CN103494038A - Chinese herbal medicine immunopotentiator for resisting white spot syndrome virus (WSSV) of procambarus clarkia, as well as preparation method and application thereof - Google Patents
Chinese herbal medicine immunopotentiator for resisting white spot syndrome virus (WSSV) of procambarus clarkia, as well as preparation method and application thereof Download PDFInfo
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a Chinese herbal medicine immunopotentiator for resisting white spot syndrome virus (WSSV) of procambarus clarkia, as well as a preparation method and an application thereof. The preparation method comprises the following steps: milling rheum officinale, epimedium, astragalus membranaceus and isatis root and fully mixing the ingredients in proportion to prepare the powdery immunopotentiator. The Chinese herbal medicine immunopotentiator disclosed by the invention is stable and wide in raw material resources, simple to prepare, and low in cost, is based on the immunomechanism of shellfish to provide Chinese herbal medicines for the shellfish, and can be used for improving the immunity and oxidation resistance of a body during application, improving the capacity of the procambarus clarkia for resisting WSSV infection, and promoting the growth of the procambarus clarkia.
Description
Technical field
The invention belongs to aquaculture field, relate to anti-white spot syndrome virus Chinese medicine immunity enhancer of a kind of Procambius clarkii and its preparation method and application.
Background technology
Procambius clarkii (Procambarus clarkii) is commonly called as " cray ", be subordinate to Crustachia (Cmstacea), Decapoda (Decapoda), Astacidae (Cambaridae), former Astacus (Procambarus) because of its delicious meat, nutritiously be subject to consumer favor, is topmost crayfish cultivation kind in the world today, its output accounts for 70%～80% of whole crayfish output, is also China's important economic aquaculture object at present.In recent years, Procambius clarkii cultivation scale enlarges year by year, and according to statistics, only the water surface of Jingzhou Prefecture, Hubei Province cultivation Procambius clarkii has just reached tens thousand of hectares.But, along with the rapid expansion of propagating area artificially, the generation of disease is also more and more frequent and serious.The diseases such as the domestic spiral shell substance disease of in succession having reported Procambius clarkii, fin rot, these diseases have brought heavy economic losses to the Procambius clarkii culturist, white spot syndrome virus (White Spot Syndrome Virus wherein, WSSV) be a kind of without Viruses With Inclusion Body, geographical distribution is wide, Epidemic Scope is large, and infectiousness is strong, there is no at present effective treatment approach.In recent years, white spot syndrome virus comes into vogue in the Procambius clarkii cultivation, and becomes the important disease in the Procambius clarkii cultivation, to aquaculture, brings massive losses.
Treat traditionally and prevent the Procambius clarkii disease mainly to rely on antibiotic, chemicals etc., but these drug abuses easily cause many negative effects such as drug resistance, immunosupress and destruction body microecological balance of environmental pollution, medicament residue, pathogenic microorganism, are seriously perplexing the cultivation of Procambius clarkii.Therefore, the immunity of raising animal self is very important to solving the farming disease harms problem.Natural Chinese medicinal herb is taken from the natural animal-plant product, the nature and the biologically active that have kept its composition, structure, as feed addictive, extra-nutrition is not only arranged, promote the effect of growing, building up health, improving premunition, add medicament with chemistry and compare, also there are the characteristics such as low toxicity, low-residual, drug resistance are not remarkable.Therefore Chinese herb feed additive becomes the additive of new generation of substitute antibiotics and chemical synthetic drug day by day.Rheum officinale, barrenwort, the Radix Astragali and Radix Isatidis are China's traditional Chinese medicine materials, have antiviral, anti-oxidant, regulate the effects such as humoral immunity of organism, the activate immunity factor, especially with the rheum officinale best results.But it remains the weak point in single use, improving shellfish immunity, anti-WSSV aspect DeGrain.
Shellfish is without the specific immunity system, take nospecific immunity as main, lysozyme (LZM), superoxide dismutase (SOD), CER (CP) and peroxidase (POD) etc. are important immune factors, resist in a different manner the invasion and attack of pathogen.Therefore, many scholars estimate shellfish immunity with above index.
Have at present some about rheum officinale, barrenwort, the Radix Astragali and Radix Isatidis a small amount of reports that use in aquaculture, as CN102389499A discloses a kind of Chinese medicine extract for aquaculture Disease control, it is mainly to be made by the raw material of following weight portion: 20～30 parts of 40～60 parts of rheum officinales, 20～40 parts of the Radixs Astragali, 15～25 parts of Radix Codonopsis, 20～25 parts of granatums, 15～25 parts of folium isatidis, 20～30 parts of Radix Isatidis, 15～25 parts of barrenwort, windproof 20～30 parts, 15～25 parts of the roots of purple-flowered peucedanum, 10～20 parts, Radix Glycyrrhizae and peppermints.But this Chinese medicine extract is mainly that immunity of organisms and the resistance against diseases to improving fish is effective, to shrimps, Procambius clarkii DeGrain particularly.Compatibility about rheum officinale, barrenwort, the Radix Astragali and Radix Isatidis is used and the Procambius clarkii impact be have not been reported at present.Therefore, research by death condition after the mensuration to Serum of Procambarus clarkii and infection WSSV, screening promotes growth and regulates the Chinese herbal medicine immunomodulator of Procambius clarkii non-specific immune function, may further be the mechanism research of Procambius clarkii immune response and improving its resistance against diseases provides scientific basis, significant to Procambius clarkii healthy aquaculture and sustainable development.
Summary of the invention
The objective of the invention is the above-mentioned deficiency for prior art, provide a kind of Procambius clarkii anti-white spot syndrome virus Chinese medicine immunity enhancer.
Another object of the present invention is to provide the preparation method of the anti-white spot syndrome virus Chinese medicine immunity enhancer of this Procambius clarkii.
Another purpose of the present invention is to provide the application of the anti-white spot syndrome virus Chinese medicine immunity enhancer of this Procambius clarkii.
Purpose of the present invention can be achieved through the following technical solutions:
The anti-white spot syndrome virus herbal composite of a kind of Procambius clarkii, composed of the following components: rheum officinale 25～45 weight portions, barrenwort 20～35 weight portions, the Radix Astragali 15～30 weight portions, Radix Isatidis 12～32 weight portions.
Described herbal composite is preferably composed of the following components: rheum officinale 30～38 weight portions, barrenwort 28～35 weight portions, the Radix Astragali 15～25 weight portions, Radix Isatidis 15～25 weight portions.
The application of herbal composite of the present invention in the preparation of the preparation anti-white spot syndrome virus of Procambius clarkii and/or raising immunity.
The anti-white spot syndrome virus Chinese medicine immunity enhancer of a kind of Procambius clarkii, contain herbal composite of the present invention.
Described immunopotentiator preferably prepares by the following method: by the first coarse crushing of each component of composition of the present invention, again through the micronizer pulverizing, after crossing 120 purpose screen clothes, in mixer, fully mix, make described immunopotentiator.
The preparation method of the anti-white spot syndrome virus Chinese medicine immunity enhancer of Procambius clarkii of the present invention, by the first coarse crushing of each component of herbal composite of the present invention, pulverize through micronizer again, after crossing 120 purpose screen clothes, fully mix in mixer, make described immunopotentiator.
The application of the anti-white spot syndrome virus Chinese medicine immunity enhancer of Procambius clarkii of the present invention in the anti-white spot syndrome virus feed of preparation Procambius clarkii.
The anti-white spot syndrome virus feed of described Procambius clarkii is to add described Chinese medicine immunity enhancer in conventional freshwater crayfish feed stuff gained according to the amount of 1wt%～1.5wt%.
Beneficial effect:
The present invention is directed to the characteristic of Procambius clarkii, immune mechanism from shellfish, filter out rheum officinale, barrenwort, the Radix Astragali and Radix Isatidis from Chinese herbal medicine, and it is composite according to specific proportioning, the one, well utilized " mutual-assistance relation " of rheum officinale and the Radix Astragali; The 2nd, rheum officinale, barrenwort and the Radix Astragali all have regulates immunity and anti-inflammatory activity, but the generation of the various factors in bidirectional modulation immune response, inflammatory reaction process; The 3rd, with Radix Astragali tonifying middle-Jiao and Qi, barrenwort is regulated immunity to hold up, and Radix Isatidis is clearing heat and detoxicating with eliminating evil, and three medicines share, and plays altogether strengthening vital QI to eliminate pathogenic factors, clearing heat and detoxicating merit.Four kinds of medicines are composite, overcome the weak point of single medicine, obtain anti-white spot syndrome virus herbal composite, and prepare on this basis the anti-white spot syndrome virus Chinese medicine immunity enhancer of Procambius clarkii.This immunopotentiator can directly be removed the free radical in the Procambius clarkii body, eliminate the negative effect of radical pair body tissue organ normal function, accelerate to repair destroyed immune system, can improve immunity, oxidation resistance and the opposing WSSV infection ability of body, promote the Procambius clarkii body growth, improve survival rate.
Raw materials enjoy stable sources of the present invention, preparation is simple, draws materials extensively, with low cost.
The specific embodiment
Embodiment 1
Get rheum officinale 30 weight portions, barrenwort 30 weight portions, the Radix Astragali 20 weight portions, Radix Isatidis 20 weight portions; By the first coarse crushing of above-mentioned each component, again through the micronizer pulverizing, after crossing 120 purpose screen clothes, in mixer, fully mix, make immunopotentiator.
Embodiment 2
Growth test: Procambius clarkii is provided by the Lu Kou of Jiangsu Fresh Water Aquatic Product Inst. base.Health, specification, the basically identical shrimp of weight are selected in test, the individual just about 7.5g of counterpoise.Random packet, be provided with control group A (basal diet of throwing something and feeding), control group B(adds 1.0% rheum officinale in basal diet), control group C(adds 0.5% rheum officinale and 0.5% barrenwort in basal diet) with test group (adding the Chinese medicine immunity enhancer of 1.0% embodiment 1 preparation in basal diet), establish 3 parallel tests for every group, every parallel group 50 tail test shrimp, totally 6 cement pits (2.50m * 2.5m * 0.8m).Shrimp domestication after 7 days, throw something and feed control group basal diet and 1.0% Chinese medicine immunity enhancer 60 days, measure the indexs such as its growth, immunity.
Table 1 basal diet formula
Annotate: compound premix (every kg feed): vitamin e1 50mg, vitamin K5 0mg, vitamin B1 80mg, vitamin B2 50mg, nicotinic acid 150mg, pantothenic acid 150mg, vitamin B6 50mg, biotin 1mg, cobalamin 0.02mg, folic acid 10mg, vitamin C 300IU, retinol1 0000IU, calciferol 000IU. copper sulphate 2.0g, ferric sulfate 25g, zinc sulfate 22g, manganese sulfate 7g, sodium selenite 0.04g, KI 0.026g, cobalt chloride 0.1g.
Table 2
* heavy-average just body weight of the average end-body of average growth rate %=()/average just body weight * 100
Result shows: with control group A, compare, test group has significantly improved: rate of body weight gain 27.55%, lysozyme (LZM) vigor 96.38%, albumin (ALB) content 66.92%, CER (CP) vigor 27.36%, total protein (TP) content 8.50%, total superoxide dismutase (SOD) vigor 23.00%, nitric oxide (NO) content 57.55%, peroxidase (POD) vigor 12.37%, substantial reduction in triglycerides (TG) content 35.37% (P<0.05); With control group, B compares, and test group has significantly improved: rate of body weight gain 25.06%, albumin (ALB) content 56.01%, nitric oxide (NO) content 17.72%, peroxidase (POD) vigor 8.50% (P<0.05); With control group, C compares, and test group has significantly improved: albumin (ALB) content 18.12%, nitric oxide (NO) content 25.38%, peroxidase (POD) vigor 9.74% (P<0.05).Therefore, immunopotentiator of the present invention can significantly improve the immunity of shrimp body and oxidation resistance than each control group, promotes the shrimp bulk-growth.
Challenge test: growth test is after 60 days, fasting 24 hours, select again every group of 30 tails of shrimp that specification is consistent and carry out the WSSV infection experiment, observe the physiological activity after shrimp is injected WSSV virus (attacking poison), and statistics control group A (basal diet of throwing something and feeding), control group B(add 1.0% rheum officinale in basal diet), control group C(adds 0.5% rheum officinale and 0.5% barrenwort in basal diet) with test group (Chinese medicine immunity enhancer that in basal diet prepared by interpolation 1.0% embodiment 1), the cumulative mortality of four groups of shrimp 0h, 12h, 24h, 36h, 48h, 60h.
Table 3
Result shows: after Procambius clarkii infects WSSV, the vigor of shrimp starts to descend, respond such as weakens at the phenomenon to external world; Each timing statistics point, the results are shown in Table 3.From table 3, with control group A, to compare, the 1.0% Chinese medicine immunity enhancer death rate has significantly reduced by 27.27%, 53.86% (P<0.05) at 48h, 60h; With control group, B compares, and the 1.0% Chinese medicine immunity enhancer death rate has significantly reduced by 18.18%, 15.38% (P<0.05) at 48h, 60h; With control group, C compares, and the 1.0% Chinese medicine immunity enhancer death rate has significantly reduced by 18.18%, 7.69% (P<0.05) at 48h, 60h.Therefore, this immunopotentiator can improve Procambius clarkii opposing WSSV infection ability, reduces the death caused by infecting WSSV.
Embodiment 3
Get rheum officinale 30 weight portions, barrenwort 25 weight portions, the Radix Astragali 15 weight portions, Radix Isatidis 15 weight portions; By the first coarse crushing of above-mentioned each component, again through the micronizer pulverizing, after crossing 120 purpose screen clothes, in mixer, fully mix, make immunopotentiator.
Embodiment 4
Get rheum officinale 33 weight portions, barrenwort 30 weight portions, the Radix Astragali 19 weight portions, Radix Isatidis 18 weight portions; By the first coarse crushing of above-mentioned each component, again through the micronizer pulverizing, after crossing 120 purpose screen clothes, in mixer, fully mix, make immunopotentiator.
Embodiment 5
Get rheum officinale 38 weight portions, barrenwort 28 weight portions, the Radix Astragali 25 weight portions, Radix Isatidis 25 weight portions; By the first coarse crushing of above-mentioned each component, again through the micronizer pulverizing, after crossing 120 purpose screen clothes, in mixer, fully mix, make immunopotentiator.
Embodiment 6
Get rheum officinale 30 weight portions, barrenwort 25 weight portions, the Radix Astragali 15 weight portions, Radix Isatidis 15 weight portions; By the first coarse crushing of above-mentioned each component, again through the micronizer pulverizing, after crossing 120 purpose screen clothes, in mixer, fully mix, make immunopotentiator.
Embodiment 7
Get rheum officinale 37 weight portions, barrenwort 32 weight portions, the Radix Astragali 23 weight portions, Radix Isatidis 23 weight portions; By the first coarse crushing of above-mentioned each component, again through the micronizer pulverizing, after crossing 120 purpose screen clothes, in mixer, fully mix, make immunopotentiator.
Embodiment 8
Get rheum officinale 25 weight portions, barrenwort 24 weight portions, the Radix Astragali 17 weight portions, Radix Isatidis 15 weight portions; By the first coarse crushing of above-mentioned each component, again through the micronizer pulverizing, after crossing 120 purpose screen clothes, in mixer, fully mix, make immunopotentiator.
Embodiment 9
Get rheum officinale 43 weight portions, barrenwort 32 weight portions, the Radix Astragali 30 weight portions, Radix Isatidis 28 weight portions; By the first coarse crushing of above-mentioned each component, again through the micronizer pulverizing, after crossing 120 purpose screen clothes, in mixer, fully mix, make immunopotentiator.
Embodiment 10
According to the growth test in embodiment 2 and challenge test method, measure respectively and throw something and feed containing the index such as the Procambius clarkii growth of the immunopotentiator of preparation in embodiment 3～9, immunity and to infect WSSV48h and 60h cumulative mortality.The control group basal diet of throwing something and feeding, each test group is added respectively the Chinese medicine immunity enhancer of 1.0% embodiment 3～9 preparations in basal diet.The results are shown in Table 4 and table 5.
Table 4
* heavy-average just body weight of the average end-body of average growth rate %=()/average just body weight * 100
Table 5
Comparative Examples 1
Prepare Chinese medicine extract 3 according to CN102389499A embodiment 3 records.
The Contrast on effect example
Growth test: Procambius clarkii is provided by the Lu Kou of Jiangsu Fresh Water Aquatic Product Inst. base.Health, specification, the basically identical shrimp of weight are selected in test, the individual just about 7.5g of counterpoise.Random packet, be provided with control group: the Chinese medicine extract 3 that adds 1.0% Comparative Examples 1 preparation in basal diet, the basal diet formula is shown in embodiment 2, test group: add the Chinese medicine immunity enhancer of 1.0% embodiment 1 preparation in basal diet, the basal diet formula is shown in embodiment 1; Parallel group 50 tail test shrimp, totally 6 cement pits (2.50m * 2.5m * 0.8m).Shrimp domestication after 7 days, throw something and feed control group and test group feed 60 days, measure the indexs such as its growth, immunity, and result data is in Table 6.Visible immunopotentiator of the present invention can significantly improve the immunity of shrimp body and oxidation resistance than the Chinese medicine extract of CN102389499A, promotes the shrimp bulk-growth.
Table 6
* heavy-average just body weight of the average end-body of average growth rate %=()/average just body weight * 100
Challenge test: growth test is after 60 days, fasting 24 hours, select again every group of 30 tails of shrimp that specification is consistent and carry out the WSSV infection experiment, observe the physiological activity after shrimp is injected WSSV virus (attacking poison), and statistics control group (adding the Chinese medicine extract 3 of 1.0% Comparative Examples 1 preparation in basal diet) and test group (adding the Chinese medicine immunity enhancer of 1.0% embodiment 1 preparation in basal diet), the cumulative mortality of two groups of shrimp 0h, 12h, 24h, 36h, 48h, 60h, the results are shown in Table 7.Have result visible, immunopotentiator of the present invention can significantly improve Procambius clarkii opposing WSSV infection ability than the Chinese medicine extract of CN102389499A, reduces the death caused by infecting WSSV.
Table 7
Claims (8)
1. the anti-white spot syndrome virus herbal composite of Procambius clarkii, is characterized in that composed of the following components: rheum officinale 25～45 weight portions, barrenwort 20～35 weight portions, the Radix Astragali 15～30 weight portions, Radix Isatidis 12～32 weight portions.
2. herbal composite according to claim 1, is characterized in that composed of the following components: rheum officinale 30～38 weight portions, barrenwort 28～35 weight portions, the Radix Astragali 15～25 weight portions, Radix Isatidis 15～25 weight portions.
3. the application of the described herbal composite of claim 1 or 2 in the preparation of the preparation anti-white spot syndrome virus of Procambius clarkii and/or raising immunity.
4. the anti-white spot syndrome virus Chinese medicine immunity enhancer of Procambius clarkii, is characterized in that containing the described composition of claim 1 or 2.
5. the anti-white spot syndrome virus Chinese medicine immunity enhancer of Procambius clarkii according to claim 4, it is characterized in that described immunopotentiator prepares by the following method: by the first coarse crushing of each component of the described composition of claim 1 or 2, pulverize through micronizer again, after crossing 120 purpose screen clothes, fully mix in mixer, make described immunopotentiator.
6. the preparation method of the anti-white spot syndrome virus Chinese medicine immunity enhancer of Procambius clarkii claimed in claim 4, it is characterized in that the first coarse crushing of each component of the described composition of claim 1 or 2, pulverize through micronizer again, after crossing 120 purpose screen clothes, fully mix in mixer, make described immunopotentiator.
7. the application of the anti-white spot syndrome virus Chinese medicine immunity enhancer of Procambius clarkii claimed in claim 4 in the anti-white spot syndrome virus feed of preparation Procambius clarkii.
8. application according to claim 7, is characterized in that the anti-white spot syndrome virus feed of described Procambius clarkii is to add Chinese medicine immunity enhancer claimed in claim 4 in conventional freshwater crayfish feed stuff gained according to the amount of 1wt%～1.5wt%.
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| CN106581041A (en) * | 2016-12-01 | 2017-04-26 | 兴化市恒威生物技术有限公司 | Immunopotentiator for preventing and treating crayfish white spot syndrome virus (WSSV) and use method thereof |
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| CN110540996A (en) * | 2019-08-29 | 2019-12-06 | 华中农业大学 | Procambarus clarkii i-type lysozyme gLysi2 gene, gLysi2 protein coded by same and application thereof |
| CN110731427A (en) * | 2019-10-11 | 2020-01-31 | 南京农业大学 | fishing gonadal development promoting additive and its prepn and application |
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