JPH09315983A - Preventing and therapeutic agent for infectious disease and feed - Google Patents

Abstract

PROBLEM TO BE SOLVED: To prepare a preventing and therapeutic agent for infectious diseases in fishes such as a yellowtail or a red sea-bream, crustaceans such as a prawn or a black tiger shrimp, comprising a milk whey (a hydrolyzates thereof) as an active ingredient, excellent in safety and substitutive for an antibiotic substance causing problems in development of resistant bacteria, remaining in the body, etc. SOLUTION: This preventing and therapeutic agent for infectious diseases comprises (A1 ) a milk whey or (A2 ) a hydrolyzate of the milk whey as an active ingredient. Furthermore, the agent is preferably administered in a daily dose of 1-6000mg/kg body weight, expressed in terms of the dried ingredient A1 or A2 and divided in 2 or 3 portions. The ingredient A1 is obtained by adding an acid to a milk or a skimmilk of a mammal such as a cow, a woman or a ewe and removing the produced coagulated substance. The ingredient A2 is prepared by dissolving, e.g. the ingredient A1 in water so as to provide 5-15% protein concentration, then thermally sterilizing the resultant solution, enzymically treating the sterilized solution with a proteolytic enzyme such as a pepsin, subsequently inactivating the enzyme and, as necessary, carrying out the ultrafiltering, concentrating and drying of the prepared solution.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a preventive / therapeutic agent for fish and shellfish infections and a feed.

[0002]

2. Description of the Related Art Fish farming technology has been rapidly developing in recent years, and the area of aquaculture (the number of farms) has been increasing year by year. However, unlike natural fish and crustaceans, in the case of aquaculture, they are bred at a high density, so many of them suffer from diseases. In particular, the damage caused by pathogenic viruses is enormous. For example, the mass mortality of shrimp in Taiwan in 1992 spread to South Korea, China, and Japan the following year, causing great damage to the shrimp aquaculture industry. It is believed that these are due to the secondary infection of baculovirus-infected shrimp with vibriopathies.

Conventionally, antibiotics have been used for infectious diseases of fish and crustaceans, but they are not used because of the occurrence of resistant bacteria and the problem of residual fish and crustaceans in the body. It is restricted. Among these diseases, lactoferrin, lactoferrin degradation products, and colostrum have been found to show remarkable effects as alternatives to antibiotics for bacteriopathies (JP-A-7-48274 and JP-A-7-274). 1
No. 45069) has been studied as a highly safe preventive / therapeutic agent, but no effective drug has been known as an alternative to antibiotics for viral diseases. Further, there is no reference to the use of milk whey and a milk whey degradation product obtained by enzymatically decomposing milk whey as a preventive / therapeutic agent for fish or crustacean infectious diseases or feed.

[0004]

SUMMARY OF THE INVENTION An object of the present invention is to provide a preventive and therapeutic agent for fish and crustacean infectious diseases and a feed, which are safe and have excellent effects which can replace antibiotics.

[0005]

As a result of intensive studies to solve the above problems, the present inventors have found that milk whey or a decomposed product of milk whey is effective for infectious diseases of fish and crustaceans. The present invention has been completed.

That is, the present invention is a preventive / therapeutic agent for infectious diseases of fish and crustaceans containing milk whey or a decomposed product of milk whey as an active ingredient. The present invention is also a feed for fish and crustaceans containing milk whey or a decomposed product of milk whey. Hereinafter, the present invention will be described in detail.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, fishes to be prevented or treated include yellowtail, red sea bream, black sea bream, eel, carp, rainbow trout, ayu, coho salmon, striped horse mackerel, tilapia, flounder, troughfish, crucian carp, guppy, Examples thereof include neon tetra, but are not limited thereto. Further, examples of the crustaceans include, but are not limited to, prawns, bull shrimp, Taisho shrimp, white shrimp and the like.

The active ingredient of the preventive / therapeutic agent for infectious diseases of the present invention is
Milk whey or a milk whey decomposition product, which is mainly separated from milk, is not limited thereto, and those derived from humans, sheep, goats, horses, dogs, cats and the like are also effective. Of the milk of these mammals,
It is suitable from the viewpoint of availability that milk, which is the most common mammalian milk, is used as a raw material, but from the viewpoint of economy, cheese, butter and other by-products produced when separating other milk components from milk, Whey or skim milk may be used as a raw material.

[0009] Milk whey refers to the remaining aqueous solution excluding the coagulated product (curd) produced by adding an acid or a milk-clotting enzyme to the above-mentioned mammalian milk or skim milk. As the milk whey, it is preferable to use a commercially available milk whey protein concentrate such as WPC or a milk whey protein isolate such as WPI, but the invention is not limited thereto.

The milk whey decomposed product is obtained by subjecting the milk whey obtained by the above method to proteolytic enzyme treatment, fermentation by microorganisms, acid hydrolysis and the like. For example, when treated with proteolytic enzymes, milk whey powder, milk whey protein concentrate is used with milk whey or water to achieve a protein concentration of 5-15.
Dissolve in the range of%, and heat sterilize at 60 to 130 ° C for 1 second to 30 minutes. Then, after adjusting the pH to 2-10 if necessary,
Enzymatic treatment is performed. Examples of the proteolytic enzyme include pepsin, pancreatin, trypsin, chymotrypsin, papain, promeline, and Aspergillus spp., And proteases derived from Streptomyces spp. Add it in portions according to the decomposition conditions, treat with an enzyme at a temperature range of 10 to 70 ° C for 3 to 24 hours, and then adjust the pH to 5.
Adjust to 5 to 7.5 and deactivate the enzyme by heat treatment at 80 to 140 ° C for 2 seconds to 20 minutes to obtain a degradation solution.

The decomposed solution may be used as it is, but may be concentrated and dried to give a milk whey decomposed product, which may be used. Alternatively, the decomposed solution was treated with an ultrafiltration membrane having a molecular weight cutoff of 3,000 to 500,000, and the decomposed solution that had permeated through the membrane was concentrated and dried to give a milk whey decomposed product, or celite filtration or a membrane pore size of 0.1 to 1 μm. A product obtained by performing filtration by microfiltration filtration and concentrating and drying the filtrate to obtain a milk whey decomposition product may be used.

The milk whey used in the present invention has a molecular weight of 100 to 800,000, and the decomposed product of milk whey has a molecular weight of 100 to 20,000.

The above milk whey or a decomposed product of milk whey is concentrated and dried, pulverized, and then thoroughly mixed with an appropriate amount of corn starch, talc, magnesium stearate, etc., and the particle size is adjusted by a conventional method to prepare a formulation. The content of milk whey or a decomposed product of milk whey in the preparation is not particularly limited, but when granulating from a powder, it may be appropriately adjusted depending on the type of fish or crustacean or according to its growth stage.

On the other hand, in order to prepare a feed, the feed raw material usually used in aquaculture is appropriately selected and blended according to the target aquatic animal, and the above milk whey or a decomposed product of milk whey is added thereto, blended and moistened. do it. As feed ingredients, generally, fish meal, shrimp meal, squid meal, wheat flour, corn gluten, brewer's yeast, cholesterol,
Vitamins and minerals are used. When the above milk whey or the decomposed product of milk whey is added to, mixed with and moistened with the feed, the content may be 0.01 to 5% by weight in the total feed.

Infectious diseases which can be treated or prevented by the preventive / therapeutic agent for infectious diseases and the feed of the present invention include fish,
It is a viral or bacterial infection of crustaceans. Specific examples of viral infections include Japanese flounder flounder melabud virus infection, trough puffer disease virus, rainbow trout infectious pancreatic necrosis, viral hemorrhagic sepsis, infectious hematopoietic necrosis, and carp spring. Viral disease, fin inflammation, catfish viral disease, salmon herpesvirus disease, Pike fry viral disease, striped horse mackerel iridovirus infection, red sea bream iridovirus infection, eel viral gills disease, coho salmon transmission Pancreatic necrosis, infectious hematopoietic necrosis, herpesvirus infection, Erythrocytic Inclus
ion Body Syndrome Virus infection, Rod of prawns
Examples include, but are not limited to, shapaedvirus-Penaeus japonicus infectious disease, bald shrimp's jaundice, lymphocystis disease that infects many seawater and freshwater fish, and the like.

Further, as the bacterial infection, specifically,
Vibrio anguillarum infection of trout, rainbow trout, Pseudotuberculosis infection of yellowtail, congenital disease, streptococcal infection, Flexibacter maritimus infection of black sea bream, Vibrio vulnificus infection of prawns, vibrio infection of prawns, vibrio infection of Thailand Infectious disease, Edwardsiel
Examples include, but are not limited to, la tardaa infection, eel paracolo disease, and the like.

The prophylactic / therapeutic agent for infectious disease as used in the present invention refers to an agent for preventing infection and onset of the above-mentioned infectious diseases, or for treating the infected one.

The dose and timing of administration of the preventive / therapeutic agent for infectious disease or feed of the present invention may be appropriately changed depending on the type of fish, crustacea, type of infectious disease, etc. to be applied. As 1 to 6,000 mg / kg
The weight may be given in two to three times a day. In addition, a preventive effect can be obtained by giving the fish or crustacean infectious disease preventive / therapeutic agent or feed of the present invention during the juvenile stage.

[0019]

EXAMPLES The present invention will be described in more detail below with reference to Reference Examples, Examples and Test Examples, but these do not limit the scope of the present invention in any way.

[Reference Example 1] Preparation of Milk Whey Protein Concentrate Sulfuric acid was added to skim milk to precipitate casein at an isoelectric point, which was then removed, and the remaining insoluble matter and various bacteria in the sulfuric acid whey were clarified. I excluded it. After that, concentrate with an ultrafiltration module until the solid content is 15% or more,
After further neutralizing the pH, the mixture was concentrated with an evaporator until the solid content became 25% or more. This concentrate was dried by a spray dryer by a conventional method to obtain a milk whey protein concentrate.

Reference Example 2 Preparation of Decomposed Product of Milk Whey Protein Concentrate (1) Commercially Available Milk Whey Protein Concentrate (WP
C: Protein content 80%) 1 kg was dissolved in 9 kg of purified water, sterilized by a plate type sterilizer at 75 ° C. for 15 seconds, and then sodium hydroxide was added to adjust the pH of the solution to 9.0. Bacillus subtilis 10 g of the protease preparation derived from the above, 4 g of trypsin, and 4 g of papain were added, and after decomposing at 55 ° C. for 5 hours, the enzyme was inactivated by heating at 90 ° C. for 20 minutes.
Then, it was concentrated by a conventional method and dried to obtain 950 g of a powdery milk whey protein concentrate hydrolyzate.

(Part 2) 1 kg of commercially available milk whey protein concentrate (WPC: protein content 80%) was dissolved in 9 kg of purified water, sterilized with a plate type sterilizer at 75 ° C. for 15 seconds, and sodium hydroxide was added thereafter. Then, the solution was adjusted to pH 8.5, and 8 g of the Bacillus subtilis-derived protease preparation was added. After 30 minutes while keeping the pH at 8.0,
4 g of trypsin was added, and the mixture was decomposed at 50 ° C. for 6 hours. Then, the enzyme was inactivated by heating at 85 ° C for 15 minutes. Then
It was concentrated by a conventional method and dried to obtain 940 g of a powdery milk whey protein concentrate decomposition product.

(Part 3) 100 kg of commercially available milk whey protein concentrate (WPC: protein content 80%) was dissolved in 900 kg of purified water, sterilized by a plate type sterilizer at 75 ° C. for 15 seconds, and then sodium hydroxide was added. Then, the pH of the solution is adjusted to 9.0, 1 kg of a Bacillus subtilis-derived protease preparation is added, and the mixture is kept at 50 ° C for hydrolysis, and after 8 hours, the enzyme is inactivated by heat treatment at 120 ° C for 2 seconds. After cooling, cooling, adjusting the pH to 6.0 with hydrochloric acid, ultrafiltration with an ultrafiltration membrane with a cut-off molecular weight of 10,000, then concentrating by the usual method, drying, and decomposing powdery milk whey protein concentrate. Got 85 kg.

[Reference Example 3] Preparation of lactoferrin Bovine lactoferrin was prepared from raw milk by the method of wax and lighter.
[BALAW & B. Reiter, Journal of Dairy Research, Vol. 44, Vol.
595-599, 1977]. Raw milk (450L)
1 mol hydrochloric acid was added thereto to adjust the pH to 6.3, and 100 g of Rennet (manufactured by Hansel) was added at a temperature of 32 ° C. After 30 minutes, the precipitated curd was removed to obtain milk whey. Concentrate milk whey approximately 20 times by ultrafiltration and concentrate it in concentrated milk whey (10 L) in advance with 0.05 M Tris-HCl buffer (pH
4 L of CM Sephadex C-50 resin (Pharmacia) equilibrated with 8.0) was added, and the mixture was stirred at 4 ° C for 1 hour, then left to stand and the supernatant was removed. The resin was packed in a column (90 × 9 cm), 0.05 M Tris-HCl buffer (pH 8.0) was passed through about 3 L to wash the resin, and then 0.05 M Tris-HCl buffer (pH 8) containing 0.2 M sodium chloride was added. .0) Approximately 5 L is passed through and finally contains 0.05M sodium chloride.
About 2 L of 0.05 M Tris-HCl buffer (pH 8.0) was passed through to elute lactoferrin. This eluate was diafiltered with an ultrafiltration device, desalted and freeze-dried to obtain about 8 g of powdered lactoferrin.

[Reference Example 4] Preparation of lactoferrin degradation product Bovine lactoferrin prepared by the same method as in Reference Example 3 was dissolved in purified water at a rate of 50 g / 950 ml, and hydrochloric acid was added to the resulting solution to adjust pH. Was adjusted to 3.0, and then commercially available pork pepsin (manufactured by Wako Pure Chemical Industries, Ltd.) was added to 1 per 50 g of lactoferrin.
It was added at a rate of g and hydrolyzed at 37 ° C. for 6 hours. Then, adjust the pH to 7.00 with 6M sodium hydroxide, and at 80 ℃
The enzyme was inactivated by heating for 10 minutes, cooled to room temperature, the insoluble matter was filtered off, and the filtrate was freeze-dried to obtain about 45 g of powdery lactoferrin degradation product.

Reference Example 5 Preparation of Cow Colostrum Powder 3 kg of bovine colostrum within 3 days after calving are filtered off with a filter cloth, and sodium hydroxide is added to adjust the titrated lactic acid content of colostrum to 0.30%. Then, the mixture was held and sterilized at 63 ° C. for 30 minutes while stirring, and spray-dried using a spray dryer to obtain about 395 g of colostrum powder. [Example 1] Preparation of prophylactic / therapeutic agent for infectious diseases 200 kg of milk whey protein concentrate hydrolyzate, 150 kg of corn starch, 80 kg of talc and 30 kg of magnesium stearate similar to those in Reference Example 2 (No. 1) were thoroughly mixed to prepare a 60 mesh powder. The particle size was adjusted by passing through a wire net to obtain the infectious disease preventive and therapeutic agent of the present invention.

Example 2 Preparation of Feed for Cultured Shrimp A basic feed for prawns [made by Hayashikane Sangyo Co., Ltd.] and milk whey protein concentrate (WPC, protein content 80%),
Alternatively, 3% by weight of the decomposed product of milk whey protein concentrate prepared in the same manner as in Reference Example 2 (No. 2) was added. The resulting formulation is steamed at 90 ° C or above for 5 minutes to modify the gluten and impart the required shape retention property, then dried with hot air at 80 ° C for several hours and pelletized (2 mm (φ × 5 mm) and contains milk whey protein concentrate or milk whey protein concentrate degradation product,
Two kinds of prawn feed were obtained for each.

[Example 3] Preparation of feed for cultured troughs [0028] Commercial feed basic powder for troughs [manufactured by Taiyo Feed Co., Ltd.]
To this, 3% by weight of milk whey protein concentrate (WPC, protein content 80%) or a decomposed product of milk whey protein concentrate prepared in the same manner as in Reference Example 2 (Part 3) was added. Mold the resulting blend using a pellet machine
(5 mmφ × 10 mm) and dried at 40 ° C. for 5 hours to obtain two types of dry pellets for troughs containing milk whey protein concentrate or milk whey protein concentrate degradation product.

[Example 4] Preparation of feed for cultured trout Commercially available feed powder for trout fry [manufactured by Taiyo Feed Co., Ltd.]
3% by weight of the milk whey protein concentrate of Reference Example 1 or the decomposed product of milk whey protein concentrate prepared in the same manner as in Reference Example 2 (Part 2) was added. The resulting blend was molded using a pellet machine (2 mmφ × 3 mm) and dried at 40 ° C for 5 hours to contain milk whey protein concentrate or milk whey protein concentrate decomposed product, 2 types each To obtain dry pellets for trout fry.

Example 5 Preparation of Cultured Flounder Feed for Commercial Flounder Dry Pellet Feed for Flounder (Taiyo Feed Co., Ltd., 5 mm φ × 5 mm) 100 g, milk whey protein concentrate (WPC, protein content 75%) ), Or 100 ml of a suspension (3 g / ml) of a decomposed product of milk whey protein concentrate prepared in the same manner as in Reference Example 2 (Part 1), and then at 30 ° C.
By drying for 3 hours, it contains milk whey protein concentrate or milk whey protein concentrate degradation product,
Two kinds of flounder feeds were obtained respectively.

[Example 6] Preparation of feed for cultured coho salmon Commercially available feed powder for coho salmon [manufactured by Taiyo Feed Co., Ltd.]
3% by weight of the milk whey protein concentrate of Reference Example 1 or the decomposed product of milk whey protein concentrate prepared in the same manner as in Reference Example 2 (Part 2) was added. The resulting blend was molded using a pellet machine (2 mmφ × 3 mm) and dried at 40 ° C for 5 hours to contain milk whey protein concentrate or milk whey protein concentrate decomposed product, 2 types each A coho salmon feed was obtained.

[Example 7] Preparation of feed for cultured striped horse mackerel Commercially available feed powder for striped horse mackerel [manufactured by Taiyo Feed Co., Ltd.]
3% by weight of milk whey protein concentrate or a decomposed product of milk whey protein concentrate prepared in the same manner as in Reference Example 2 (Part 3) was added. The obtained compound was molded using a pellet machine (8 mmφ × 10 mm) and dried at 40 ° C. for 5 hours to contain milk whey protein concentrate or milk whey protein concentrate decomposed product, each of two types. A feed for striped horse mackerel was obtained.

[Example 8] Preparation of feed for cultured red sea bream Commercially available feed basic powder for red sea bream [manufactured by Taiyo Feed Co., Ltd.]
3% by weight of the milk whey protein concentrate prepared in the same manner as Reference Example 1 or the decomposed product of milk whey protein concentrate prepared in the same manner as Reference Example 2 (Part 3) was added. The obtained mixture was molded (2 mmφ × 3 mm) using a pellet machine, dried at 40 ° C. for 5 hours, and contained milk whey protein concentrate or milk whey protein concentrate decomposed product. We obtained red sea bream feed.

[Test Example 1] Shrimp Rod shaped virus-Pen
Preventive effect against infection with aeus japonicus (RV-PJ) virus The midgut gland around the gut of prawns infected with RV-PJ was excised, and a homogenized extract containing 10 volumes of buffer was added to a 0.2 μm pore size filter. When it was injected into the abdominal muscle of healthy shrimp, it died with the symptoms peculiar to this infection such as vitiligo on the instep. Furthermore, electron microscopic observation revealed that this infection was caused by a virus that seems to belong to the baculovirus. Therefore, two kinds of prawn feed containing milk whey protein concentrate or milk whey protein concentrate degradation product and a preventive and therapeutic agent for infectious diseases were given to prawns, and the preventive effect against RV-PJ infection was examined.

Healthy prawns (average body weight: 22 g) were thoroughly mixed twice a day with the therapeutic agent for infectious disease obtained in the above Example 1 to 6% by weight of mussel minced meat, in the above Example 2. The obtained milk whey protein concentrate or two kinds of shrimp samples containing milk whey protein concentrate degradation product, for comparison, lactoferrin, lactoferrin degradation product, colostrum powder 3% feed, 1% of body weight per day Given the amount. Each administration group had 10 fish per group. In addition, a group fed a feed to which no additive was added was used as a control. On the 14th day after the first administration of the feed, 100 μl of the virus solution obtained in the same manner as above was diluted 80-fold to 100 prawns per group, or a buffer solution was injected as a blank, and the mortality rate after that I checked. The results are shown in Table 1.

[0036]

[Table 1]

When healthy shrimp were infected with the RV-PJ virus, the mortality rate after death was 60% after 7 days, but the preventive and therapeutic agent for infectious diseases of the present invention, milk whey protein concentrate, milk whey protein concentrate degradation A reduction in the mortality rate was observed in those fed the supplemented diet. In particular, administration of a prophylactic / therapeutic agent for infectious disease and a feed containing a decomposed product of a milk whey protein concentrate could suppress the mortality to 30% and 20%, respectively, and the effect was remarkable. On the other hand, in the lactoferrin, the lactoferrin degradation product, and the colostrum powder-added feed, the mortality rate was 60%, which was the same as the control, and no effect was observed. The shrimp (blank) not infected with the virus did not die at all during the test.

Test Example 2 Takifugu rubrip
es) to prevent infection of the disease of white-spots virus using healthy troughfish (average weight of 50 g) containing two types of milk whey protein concentrate or a milk whey protein concentrate degradation product obtained in Example 3 above. Feed for troughs,
For comparison, lactoferrin, a lactoferrin degradation product, and a diet containing 3% colostrum powder were given in an amount of 5% of the body weight per day.
Each feed-administered group had 10 fish per group. In addition, a group fed a feed to which no additive was added was used as a control. On the 14th day after the first administration of the feed, 0.1m of the oral virus of the leukoporosis having a titer of 1 × 10 ^3.7 TCID ₅₀ / ml was applied to 10 troughs per group.
l of each was subcutaneously inoculated on the back, and the mortality rate after that was examined. Table 2 shows the results.

[0039]

[Table 2]

Infection of a healthy troughfish with the oral disease virus results in 80% mortality after 7 days. However, the milk whey protein concentrate and the milk whey protein concentrate degradation product-added feed of the present invention were fed. A decrease in the mortality rate was observed, and particularly in the milk whey protein concentrate hydrolyzate-added feed, the mortality rate could be suppressed to less than 10%, and the effect was remarkable. Further, the mortality rate in the group administered with the milk whey protein concentrate-added feed was 50%. On the other hand, in the lactoferrin, lactoferrin degradation product, and colostrum powder-added feed, the mortality rate was 8
However, no effect was observed. The mortality rate of the tiger pufferfish (blank) that was not infected with the virus was 10%.

Test Example 3 Rainbow trout (Oncorhynchus my
Infection prevention effect against infectious pancreatic necrosis virus (IPNV) using kiss) Healthy rainbow trout (average body weight: 4 g) was added to the milk whey protein concentrate or the milk whey protein concentrate degradation product obtained in Example 4 above. Two species of rainbow trout feed containing
For comparison, lactoferrin, a lactoferrin degradation product, and a diet containing 3% colostrum powder were given in an amount of 20% of the body weight per day.
Each feed-administered group had 10 fish per group. In addition, a group fed a feed to which no additive was added was used as a control. On the 14th day after the first administration of the feed, 50 μl of IPNV solution having a titer of 5 × 10 ^4.8 TCID ₅₀ / ml was intravenously administered to 10 rainbow trouts per group. The results are shown in Table 3.

[0042]

[Table 3]

When a healthy rainbow trout was infected with IPNV, 10
After a day, the mortality rate is 80%, but the milk whey protein concentrate of the present invention and the feed containing the decomposed product of the milk whey protein concentrate show a decrease in the mortality rate, especially the milk whey protein concentration. It was possible to reduce the mortality rate to 10% or less with the feed containing the decomposed product, and the effect was remarkable. On the other hand, in the lactoferrin, the lactoferrin degradation product, and the colostrum powder-added feed, the mortality rate was 80% as in the control, and no effect was observed.
Rainbow trout (blank) not infected with the virus did not die at all during the test period.

Test Example 4 Flounder (Paralichthys oliv)
(Aceus) against rhabdovirus (HRV) infection prevention effect The flounder feed containing the milk whey protein concentrate or the milk whey protein concentrate degradation product obtained in Example 5 is fed to flounder to prevent infection against HRV. I checked. Two types of flounder feed containing healthy flounder (average weight 80 g) containing the milk whey protein concentrate obtained in Example 5 above or a decomposed product of milk whey protein concentrate,
For comparison, lactoferrin, a lactoferrin degradation product, and a colostrum powder 3% -added diet were given in an amount of 5% of the body weight per day in two divided doses. Each feed-administered group had 10 fish per group. In addition, a group fed a feed to which no additive was added was used as a control. On the 14th day after the first administration of the feed, HRV solution having a titer of 5 × 10 ^6.8 TCID ₅₀ / ml was applied to 10 flatfish per group.
Each 0.1 ml was intravenously administered. The results are shown in Table 4.

[0045]

[Table 4]

When healthy flounder was infected with HRV, the mortality rate was 40% after 14 days. However, those fed the feed containing the milk whey protein concentrate and milk whey protein concentrate hydrolyzate of the present invention A decrease in the mortality rate was observed, and the mortality rate could be suppressed to less than 10% with the milk whey protein concentrate hydrolyzate-added feed, and the effect was remarkable. On the other hand, in the lactoferrin, the lactoferrin degradation product, and the colostrum powder-added feed, the mortality rate was 40% as in the control, and no effect was observed.
Flounder (blank) not infected with the virus did not die at all during the test period.

Test Example 5 Coho salmon (Oncorhynchus ki)
sutch) Erythrocytic inclusion body syndrome
(EIBS) infection preventive effect The coho salmon feed containing the milk whey protein concentrate obtained in Example 6 or the milk whey protein concentrate degradation product was fed to coho salmon, and the infection preventive effect against EIBS virus was examined. Healthy whey salmon (average weight 180 g) on milk whey protein concentrate obtained in Example 6 above,
Alternatively, two kinds of diets for coho salmon containing a decomposed product of milk whey protein concentrate, for comparison, lactoferrin, a decomposed product of lactoferrin, and a diet containing 3% of colostrum powder were given in an amount of 10% of the body weight per day. Each feed-administered group had 10 fish per group. In addition, a group fed a feed to which no additive was added was used as a control. On the 14th day after the first administration of feed, 500 μ of EIBS virus solution (10% coho salmon per group filtered with 0.45 μm filter of head kidney extract of infected coho salmon)
was administered intraperitoneally. Table 5 shows the results.

[0048]

[Table 5]

Infection of healthy coho salmon with EIBS virus resulted in 80% mortality after 10 days, but those given the milk whey protein concentrate or milk whey protein concentrate degradation product of the present invention died. A decrease in the rate was observed, and the mortality rate of mortality could be suppressed to less than 10% particularly with the milk whey protein concentrate hydrolyzate-added feed, and the effect was remarkable. On the other hand, in the lactoferrin, the lactoferrin degradation product, and the colostrum powder-added feed, the mortality rate was 80% as in the control, and no effect was observed.
Coho salmon (blank) not infected with the virus did not die at all during the test period.

Test Example 6 Striped jack (Pseudocaranx de
tex) for preventing infection against iridvirus infections The milk whey protein concentrate obtained in Example 7 or a feed for striped horse mackerel containing the decomposed product of milk whey protein concentrate is given to striped horse mackerel, and the infection preventive effect against ylide virus is obtained. I checked. A healthy striped horse mackerel (average weight: 12.5 g) containing the milk whey protein concentrate obtained in Example 7 or a milk whey protein concentrate degradation product 2
Five kinds of diets for striped horse mackerel, for comparison, lactoferrin, lactoferrin degradation product, and 3% colostrum powder-added diet were given at 5% of body weight per day. 10 for each feed administration group
It has a tail. In addition, a group fed a feed to which no additive was added was used as a control. On the 14th day after the first administration of the feed, 200 μl of the ylidovirus solution (the spleen extract of the infected sardine filtered with a 0.45 μm filter) was intraperitoneally administered to 10 Japanese horse mackerel per group. Table 6 shows the results.

[0051]

[Table 6]

Infection of healthy striped horse mackerel with iridovirus resulted in 80% mortality after 10 days. However, milk whey protein concentrate of the present invention and milk whey protein concentrate degradation product-added feed were given. A reduction in the mortality rate was observed, and particularly in the milk whey protein concentrate hydrolyzate-added feed, the mortality rate could be suppressed to less than 10%, and the effect was remarkable. On the other hand, lactoferrin,
In the lactoferrin degradation product and the colostrum powder-added feed, the mortality rate was 80% as in the control, and no effect was observed. The horse mackerel (blank) not infected with the virus did not die at all during the test period.

[Test Example 7] Infection prevention effect against iridovirus infection using red sea bream (Pagrus major) The milk whey protein concentrate obtained in Example 8 or a feed for red sea bream containing the decomposed product of milk whey protein concentrate. Was given to the red sea bream and its preventive effect against the iridvirus was examined. Two types of red sea bream feed containing healthy whale (average weight 15.3 g) containing the milk whey protein concentrate obtained in Example 8 or the milk whey protein concentrate degradation product, as a comparison, lactoferrin, lactoferrin degradation product, 5% of body weight per day with 3% milk powder added feed
Given the amount. Each feed-administered group had 10 fish per group.
In addition, a group fed a feed to which no additive was added was used as a control. 10 days per group 14 days after the first feeding
The tail red sea bream was intraperitoneally administered with 200 μl of an iridovirus solution (infected red sea bream spleen extract filtered through a 0.45 μm filter). Table 7 shows the results.

[0054]

[Table 7]

Infection of healthy red sea bream with iridovirus results in 80% mortality after 10 days. However, the milk whey protein concentrate and the milk whey protein concentrate hydrolyzate-added feed of the present invention were fed. A reduction in the mortality rate was observed, and in particular, the degradation rate of the milk whey protein concentrate could be suppressed to less than 10%, and the effect was remarkable. On the other hand, in the lactoferrin, the lactoferrin degradation product, and the colostrum powder-added feed, the mortality rate was 80% as in the control, and no effect was observed.
The red sea bream not infected with the virus (blank) did not die at all during the test period.

[Test Example 8] Bullfish (Penaues monodo)
n) was used to prevent infection of Vibrio vulnificus against healthy bovine shrimp (average body weight: 20 g) twice a day containing the milk whey protein concentrate or the milk whey protein concentrate degradation product obtained in Example 2 above. 5% of the body weight per day was given to the cultivated shrimp feed, for comparison, lactoferrin, lactoferrin degradation product, and colostrum powder 3% feed. Each feed-administered group had 10 fish per group. In addition, a group fed a feed to which no additive was added was used as a control. On the 14th day after the first administration of the diet, 10 Shrimp per group were soaked in Vibrio vulnificus 5 × 10 ⁷ CFU / ml seawater for 12 hours, and the mortality rate after that was examined. Table 8 shows the results.
Shown in

[0057]

[Table 8]

Infection of healthy bull shrimp with Vibrio vulnificus resulted in 60% mortality after 7 days. However, milk whey protein concentrate of the present invention and milk whey protein concentrate degradation product-added feed were given. A reduction in the mortality rate was observed, and particularly in the milk-whey protein concentrate hydrolyzate-added feed, the mortality rate could be suppressed to 20% or less, and the effect was remarkable. On the other hand, in the lactoferrin, the lactoferrin degradation product, and the colostrum powder-added feed, the mortality rate was 60%, which was the same as the control, and no effect was observed. The shrimp (blank) not infected with the virus did not die at all during the test period.

Test Example 9 Rainbow Trout (Oncorhynchus my
infection prevention effect against Vibrio anguillarum using kiss) Two kinds of rainbow trout feeds containing the milk whey protein concentrate obtained in the above Example 4 or the milk whey protein concentrate degradation product in healthy rainbow trout (average weight 4 g) For comparison, lactoferrin, lactoferrin degradation product, and colostrum powder 3% -added feed were given in an amount of 20% of the body weight per day. Each feed-administered group had 10 fish per group. In addition, a group fed a feed to which no additive was added was used as a control. On the 14th day after the first feed administration, 10 rainbow trouts per group
Vibrio anguillarum solution having a titer of 2.8 × 10 ⁷ CFU was intravenously administered in an amount of 50 µl each. The results are shown in Table 9.

[0060]

[Table 9]

Infection of healthy rainbow trout with Vibrio anguillarum resulted in 80% mortality after 10 days. However, milk whey protein concentrate of the present invention and milk whey protein concentrate degradation product-added feed were given. A reduction in the mortality rate was observed, and particularly in the milk whey protein concentrate hydrolyzate-added feed, the mortality rate could be suppressed to less than 10%, and the effect was remarkable. On the other hand, in the lactoferrin, the lactoferrin degradation product, and the colostrum powder-added feed, the mortality rate was 80% as in the control, and no effect was observed. Rainbow trout (blank) not infected with the virus did not die at all during the test period.

[0062]

INDUSTRIAL APPLICABILITY According to the present invention, a preventive / therapeutic agent for infectious diseases and a feed effective for infectious diseases of fish and crustaceans are provided.

The infectious disease preventive / therapeutic agent and feed of the present invention have no problem in terms of safety in that they contain milk whey or a decomposed product of milk whey as an active ingredient.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification number Office reference number FI technical display location A23K 1/18 A23K 1/18 B A61K 38/00 AFF A61K 37/02 AFF (72) Inventor Nakajima Hiroshi Maruha Co., Ltd. 16-2, Wadai, Tsukuba, Ibaraki Prefecture (72) Inventor Nobuyuki Sato 16-2 Wadai, Tsuruba, Ibaraki, Maruha Co., Ltd. Central Research Institute

Claims (2)

[Claims] 1. A preventive / therapeutic agent for infectious diseases of fish and crustaceans, which comprises milk whey or a decomposed product of milk whey as an active ingredient. 2. A feed for fish and crustaceans containing milk whey or a decomposed product of milk whey.