CN1583792A - Vitellus immune globulin for preventing prawn virus, tis preparing method and use thereof - Google Patents

Abstract

The invention belongs to the field of biotechnology, relates to the technical field of aquatic animal breeding, and in particular relates to a yolk immunoglobulin (IgY) for preventing and treating prawn virus disease and a preparation method thereof. It is characterized in that one or more inactivated strains/attenuated strains/gene subunit immunogens/nucleic acid immunogens of the prawn white spot syndrome virus are made into a single antigen or a composite antigen, and egg-laying poultry is used as a biological reaction Specific IgY against white spot syndrome of prawns can be obtained by immunization, and this IgY can be used alone or in combination with peptidoglycan, prebiotics and Chinese herbal medicines as feed additives, medicinal bathing agents or active ingredients of shrimp pond spraying agents to apply to prawn diseases Prevention and treatment. The effect and benefit of the present invention are that the IgY plays a role through the passive immune mechanism, directly suppresses and kills the virus, enhances the immunity of the shrimp body, and has the advantages of significant effect, safety, environmental protection, convenient use and the like in the prevention and treatment of white spot syndrome of prawns.

Description

A kind of yolk immunoglobulin for preventing and treating prawn virus disease and its preparation method and application

technical field

The invention belongs to the field of biotechnology, relates to the technical field of aquatic animal breeding, and in particular relates to a yolk immunoglobulin (IgY) for preventing and treating prawn virus disease and a preparation method thereof. The IgY can be used as a feed additive or a medicine bathing agent or the active ingredient of the splashing agent to control the white spot syndrome of prawns.

Background technique

my country's prawn farming started in the late 1970s. The continuous improvement and perfection of aquaculture production technology made it quickly become one of the pillar industries of the mariculture industry. The density of stocking, coupled with the continuous deterioration of the ecological environment for farming, has made shrimp diseases, especially viral diseases of shrimps, an important factor hindering the development of this industry. From 1993 to 1994, prawn virus disease ravaged all over my country and swept across all prawn farms, causing more than 70% of the cultured prawns to die. According to incomplete statistics, the direct economic loss caused by this amounted to tens of billions of yuan, seriously affecting the development of fishery economy in coastal areas. In the study of the etiology of large-scale mortality of cultured prawns, it was found that viral diseases are often the culprit. There are no less than 15 kinds of viral diseases that have been reported, and new viral diseases are constantly being discovered and reported. According to a research report published in the "Nature" magazine, the results obtained by applying a new quantitative method show that the number of virus particles in natural water bodies is much higher than expected, as high as 2.5×10 ⁸ per liter.

White spot syndrome is one of the most important diseases in prawn farming, and its pathogen is a kind of white spot syndrome virus (WSSV-type, White Spot Syndrome Virus group). Including white spot baculovirus (WSBV), subcutaneous and hematopoietic baculovirus (HHNBV), shrimp nuclear baculovirus (RV-PJ), systemic ectoderm and mesoderm baculovirus (SEMBV), noninclusion Somatic Shrimp Virus (NOSV). The virus is highly contagious, and the mortality rate of infected prawns is as high as 90-100% within a week. In the prevention and treatment of white spot syndrome, prevention-oriented measures are mainly adopted: on the one hand, by transforming shrimp ponds, improving the breeding environment, and improving the breeding technology to play an indirect preventive role; on the other hand, mainly using peptidoglycan, microbial polysaccharide, Vitamin C, Chinese herbal medicine, probiotics, and fly maggots can improve the self-immunity of shrimp, in order to enhance the ability of shrimp to resist virus infection, but there is no specific therapeutic drug preparation to prevent and treat white spot syndrome.

These above-mentioned pharmaceutical preparations mainly enhance the ability of the shrimp to resist virus infection by improving the non-specific immunity of the shrimp, but they are helpless against the virus itself. The activity of peptidoglycan and microbial polysaccharides is easily affected by factors such as strains and fermentation conditions, which limits their application; the reduced state of vitamin C is extremely unstable, and the efficacy of the medicine is affected; the resources of Chinese herbal medicine are very limited, the efficacy of the medicine is slow, and the side effects are toxic. Further research is needed, and it is restricted by factors such as raw material quality, processing methods, and compatibility ratios; probiotics (live bacteria preparations) are easily affected by external conditions, and are easily inactivated during processing and storage, and will also be absorbed by the digestive juice after entering the digestive tract. Inactivated, it is difficult to have a sufficient number of beneficial bacteria to play a role, and there are fewer types of live bacteria.

IgY is an avian-derived antibody similar in structure to mammalian IgG. It has high immune activity, and its specific activity can still be detected at a content of 50ng/ml. It has stable performance and strong heat resistance. Under certain conditions, the activity of IgY can be maintained for at least 24 hours, and the activity remains unchanged after pasteurization; the range of acid and alkali resistance is wide, and it is stable at pH 4.0-11.0, and its stability can be further improved by microencapsulation, and there is no Toxic, no residue, no side effects, green and environmental protection. According to the needs, different antigens are used to immunize poultry to produce specific IgY against the antigen, and the use of poultry to produce specific IgY has the advantages of large-scale production, low cost, high content of specific antibodies (100-400mg/egg) and It has the advantages of good safety, etc., and then it can be added to feed or bait after being dried and processed into powder, which is convenient to use and suitable for popularization.

IgY can specifically bind to the corresponding antigen, thereby inhibiting or changing the state or activity of the antigen. Oral administration of IgY can enable the body to obtain passive immunity. IgY can block the epitope of the virus-adsorbed susceptible cell receptor, thereby preventing the virus from adsorbing and penetrating into the cell to proliferate, thus limiting the spread of the virus between cells and tissues and in the bloodstream. spread. Immune poultry eggs have a variety of immune enhancing factors, which can play a synergistic effect after the body ingests them, enhance the body's non-specific immunity, and improve disease resistance.

Because IgY and immune poultry eggs have the above-mentioned excellent characteristics, that is, to overcome the shortcomings of current immune enhancers and other drugs that have little actual killing effect on the virus and limited practical production and application, it can prevent and treat the disease caused by white spot syndrome virus from the root. Shrimp viral diseases caused by pathogens.

At present, although published WO03070258, KR2003069506-A and WO2004004764-A1 patent documents provide a method for preventing or treating prawn white spot syndrome using yolk immunoglobulin, the above invention still has the following deficiencies:

1. The pathogen of prawn white spot syndrome is a type of virus. There are differences in the white spot disease virus infected by different hosts in different regions. Only one kind of white spot syndrome virus is selected to prepare the antigen. It cannot be applied to infections caused by other virus strains, the practical application range is narrowed, and the effect of suppressing and killing viruses is weakened.

2. The nucleic acid immunogen is made without using the white spot syndrome virus gene or its antigenic determinant gene, which brings disadvantages such as uneven antibody titer produced by immunized poultry, unstable yield, and reduced practical application effect.

3. Only IgY is used as the only active ingredient for the prevention and treatment of white spot syndrome in prawns. Not only the dosage is increased, but the immunity of the infected prawns is reduced, which leads to the entry of other pathogenic microorganisms, but the IgY cannot effectively inhibit the water body or These pathogenic microorganisms existing inside the shrimp body cannot achieve the best therapeutic effect.

Contents of the invention

The object of the present invention is to provide a kind of yolk immunoglobulin (IgY) of preventing and treating prawn white spot syndrome, and further improve the preparation method of this IgY, use it as the main activity of feed additive, medicinal bathing agent or shrimp pond splashing agent The ingredients are used in the prevention and control of prawn diseases to achieve the dual effects of inhibiting and killing viruses and enhancing the immune function of prawns, significantly reducing the probability of prawn infection, improving the survival rate, and solving the problem that there is no specific drug preparation for the prevention and treatment of prawn white spot syndrome.

The purpose of the present invention is to realize through following technical scheme:

1. Antigen Preparation

Taking WSBV as the representative strain of white spot syndrome virus, the antigen preparation process is described in detail:

(1) Preparation of WSBV inactivated strain

Take healthy crayfish without WSBV infection, wash the body surface, soak in 0.01% potassium permanganate solution for 5 minutes, rinse with sterile water, and disinfect the junction of the carapace and abdomen with alcohol cotton balls. Prepare crayfish primary cells, gently and evenly resuspend them with whole culture medium, inoculate into culture flasks or culture plates, culture overnight at 27°C, and set aside. Another WSBV-infected shrimp tissue was collected, and the WSBV virus liquid was obtained by sterile filtration, and intramuscularly injected into the third and fourth abdominal segments of healthy crayfish (60-120 μl/head). After 4 to 6 days of infection, the muscle tissue of the infected crayfish was stripped, and the L-15 medium was used as a homogenization buffer to extract the virus. Select cells that spread evenly and in good condition, add freshly prepared virus infection solution at a ratio of 1/10, incubate for 1 hour, wash cells with L-15 for 3 times, add full culture solution, continue culturing at 27°C and harvest virus solution, 15 %NaCl inactivated virus solution for 18-24 hours, and stored at 4°C or -20°C.

(2) Preparation of WSBV gene subunit immunogen

Remove the carapace of the diseased shrimp, remove the eyes and appendages, add an equal volume of TN (Tris-NaCl) buffer, and homogenize for 5 minutes with a high-speed tissue homogenizer at 12000 r/min. The homogenate was centrifuged at 5000g for 1h, the supernatant was taken, centrifuged at 8400g for 20min, then the supernatant was centrifuged at 30000g for 1h, the precipitate was taken, resuspended by adding 3-5 times of TN buffer, and placed at the top of the 15-16% sucrose density gradient. After centrifugation at 75,000 g for 2 h, 1 ml of the virus layer was collected, diluted with TN buffer, and then centrifuged at 8,400 g for 0.5 h, and the supernatant was taken, resuspended with an equal amount of TN buffer to obtain purified virus. All centrifugation was performed at 4°C. Design and synthesize primers, extract the genomic DNA of the Chinese strain of WSBV (GeneBank accession number AF332093) as a template, clone the VP24, VP26, and VP28 genes by PCR amplification, and subclone them into the multiple cloning site of the vector pUC118 to obtain VP24, The recombinant cloning plasmids of VP26 and VP28 genes were connected with the expression plasmid pGEX-2T respectively, transformed into competent cells BL21, and positive colonies were screened. Pick a single positive colony and culture overnight at 37°C in LB medium, transfer 1/50 or 1/100 of the bacterial solution to 5L liquid medium containing Amp, shake until A(OD) ₅₅₀ =0.4～0.6, add 1.5ml of IPTG (1M) induced expression, after 1-4 hours, centrifuged to collect the bacteria, SDS-PAGE and Western Blot detection showed that the target protein was induced, and the fusion protein was purified with glutathione-activated Sepharose4B affinity chromatography column, and the purification was determined. The protein concentration was stored at -20°C for future use.

(3) Preparation of nucleic acid immunogen

The VP24, VP26, and VP28 genes obtained by the method described in (2) were respectively connected to the expression plasmid pVAX1, transformed into competent cells DH5α, and positive colonies were screened. A large number of recombinant plasmids were extracted, the plasmids were purified by Sepharose-CL4B column chromatography, and the bacterial endotoxin contained in the plasmid DNA was detected by the Limulus reagent method, and the endotoxin content was controlled within the range of less than 20EU/mg plasmid, in sterile PBS Or store in sterile water at 4°C or -20°C for later use.

2. Poultry Immunization

The single antigen or compound antigen prepared by the above method is mixed with Freund's complete adjuvant or Freund's incomplete adjuvant respectively in equal amounts, and the healthy disease-free chickens to be laid or newly opened are respectively primed and boosted. A total of three times, after 14 days, blood was collected every 3 days to measure the antibody titer. When it reached log ₂ 11 or more, the immunized eggs were collected and stored at 4°C.

The above immunization methods and injection frequency can be appropriately adjusted and changed according to the actual situation; the same immunization technique as above can also be used to immunize different egg-laying poultry such as laying ducks or gooses or turkeys or ostriches with the above-mentioned antigens to obtain Corresponding IgY.

3. Preparation of biological agents with immunological activity

(1) Separation and purification of specific IgY

When the number of immunized poultry eggs reaches a certain amount, wash the eggshell surface with 0.2-0.5% bromogeramine or potassium permanganate aqueous solution, then disinfect with 5% tincture of iodine, and finally deiodine twice with 75% alcohol. The egg beater breaks the immune eggs and separates the yolk. Dilute the immunocompetent egg yolk liquid 6-10 times with sterilized water or buffer solution, adjust the pH to 5-5.2 with NaOH, let it stand at 4°C for 12-18 hours, and centrifuge to harvest the supernatant, (NH ₄ ) ₂ The SO ₄ solution was salted out, and after standing for several hours, the crude IgY liquid was obtained by ethanol precipitation or sodium sulfate precipitation, and the crude extract was purified by Sephadex G-200 column chromatography and concentrated.

(2) Preparation of immune active whole egg powder/yolk powder/antibody powder

Whole egg liquid or egg yolk liquid or purified IgY solution with immune activity is stirred and emulsified into an emulsion, and spray-dried or freeze-dried to make powder.

The effect and benefit of the present invention are that the specific IgY against prawn white spot syndrome prepared by applying the technical scheme directly acts on the pathogen of prawn white spot syndrome, exerts a specific inhibitory effect, and overcomes peptidoglycan, prebiotics, Chinese herbal medicine, etc. The use of preparation does not have the shortcoming that kills effect to virus itself; Poisoned prawns, poisoned prawns and cultured shrimp ponds all have the characteristics of preventing infection, suppressing and killing viruses, purifying water quality, and improving the environment, so that the invention has the advantages of direct effect, permanent cure and obvious effect.

What needs to be pointed out in particular is that the specific IgY obtained by using the compound antigen made by different strains of white spot syndrome virus is a polyclonal antibody, which can simultaneously produce inhibitory and killing effects on a variety of different pathogens, thus greatly improving the immunity of prawns. Anti-infection ability, the survival rate of prawns in the compound antigen test group can reach 95-100%, and the survival rate of prawns in the single-antigen test group is 90-93%. Due to the differences in the white spot syndrome virus infected by different hosts in different regions, the compound antigen prepared The dosage of specific IgY is small, the cost is low, and the effect is particularly remarkable.

And the nucleic acid immunogen made by the white spot syndrome virus gene or its antigenic determinant gene, the specific IgY yield obtained by immunization is large and stable, each egg contains 150-200mg of IgY, the antibody titer is uniform, and the highest can be obtained up to 1:1024, thus further improving the disease resistance of prawns and greatly reducing the production cost.

Furthermore, in addition to using specific egg yolk antibody powder or egg yolk powder or whole egg powder containing the egg yolk antibody, active ingredients such as prebiotics, peptidoglycan or Chinese herbal medicines are added to the pharmaceutical composition, and the synergistic effect of various immune enhancers, Completely block the invasion of various pathogenic microorganisms, enhance the physique of shrimp, activate the body's non-specific immune defense function, and enhance its ability to resist and prevent diseases.

The anti-prawn white spot syndrome specific IgY prepared by the present invention is a unique pure natural biological preparation, which is safe to use, has no toxic and side effects, is green and environmentally friendly, and has no drug residues; meanwhile, it will not induce the generation of drug-resistant strains at all, It will not kill beneficial bacteria and cause "flora imbalance". And prawns belong to lower invertebrates, lack of specific antigen/antibody immune response mechanism, so that it can only resist external invasion through simple ways such as phagocytosis of blood cells instead of producing immunoglobulin, and the IgY prepared by the present invention is An IgG-type immunoglobulin, which prevents and treats shrimp virus diseases by exerting a passive immune mechanism, thus indirectly supplies shrimp immunoglobulins to make up for the immune function defects of shrimps, and achieves the dual effects of inhibiting virus killing and enhancing shrimp immunity. Significantly reduce the probability of prawn infection with viruses, promote the growth and reproduction of prawns, and then solve the urgent needs of the prawn farming industry and fill the gap in the lack of prawn disease control drugs.

Detailed ways

Example 1

Preparation of Crawfish Primary Cells

Take healthy crayfish without WSBV infection, wash the body surface, soak in 0.01% potassium permanganate solution for 5 minutes, rinse with sterile water, and disinfect the junction of the carapace and abdomen with alcohol cotton balls. Draw 2-3mL anticoagulant (0.14mol/L NaCl, 0.1mol/L glucose, 30mmol/L citric acid, 26mmol/L sodium citrate, 10mmol/L EDTA, pH4.6) with a sterile syringe, and collect blood from the sinus , anticoagulant and hemolymph were mixed and centrifuged, centrifuged at 1500r/min for 10min, the supernatant was discarded, the precipitate was rinsed twice with L-15 medium, and washed with whole culture solution (15% calf serum, 100U/ml penicillin and 100 μg/ml streptomycin in L-15 medium), gently and uniformly resuspend lymphocytes, inoculate into culture flasks or culture plates, culture overnight at 27°C, and set aside.

Example 2

Target gene cloning

The VP28 gene was amplified by PCR method, and the sequencing results showed that the gene contained 615 nucleotides, and the homology of sequence fragments of WSSV from different sources in GeneBamk was 100%.

Example 3

protein induced expression

The results of SDS-PAGE electrophoresis showed that the strain carrying the recombinant plasmid pGEX-2T produced a specific protein band at the molecular weight of 24KDa, 26KDa, and 28KDa respectively, and the expression amount accounted for more than 40% of the total bacterial protein. , while no corresponding protein band was produced in the control strain.

Example 4

Separation and purification of recombinant protein

Use PBS to balance the pH value of the Sepharose4B affinity chromatography column, take 25ml of bacterial lysate, centrifuge to remove cell debris, take the supernatant with a pipette, add the affinity gel column, let the liquid flow out and discard; When the top of the column is even, add 25ml of PBS; then add 2.5ml of glutathione eluent (Glutathion Elution Buffer, glutathione 10mmol/L, pH8.0 Tris-HCl50mmol/L), act for 10min at room temperature and collect The liquid flowing out; repeat the elution 4 times, and collect the eluate.

Example 5

Immunization with Nucleic Acid Vaccines

The optical density ratio of the recombinant plasmid obtained as described in the technical scheme was 1.9 measured at 260 nm and 280 nm. Calculate the plasmid content according to 1D = 50 μg, dissolve it in 100 μl of sterile PBS, and inject 100 μl/point of Agkistrodon venom into chicken breast and chicken hind leg muscles one week before the immunization of 3-week-old Roman chickens (20 μg/ml dissolved in normal saline ), and marked, and then injected 50 μg/point into the chicken breast, chicken hind leg muscle, subcutaneously, intraperitoneally and intravenously, and performed the second immunization with the same dose and the same position one week later, and increased the immune dose to 60 μg/point at the same position after two weeks Three exemptions.

Example 6

Determination of antibody titer

Prepare single inactivated virus antigen, gene subunit antigen, and nucleic acid antigen from WSBV from Penaeus monodon as described in the technical scheme, and then use the antigen to immunize laying hens according to the method of immunizing poultry described in the technical scheme to prepare anti-prawn white spot Syndrome-specific IgY. Then, using the isolated and purified WSBV from the prawn as a detection antigen, the binding titer of the prepared anti-prawn virus disease-specific IgY was detected by ELISA. The results are as follows:

IgY Antigen for Detection Titer

I 1:1024

II 1:512

III 1:256

IV WSBV 1:256

V 1:1024

VI 1:512

VII 1:512

I: anti-inactivated virus-specific IgY

II: Anti-VP28 protein-specific IgY V: Anti-pVAX1-VP28-specific IgY

III: Anti-VP26 protein-specific IgY VI: Anti-pVAX1-VP26-specific IgY

IV: Anti-VP24 protein-specific IgY VII: Anti-pVAX1-VP24-specific IgY

Example 7

Prepare the inactivated strains of WSBV, HHNBV, HHNBV, SEMBV, and NOSV according to the preparation method of the inactivated strains described in the technical scheme, mix them in a ratio of 1:1:1:1:1 to make a composite antigen, and press the technical scheme The method for immunizing poultry immunizes laying hens to prepare anti-prawn white spot syndrome specific IgY. Then WSBV, HHNBV, RV-P, SEMBV and NOSV were used as detection antigens respectively, and the binding titer of the prepared anti-prawn virus disease-specific IgY was detected by ELISA method. The results are as follows:

IgY Antigen for Detection Titer

                                                                       

Hhnbv 1: 1024

against shrimp white spot syndrome

RV-P 1: 512

Hetero-IgY

SEMBV 1:512

NOSV 1: 512

Example 8

Mix the gene subunit vaccines VP28, VP26, and VP24 from Penaeus monodon WSBV prepared as described in the technical method at a ratio of 1:1:1 to prepare a composite antigen, and then use the antigen to immunize laying hens, and prepare as described in the technical plan Specific IgY against white spot syndrome in shrimp. Then, using the WSBV isolated and purified from the prawn as the detection antigen, the binding titer of the prepared anti-prawn virus disease-specific IgY detected by ELISA was 1:1024.

Example 9

100 tails of Penaeus monodon were randomly divided into 10 groups, one of which was the control group, and the other was the treatment group, 20 tails in each group, and the crude extract of WSBV was injected intramuscularly into the superficial layer of the heart between the second and third abdominal segments of the prawns. tail 0.05ml, the control group died after 2 days, and the mortality rate reached 100% after 3 days; the treatment group was fed with specific IgY bait containing anti-prawn virus disease twice a day, no death cases occurred, and the treatment group was randomized. 6-8 tails were taken for PCR virus detection, and the test results were all negative.

Example 10

Prepare a single inactivated virus antigen from WSBV from Penaeus monodon as described in the technical method, immunize laying hens, harvest some eggs, separate and obtain 200ml of yolk liquid, dilute it 5 times with normal saline, add peptidoglycan and 100 mg each of Chinese herbal medicine powder, mixed evenly, soaked in 6hm ² ponds of Penaeus monodon, after 110 days, the incidence rate of shrimp ponds soaked in egg yolk liquid was between 0% and 10%, while that of the control shrimp ponds (not soaked in egg yolk liquid) The incidence rate is between 90 and 100%.

                                             

Shrimp Farming Chifa Shrimp Farming Chifa

                                            

Shrimp Farm Pond Days Sickness Rate Pond Days Sickness Rate

Product (hm ² ) Product (hm ² )

Number (d) (%) Number (d) (%)

Wafangdian 5 1.5 110 ＜10% 5 1.5 110 90

Zhuanghe 3 1 110 0 3 1 110 100

Example 11

The WSBV from Penaeus monodon was prepared as described in the technical method to immunize laying hens with a single inactivated virus antigen prepared as described in the technical method, harvest some eggs, separate and obtain egg yolk liquid, prepare egg yolk powder by spray drying method, combine egg yolk powder with prebiotics, peptidoglycan Prepare prawn feed additive with Chinese herbal medicine powder: add each 100g of probiotics, peptidoglycan and Chinese herbal medicine powder to 1 ton of egg yolk powder, and stir evenly.

Purchase feed specially used for raising prawns, add egg yolk powder, probiotics, peptidoglycan and Chinese herbal medicine prepared additives to the basic feed in an amount of 10%, stir with a mixer, and then dry and pack in a blower box at 35°C.

In a 50L aquarium, feed 25 each of Penaeus monodon, Penaeus chinensis, Penaeus japonicus, and Penaeus vannamei, and feed them with basic feed for 3 days, artificially infect each aquarium with WSBV, and treat the group for 1 day The synthetic feed containing the above-mentioned feed additives was fed twice for 5 days, and the control group continued to be fed with basic feed.

Example 12

The area of the shrimp pond is 20 mu, the water depth is 1 meter, and 220,000 seedlings (Penaeus monodon) are released. After 40 days, an outbreak of viral white spot occurs. There were more than 90 dead shrimp on the first day, and nearly 300 on the second day. Take the following measures: external use of immune egg yolk liquid (2mg/L) is splashed in the whole pool, and artificial compound feed containing 15% egg yolk powder is taken orally for 8 days. The number of dead shrimps decreased on the 3rd day after the medication, and only more than 10 dead shrimps were found on the 5th day, and returned to normal on the 7th day.

Claims (8)

1. A yolk immunoglobulin for preventing and treating prawn virus disease and its preparation method and application, preparing specific yolk immunoglobulin IgY for resisting prawn virus disease or a composition containing the IgY, characterized in that the IgY will cause prawn virus disease One or more of a class of viruses of viral diseases are used as artificial antigens to immunize egg-laying poultry, and the poultry eggs produced by the immunized poultry are collected, and the anti-prawn virus disease-specific yolk immunoglobulin extracted from the yolk , and use it as a biological product for the prevention and treatment of shrimp virus diseases. 2. a kind of yolk immunoglobulin for preventing and treating prawn virus disease according to claim 1 and its preparation method and application, is characterized in that the egg-laying poultry that prepares this IgY adopts comprises hen, female duck, hen goose, turkey Chicken, ostrich. 3. A kind of yolk immunoglobulin for preventing and treating prawn virus disease according to claim 1 and 2 and its preparation method and application, it is characterized in that the virus that causes prawn virus disease is a class of white spot syndrome virus WSSV: comprising white spot rod WSBV, subcutaneous and hematopoietic tissue necrosis baculovirus HHNBV, Japanese shrimp karyotype baculovirus RV-PJ, systemic ectoderm and mesoderm baculovirus SEMBV, non-inclusion body shrimp virus NOSV. 4. according to claim 1 or 2 described a kind of yolk immunoglobulin for preventing and treating prawn virus disease and its preparation method and application, it is characterized in that the main antigen of preparing this IgY is the inactivated strain of white spot syndrome virus class or Its attenuated strain or its gene subunit immunogen or its nucleic acid immunogen. 5. according to claim 1 or 2 described a kind of yolk immunoglobulin for preventing and treating prawn virus disease and its preparation method and application, it is characterized in that the prawn virus disease single antigen or compound antigen method described in the IgY preparation method is as follows Cultivate preferably one or more viruses according to claim 3 at first, when selecting a kind of pathogenic virus, directly add immune adjuvant to make single antigen; More than one white spot syndrome virus according to 1-10: 1-10 or 1-10: 1-10: 1-10 or 1-10: 1-10: 1-10: 1-10 or 1-10: 1-10 : 1-10: 1-10: 1-10 and so on, mix evenly, and then add immune adjuvant to make composite antigen. 6. A kind of yolk immunoglobulin IgY for preventing and treating prawn virus disease according to claim 1 and 2 and its preparation method and application, it is characterized in that the host of white spot syndrome virus infection is the white shrimp, South American White prawns, monodon prawns, Japanese prawns, Moji prawns, oriental white prawns, long haired prawns, Chinese prawns, Indian prawns, pink prawns, blue prawns, brown prawns, Zhoushi new prawns, Japanese cherry shrimps, and related New prawns , Long Arm Shrimp, Penaeus prawn, Mantis Shrimp, Hairy Shrimp. 7. A kind of egg yolk immunoglobulin for preventing and treating prawn virus disease according to claim 1 or 2 and its preparation method and application, is characterized in that the composition containing the IgY also contains any combination of prebiotics or peptidoglycan or Chinese herbal medicine. 8. A kind of yolk immunoglobulin for preventing and treating prawn virus disease according to claim 1 and its preparation method and application, is characterized in that the IgY can be used as feed additive or medicine bathing agent or the active ingredient of shrimp pond spraying agent: Each ton of feed additive contains 1-100g of specific IgY against prawn white spot syndrome or egg yolk powder/whole egg powder with immune activity; Each liter of medicinal bath contains 10-200ml of specific IgY against prawn white spot syndrome or egg yolk liquid/whole egg liquid with immune activity; Each liter of shrimp pond spray contains 2%-10% of specific IgY against prawn white spot syndrome or egg yolk liquid/whole egg liquid with immune activity.