KR20180096201A - A composition having antiviral activity for white spot syndrome virus - Google Patents

Abstract

Provided is a novel antiviral substance to prevent and treat infection by white spot syndrome virus in shrimp. According to the present invention, an extract mixture composed of Dryopteris crassirhizoma Nakai, Cleistocalyx operculatus, Aesculus turbinata Blume, licorice, turmeric, Angelica dahurica root, and camellia has shown excellent antiviral activities against white spot syndrome virus. In addition, the plant extract mixture is supplied to shrimp by coating a surface thereof with a polymeric substance in an attempt to increase feeding rate in the shrimp.

Description

[0001] The present invention relates to a composition having an antiviral activity against a white spot virus,

The present invention relates to an antiviral composition for preventing or treating diseases caused by white spot syndrome virus (WSSV) infection.

White spot syndrome virus (WSSV) is a virus widely distributed throughout the world. It has a tail-like attachment at the end of the viaon, and a rod-shaped capsid and envelope exist Which is similar to that of egg-shaped bacillus. Due to this structural feature, it has been called baculovirus or bacillus-like formed virus, but recently it has been named as a genetically new virus group, whispovirus (van Hulten MC, et al., J. Gen. Virol., 81 (Pt2), pp307-16, 2000). The white spot virus is composed of double-stranded DNA with a length of about 275 nm and a diameter of about 120 nm and a size of about 290 kb. WSSV is mainly infected with crustaceans such as shrimp, crayfish, and crabs. Especially, when the shrimp is infected, white spots appear on the carapace, appendage and cuticle of shrimp, and hepatopancreas of shrimp are red Color. White spots are accumulations of calcium salts and are most clearly formed on the inner surface of the carapace. Infected shrimps become lethargic, stop feeding, move slowly on the surface of water, and die. It is an extremely infectious disease in shrimp such as 80 ~ 100% dead within 3 ~ 10 days after infection and all dead within 20 days. Currently, WSSV is occurring all over the world, and although it does a lot of damage to aquaculture shrimp, there is no clear measure yet.

Although there are more than 100 kinds of diseases related to shrimp, it is reported that the damage caused by WSSV is the most serious. In many countries, white spot viruses have been studied, and studies have been actively conducted to identify viruses and their biological properties. However, the immune system of shrimp is different from vertebrate animals, and there are many difficulties in vaccine development. Most of the pathogens infected with vertebrate animals show a tendency to recede after antibody production in the host, but there is no report that shrimps resistant to WSSV have appeared. In Korea, glucan (Glucan) is added to feed to enhance immunity, or as a chlorine disinfectant, it disinfects the storage space before shrimp ingestion, but it does not have any clear effect. In addition, Korean Patent Registration No. 10-0824106 discloses that Zygomycetes japonica extract and Saururus chinensis extract have antiviral activity against white spot virus. In addition, licorice, licorice, peanut, buckwheat, hearing, and Kim extract have been reported to have inhibitory activity against white spot viruses. However, epoch-making methods for the prevention and treatment of white spot virus have not been developed to date.

Korean Patent Application No. 10-2014-0128540 Korean Patent Application No. 10-2004-0088637 Korean Patent Application No. 10-2006-0048317 Korean Patent Application No. 10-2006-0048780

Artificial infection of native Korean freshwater shrimp with white spot syndrome virus (WSSV). Experimental infection of white spot baculovirus in some cultured and wild decapods in Taiwan.

It is an object of the present invention to provide a novel antiviral composition having inhibitory activity against white spot syndrome virus (WSSV).

Another object of the present invention is to provide a mixture of plant extracts as a novel antiviral composition with inhibitory activity against white spot viruses.

Another object of the present invention is to provide a method for preventing or treating an infection caused by white spot virus in crustaceans.

In order to achieve the object of the present invention, the present invention provides an antiviral composition comprising a mixture of plant extracts, which is a mixture of plant extracts for preventing or treating infections caused by white spot viruses in crustaceans such as shrimp and crayfish .

The mixture of the above plant extracts may be a mixture of extracts for one of the crows, Craystokarix auxercuratus, Chilobar, Licorice, Ulvae, White paper and Camellia, or a mixture of the above-mentioned crowns, Craistocarix auxercutatus , Chickwood, licorice, kelp, white paper, and camellia. That is, it is possible to mix the extracts for each of the above-mentioned crows, Craystokarix operkurtus, Chilobar, Licorice, Ulgum, White paper and Camellia, and the above-mentioned crows, Craystokarix auxercutus, , White paper, and camellia. When each of the extracts is mixed, each component of each extract may be a pure extract in the process of production. On the other hand, in the case of extracting in a batch, there may be a slight difference in the effect of extracting various components together at the time of extracting each component.

1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 3.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0. 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0, and it is preferable that they are collectively extracted. Further, when extracting the component from the natural product, any one of purified water, ethyl acetate, butyl acetate, methyl alcohol, propanol, isopropyl alcohol and butyl alcohol, or a mixture of two or more thereof is used as an extraction solvent, Can be produced. At this time, the mixture of the plant extracts can be mixed with each other after extracting one of the crows, Craystokarix operculus, Chilobus, Licorice, Ulgus, White paper and Camellia, have.

The present invention also provides a method for preventing or treating infections caused by white spot viruses in crustaceans such as shrimp and lobster using a mixture of plant extracts. In the present invention, a mixture of plant extracts supplied for the purpose of preventing and treating infections caused by white spot virus on shrimp, etc., may be contained in an amount of 100 to 1,000 g per 1 ton of feed.

In one example of the present invention, the present inventors have examined the virus-inhibiting activity of the plant extract mixture after infecting the shrimp with WSSV, which is a native shrimp of Korean native shrimp. Specifically, the virus used for infecting shrimp in the present invention is a virus isolated from P. vannamei-infected P. vannamei. The virus isolated from the P. vannamei is inoculated with a syringe and infected with a flower or a shrimp. The amount of WSSV inoculated to infect shrimp is 5.8 × 10 ³ copy / ㎖, and 100 μl of virus is inoculated per 1 g of shrimp, which is the amount of virus in which the cumulative mortality rate of shrimp is about 80%. In order to confirm the viral inhibitory activity of the plant extract mixture, a mixture of the plant extract of the present invention is supplied together with the general shrimp feed before the WSSV inoculation.

In another example of the present invention, a mixture of plant extracts may be used by coating with commercially available shrimp feeds. In the present invention, the antiviral activity against the shrimp was used with coating of the extract, while the antiviral activity against the shrimp was used without coating the mixture of the extract. The coating treatment of the plant extract mixture can significantly reduce the ingestion of shrimp and the like by blocking the fragrance peculiar to the extract.

As a coating method, antiviral extract was mixed with polyvinyl alcohol (Sigma, P8136) at a ratio of 7: 1 to 11: 1 and then coated on a general feed (Daeha Plus No. 4, dominant feed) ≪ / RTI > to 2 days. After completely dissolving ethyl cellulose (Duksan, 5148) in a mixed solution of alcohol and acetone in a completely dried state, it is possible to spray 5-15% of the total weight of the general feed coated with the antiviral extract. The two-step coating treatment can not only completely block the unique flavor of the plant extract mixture but also help improve the stability of the feed.

In one example of the present invention, a mixture of plant extracts having an antiviral activity of the present invention is a product requiring a virus-suppressing activity and can be used for the production of a feed additive composition, a cosmetic composition, a disinfectant composition and the like. When used as a disinfecting composition, it is of course possible to include a pharmacologically, veterinarily or agriculturally acceptable carrier or diluent.

In the present invention, the effectiveness of the mixture of seven plant extracts in the laboratory was evaluated using Trachypenaeus curvirostris and Macrobrachium nipponense. As a result, the relative survival rate was confirmed to be 60% or more of the attacked group.

According to the present invention, the plant extract mixture of the present invention can be effectively used in combination with shrimp feed to prevent or treat white spot virus infection on shrimp. In addition, the plant extract mixture of the present invention can be prepared by first coating with polyvinyl alcohol, followed by secondary coating with ethylcellulose, which can improve shrimp feeding by shielding the odor of the plant extract.

Fig. 1 is a photograph of a giant tortoise ( Trachysalambria curvirostris ) used in the present invention.
Fig. 2 is a photograph of a shrimp ( Macrobrachium nipponense ) used in the present invention.
FIG. 3 is a photograph of a plant extract mixture of the present invention before and after coating. FIG.
FIG. 4 is a graph showing the relative survival rate of the white spot virus-infected female shrimp in the antiviral activity test using the plant extract mixture of the present invention.
FIG. 5 is a graph showing the relative survival rate of P. japonica shrimp infected with white spot virus in the antiviral activity test using the plant extract mixture of the present invention.
FIG. 6 shows the results of confirming the presence of WSSV infection by PCR. Lanes 1 and 2 are WSSV vaccinated shrimp, and lanes 3 and 4 are pre-WSSV vaccinated populations.
FIG. 7 shows PCR results of WSSV infection of P. japonica after the inoculation with WSSV, showing the results of the early (2-5 days), middle (6-10 days), and late (11-14 days) .

Hereinafter, the present invention will be described in more detail based on specific examples. The following examples are provided to aid understanding of the present invention and are not to be construed as limiting the scope of the present invention.

Materials and Experiments

Isolation of white spot virus for attack inoculation

WSSV-infected P. vannameiLitopenaues vannamei), And obtained from Pukyong National University Department of Microbiology, The virus titer of WSSV was 1.16 x 10 < RTI ID = 0.0 >⁶copy / ml. Was diluted 20-fold with TNE buffer (50 mM Tris-HCl, 400 mM NaCl, 5 mM EDTA, 1 mM proteinase inhibitor PMSF, pH 8.5) and inoculated with 100 μl per 1 g of shrimp body weight using a 1 ml (26 gauge) syringe. The final concentration of shrimp inoculated WSSV was 5.8 X 10³copy / 100 μl, and the virus is about 80% dead when attacked on the shrimp.

An antiviral plant extract mixture sample

The plant extracts of Table 1 below were obtained from the Central Vaccine Research Center. The plant extracts were extracted with 80 ~ 100% ethanol and concentrated. Each of the extracts was suspended in water and then fractionated with EtOAC (ethyl acetate) or n-BuOH (butanol) according to the respective samples to prepare a plant extract solution. The extracts of 7 kinds of plant extracts were 1: 1 or 1.0-1.5: 1.0 to 3.0 relative ratios. Table 1 shows the plant species and the extraction conditions used in the experiment, and Table 2 shows the relative weight ratios of the plant extracts, and it can be manufactured to have various weight ratios.

extract Scientific name Extraction condition crowd Dryoperis crassirhizoma Nakai Ethyl acetate Creastocarix Operkuratus Cleistocalyx opercalatus Chicory Aesculus turbinata (horse chestnut) licorice Glycyrrhiza uralensis Kool Curcuma longa blank Angelica dahurica Camellia Camellia japonical n-butanol

Weight ratio of plant extract Production Example 1 Production Example 2 Production Example 3 Production Example 4 Production Example 5 crowd 1.0 1.0 1.5 1.5 3.0 Creastocarix Operkuratus 1.0 1.5 One 2.0 1.0 Chicory 1.0 1.0 1.5 1.5 3.0 licorice 1.0 1.5 One 2.0 1.0 Kool 1.0 1.0 1.5 1.5 3.0 blank 1.0 1.5 One 2.0 1.0 Camellia 1.0 1.0 1.5 1.5 3.0

Setting the concentration of the antiviral plant extract mixture

In order to evaluate the efficacy of the plant extract mixture, the maximum concentration of polyvinyl alcohol was set. The polyvinyl alcohol to plant extract mixture could be included at a ratio of up to 7: 1 to 11: 1.

Acquisition of the giant prawns and prawns

The shrimp used in the experiment in the present invention are those in which WSSV artificial infection is confirmed (Experimental infection of white spot baculovirus in some cultured and wild decapods in Taiwan). (Trachysalambria curvirostris) weighing about 2 ~ 3 g and about 500 eggs were sampled in Haenam, Jeollanam-do, and then moved to Clarendon, Gyeongsangnam-do where seawater circulation system was maintained at 21 ℃ (± 1 ℃) . The water tank used for the test was 60 cm in length, length and height, and the water tank was filled with two-thirds of the seawater, and the water temperature was maintained at 21 ° C. (± 1 ° C.) using a temperature holding device (Amazon, AH 200) Commercially available shrimp feeds (Daeha Plus 4, Woosung Feed) were stabilized in a changing environment for one week while feeding 1% of shrimp average weight per day. The water in the tank was changed from 1/3 to 1/4 of the total amount of water once a day while monitoring the condition. Fig. 1 is a photograph of a flower prairie used in the present invention.

Macrobrachium nipponense is a native Korean freshwater shrimp that has been confirmed to be infected with WSSV (Artificial infection of native Korean freshwater shrimp with white spot syndrome virus). Approximately 500 shrimp weevils weighing about 2 ~ 4 g were sampled at Muju, Jeonbuk, and were transferred to the Muju test site while maintaining a temperature of 21 ℃ (± 1 ℃). The water tank used for the test was 60 cm in length, length and height, and the water tank was filled with two-thirds of the fresh water, and the water temperature was maintained at 21 ° C. (± 1 ° C.) using a temperature holding device (Amazon, AH 200) , Commercial shrimp feed (Daeha Plus 4, Woosung Feed) were stabilized in the changed environment for one week while feeding 1% of the average weight of shrimp per day. The water temperature was maintained at 21 ° C (± 1 ° C) to ensure optimal health of the shrimp before challenge with WSSV. This takes into account that replication of WSSV occurs at about 20-25 ° C. One-third to one-fourth of the total amount of water was changed once a day while monitoring the water condition. Shrimp feed containing the plant extract mixture was fed 1% of the shrimp average weight from 1 week prior to inoculation with WSSV. Fig. 2 is a photograph of a shrimp shrimp used in the present invention.

WSSV inoculation against shrimp

Inoculation with WSSV was carried out by infecting a female shrimp and a shrimp with a 1 ml syringe (needle diameter 26G). The viral load of the injected WSSV was 1.16 × 10 ⁶ copy / ml and diluted 20-fold in TNE buffer (50 mM Tris-HCl, 400 mM NaCl, 5 mM EDTA, 1 mM protease inhibitor PMSF, pH 8.5) ml < / RTI > per gram of shrimp weight using a syringe Lt; / RTI > The final concentration of WSSV in shrimp was 5.8 × 10 ³ copy / ㎖ per 1 g of shrimp, which is about 80% mortality when attacked on the shrimp.

Experimental Example

Experimental Example 1: Inoculation with WSSV

50 shrimp or shrimp were added to each water tank, and general food (Daeha Plus 4, dominant diet) was supplied for 7 days to adapt to the environment. After 7 days, the number of surviving shrimp in each tank was checked, and the weight of the surviving shrimp was measured. The WSSV for the inoculation was 1.16 × 10 ⁶ copies / ㎖, and 100 ㎕ of WSSV diluted 20 times with TNE buffer was used for the shrimp. Needle diameter 26G). The needle of the syringe was not inserted more than 1/3. Both the test group and the inoculation control group (positive control group) were administered WSSV in the same manner. In the negative control group, only 100 μl of TNE buffer solution was administered per 1 g of shrimp. All test groups, positive control group and negative control group were observed for 14 days after inoculation. In the space where the flower shrubs and ginseng shrimp are raised, a filter which can purify the water and an aeration stone were put in, and the water was circulated to remove the foreign substances inside. 1/3 to 1/4 of the total amount of water was changed once every 2-3 days.

Experimental Example 2: Feed additives for shrimp including plant extract mixture article

There was a need for a way to mask the effects of specific components of the plant extract mixture on eating habits. To increase the feeding power of the plant extract mixture, the mixture ratio of polyvinyl alcohol (Sigma, P8136) to plant extract mixture was adjusted to 7: 1 to 11: 1, and the mixture was coated on a general feed (Daeha Plus No. 4, dominant feed) The plant extract mixture is secondarily coated with ethyl cellulose (Duksan, 5148) so that the plant extract mixture does not dissolve in water, so that the plant extract mixture is slowly dissolved in water to minimize the effect of the plant extract mixture on the test shrimp Respectively. The polyvinyl alcohol solution was prepared by dissolving 100 g of polyvinyl alcohol in 1,000 ml of purified water or water for injection. The ethylcellulose solution was prepared by dissolving 49.5 g of ethylcellulose in 250 mL of acetone and 250 mL of 100% alcohol. The polyvinyl alcohol solution and the plant extract mixture were mixed at a ratio of 7: 1 to 11: 1, 10% of the total weight of the feed was applied, and the mixture was thoroughly dried for 24 hours at a temperature of 24 to 25 ° C., Of the total weight of the primary coated feed was added to make a secondary coating.

The coated shrimp feed of the present invention was able to substantially block the unique flavor of the plant extract mixture. Next, it was confirmed whether the shrimp feeds the coated shrimp feed well. For this purpose, 1% of the total weight of the shrimp was added to the shrimp feed without shrimp feeding during the day, and the coated shrimp feed was added at the evening of the active shrimp (6-7 pm). As a result, the supplied shrimp feed was consumed at about 1/3 of the first day and about 1/2 of the second day. As a result of continuous observation for 7 days, it was confirmed that the coated shrimp feeds were consumed on an average of about ½ or more daily per day. This was a very good intake compared to normal feeds.

Experimental Example 3: Confirmation of WSSV infection in the shrimp shrimp

Identification of WSSV infection in shrimp was confirmed through our company. PCR (Polymerase chain reaction) was performed to determine whether the cause of the shrimp was due to WSSV.

DNA was first extracted for PCR. Extracts from shrimp gills, body, and pontoon were obtained, homogenized directly without buffer, and placed in 1.5 ml tubes. And homogenized using 500 μl extraction buffer (400 mM NaCl, 20 mM Trizma base, 5 mM EDTA, 1% SDS, pH 8.0). Next, 500 μl of phenol (cholloform: isoamylalcohol) was added to the homogenized extract and mixed well. After centrifugation at 15,800 x g (about 12,000 rpm) for 5 minutes, the supernatant was transferred to a 1.5 ml tube and 3 M NaOAC (0.1 vol) and 100% ethanol (2 vol) were added. After mixing again, it was stored at -80 ° C for 10 minutes. After centrifugation at 18,506 x g for 30 minutes, the supernatant was removed and dried at room temperature for 1 to 2 minutes. The obtained DNA pellet was resuspended in 20 to 50 mu l of elution buffer. The WSSV-F primer (SEQ ID NO: 1: 5'-CTTTCACTTTCGGTCGTGTC-3 ') and the WSSV-R primer (SEQ ID NO: 2'5'-TACTCGGTCTCAGTGCCAGA-3') and the targeted WSSV vp28 gene (604 bp) were used. PCR was performed by repeating 35 cycles under the conditions of denaturation (95.0 ° C.), annealing (57.0 ° C.), and elongation (72.0 ° C.) using 1 pmol of each of the above primers, 1 μl of DNA extract and 7 μl of distilled water Respectively.

The shrimp was sampled before the WSSV attack and confirmed by WSSV infection by PCR. The size of the PCR product for WSSV was 604 bp, confirming that the shrimp was not infected with WSSV before inoculation. In FIG. 6, lanes 1 and 2 are the group inoculated with the WSSV and the lanes 3 and 4 are the shrimp before WSSV inoculation. In addition, after the inoculation with WSSV, sampling of the plant extract mixture (2 ~ 5 days), midterm injection (6 ~ 10 days) and later injected shrimp (11 ~ 14 days) Respectively. It was also confirmed that the WSSV replicated in the shrimp was not amplified and maintained for 14 days even when the plant extract mixture was added. This suggests that the plant extract mixture inhibits WSSV proliferation. Figure 7 shows the result.

Experimental Example 4: Mortality and Survival Rate Before and After Inoculation with WSSV

(1) Relative survival rate of ginseng to seven plant extract mixtures

The shrimp survived until the inoculation of WSSV was considered to be well adapted to the environment and 5.8 X 103 copy / ㎖ WSSV was inoculated 100 ㎕ per 1 g of the shrimp body weight and observed for 14 days. As a result, the cumulative mortality rate in the positive control group (WSSV inoculation, general feeding) was the highest at 62.50%, and the cumulative mortality rate of the plant extract mixture of the present invention was 9.52% to 27.78%. The test group that died on the first day of the attack inoculation was excluded from the test results. Shrimp feeds, each containing a mixture of plant extracts of Table 1 and Table 2, were fed for 7 days before WSSV inoculation followed by inoculation with WSSV and shrimp feed containing the plant extract mixture was continuously fed after inoculation.

The antiviral efficacy of the plant extract mixture was evaluated four times by calculating the relative survival rate through the cumulative mortality of the flower fowl for 14 days after the inoculation of WSSV. FIG. 4 shows the average value of five tests for each mixture having the ratios according to the preparation examples in Table 2. [ As a result, it was confirmed that the relative survival rate to the cumulative mortality rate exceeded about 76% on average, and the mortality rate was remarkably decreased due to the effect on the plant extract mixture. Relative survival rate is [1- (cumulative mortality of plant extract mixture group / cumulative mortality of positive control group)] X100.

(2) Relative survival rate of P. japonicus shrimp on seven plant extract mixtures

As with the experiment examples above (1), WSSV attack inoculation before the initial injection of the surviving shrimp except cumulative mortality against shrimp, survival until WSSV attack vaccinated shrimp is determined by the objects adapted to the environment, 5.8 X 10 ³ The copy / ml WSSV was inoculated 100 당 per 1 g of the test shrimp body weight and observed for 14 days. As a result, the cumulative mortality rate in the positive control (WSSV inoculation, general feed) was the highest at 80.00%, and the cumulative mortality rate of the plant extract mixture was found to be 22.86% to 24.12%. In the test results, the test group that died on the first day of the attack inoculation was excluded from the test results. The shrimp feeds, each containing the plant extract mixture of Table 1, were fed for 7 days before WSSV inoculation followed by inoculation with WSSV and shrimp feeds containing the plant extract mixture were continuously fed after challenge. The feeds were prepared by mixing 10% of the total weight of the feed with the polyvinyl alcohol solution and the plant extract mixture at a ratio of 7: 1 to 11: 1, After drying, about 10% of the total weight of the primary coated feed was added to the ethyl cellulose solution, and the secondary coated feed was used. In addition, the relative survival rate was calculated by multiplying the cumulative mortality of shrimp shrimp for 14 days after the inoculation of WSSV to evaluate the antiviral efficacy of the plant extract mixture four times. 4 shows the average value obtained by four experiments for each mixture having the ratio according to the preparation example of Table 2. [ As a result, the relative survival rate of the plant extract mixture of the present invention exceeded about 71%, and it was confirmed that the mortality rate was remarkably decreased due to the effect on the plant extract mixture. Relative survival rate is [1- (cumulative mortality of plant extract mixture group / cumulative mortality of positive control group)] X100.

(3) Relative survival rate of P. japonicus shrimp to plant extract mixture coating concentration

The relative survival rate of each plant was determined by varying the concentration of the plant extract mixture.

As with the experiment examples above (1), WSSV attack inoculation before the initial injection of the surviving shrimp except cumulative mortality against shrimp, survival until WSSV attack vaccinated shrimp is determined by the objects adapted to the environment, 5.8 X 10 ³ The copy / ml WSSV was inoculated 100 당 per 1 g of the test shrimp body weight and observed for 14 days. As a result, the cumulative mortality rate in the positive control group 1 (WSSV inoculation, general feeding) was the highest at 68.00%, and the plant extract mixture showed the lowest cumulative mortality at the concentration of 30 ~ 300 ㎎ / ㎖. The mortality rate was higher than the control level.

The present invention provides antiviral agents that are active against white spot viruses that can cause serious damage when infected with crustaceans. Therefore, the present invention can be widely used in crustacean farms including shrimp farms.

<110> Choong Ang Vaccine Lab. <120> A composition having antiviral activity for white spot syndrome          virus <130> CVA17P-0008-KR <160> 2 <170> KoPatentin 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> WSSV-F primer <400> 1 ctttcacttt cggtcgtgtc 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> WSSV-R primer <400> 2 tactcggtct cagtgccaga 20

Claims (17)

 For preventing, treating, or preventing infection with crustaceans caused by white spot syndrome virus, which is a mixture of a cigarette, a crayfish, a crayfish, a persimmon, a licorice, Composition. The method according to claim 1,
1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 3.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: Wherein the composition has a weight ratio of 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0, to prevent, treat or prevent the infection of crustaceans caused by white spot virus. 3. The method according to claim 1 or 2,
A composition for preventing, treating, or preventing and treating a crustacean infection caused by white spot virus, wherein a mixture of said extract is contained in an amount of 30 to 300 mg / ml based on said composition. 3. The method according to claim 1 or 2,
A composition for preventing, treating, or preventing and treating a crustacean infection caused by a white spot virus, characterized in that the mixture of the extract is coated with a polymer material. 5. The method of claim 4,
A composition for preventing, treating or preventing and treating infections of crustaceans caused by white spot virus, characterized in that the coating consists of a primary coating with a hydrophilic polymer and a secondary coating with a non-hydrophilic polymer. 3. The method according to claim 1 or 2,
A composition for preventing, treating, or preventing and treating infection of crustaceans by white spot virus, characterized in that said crustaceans are shrimp.  A method for preventing, treating or preventing photophobia of a crustacean caused by a white spot virus by using a mixture of a crowd, a craystalkalix operkurtus, a chrysanthemum, a licorice, a curd, a white paper and a camellia extract. 8. The method of claim 7,
1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 3.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0. A method for preventing, treating or preventing photophobia of a crustacean caused by white spot virus. A mixed solution of a mixture of a crowd, Craystokarix auxercuratus, cherry tree, licorice, kelp, white paper and camellia extract and a polymer substance was prepared.
The mixed solution is mixed with a feed and dried to obtain a feed composition in which the mixture and the feed are coated with a polymer material,
Comprising feeding said feed composition to a crustacean which is required to prevent, cure or prevent and treat infections caused by white spot viruses,
A method for preventing, treating or preventing an infection of a crustacea caused by a white spot virus. 10. The method of claim 9,
1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 3.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0: 1.0 to 3.0. A method for preventing, treating, or preventing and treating infection of crustaceans by white spot virus. 10. The method of claim 9,
Characterized in that the coating comprises a primary coating with a hydrophilic polymer and a secondary coating with a non-hydrophilic polymer.
A method for preventing, treating or preventing an infection of a crustacea caused by a white spot virus. 9. The method according to claim 7 or 8,
Characterized in that a mixture of said extracts comprises 30 to 300 mg / ml, based on said composition.
A method for preventing, treating or preventing an infection of a crustacea caused by a white spot virus. 12. The method according to any one of claims 9 to 11,
Characterized in that a mixture of said extracts comprises 30 to 300 mg / ml, based on said composition.
A method for preventing, treating or preventing an infection of a crustacea caused by a white spot virus. A food composition for preventing or ameliorating a disease caused by a white spot virus, which comprises a mixture of a crowd, Craystokarix operkurtus, a cherry tree, licorice, kelp, white paper and camellia extract. A feed additive composition for preventing or ameliorating a disease caused by a white spot virus, which comprises a mixture of a crowd, Craistocarix operculus, a cherry tree, a licorice, a herb, a white paper and a camellia extract. A cosmetic composition having an inhibitory activity against a white spot virus, comprising a mixture of a crowd, Craystokarix operkurtus, chiller, licorice, kelp, white paper and camellia extract. The inhibitory activity against white spot viruses, including phlebotomists, a mixture of Crestor Carrícís auxercuratus, cherries, licorice, ugly, white and camellia extracts, and pharmacological, veterinary or agriculturally acceptable carriers or diluents &Lt; / RTI &gt;

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