-dose group in the comparative example 1 is obvious. The results show that the high-dose group is equivalent to the azithromycin group, and the apoptosis rate of uterine tissue cells is reduced most obviously, which shows that the astragalus membranaceus and acanthopanax senticosus soluble powder prepared in the embodiment 1 can resist the infection of ureaplasma urealyticum and can effectively reduce the apoptosis rate of the uterine tissue cells of the female chicks. In the same manner, when the intermediate dose is administered, example 1 is better than comparative example 1, which shows that the extract obtained by subcritical extraction using cherokee rose root and toosendan fruit has stronger anti-infection property.
Compared with the model group, the mean value shows that the vaginal tissue apoptosis rate interference of each group has a tendency to be reduced compared with the model group, particularly the high-dose group, the medium-dose group and the azithromycin group are most obvious, and the lower-dose group of the comparative example 1 is obvious. The results show that the high-dose group is equivalent to the azithromycin group, and the apoptosis rate of vaginal tissue cells is reduced most obviously, which shows that the radix astragali acanthopanacis soluble powder prepared in the embodiment 1 can resist the infection of ureaplasma urealyticum and can effectively reduce the apoptosis rate of uterine tissue cells of the female chick. In the same manner, when the intermediate dose is administered, example 1 is better than comparative example 1, which shows that the extract obtained by subcritical extraction using cherokee rose root and toosendan fruit has stronger anti-infection property. The results are shown in Table 9.
TABLE 9 comparison of apoptosis (%)
The results show that after Uu bacterial liquid is infected, mycoplasma can stimulate human monocytes, macrophages and the like to secrete tumor necrosis factors I L-a, I L-1 β and I L-6, while TNF-a, I L-1 β and the like are all proinflammatory cytokines, are key cytokines for starting acute inflammatory response and important cytokines for starting apoptosis, are closely related to apoptosis, and can reduce inflammatory response after being used, so that apoptosis is inhibited, and the apoptosis inhibiting effect of the uterine tissue cells is most obvious in high-dose group.
In conclusion, the soluble powder of acanthopanax astragali provided by the invention can inhibit the generation of I L-1 β and I L-6, so that the pathological damage caused by mycoplasma infection inflammatory reaction is relieved, the germ cell apoptosis caused by mycoplasma infection is reduced, and the effect is enhanced along with the increase of the dosage of the drug
And (4) testing: immunity test of clinical trial
1. Test drugs: example 1 soluble powder of radix astragali and Acanthopanax gracilistylus (100 g/bag, 1.6g crude drug/g), produced by Jiangxi Chinese patent medicine raw materials GmbH
2. Grouping and administration:
1 day old Holland brown cockling purchased from chicken farm of agricultural science research institute in Changchun city, 450 chicks are selected when being fed to 7 days old, the chicks are randomly divided into 9 groups, 50 groups are fed separately by blank control groups without any treatment, a pure drug group only takes the soluble powder of radix astragali and acanthopanax senticosus without vaccine immunization, the rest 3 drug test groups begin to take the soluble powder of radix astragali and low dose, the drug control group begins to take astragalus polysaccharide powder, 2 immunosuppression groups take 4mg of dexamethasone sodium phosphate powder orally by each chick every day, the administration dose is accurately controlled, the administration is carried out by gastric perfusion, the drugs of different doses in each group are dissolved in warm water and diluted into solution with the same volume, the vaccine control group is not administered, when the day is 10, the rest groups are subjected to nasal dropping eye immunization by live vaccine (L a Sota strain), 2 parts of the chickens are subjected to each other group, and the intramuscular injection of the inactivated vaccine (L a Sota strain L) is specifically treated according to the treatment table 10.
Table 10 test grouping and handling
3. Detecting the index
3.1 newcastle disease antibody titer assay:
at the ages of 14, 21, 28, 35, 42 and 49 days, respectively, 8 jugular veins are randomly drawn from each group for blood collection, serum is separated, and dynamic changes of the antibody titer of the Newcastle disease are determined by a hemagglutination inhibition method according to the technical standard of Newcastle disease diagnosis (GB/T16550-2008).
3.2 peripheral blood lymphocyte proliferation assay
The proliferation of peripheral blood lymphocytes was measured by the MTT method at 28, 35, 42 and 49 days of age, respectively.
3.3 erythrocyte immune function assay
The red blood cell C3bR yeast rosette test was performed at 28, 35, 42, 49 days of age, respectively.
3.4 immune organ index assay
At 28, 35, 42, 49 days old, thymus, spleen and bursa of fabricius were isolated, weighed, and immune organ index, immune organ weight (g)/body weight (g) × 100 was calculated.
3.5 data statistics
The test data is statistically processed by SPSS18.0 software, the result is represented by mean (x) + -Standard Deviation (SD), the difference is significant when P is less than 0.05, and the difference is extremely significant when P is less than 0.01.
4. Results
4.1 Change in Newcastle disease antibody Titers
After 7 days of immunization of the newcastle disease vaccine, the antibody level of the newcastle disease virus of the chicks is obviously increased and reaches a peak value after 2 weeks.
The anti-newcastle disease virus antibody level of the chicks immunized by the newcastle disease vaccine is increased more quickly than that of the chicks immunized by the vaccine in 3 days in advance, and the higher level is maintained for a longer time, wherein the effect of a high (0.6g per 1kg body weight) dose group and a medium (0.4 g per 1kg body weight) dose group is more obvious.
Dexamethasone can obviously inhibit the immune function of chicks and reduce the immune response of the chicks to newcastle disease vaccines. The soluble powder of radix astragali and acanthopanax is taken orally according to the dose of 0.4g per 1kg body weight, the immunosuppression effect of dexamethasone can be resisted to a certain degree, the immune response capability of chicks is protected, and the result is shown in a table 11.
TABLE 11 dynamic Change of the Newcastle disease HI antibodies (log2)
Note: capital letters differ to indicate a very significant difference (P <0.01), lower case letters differ to indicate a significant difference (P <0.05), the same letters indicate no significant difference, and the following table is the same.
4.2 changes in lymphocyte proliferation
The oral soluble powder of radix astragali and radix Acanthopanacis Senticosi has effects of promoting proliferation of chicken lymphocyte, and resisting immunosuppression of dexamethasone, and has good effect in high and medium dosage groups. The results are shown in Table 12 below,
TABLE 12 dynamic changes in lymphocyte proliferation for each group (A)570Value)
4.3 erythrocyte C3bR Yeast rosette Change
The application of the soluble powder of radix astragali and acanthopanax can increase the forming rate of erythrocyte C3bR yeast rosette, which shows that the soluble powder can improve the immune function of erythrocyte to a certain extent, and the result is shown in table 13.
TABLE 13 influence of Eleutherococcus astragalis on the Ring ratio of chick Red blood cell C3bR (%)
4.4 changes in immune organ index
The thymus, spleen and bursal index of tested chicks are detected to prove that the oral administration of the soluble powder of radix astragali and acanthopanax can promote the development of immune organs, and the results are shown in tables 14, 15 and 16.
TABLE 14 change in thymus index (%)
TABLE 15 change in spleen index (%)
TABLE 16 change in bursa of Fabricius index (%)
4.5 Change in production Properties
The indexes of the tested chicken such as weight, feed intake, feed-to-weight ratio and the like are detected, so that the orally-taken soluble powder of radix astragali and acanthopanax has certain effects of promoting growth and development and improving production performance, and the results are shown in tables 17, 18 and 19.
TABLE 17 Effect of soluble powder of radix astragali and Acanthopanax gracilistylus on the body weight of test chickens (g/d. only)
TABLE 18 dynamic change of daily food intake (g/d. only)
TABLE 19 dynamic change of the material weight ratio (%)
The above results show that: the orally taken soluble powder of radix astragali and acanthopanax has better effect of enhancing the immune function of chicken, can improve the level of antibodies of the chicken against newcastle disease virus by being matched with newcastle disease vaccine, resist the immune suppression effect of dexamethasone, promote the lymphocyte proliferation of chicken, increase the forming rate of erythrocyte C3bR yeast rosette, and promote the development of immune organs of chicken such as thymus, spleen, bursa of fabricius and the like.
The recommended clinical dose of the soluble powder of radix astragali and acanthopanax is 0.4g/kg body weight d, the administration is started 3 days before the immunization of the vaccine, and the administration is continuously carried out for 5 days. The high-dose soluble powder (0.6g/kg body weight. d) of radix astragali and acanthopanax is orally taken, no adverse reaction is found after continuous administration for 5 days, and the preparation is proved to have higher clinical medication safety.
Claims (7)
1. The soluble powder of radix astragali and acanthopanax for poultry is characterized by comprising the following raw materials in parts by weight: 20-30 parts of astragalus membranaceus, 20-30 parts of acanthopanax senticosus, 10-20 parts of schisandra chinensis, 15-25 parts of cherokee rose root, 10-20 parts of radix sophorae flavescentis, 10-20 parts of szechwan chinaberry fruit, 8-15 parts of lygodium japonicum, 8-15 parts of Chinese knotweed herb and 20-30 parts of glucose.
2. The soluble powder of radix astragali and acanthopanax for poultry according to claim 1, which is characterized by comprising the following raw materials in parts by weight: 23-27 parts of astragalus membranaceus, 23-27 parts of acanthopanax senticosus, 13-17 parts of schisandra chinensis, 18-22 parts of cherokee rose root, 13-17 parts of radix sophorae flavescentis, 13-17 parts of szechwan chinaberry fruit, 10-13 parts of lygodium japonicum, 10-13 parts of Chinese knotweed herb and 23-27 parts of glucose.
3. The soluble powder of radix astragali and acanthopanax for poultry according to claim 1, which is characterized by comprising the following raw materials in parts by weight: 25 parts of astragalus, 25 parts of acanthopanax, 15 parts of schisandra, 20 parts of cherokee rose root, 15 parts of sophora flavescens, 15 parts of szechwan chinaberry fruit, 12 parts of lygodium japonicum stem, 12 parts of Chinese knotweed herb and 25 parts of glucose.
4. The soluble powder of radix astragali acanthopanacis senticosi for poultry according to any one of claims 1 to 3, wherein the preparation method of the soluble powder of radix astragali acanthopanacis senticosi comprises the following steps:
s1, weighing astragalus membranaceus, acanthopanax senticosus, schisandra chinensis, sophora flavescens, lygodium japonicum and Chinese knotweed, adding water for decocting twice, adding 8-12 times of water by weight for each time, decocting for 1-2 hours, filtering decoction solutions, combining the filtrate, and concentrating to obtain an extract;
s2, weighing cherokee rose root and szechwan chinaberry fruit, crushing, sieving, performing subcritical extraction by using ethyl acetate as a solvent, wherein the feeding ratio of the cherokee rose root to the ethyl acetate is 1g:1m L-1 g:5m L, the subcritical extraction temperature is 35-50 ℃, the extraction time is 60-80 min, the extraction pressure is 18-22 MPa, the volume flow is 2-4 kg/h, and ethyl acetate is removed by reduced pressure concentration to obtain a mixed extract;
and S3, heating and mixing the extract and the mixed extract, carrying out spray drying at the temperature of 60-70 ℃, adding the glucose, and uniformly mixing to obtain the glucose-.
5. The method for preparing the soluble powder of radix astragali and acanthopanax for poultry according to claim 4, wherein the relative density of the thick paste in the step S1 is 1.25-1.40.
6. The method for preparing the soluble powder of radix astragali and acanthopanax for poultry according to claim 4, wherein the spray drying process conditions of the step S3 are as follows: the air temperature of the inlet tower is 170 ℃, the flow rate of the peristaltic pump is 400 ml/min-600 ml/min, and the centrifugal speed is 200 r/min-400 r/min.
7. The soluble powder of radix astragali and acanthopanax for poultry according to any one of claims 1-3, wherein the soluble powder of radix astragali and acanthopanax has the effects of resisting infection, enhancing lymphocyte immunity and promoting growth, and is applied to the anti-infection treatment of genitourinary tract when the immunity of chicks is low.
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