JPH05255113A - Specific antibody and shrimp infectious disease-controlling composition compounded with the same - Google Patents

Abstract

PURPOSE:To provide a specific antibody having a specified bonding activity to shrimp infectious disease pathogens used for passive immunity, and also to provide a shrink infectious disease control composition compounded with the antibody and effective for the control of the shrimp infectious diseases. CONSTITUTION:The specific antibody has a specified bonding activity to the pathogens of shrimp infectious diseases, e.g. bacterial pathogen such as vibrio bacteria or viral pathogen such as Baculoviridae. Preferably, an egg antibody or milk antibody obtained from the egg of a laying hen hyperimmunized with the above antibody or from the milk of a mammalian hyperimmunized with the antibody. And, the shrimp infectious diseases control composition, preferably a shrimp feed, is compounded with the antibody as an active ingredient for controlling the shrimp infectious diseases. The administration of the antibody or the composition compounded with the antibody into the shrimp permits to effectively control the shrimp infectious diseases, especially the shrimp viral infectious diseases having not had any controlling method as well as the shrimp bacterial infections diseases.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to the control of shrimp infectious diseases containing a specific antibody having a specific binding activity against a pathogen of shrimp infectious diseases and the antibody having a control effect against shrimp infectious diseases. It relates to a composition.

[0002]

2. Description of the Related Art In recent years, shrimp aquaculture technology has been established, and shrimp aquaculture centering on the genus Kuruma shrimp has been actively conducted in various countries around the world. Under such circumstances, the damage caused by shrimp infectious diseases is increasing, which is a serious problem. Shrimp infectious diseases are mainly vibrio diseases caused by pathogenic bacteria and baculovirus midgut necrosis caused by pathogenic viruses.

As control measures against these infectious diseases,
Generally, antibacterial agents (antibiotics) are used for bacterial infections including Vibrio disease. However, in recent years, with the frequent use of antibacterial agents, the emergence of resistant bacteria has become a problem, and the control effect by administration of antibacterial agents cannot be expected. In addition, for viral infections, effective drugs have not yet been developed, and there is currently no effective control measure.

Active immunity (vaccination) has been put to practical use in mammals such as humans and livestock as a control measure against viral or bacterial infections. In active immunization, inactivated or attenuated pathogens such as viruses and bacteria are administered to individuals such as humans and livestock, and the autoimmune power is stimulated to produce antibodies against the pathogens to each individual and infectious diseases. Is being controlled.

In the field of aquaculture as well, active immunity research is actively underway for the purpose of controlling infectious diseases. In Japan, application research of active immunity against vibrio disease of cultured ayu is most advanced (Japanese Patent Publication No. 56-53286, Japanese Patent Publication No. 56-53287, Japanese Patent Publication No. 58-500).
(Publication No. 026, etc.) Only in practical use.

It is said that invertebrates such as shrimp do not have a specific immune function mediated by antibodies unlike vertebrates. Therefore, active immunity against shrimp infections is generally denied its infection control effect. However, in recent years, it has been reported that the effect of controlling the infection of Vibrio disease by the action of promoting phagocytic activity of shrimp blood cells using the inactivated bacterium of Vibrio peneus, which is a bacterium causing vibrio disease of shrimp (JP-A-3-204820). .. In other words, in invertebrates such as shrimp, the biological defense function by phagocytosis of blood cells is not the specific immune function mediated by antibodies, but the details of these mechanisms are still unclear. Are left.

In recent years, passive immunity has been attracting attention as a control measure against infectious diseases, instead of conventional active immunity. Passive immunity is an attempt to administer antibodies against pathogens to humans and livestock to help control infectious diseases.

As a known example of passive immunization, prevention of dental caries caused by adherent infection of Streptococcus mutans bacteria on teeth (J. Dent. Res., 70, 162-166, 1991).
, Rotavirus adheres to intestinal epithelial cells and infects them to prevent rotavirus diarrhea (Microbiol. Immunol., 34,
617-629, 1990) has been reported.

Known examples of passive immunity in the field of aquaculture have been reported on the effectiveness of passive immunization using antibodies against fish infectious diseases, such as prevention of yellowtail nodule disease, prevention of flounder fry intestinal tract opacification, etc. (Japanese Patent Laid-Open No. 1-1682
46).

However, as described above, the biological defense mechanism of invertebrates such as shrimp is remarkably different from the immune mechanism of vertebrates which has been well known in the past, and therefore, the control of infectious diseases of invertebrates such as shrimp is controlled. The fact is that no specific report has been made regarding the effectiveness of passive immunization against. Therefore, it is difficult to easily infer the application of passive immunity to shrimp, etc. from the result of the effectiveness of passive immunity against vertebrates, and practical control measures against shrimp infections by passive immunization using antibodies. It was completely unclear whether or not it was possible.

[0011]

[Problems to be solved] Shrimp infectious diseases have become a frequent problem. As a control method, a large amount of antibacterial agent is administered, and at present, there is a problem that the antibacterial agent remains in the shrimp body. In addition, the emergence of resistant bacteria against antibacterial agents occurs quite often, and the development of effective antibacterial agents against the resistant bacteria cannot be kept up. Furthermore, there has been no success in developing effective agents for controlling shrimp virus diseases. Regarding active immunity (vaccination) against shrimp infections, it has been shown to be effective for promoting the phagocytosis of blood cells against the Vibrio disease of prawns, and has not yet been put to practical use. On the other hand, the effectiveness of passive immunity against shrimp infectious diseases has not been proved as described above, and development of a practical control method for shrimp infectious diseases is eagerly desired in shrimp aquaculture.

The object of the present invention is to provide a specific antibody having a specific binding activity against a pathogen of shrimp infection used for passive immunization, and a shrimp effective for controlling shrimp infection containing the antibody. An object is to provide a composition for controlling infectious diseases.

[0013]

[Means for Solving the Problems] In order to solve the above problems, the present inventors have focused their attention on passive immunity as a method for controlling shrimp infectious diseases, and as a result of the research, as a result, specific pathogens of shrimp infectious diseases Successful large-scale preparation of antibody. Furthermore, when a shrimp feed was prepared as a composition containing the specific antibody and tested for the passive immunity effect of shrimp infection by oral administration of the specific antibody, a surprisingly remarkable infection control was obtained. The effect can be found, and the present invention has been completed. That is, the present invention relates to a shrimp infection control composition comprising a specific antibody against a pathogen of shrimp infection used for the control of shrimp infection and the antibody. In the present invention, the control of shrimp infectious diseases means preventing infection by a pathogen of shrimp infectious diseases and curing the infection by exerting a therapeutic effect on the infection.

The shrimp in the present invention means a prawn (Pe)
naeus japonicus), bull shrimp (Penaeus monodon), black shrimp (Penaeus chinensis), banana shrimp (Penaeus
Shrimp of the genus Kuruma prawn such as morgui-ensis) and other genera.

The shrimp infectious disease in the present invention refers to an infectious disease caused by a bacterial pathogen belonging to the genus Vibrio and a genus of Vibrio, and an infectious disease caused by a baculovirus and other viral pathogens.

The specific antibody of the present invention means an antibody having a specific binding activity to a pathogen of shrimp infectious disease, and the kind of the antibody is that of laying hens hyperimmunized with the pathogen of shrimp infectious disease. It refers to a chicken egg antibody obtained from an egg or a milk antibody obtained from milk of a mammal hyperimmunized with a pathogen of a shrimp infection, and the antibody purity is not particularly limited. That is, it may be a pure antibody,
In the case of chicken egg antibody, whole egg powder, egg yolk powder containing it,
Alternatively, it may be a water-soluble protein fraction powder of egg yolk. In the case of a milk antibody, it may be powdered whole milk powder, skim milk powder or whey protein powder containing it.

The animal hyperimmunized with the pathogen of shrimp infection may be an animal capable of producing an antibody having a specific binding activity to the pathogen, but the shrimp infection which is the object of the present invention Considering from a practical viewpoint of providing a specific antibody used for disease control and a shrimp infectious disease control composition comprising the antibody, a laying chicken or a cow, a goat, which produces a large amount of specific antibody, It is preferable to use mammals such as sheep. Among these, the workability of hyperimmunity,
The method of immunizing a laying hen and obtaining the antibody from the hen's egg is the most desirable from the viewpoints of antibody production ability, animal breeding cost and the like.

As a method of hyperimmunizing the laying hen, the amount of specific antibody against the pathogen appearing in the hen's egg may be increased by repeatedly administering to the laying hen the pathogen of shrimp infection as an antigen. As a method of hyperimmunizing a mammal, if the antigen is repeatedly administered to mammals such as cows, goats, and sheep whose pregnancy has been confirmed, the amount of specific antibody against the pathogen appearing in milk can be increased. Good.

The antigen used in this case may be prepared by a known method. For example, a large amount of antigens can be prepared by culturing a large number of pathogenic bacteria or viruses of shrimp infectious diseases and then treating them with a known inactivating agent such as formalin. Alternatively, it is also possible to collect shrimp killed by an infectious disease, homogenize the affected part by a method such as homogenate, and use it as an antigen. Alternatively, for the purpose of increasing the purity of the pathogen contained in the affected part homogenate, the pathogen can be purified by a combination of known methods such as centrifugation, column chromatography, and membrane filter filtration, and used as an antigen.

As the method for administering the antigen to the laying hen or mammal, general administration methods such as intramuscular injection, subcutaneous injection, intravenous injection, intraperitoneal injection, and oral administration by drinking water are used. Intramuscular injection is preferable from the viewpoint of the production effect of specific antibodies. The antigen used for administration is a suspension of a pathogen of shrimp infection, or
A mixture of the solution with an aluminum hydroxide gel or Freund's adjuvant is generally used.

The administration of the antigen is carried out by investigating the change in the amount of the specific antibody (antibody having a specific binding activity to the corresponding antigen) appearing in chicken egg or milk by a method such as enzyme immunoassay. Is repeated until the maximum value is reached. It should be noted that the amount of the antibody can be maintained at a certain level or more of the specific antibody throughout the egg-laying period or the feeding period by repeatedly performing the antigen administration to the laying hen or the mammal at appropriate intervals.

Since the amount of the antigen to be administered depends on the type of animal to be administered, the type of antigen, etc., it is necessary to select it in a timely and preliminary test, but in the case of administration to laying hens, it is generally , In the case of bacterial antigen, 10 ⁷ to 10 ⁹ / bird, and in the case of viral antigen, the amount of protein is 10 μg ~
An antigen dose of 1 mg / bird is selected.

After administration of a pathogen of a shrimp infection as an antigen to laying hens or mammals, chicken eggs or milk containing a specific antibody against the antigen are collected, and the shrimp used for controlling the shrimp infection from this is collected. Specific antibodies of the invention to infectious agents are prepared. When using hen's eggs, separate whole egg liquid or yolk liquid after splitting, homogenize with a homogenizer, etc., and sterilize to obtain whole egg powder or yolk powder containing specific antibody against the used antigen by hot air drying or freeze drying. be able to. Further, a known method for purifying chicken egg antibody from the egg yolk liquid or egg yolk powder (Japanese Patent Laid-Open No. 64-38).
098, Agric. Biol. Chem. 54 (10), 2531-2535, 1990.
Year), the egg yolk water-soluble protein powder or the pure powder of the specific antibody having a high specific antibody purity against the used antigen can be prepared. When using milk, after sterilizing the milk or skim milk separated lipid components in the milk with a cream separator or the like, hot air drying or freeze drying,
A whole milk powder or skim milk powder containing a specific antibody against the used antigen can be obtained. From the milk or the skim milk, a whey protein powder or a pure powder of a specific antibody having an increased purity of a specific antibody against an antigen used by a known method is prepared.

The shrimp infection control composition of the present invention refers to whole egg powder, egg yolk powder, egg yolk water-soluble protein powder, chicken egg antibody obtained from eggs of a hyperimmune chicken as a specific antibody against a pathogen of shrimp infection. A pure powder, or a powder of whole milk powder, skim milk powder obtained from milk of a hyperimmune mammal, whey protein powder, milk antibody pure powder, etc., is added as an effective component for controlling shrimp infections. To tell. The content of specific antibody is usually 0.01% as pure antibody.
The range of 10%, preferably 0.1% to 1% is selected, but the effective compounding amount varies depending on the titer of the specific antibody.
More preferably, it is most desirable to determine the effective amount of the shrimp in a timely manner using the control effect of the infectious disease as an index by an experimental system in which shrimp is artificially infected with the infectious disease pathogen. As other components of the composition, sugar, protein, lipid, etc., as a voluminous agent,
As other auxiliary components, vitamins, minerals, food attractants for shrimp, antibiotics, etc. can be mixed at the same time.

The shrimp infection control composition of the present invention may have any shape such as powder, granules, pellets and the like, but when it is intended for oral administration to shrimp, Most preferably, it is formed into pellets. When it is intended for oral administration to shrimp larvae, it is desirable to process it into a fine particle powder by a spray drying method or the like.

In the present invention, the active ingredient used for controlling shrimp infectious diseases is a specific antibody against the pathogen of the infectious diseases, which is a protein that also serves as a nutritional ingredient of shrimp. Therefore, the shrimp infection control composition of the present invention, the specific antibody or the specific antibody-containing whole egg powder, egg yolk powder, egg yolk water-soluble protein powder, whole milk powder,
Non-fat dry milk, whey protein, etc. can be blended with known feed ingredients of shrimp as a basic feed composition and processed into pellets for adult shrimp and fine powder feed for young shrimp. desirable.

According to the present invention, a specific antibody against a pathogen of shrimp infection, a feed containing the antibody, a drug containing the antibody and the like are administered to shrimp by an oral or dipping method, etc. In addition, shrimp infections can be effectively controlled. Regarding its control mechanism, the biological defense mechanism of shrimp is significantly different from that of vertebrates, so there are still many unclear points, but in the shrimp body, the administered specific antibody is specific to the pathogen of shrimp infection It is considered that they are bound to each other to eliminate the pathogenicity, adhesive infectivity and the like.

[0028]

EXAMPLES The present invention will be described in more detail with reference to Examples and Test Examples, but the present invention is not limited to these Examples. Example 1. Preparation of chicken egg antibody against Vibrio parahaemolyticus (antigen) 50% natural filtered seawater 1 liter (natural filtered seawater 50
Meat extract (3 g), peptone (10 g) and sodium chloride (5 g) were dissolved in 0 ml + deionized water (500 ml), and the pH was adjusted to 7.2 to 7.
After adjusting to 6, it was sterilized (121 ° C, 15 minutes) to obtain a liquid medium. The liquid medium was inoculated with Vibrio sp., Which was isolated and identified as a vibrio pathogen of Kuruma prawn, and was inoculated.
The cells were shake-cultured at ℃ for 24 hours. The number of bacteria at this time reached 10 ⁹ cells / ml. After formalin was added to the culture solution at a rate of 0.5% to inactivate the cells, the dead cells were washed with sterile physiological saline (sterile saline) by centrifugation. The washed dead cells were suspended in sterile saline and adjusted to a concentration of 10 ⁸ cells / ml to serve as an immunizing antigen for egg laying eggs. (Immunity) 100 hens (Rhode Island Red) were immunized intramuscularly with 1 ml of the antigen solution per bird.
The immunization injection was repeated once a week for a total of 4 times to increase the specific antibody titer against the same antigen appearing in the egg yolk. After that, the same immunization injection was repeated once every two months for the purpose of maintaining the specific antibody titer. (Preparation of hen egg antibody) Eggs with increased titer of specific antibody against the antigen (for 2 months from 1 month after immunization) were laid to obtain 205 kg of whole egg solution. After sterilizing the whole egg liquid for 3 minutes at 63 ° C., it was pulverized by a spray drying method (air temperature of 150 ° C., air temperature of exhaust air of 80 ° C.) to obtain 49.0 kg of chicken egg antibody (whole egg powder) against Vibrio spp.

Example 2. Preparation of Chicken Egg Antibody against Bovine Shrimp Vibrio Species (Antigen) An immunoantigen solution for laying chickens was prepared by the same method as in Example 1 using Vibrio sp. (Immunity) The immunization was carried out by the same immunization method as in Example 1 using an immunizing antigen prepared by using the pathogenic bacterium of Vibrio genius of bovine shrimp. (Preparation of hen egg antibody) Eggs with increased specific antibody titer against the antigen (Bovine shrimp vibrio bacterium) (for 2 months from 1 month after immunization) were laid to obtain 198 kg of whole egg solution. The whole egg liquid
After sterilizing at 63 ℃ for 3 minutes, spray dry method (air temperature: 150
℃, the temperature of the exhaust air 80 ℃) to obtain 46.5 kg of egg egg antibody (whole egg powder) against Vibrio parahaemolyticus.

Example 3. Preparation of hen's egg antibody against baculovirus of Kuruma prawn larvae (Antigen) Kuruma prawns killed by baculovirus midgut necrosis of the intestine (BMN) were homogenized in sterile saline. Centrifuge the homogenate (12,000 xg, 30 minutes)
The insoluble material was separated as a precipitate. 0.45 μm of the supernatant
It filtered with the membrane filter of. The filtrate (protein concentration 2.30 mg / ml: Lowry method) was mixed and emulsified in the same amount as Freund's complete adjuvant to prepare an immunizing antigen solution for laying hens. (Immunity) Ten laying hens (Rhode Island Red line) were used, and 1 ml of the antigen solution was intramuscularly injected per bird.
The immunization injection was repeated once a week for a total of 4 times to increase the specific antibody titer against the same antigen appearing in the egg yolk.
Thereafter, similar immunization injections were repeated once every two months for the purpose of maintaining the specific antibody titer. (Preparation of hen's egg antibody) Eggs (within 2 months from 1 month after immunization) whose specific antibody titer against the antigen has increased are divided into whole egg liquids.
23.2 kg was obtained. The whole egg solution was sterilized at 63 ° C. for 30 minutes, and then pulverized by a freeze-drying method to obtain 5.1 kg of an egg egg antibody (whole egg powder) against baculovirus of a prawn larva.

Example 4. Preparation of Egg Antibody to Baculovirus of Bovine Shrimp Larvae (Antigen) Prepared in the same manner as in Example 3 from Shrimp killed by baculovirus midgut necrosis of the intestine (BMN). (Immunity) Ten laying hens (Rhode Island Red) were used, and 1 ml of the above antigen solution was intramuscularly injected per chick. The immunization injection was repeated once a week for a total of 4 times to increase the specific antibody titer against the same antigen appearing in the egg yolk. Thereafter, similar immunization injections were repeated once every two months for the purpose of maintaining the specific antibody titer. (Preparation of hen's egg antibody) Eggs (within 2 months from 1 month after immunization) whose specific antibody titer against the antigen has increased are divided into whole egg liquids.
21.5 kg was obtained. The whole egg solution was sterilized at 63 ° C. for 30 minutes and then pulverized by a freeze-drying method to obtain 5.5 kg of chicken egg antibody (whole egg powder) against baculovirus of bovine shrimp larvae.

Example 5. Preparation of shrimp vibrio disease control feed <Shrimp basic feed powder> Squid meal powder 30% Fish meal powder 20% Salmon Shirame meal powder 10% Shrimp meal powder 10% α-starch 10% Yeast powder 10% Mineral mixture 5% Vitamin mixture 3% cholesterol powder 1% squid oil 1% shrimp basic feed powder 9.75 kg of chicken egg antibody (whole egg powder) against the Vibrio genus prawn prepared in Example 1 and 0.25 kg of water and 3 kg of water were added and mixed, and then the meat chopper was mixed. Was molded into pellets with a diameter of 2.5 mm and a length of about 10 mm. Moisture content of the pellets was 10% in an air dryer at 60 ° C.
Dry until shrimp Vibrio disease control feed 10.4k
g was obtained.

Example 6. Preparation of fine particle feed for controlling viral diseases of shrimp Formula 1 Formula 2 Formula 3 Control feed Egg powder 60% 61% 61.5% 62.0% Squid meal powder 10% 10% 10% 10% Shrimp meal powder 5% 5% 5 % 5% Vitamin / Mineral Mix 10% 10% 10% 10% Soy Lecithin 2% 2% 2% 2% Cod Liver Oil 5% 5% 5% 5% Shrimp Oil 2% 2% 2% 2% Natural Polysaccharides 1% 1% 1% 1% Fish meal powder 3% 3% 3% 3% Chicken egg egg antibody (whole egg powder) of Example 3 2% 1% 0.5% 0% 400 g of powder mixed with the above formulation was added to water 600 dissolved in 65
After sterilizing at ℃ for 30 minutes, spray dryer (air temperature
A fine powder for controlling viral diseases of shrimp was obtained as a dry powder at 150 ° C and an exhaust air temperature of 80 ° C.

Test Example 1. Antibody titer measurement of chicken egg antibody The antibody titer of chicken egg antibody was measured by enzyme-linked immunosorbent assay (ELISA). The basic procedure is shown below. (1) ELISA plate: 96-well polystyrene plate (manufactured by Nunc: high adsorption capacity) (2) Antigen coating: 0.05 of the antigen solution used for immunization
Dilute 10 times with M carbonate buffer (pH 9.6) and add 100 μl
To each well of the ELISA plate and left at 5 ° C overnight. (3) Washing: 20 mM phosphoric acid 0.15 containing 0.5% Tween 20
MNaCl buffer pH7.4 (PBS-Tween) 20
Add 0 μl / well and wash each well of the plate 4 times. (4) Plate blocking: Ovalbumin in PB
Dissolved in S-Tween (10 mg / ml). This liquid 100 μ
Add 1 to each well and leave at 37 ° C for 1 hour. (5) Washing: Same as (3). (6) Addition of sample: 100 μl / well of the sample solution suspended in PBS-Tween was added (n = 4), 37
Leave at ℃ for 1 hour. (7) Washing: Same as (3) (8) Addition of secondary antibody: anti-chicken IgG rabbit IgG-
Alkaline phosphatase conjugate (Zymet) was diluted 2000 times with PBS-Tween, and this solution was diluted.
Add 100 μl / well and leave at 37 ° C for 1 hour. (9) Washing: Same as (3). (10) Enzymatic reaction: P-nitrophenyl phosphate disodium salt manufactured by Sigma was used as a substrate. Substrate
Dissolved in 0.1 M ethanolamine buffer pH 9.8 (1 mg
/ Ml). Add 100 μl / well of this substrate solution, 37
After performing the enzyme reaction for 30 minutes at ℃, add 2M NaOH solution to 50
Stop the reaction by adding μl / well. (11) Absorbance measurement: The absorbance at 405 nm of each well was measured and a blank (instead of the sample solution, PBS-Tw was used).
The value obtained by subtracting the absorbance indicated by the well added with een) was defined as the ELISA value.

1) Transition of antibody titer of hen egg antibody after immunization With respect to the hen egg antibody obtained in Examples 1 to 4, the specific antibody titer against each antigen was measured by ELISA.
After immunization, egg yolk was separated from the eggs of each week and homogenized, and used as representative samples. Egg yolk liquid is PBS-Twe
It was diluted 2,000 times with en to obtain an ELISA measurement sample solution. Figure 1 shows the changes in the specific egg yolk antibody titer (ELISA value) of the prawn prawn against Vibrio, and Figure 2 shows the specific egg yolk antibody titer (EL) of the prawn prawn against baculovirus.
The transition of the ISA value) is shown. In each case, after the first immunization, the specific antibody titer in egg yolk increased from the second week and reached the maximum value at the fourth week. After that, the antibody titer tended to decrease gradually, but with booster injections every two months,
The antibody titer recovered to the highest level. This immunization schedule confirmed that high titer egg egg antibodies were obtained throughout the egg laying period. The titer egg antibody titer against Vibrio baculovirus and baculovirus of bovine shrimp also showed the same antibody titer transition as that of Kuruma prawn.

2) Recovery rate of specific antibody titer in hen egg antibody-containing feed The specific antibody titer recovery rate of the mixed hen egg antibody (whole egg powder) in the feeds prepared in Examples 5 and 6 was determined by ELIS.
Determined by A. The feed and chicken egg antibody (whole egg powder) were each dissolved in PBS-Tween to prepare a chicken egg antibody (whole egg powder) at a concentration of 1 mg / ml. This solution was measured as an ELISA sample solution, and the recovery rate of the antibody titer was calculated from the ELISA values of the feed and the chicken egg antibody mixed therein. As a result, the shrimp vibrio disease control feed of Example 5 had an antibody titer recovery rate of 92%, and the shrimp virus disease control fine particle feed of Example 6 had an antibody titer recovery rate of 94%.

Test Example 2. Effect of Chicken Egg Antibody on Vibrio Disease of Shrimp (Infection Experiment) 30 Shrimp with an average body weight of 20 g were divided into two groups of 30 fish each. The chicken egg antibody-containing feed prepared in Example 5 was administered to the test group, and the shrimp basal feed containing no egg egg antibody was administered to the control group at 10 g / kg of shrimp body weight per day. From the 8th day of administration, the muscle of shrimp dying by intramuscular inoculation of Vibio sp., A pathogenic bacterium of Vibrio parahaemolyticus, was given to each test shrimp at 0.1 g / day for 3 days to try oral infection of Vibrio disease. The number of orally-infected bacteria at this time was 1 x 10 ⁵ per day for each shrimp tested.
It was a cell. For 10 days from the last infection day, we examined the presence or absence of mortality in each group and attempted to isolate Vibrio sp.

FIG. 3 shows changes in the survival rate after infection in the chicken egg antibody-administered group and the control group. The control group had 28 fish (30
Onaka) Death occurred and the survival rate was 6.7%. On the other hand, the hen egg antibody-administered group had a small mortality (3/30) and had a survival rate of 90%, which was significantly different from that of the control group (P <0.01). ) Was recognized. In addition, bacteria showing the same characteristics as the bacteria used for infection were isolated from the heart and lymphoid organs of all dead shrimp. By the above test, the infection prevention effect of the chicken egg antibody (whole egg powder) -containing feed against the bacterium was confirmed in the Vibrio disease infection experiment of the prawn.

Test Example 3. Effect of hen egg antibody on baculovirus midgut necrosis of Shrimp larvae (infection experiment) Zoeer stage larvae of Shrimp prawns were divided into 4 groups of 500 larvae.
In the test groups 1, 2, and 3, the fine particle feed prepared in Example 6 and containing the egg egg antibody in the proportions of 0.5%, 1.0%, and 2.0%, respectively, was added from the zoea period to the post-larber period (PL10).
Larvae were fed daily with a daily dose of about 0.2 mg per fish. The control group was similarly fed with the control fine particle feed of Example 6 (without addition of the egg egg antibody). 3 g of shrimp killed by baculovirus midgut necrosis (BMN) stored at -80 ° C was homogenized in 30 ml of sterilized seawater, and then centrifuged (12,000 xg for 30 minutes) to obtain 0.45 μm of supernatant. The filtrate filtered with a Millipore filter was put into a water tank containing Zoea stage larvae in an amount of 10 ml to try infection. 15 days after infection,
The survival rate was determined by examining the number of dead tails in each plot. Fig. 4 shows changes in the survival rate of the prawn larvae of the egg egg antibody-administered group and the control group. The survival rate of the control group was 0%, while the egg egg antibody
47.1% for 0.5% formulation, 69.8% for 1%, 83.2 for 2%
%, And there was a significant difference (P <0.01) in the survival rate between the egg egg antibody-containing group and the control group.

[0040]

The control of shrimp infections has heretofore been carried out by administration of drugs such as antibiotics. In recent years, drug administration has been carried out due to the problem of residue in the shrimp body due to large dose administration of drugs or the emergence of resistant bacteria. There has been a strong demand for the development of a method for controlling shrimp infectious diseases as an alternative. In addition, there was no effective drug against the viral infection of shrimp, which caused great damage in shrimp aquaculture. Under such circumstances, for the first time, the present invention demonstrated the control effect of shrimp infection by passive immunization practically using a specific antibody against the pathogen of shrimp infection. By using the specific antibody of the present invention or the shrimp infection control composition containing the antibody, shrimp infection can be effectively controlled. The specific antibody of the present invention is a component of hen's egg or milk, and it is safe to use without any concern about its persistence in shrimp. Further, by using the specific antibody of the present invention, or the shrimp infection control composition containing the antibody,
Not only bacterial infections of shrimp but also viral infections can be controlled.

[Brief description of drawings]

FIG. 1 is a diagram showing changes in the titer of a specific hen egg antibody against Vibrio parahaemolyticus infected with Kuruma prawn.

[Fig. 2] Fig. 2 is a graph showing changes in the titer of specific hen egg antibody against baculovirus infecting Kuruma prawn.

[Fig. 3] Fig. 3 is a diagram showing changes in the survival rate of a chicken egg antibody-administered group and a control group in a Kuruma shrimp Vibrio disease infection experiment.

FIG. 4 is a diagram showing changes in survival rate of a chicken egg antibody-administered group and a control group in a Kuruma prawn virus disease infection experiment.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification number Reference number within the agency FI technical indication location A61K 39/42 AFF 8413-4C (72) Inventor Kenya 9th, Akahori Shinmachi, Yokkaichi-shi, Mie Prefecture No. 5 In Taiyo Kagaku Co., Ltd. (72) Inventor Shigemitsu Akachi 9-5 Akahori-shinmachi, Yokkaichi-shi, Mie Prefecture Solar Chemistry Co., Ltd. (72) Yu Fujiki No. 5-5 Akahori-shinmachi, Yokkaichi-shi, Mie Solarization Gaku Co., Ltd. (72) Inventor Kin Taken 9-5 Akahori-shinmachi, Yokkaichi-shi, Mie Taiyo Kagaku Co., Ltd. (72) Inventor Yukinori Takahashi 5-43-304 Hianaka, Shimonoseki, Yamaguchi Prefecture (72) Inventor Toshiaki Itami 1-33-503 Shinaraseki Araki Shinmachi, Yamaguchi Prefecture (72) Inventor Miya ▲ saki ▼ Teruo 2-66 Edobashi, Tsu City, Mie Prefecture 5 University dormitory (72) Inventor Zenzo Iida Kanagawa Prefecture side 3-32-13 Tsuruya-cho, Kanagawa-ku, Hama-shi Within Japan Formula Feed Co., Ltd.

Claims (9)

[Claims] 1. A specific antibody against a pathogen of shrimp infection used for controlling shrimp infection. 2. The specific antibody according to claim 1, wherein the specific antibody is a chicken egg antibody obtained from eggs of laying hens hyperimmunized with a pathogen of shrimp infection. 3. The specific antibody according to claim 1, wherein the specific antibody is a milk antibody obtained from milk of a mammal hyperimmunized with a pathogen of shrimp infection. 4. The specific antibody according to claim 1, wherein the pathogen of shrimp infection is a bacterial pathogen. 5. The specific antibody according to claim 4, wherein the bacterial pathogen is Vibrio bacterium. 6. The specific antibody according to claim 1, wherein the pathogen of shrimp infection is a viral pathogen. 7. The specific antibody according to claim 6, wherein the viral pathogen is baculovirus. 8. A shrimp infection control composition comprising a specific antibody against a pathogen of shrimp infection. 9. The shrimp infection control composition according to claim 8, wherein the composition is a shrimp feed.