JP4608255B2 - Composition having anti-parasitic activity for fish and method for preventing / treating parasitic disease - Google Patents

Description

  The present invention relates to a composition having anti-parasitic activity for fish. Specifically, the present invention relates to a composition having an antiparasitic action for fish having an effect by oral administration.

  In the fish farming industry, in recent years intensive aquaculture of various marine fish has been carried out due to improved seed production and breeding techniques, and industrial damage due to infectious diseases caused by viruses, bacteria, protozoa and parasites has increased. ing. Among them, parasitic diseases are increasing, and the reason for this is the delay in establishing an effective control method. For example, leukoderma occurs frequently in cultivated fields such as Japanese flounder, red sea bream, and tiger puffer fish mainly in western Japan, resulting in massive drowning of cultured fish. . White spot rot is caused by the parasitic white worm, Cryptocaryon irritans, belonging to the parasitic protozoan ciliate, infesting fish sharks and skin. The optimum temperature for growth is 18-30 ° C. Infection with white spotted caterpillars has been observed in many marine fish such as red sea bream, tiger puffer, amberjack, and flounder.

The only way to deal with sea surface ginger is to move the ginger to a well-tided sea area where white spotworm is not occurring. By this measure, it is possible to reduce the infection caused by the bottom cyst and the establishment of the infection caused by the worms present in the fish body. Moreover, it is effective to repeat the movement to the aquarium where the worm does not exist as a countermeasure at the land facility. In other words, when the water temperature is around 25 ° C, the worms leave the host 3-4 days after infection and become cysts, and the release of hatched larvae from the cysts requires more than 4 days. Transfer to a new aquarium can prevent the spread of infection. However, this method requires a lot of labor.
Patent Documents 1 and 2 list kudin as an active ingredient, but these patents show effects on microbial infections such as Aeromonas salmonicida or Streptococcus sp., And show effects on parasites. is not. Patent Document 3 discloses that some herbal medicines have antiparasitic effects on fish.

Kujin and goat root are known as herbal medicines.
<Kugin (Sophorae Radix)>
Motohara: Legumes bitter Sophora flavescens Ait. (Clara) dried roots.
Taste: The taste is bitter and the sex is cold. (Return: heart, liver, small intestine, large intestine, stomach)
Main component: Alkaloid; flavonoids such as matrine, oxymatrine, sophoranol, xanthohumol,
kurarinone, kuraridin, kuraridinol, kurarinol, etc. are known.
Pharmacological action: shampooing / drying insecticide, diuresis / antipyretic action, antifungal action, antitrichomonas action, blood pressure lowering action, anti-peptic ulcer action, central inhibitory action, phosphodiesterase inhibitory action, liver damage inhibitory action, Hemostatic action, immunostimulatory action, anti-complexity action and the like are known.
Medicinal effects: nourishment tonic, diarrhea, urinary penalties, rashes, athlete's foot, jaundice, scabies, septic medicinal properties / uses: bitterness, bitterness stomach, tonic, anti-inflammatory, diuresis, diarrhea, analgesic or dysentery It has an effect on bacterial diarrhea, bloody stool, tumor poisoning, etc., and it has the effect of enhancing the appetite by strengthening the internal organs, but it contains the strong alkaloids matrine, oxymatrine, etc., side effects in the case of frail constitution Appears.
Add 6 liters of bitter ginseng and add 0.3 liters of water, and apply the decoction in the case of athlete's foot, ash, and silkworm. It is effective for itching and inflammatory swelling. Apply the raw root juice to the silkworm.
About 300 grams of dried shoots of Clara and decocted with about 5 liters of water are effective in combating animal skin parasites such as agricultural pests and cows and horses. Moreover, when dried foliage is used in an appropriate fineness, worms are exterminated. In China, kujin is sold as a bath salt for fish parasitic diseases.
Dose: 3-15g.
Note: Whole plants (stems and leaves) contain toxic matrine, alkaloids such as oxymatrine. Matrine should never be taken because it paralyzes the cerebrum, causing convulsions, and respiratory motor nerve palsy stops breathing and leads to death.

<Yamazu root>
Motohara: Dried root of Sophora subprostrata.
Taste: The taste is bitter and the sex is cold. (Return: heart, lungs)
Main components: matrine, matrine-n-oxide, anagyrine, methylcytisine, genistin, phenolic substances
Pharmacological action: Kiyomizu Ryo, anti-inflammatory, antitumor
Clinical application: Ingredients such as ginseng and bellflowers are added to the swelling pain of the actual fever of the throat and gingiva. For example, a sore throat may be used by blending an easteris and beef eggplant. However, it is not suitable for swelling pain due to throat fire. As an adjunct to cancer (especially in the early stages of lung cancer and laryngeal cancer), white flower serpentine grass, fish licorice, etc. are used in combination. However, it is necessary to continue to observe the therapeutic effect. External application of goat root powder for cervical inflammation and stomatitis has an anti-inflammatory effect.
Dose: 6-9g for internal use. 2-6g for powder, appropriate amount for external use.

Japanese Patent No. 2535555 Japanese Patent No. 2535559 Japanese Patent No. 3366991

  In the field of aquaculture, there has been a demand for substances effective against parasites such as white spot insects. An object of the present invention is to provide a composition that has a high antiparasitic activity against parasites such as white spotworms and is extremely less affected by side effects of drugs and drug residues on fish. In other words, unlike fish species that are cultivated in closed ponds or onshore water tanks, it is highly necessary to consider the impact on the surrounding environment in aquaculture at sea, and because it is not closed, it is a medicinal bath in the culvert. It is impossible to maintain the concentration of the agent for a long time, and it is also difficult to bathe in the closed water for a long time. Therefore, an object of the present invention is to provide a composition effective by oral administration so that it can also be used for marine fish cultivated with marine fish.

  As a result of extensive search for natural substances in search of ingredients that have less adverse effects on fish and the environment due to side effects and drug residues, we found an excellent control effect against marine fish white spotworms in the extract of kujin, a herbal medicine. As shown in the test example of Example 6, the antiparasitic action of various herbal medicines other than kudin was also examined. Some herbal medicines were effective in vitro, but only oral cucumbers and goat roots were effective.

The gist of the present invention is the following composition (1) to (7) for preventing and / or treating parasitic diseases.
(1) An orally administrable composition for preventing and / or treating parasitic diseases for fish containing matrine.
(2) The composition for prevention and / or treatment of parasitic diseases according to (1), wherein the composition containing matrine is kujin, ground bean root extract or extract.
(3) The composition for prevention and / or treatment of parasitic diseases according to (2), wherein the extract of kujin is water and / or an organic solvent extract.
(4) The composition for prevention and / or treatment of parasitic diseases according to (1) to (3), wherein the fish is a marine fish.
(5) The fish is classified as perch, puffer, flounder, salmon, catfish, carp, or (1) to (3) for prevention and / or treatment of any parasitic disease Composition.
(6) The composition for prevention and / or treatment of any of the parasitic diseases according to any one of (1) to (5), wherein the parasitic disease is caused by a protozoan ciliate or a parasite belonging to a flat animal monopod .
(7) White spotted caterpillars that belong to protozoan ciliates, blue clinella, kilodonella, epistylis, sukutika, trichodina, neobenedenia, or neobenedenia that belong to the flat-animal monoplasia, dactyrogillus, pseudodactyrogillus, microcotile, The composition for prevention and / or treatment of any parasitic disease according to (1) to (5), which is caused by bibagina, heteraxine, heterobotulinum, or neoheterobotlium.

The gist of the present invention is a method for preventing and / or treating the following parasitic diseases (8) to (16).
(8) A method for preventing and / or treating fish parasitic diseases by orally administering a composition containing matrine.
(9) Oral administration of 1 to 50 mg / day of fish weight per kg of matrine, 10 to 2000 mg / day of fish weight per kilogram of Kuzin bulk, or 10 to 2000 mg / day of fish body weight converted to yam root (8) A method for preventing and / or treating a parasitic disease.
(10) A method for preventing and / or treating fish parasitic diseases by feeding a feed containing a composition containing matrine.
(11) The feed contains a composition containing matrine as a feed for fish farming in an amount of 0.002 to 0.1% in terms of matrine, 0.1 to 5% in terms of raw material of kudin, or 0. The method for preventing and / or treating parasitic diseases according to (10), wherein 1 to 5% is added.
(12) The method for preventing and / or treating the parasitic disease of (10) or (11), wherein the composition containing matrine is processed so that a bitter component is hardly eluted.
(13) The process for making the bitterness component of the composition containing matrine difficult to dissolve is encapsulated, microencapsulated, coated tableted, or powdered with a water-insoluble component. A method for preventing and / or treating parasitic diseases.
(14) The method for preventing and / or treating the parasitic disease according to any one of (8) to (13), wherein the composition containing matrine is kujin, ground soybean extract or extract.
(15) The method for preventing and / or treating the parasitic disease of (14), wherein the extract is a water and / or organic solvent extract.
(16) The method for preventing and / or treating the parasitic disease of any one of (10) to (13), wherein the feed is a moist pellet, EP (Extruded Pellet).

  Since the composition of the present invention is known as a crude drug, it is a composition that has been used in humans and animals, and is easy to use with little concern about safety and environmental impact. It can be used as an antiparasitic agent for marine fish, etc., which is not suitable for medicinal baths, because it exerts an antiparasitic effect by oral administration to fish. The composition of the present invention has an effect of suppressing infection of parasitic diseases and a slightly weaker effect of extinguishing the parasites, and can cure the parasitic diseases by an appropriate method of use.

  The matrine of the present invention is a compound having the structure shown in Formula 1. Alkaloids contained in the roots of Sophora Flavescens (this is called bitter in herbal medicine) and have a strong bitter taste.

As herbal medicines containing matrine, bitter and yam roots are known. Matrine is water soluble and can be extracted from these herbal medicines. Synthetic products are also sold.
Kujin is the root of Sophora Flavescens, and Yamazu-root is the root of Sofa subprostrata. In the present invention, kujin and yam root can be used in any form such as raw, dried product, pulverized product, powder, extract and the like. What is necessary is just to select suitably according to the condition at the time of oral administration to a fish. In order to obtain a stable effect, it is preferable to use an extract capable of keeping the content of the active ingredient constant in a liquid or powder form.

  In the present invention, fish includes both freshwater fish and seawater fish. In the case of freshwater fish, the aquaculture ponds and aquariums are often closed systems and are subject to medicinal baths, so it may not always be necessary to use the orally administrable composition of the present invention. Can also be used for freshwater fish if there are merits of oral administration. Since the compositions of the present invention are believed to be effective against a wide range of parasitic diseases, they can be used on any fish with known parasitic diseases. In terms of classification, examples include fish species belonging to perch, puffer, flounder, salmon, catfish, and carp. Specifically, among the fishes of the perch, red sea bream (Thailand), sea bream (Issidae family), sea bream (Salphidae family), black sea bream (Thailand family), scorpionfish (Scorpaenidae), tilapia (Popularidae), sea bass (Suzuki family) Family), red turtle (red-eyed family), mahata (grouper), cued (grouper), chairomaru grouper (grouper), pheasant grouper (grouper), white-faced grouper (grouper), red-faced grouper (grouper), yellowtail ), Amberjack (Acariaceae), flounder (Acariaceae), Shimaji (Acariaceae), etc. For the pufferfish, the tiger pufferfish (Pufferaceae) etc. In salmonids, rainbow trout (salmonids), coho salmon (salmonids), Atlantic salmon (salmonids), etc., and in freshwater fish, catfish (Cutaceae) , Cypriniformes goldfish (Cyprinidae), carp (Cyprinidae), etc., etc. Perciformes Discus (cichlid (cichlid) family) are exemplified.

The parasitic disease in the present invention is a fish disease caused by parasite infestation. Examples of parasitic diseases in cultured fish include white spotted insects belonging to protozoan ciliates, blue clinella, kilodonella, epistilis, sukutika, trichodina. Neobenedenia, Neobenedenia, Dactyrogillus, Pseudodactyrogillus, Microcotile, Vivagina, Heteraxine, Heterobotulinum, Neoheterobotulium, etc. belonging to the flat-animal monopoda are known. The compositions of the present invention are useful for these parasitic diseases.
In the present invention, “prevention and / or treatment” means prevention of the action of the parasite infesting the fish, and treatment means the action of removing the parasitic parasite and curing it.
In the present invention, oral administration means that the composition of the present invention is directly administered to fish as a tablet, granule, capsule, or the like, or added to feed (mixed into a raw material, liquefied, immersed, coated, etc.). Meaning that it is taken together with feed.

The composition containing matrine of the present invention means an extract of a crude or dried crude drug containing matrine, a pulverized product, or an organic solvent such as water, hot water or alcohol. It is possible to use a synthetic product of matrine, but the price is high.
In Kujin, it was shown that antiparasitic action is enhanced by using not only components extracted with hot water but also components extracted with alcohol. Since matrine is water-soluble, it is considered that components other than matrine are functioning. As shown in the examples, after hot water extraction, the residue may be extracted with alcohol or with a mixed solution of water and alcohol. For example, the component of this invention can be extracted with the liquid mixture which added about 20 to 50% of ethanol to water. Ingredients other than matrine are also dissolved in a chloroform solvent to which alcohol has been added. A highly polar organic solvent such as ethyl acetate or acetone may be used as the extraction solvent.

The present invention is a method for preventing and / or treating fish parasitic diseases by orally administering a composition containing matrine. The composition containing matrine of the present invention is 1 to 50 mg / day of fish body weight in terms of matrine, 10 to 2000 mg / day of fish body weight in terms of kuzin bulk, or 1 kg of fish weight in terms of yam root powder. 10-2000 mg / day is administered orally. Preferably, 3-15 mg / day of fish weight per kilogram of matrine, 30-500 mg / day of fish body weight per kilogram of kudin, or 30-500 mg / day of fish body weight per milligram of mazone root To do.
Parasites have different life cycles depending on the parasite. For example, in the case of white spotworms, when a hatched larva infects a host (fish), a bag-like enclosure is formed in the epithelium of the wing or skin and grows in the bag. At 25 ° C., it leaves the host in 3-4 days and floats as cyst in water. At 25 ° C., larvae hatch again from 4 days to 2 weeks and infect the host. Therefore, even if the parasites attached to the fish are removed, the larvae from the cysts in the water will again infest and will not completely treat the parasitic disease. Aside from isolated terrestrial aquaculture, there is always a risk of parasitic diseases in sea pens. For example, in the case of white spot insects, there is a risk of infection when the seawater temperature is 18 ° C. or higher. Therefore, the composition of the present invention is preferably used continuously when the seawater temperature reaches 18 ° C. or higher. Even if it is impossible, it is necessary to use it continuously for at least one month in consideration of the life cycle as described above when infected. In the case of using for other parasitic diseases, it is preferable to continue to use it for a period in which the cycle completes in consideration of each life cycle.
When the composition containing matrine of the present invention is added to the feed and fed, the concentration in the feed may be adjusted so that the required matrine is consumed according to the daily amount of the fish species consumed. . The amount of food consumed per body weight varies depending on the fish species, and also varies depending on the growth stage. Therefore, it is necessary to adjust the amount appropriately. However, for many farmed fish, a feed of about 3% of the fish body weight per day is usually given. It is done. In that case, the composition containing matrine is 0.002 to 0.1% in terms of matrine, 0.1 to 5% in terms of raw material of kudin, or 0.1 to 5 in terms of yam root powder. %Added. Preferably, the composition containing matrine is 0.005-0.05% in terms of matrine, 0.3-2% in terms of kujin bulk, or 0.3-2 in terms of yam root %Added.

The feed material may be anything as long as it is normally used as a feed for the fish species, and the effects of the present invention are not affected by the feed material. In the case of moist pellets, frozen products such as high-catch fish are used as the main raw material, and fish meal, sea bream, vitamins, minerals, etc. are added and molded. In the case of EP, fish meal, salmon, vitamins, minerals, etc. are formed into pellets using an extruder or the like. The composition containing matrine of the present invention can be mixed with these raw materials and finished into pellets.
Since the composition of the present invention has a bitter taste, a decrease in food intake may be observed depending on the fish species and the degree of growth. In such a case, an addition method in which the bitterness of the surface is reduced by adding a large amount to the inside of the feed, such as adding pellets to the raw material to produce pellets, is more suitable than an addition method of spraying on the surface of the feed. . Further, the composition of the present invention may be processed so that bitterness is masked. Specifically, capsules, microencapsulation, tableting with a coating such as sugar coating, coating granulation, or powder together with water-insoluble components such as casein and carrageenan, which are generally performed on drugs with a bitter taste For example, the water-soluble component is not easily released. In any case, since fish swallow the feed, the bitterness should not be easily dissolved from the surface of the feed.
Examples of the present invention will be described below, but the present invention is not limited thereto.

<Preparation 1 of kujin extract>
Dry kujin ground product (Daicel Chemicals) and hot water were mixed at a ratio of 1:10 (w / v), and the components of kudin were eluted with a disperser (Biomixer; manufactured by Nippon Seiki Seisakusho) ( 30 minutes). The extract was centrifuged at 8000 rpm for 15 minutes, and the supernatant was filtered through a Millipore filter (0.22 μm, manufactured by Millipore) to obtain a hot water extract. Further, ethanol (manufactured by Wako Pure Chemical Industries, Ltd.) was mixed with the remaining precipitate at a ratio of 1: 5 (w / v), heated to 80 ° C. in a thermostatic bath and eluted with a disperser (30 minutes). The extract was centrifuged at 8000 rpm for 15 minutes, and the supernatant was filtered through a Millipore filter to obtain an ethanol extract of hot water extraction residue.

<Preparation 2 of kujin extract>
The dried cucumber pulverized product (manufactured by Daicel Chemicals) and hot water were mixed at a ratio of 1:10 (w / v) and allowed to stand in a thermostatic bath heated to 100 ° C. for 1 hour to elute the kudin component. . The extract was centrifuged at 8000 rpm for 15 minutes, and the supernatant was filtered through a Millipore filter (0.22 μm, manufactured by Millipore) to obtain a kujin extract.

<Example 1 of additive feed production>
A solid feed was produced using the kujin extract obtained in Example 1. That is, 1 kg of EP feed (Nihon Suisan Co., Ltd., D-1, D-2 for fry, D series for breeding red sea bream), 50 ml of kujin hot water extract obtained in Example 1 and ethanol extract of hot water extraction residue After spraying 25 ml, it was dried with a dryer at 40 ° C. for 5 hours. In this way, a solid feed containing 0.5% in terms of bulk powder was obtained.

<Example 2 of additive feed production>
A solid feed was produced using the kujin extract obtained in Example 2. That is, after spraying 50 ml of the kujin extract obtained in Example 2 on 1 kg of EP feed (Nihon Suisan Co., Ltd., D-1, D-2 for fry, D series for breeding red sea bream), Dry for 5 hours. In this way, a solid feed containing 0.5% in terms of bulk powder was obtained.

<Effects of kujin extract against marine white spot worms (in vitro test)>
A solution of 1 μl of the hot water extract of Example 1 in 1 ml of seawater (the white spotted caterpillars were immersed in a 0.01% solution to confirm the anti-white spotting effect.
Two hours after immersion, the number of deaths of hatching larvae was measured and the effect was judged. More than 200 parasites were tested. The water temperature during immersion was 20 ° C and 25 ° C. The control zone was only seawater.
As a result, more than 96% of white spotted larvae survived in the control group, whereas the death of all parasites was confirmed in the Kujin group, indicating that this extract has an excellent effect on white spotted insects. Indicated.

<Effects of kujin extract on marine neobenedenia (in vitro test)>
The same test as in Example 5 was performed using hatched larvae and larvae of Neo Benedenia (stomachworms). Two hours after soaking, neobenedenia hatchling larvae were counted for the number of deaths, and for adults, the number of atrophy was measured to determine the effect. More than 30 parasites were tested, and the water temperature during immersion was 20 ° C and 25 ° C. The control zone was seawater only.
As a result, more than 95% of neobenedenia survived in the control group, whereas the death of all parasites was confirmed in the Kujin group, indicating that this extract has an excellent anthelmintic effect on neobenedenia. .
From the above results, it was shown that the cucumber extract of the present invention has an anthelmintic effect on parasites whose classifications are greatly different from those of flat animals (white spotted caterpillars) and protozoa (Neobenedenia). Therefore, its effectiveness is considered to be fish parasites in general.
<Test example>
Moreover, the effect with respect to a parasite was tested by the test system similar to Example 5 about the various crude drugs shown in Table 1 as crude drugs other than a kujin. In this in vitro test system in which each extract was prepared in the same manner as in Example 1, several herbal medicines showed antiparasitic activity as shown in Table 1. However, these were ineffective or weak when administered orally, and only Kujin had a sufficient effect with oral administration.

<Anti-marine white spotted insect effect of kudin extract-added feed in red sea bream>
Using red sea bream (average fish body weight 39.3g) in which no disease was recognized, the effect of feed addition of kudin extract against white spotted insects was examined. The test plots were fed with feed containing 0.0%, 0.05%, 0.1%, 0.5%, and 1.0% of kujin extract (raw, kujin 0.05%, kudin 0.1%, kujin 0.5 % Ward and Kujing 1.0% ward). The amount of feeding was 3% / day with respect to the fish weight of each ward.
The kudin extract-added feed was produced by adjusting the amount of hot water extract and ethanol extract to be sprayed in the same manner as in Example 3.
In the attack test, each fish is placed in a 100 liter aquarium and fed with each test feed. After 3 weeks from the start of feeding, 40,000 white spotted caterpillars are placed in each aquarium and allowed to infect for 2 hours with water still. Was done. Thereafter, the animals were bred continuously on the test feed for 2 days. The water temperature during the test period was 24-25 ° C. Water exchange during the test period was 24 rotations / day.
The evaluation was performed by sampling all test fish 3 days after the start of the attack, and comparing the number of parasites of the worms parasitizing the fish and the correction values. The correction value is expressed by “correction value = number of parasites / ^2/3 fish weight” in order to correct the difference in fish size in each test section.
As a result of the test, the number of white spotted insects in the control group, 0.05% group of kudin, 0.1% group of kudin, 0.5% group of kudin and 1.0% group of kudin, respectively, was 169.1 ± 62.3 individuals / tail (mean ± standard deviation), They were 201.9 ± 56.3 individuals / tail, 144.4 ± 36.1 individuals / tail, 80.3 ± 44.2 individuals / tail, and 72.4 ± 17.4 individuals / tail. The correction values for these values are shown in FIG. The number of parasites on the moths in the 0.5% kudin and 1.0% kujin was significantly smaller than that in the control group (P <0.01). Therefore, it was shown that the infection of white spotted caterpillar can be prevented by administering the kujin extract-added feed to red sea bream. It was shown that the feed to which the extract of the present invention was added in an amount of 0.5 to 1% by weight in terms of bulk powder had an antiparasitic effect by oral administration.

<Anti-marine white spotworm effect of kudin extract-added diet in red sea bream (short-term administration)>
In Example 7, this extract-added feed was fed in advance for 3 weeks before challenge with white spotted caterpillars. In this example, the feed supplemented with this extract was fed for 4 days before challenge by white spotted caterpillars, and the anti-white spotted insect effect after short-term administration was examined.
An attack test using white spotted worms was carried out using red sea bream (average fish weight 31.1 g) in which no disease was recognized. The test plots consisted of two plots: a plot fed with feed containing 0.5% of kujin extract in terms of bulk powder (crude 0.5% plot) and a group fed with additive-free feed (control plot). The feeding amount was 3% / day with respect to the fish weight of each section. The feed produced by the production method of Example 3 was used as the feed for adding kujin extract.
In the attack test, each fish is placed in a 100 liter aquarium, and each test feed is fed. After 4 days from the start of feeding, 40,000 white spotted larvae are placed in each aquarium, and the fish are infected for 2 hours. I went there. Thereafter, the animals were continuously raised on the test feed for 2 days. The water temperature during the test period was 24-25 ° C. The water exchange during the test period was 24 rotations / day.
The evaluation was performed by sampling all test fish 3 days after the start of the attack, and comparing the number of parasites of the worms parasitizing the fish and the correction values.
The numbers of white spotted insects in the control group and the 0.5% kudin group were 313.3 ± 111.5 individuals / tail (mean value ± standard deviation) and 179.6 ± 69.6 individuals / tail, respectively. The correction values for these values are shown in FIG. The number of parasites of the worms in the 0.5% Kujin group was significantly lower than that in the control group (P <0.01). Therefore, it has been shown that even short-term administration prevents white spotworm infection.

<Anthelmintic anthelmintic effect of kudin extract-added diet in red sea bream (Anthelmintic effect on infected insects)>
As shown in Examples 7 and 8, it was found that feeding the red sea bream extract-added feed to red sea bream can prevent white spot insect infection. In this example, it was examined whether the feed of the present invention has an anthelmintic action against white spotted insects infected with red sea bream.
An attack test using white spotted worms was carried out using 20 fish of red sea bream (average fish weight 13.3 g) in which no disease was recognized. The test plots consisted of two plots: a plot fed with feed containing 0.5% of kujin extract in terms of bulk powder (crude 0.5% plot) and a group fed with additive-free feed (control plot). The feeding amount was 3% / day with respect to the fish weight of each section. The feed produced by the production method of Example 3 was used as the feed for adding kujin extract.
The attack test is carried out by placing each fish in a 100 liter aquarium, acclimatizing them with normal feed for one week, and then injecting 100,000 white spotted larvae into each aquarium and infecting them for 2 hours in a stationary state. It was. Each test feed was fed for 3 days from 3 hours after infection. The water temperature during the test period was 18.5 to 19.5 ° C. The water exchange during the test period was 24 rotations / day.
The evaluation was performed by sampling all test fish 3 days after the start of the attack, and comparing the number of parasites of the worms parasitizing the fish and the correction values.
The numbers of white spotted caterpillars in the control group and 0.5% kudin group were 179.8 ± 78.9 individuals / tail (mean value ± standard deviation) and 133.6 ± 48.7 individuals / tail, respectively. The correction values of these values are shown in FIG. The number of parasites of worms in the 0.5% kudin group was significantly lower than that in the control group (P <0.05), but there was a significant difference (P = 0.072) in both groups in the corrected value. I couldn't. From these results, it was considered that the anti-parasitic effect of the cucumber extract-added feed was more related to the infection-inhibiting action than the anthelmintic action.

<Treatment / survival improvement effect of white-spotted insect infections of kudin extract-added diet in red sea bream>
In the tests of Examples 7 to 9, the number of white spotted insects on the culm of the test fish was examined three days after attacking with white spotted insects, and the effect was judged. In this test, the test feed was continuously bred for about 2 months after being challenged by white spot insects, and the anti-white spot effect (treatment / survival improvement effect) of this extract-added feed after long-term administration was examined.
An attack test using white spotted worms was conducted using 50 fish of red sea bream (average fish weight 8.0 g) in which no disease was recognized. The test plots consisted of two plots: a plot fed with feed containing 0.5% of kujin extract in terms of bulk powder (crude 0.5% plot) and a group fed with additive-free feed (control plot). Feeding amount was satiation in each ward. The feed produced by the production method of Example 3 was used as the feed for adding kujin extract.
The attack test was carried out by placing each fish in a 100 liter aquarium, acclimatizing each test feed for 3 weeks, and then injecting 70,000 white spotted larvae into each aquarium and infecting them with water still for 2 hours. . Thereafter, each test feed was continuously raised for 63 days. The water temperature during the test period was 17.5 to 24.5 ° C., and its transition is shown in FIG. The water exchange during the test period was 24 rotations / day.
Evaluation was made by recording the number of deaths every day and comparing the survival rate during the breeding period, and sampling 5 fish in each section 20 days and 30 days after the start of the attack, and parasitizing the parasites that parasitize the moths. This was done by comparing the number and the correction value.
The numbers of white spotted insects in the control group and the Kujin 0.5% group 20 days after the start of the attack are 206.2 ± 17.5 individuals / tail (average value ± standard deviation) and 28.7 ± 59.4 individuals / tail, respectively. 5 is shown in FIG. 5 in a ratio where the value of the control group is 1. The number of parasites on the moths in the 0.5% kudin group was significantly lower than that in the control group (P <0.01). 30 days after the start of the attack, the number of white spotted insects in the control group and the 0.5% kujin is 1302.0 ± 577.3 individuals / tail (mean ± standard deviation) and 5.0 ± 11.2 individuals / tail, respectively. 6 is shown in FIG. 6 at a ratio where the value of the control group is 1. The number of parasites on the moths in the 0.5% kudin group was significantly lower than that in the control group (P <0.01). Furthermore, the feeding of fish in the control group was reduced due to white spot infection, and the average fish weight of the sampling fish at this time was 13.3 g in the control group and 14.6 g in the 0.5% kudin. The control area began drowning 31 days after the start of the attack and was annihilated 34 days later (Figure 7). On the other hand, no moribundity was observed in the 0.5% kudin group until the end of the study. At the end of the experiment, we observed the culm of the surviving fish in the 0.5% kudin, but no white spot insects were observed. Therefore, it was shown that the cucumber-added feed has an effect on the survival of fishes infected with white spotted insects and has a therapeutic effect.

<Comparison of anti-white spotted insect effect of hot water extract and ethanol extract of hot water extract residue and examination of synergistic effect by adding both extracts>
A common method for extracting crude drugs is hot water extraction, and kujin is no exception. In Examples 7 to 10, both the hot water extract and the ethanol extract of the hot water extraction residue were added to the feed for evaluation. In this study, hot water extract, ethanol extract, and combined antiparasitic activity were examined.
An attack test using white spotted worms was carried out using nine fish of red sea bream (average fish weight 55 g) in which no disease was recognized. The test plots were fed with feed containing 0.5% or 1.0% of kujin hot water extract (raw kujin hot water extraction 0.5%, kujin hot water extraction 1%), and kujin hot water extraction residuals. Feeding a feed supplemented with 0.5% or 1.0% ethanol extract (0.5% kudin ethanol extraction, 1.0% kudin ethanol extraction), 0.5% kujin hot water extract and residual ethanol extraction from kujin hot water extraction A total of 6 wards were fed: 0.5% food feed (kujin hot water extraction 0.5% + ethanol extraction 0.5% ward) and non-additive feed fed (control ward). The feeding amount was 3% / day with respect to the fish weight of each section. A hot water extract and a hot water extraction residual ethanol extract were produced by the method of Example 1. The feed to which these extracts were added was produced by the production method of Example 3.
In the attack test, each fish is placed in a 100 liter aquarium and fed with each test feed. Three weeks after the start of feeding, 50,000 white spotted caterpillars are placed in each aquarium, and the fish are infected for 2 hours with water stopped. Was done. Thereafter, the animals were bred continuously on the test feed for 2 days. The water temperature during the test period was 20-23 ° C. Water exchange during the test period was 24 rotations / day.
The evaluation was performed by sampling all test fish 3 days after the start of the attack, and comparing the number of parasites of the worms parasitizing the fish and the correction values.
White spotted insects in the control group, 0.5% kujin hot water extraction, 1.0% kujin hot water extraction, 0.5% kudin ethanol extraction, 1.0% kudin ethanol extraction and 0.5% kujin hot water extraction + 0.5% ethanol extraction The numbers are 259.6 ± 57.0 individuals / tail (mean value ± standard deviation), 111.9 ± 48.1 individuals / tail, 125.8 ± 41.1 individuals / tail, 210.4 ± 47.2 individuals / tail, 201.0 ± 54.8 individuals / tail, 76.0 ± 48.5 individuals / It was a tail. The correction values for these values are shown in FIG. The number of parasites of the worms in the 0.5% kujin hot water extraction, 1.0% kujin hot water extraction and 0.5% kujin hot water extraction + 0.5% ethanol extraction is significantly lower than that in the control group. (P <0.01). Accordingly, it has been found that the hot water extract contains a large amount of active ingredients that provide an anti-white spot effect. However, the number of parasites in the 0.5% kujin hot water extraction + 0.5% ethanol extraction tends to be smaller than that in the 0.5% kujin hot water extraction (P = 0.16), compared with the 1.0% kujin hot water extraction. Resulted in significantly fewer results (P <0.05). Therefore, it was thought that the effect was enhanced by adding both the kujin hot water extract and the ethanol extract to the feed as compared with the hot water extract alone. That is, it was found that the active ingredient is contained in both the heat extract and the ethanol extract.

<Examination of the synergistic effect of anti-white spotted insects by adding hot water extract and hot water extract residue ethanol extract>
An attack test with white spotted insects was performed using five fish of red sea bream (average fish weight 6 g) in which disease was not recognized. The test group is fed with feed containing 0.5% of kujin water extract in raw material equivalent (0.5% of kujin hot water extraction), 0.5% of kujin hot water extract and ethanol extract of kujin hot water extraction residue. A total of 3 sections were fed, 0.5% fed feed (0.5% kujin hot water extraction + 0.5% ethanol extraction) and a group fed non-added feed (control). The feeding amount was 3% / day with respect to the fish weight of each section.
The kudin extract was obtained in the same manner as in Example 1. A kujin-containing feed was obtained in the same manner as in Example 3. In the control group, only commercially available initial sample pellets were fed.
In the attack test, each fish is placed in a 100 liter aquarium and fed with each test feed. Three weeks after the start of feeding, 50,000 white spotted caterpillars are placed in each aquarium, and the fish are infected for 2 hours with water stopped. Was done. Thereafter, the animals were bred continuously on the test feed for 2 days. The water temperature during the test period was 24-25 ° C. Water exchange during the test period was 24 rotations / day.
The evaluation was performed by sampling all test fish 3 days after the start of the attack, and comparing the number of parasites of the worms parasitizing the fish and the correction values.
The number of white spotted insects in the control group, 0.5% kudin hot water extraction, and 0.5% kujin hot water extraction + 0.5% ethanol extraction is 146.4 ± 33.2 individuals / tail (average ± standard deviation), 31.0 ± 17.9, respectively. Individuals / tail, 10.4 ± 4.3 individuals / tail. The correction values for these values are shown in FIG. The number of parasites of the worms in the 0.5% kujin hot water extract and 0.5% kujin hot water extract + 0.5% ethanol extract was significantly lower than that in the control group (P <0.01). In addition, the number of parasites in 0.5% kujin hot water extraction + 0.5% ethanol extraction was significantly smaller than that in 0.5% kujin hot water extraction (P <0.05). By adding both hot water extract of hot water extract and ethanol extract of hot water extract residue to the feed, it was reproduced that the effect was enhanced compared with the addition of hot water extract alone.

<Examination of active ingredients-Anti-white spot insect effect of matrine-added feed>
One of the active ingredients of kudin is alkaloid matrine. Therefore, the anti-white spot insect effect of matrine-added feed was examined.
An attack test using white spotted worms was conducted using 10 fish of red sea bream (average fish weight 55 g) in which no disease was recognized. The test plot is a commercially available matrine (ICN
Biomedical Inc) was added to 0 kg, 20 mg, 100 mg, and 500 mg of feed added to 1 kg of feed as an external additive (control group, matrine 20 mg group, matrine 100 mg group, matrine 500 mg group). The feeding amount was 3% / day with respect to the fish weight of each section.
The production of matrine-containing feed was performed by dissolving matrine in an organic solvent and spraying it on commercially available initial feed pellets. Matrine was dissolved in ethanol at a ratio of 100 mg / 100 mL, and this dissolved solution was sprayed to 1 kg of commercially available pellets for initial feed, 20 mL for matrine, 100 mL for 100 mg matrine, and 500 mL for 500 mg matrine. After spraying, the feed was dried at 40 ° C. for 5 hours.
In the attack test, each fish is placed in a 100 liter aquarium and fed with each test feed. Three weeks after the start of feeding, 50,000 white spotted caterpillars are placed in each aquarium, and the fish are infected for 2 hours with water stopped. Was done. Thereafter, the animals were bred continuously on the test feed for 2 days. The water temperature during the test period was 24-25 ° C. Water exchange during the test period was 24 rotations / day.
The evaluation was performed by sampling all test fish 3 days after the start of the attack, and comparing the number of parasites of the worms parasitizing the fish and the correction values.
The numbers of white spotted worms in the control group, matrine 20 mg group, matrine 100 mg group, and matrine 500 mg group are 345.9 ± 134.1 individuals / tail (mean ± standard deviation), 380.6 ± 108.0 individuals / tail, 281.6 ± 90.9 individuals / tail, respectively. 153.4 ± 78.4 individuals / tail. The correction values for these values are shown in FIG. The number of parasites on the moths in the matrine 500 mg group was significantly smaller than that in the control group (P <0.01). Therefore, it was found that one of the active ingredients of anti-white spot insects in the kujin extract is matrine.

<Anti-white spotworm effect of diet supplemented with matrine (Follow-up)>
An attack test using white spotted worms was carried out using 16 fish of red sea bream (average fish weight 55 g) in which no disease was recognized. The test group was divided into two groups, a group for feeding a diet in which 500 mg of matrine was externally added to the feed (500 mg group for matrine) and a group for feeding an additive-free feed (control group). The feeding amount was 3% / day with respect to the fish weight of each section.
Matrine-added feed was produced in the same manner as Example 14.
In the attack test, each fish is placed in a 100 liter aquarium and fed with each test feed. Nine days after the start of feeding, 50,000 white spotted larvae are placed in each aquarium, and the fish are infected for 2 hours with water stopped. Was done. Thereafter, the animals were bred continuously on the test feed for 2 days. The water temperature during the test period was 24-25 ° C. Water exchange during the test period was 24 rotations / day.
The evaluation was performed by sampling all test fish 3 days after the start of the attack, and comparing the number of parasites of the worms parasitizing the fish and the correction values.
The numbers of infestation in the control group and matrine 500 mg group were 38.7 ± 20.6 individuals / tail (mean ± standard deviation) and 7.6 ± 5.9 individuals / tail, respectively. The correction values for these values are shown in FIG. The number of parasites of moths in the 500 mg section of Matrine was significantly less than that in the control section (P <0.01). The results of Example 14 were reproduced.

<Anti-white spotted insect effect of herbal extracts containing matrine other than kujin>
It is known that the extract of yam root which is a crude drug contains matrine as well as kujin. Then, the control effect with respect to white spotted insects of the diet with added soybean root extract was examined.
An attack test using white spotted worms was conducted using 8 fish of red sea bream (average fish weight 65 g) in which disease was not recognized. The test area is fed with feed containing 0.5% or 1.0% of yam-root 20% ethanol extract in terms of raw powder (0.5% of bean-root or 1.0% of bean-root), and is fed with no additive. A total of three wards (control wards). The feeding amount was 3% / day with respect to the fish weight of each section.
A 20% ethanol extract of goat root was obtained in the same manner as in Example 2, and a feed containing goat root was prepared in the same manner as in Example 4.
In the attack test, each fish is placed in a 100 liter aquarium and fed with each test feed. After 18 days from the start of feeding, 110,000 white spotted larvae are placed in each aquarium and allowed to infect for 2 hours with water still. Was done. Thereafter, the animals were bred continuously on the test feed for 2 days. The water temperature during the test period was 24-25 ° C. Water exchange during the test period was 24 rotations / day.
The evaluation was performed by sampling all test fish 3 days after the start of the attack, and comparing the number of parasites of the worms parasitizing the fish and the correction values.
The number of white spotworms in the control group, 0.5% of bean roots, and 1.0% of bean roots is 444.3 ± 131.2 individuals / tail (mean ± standard deviation), 235.9 ± 73.0 individuals / tail, 146.3 ± 55.6 individuals, respectively. / It was a tail. The correction values of these values are shown in FIG. The number of parasites on the moths in 0.5% of the bean roots and 1.0% of the bean roots was significantly less than that in the control group (P <0.01). Therefore, it was confirmed that the herbal extract-added feed containing matrine is effective against white spot infection.

<Effects of cucumber extract-added feed on red sea bream feeding and its improvement-appetite improvement by coating of kudin extract>
When kujin extract-added feed (0.5% or more in raw material equivalent) is administered to red sea bream, feeding tends to be inactive. This phenomenon is not noticeable in several grams of fry, but is often observed in fish over 50 g. Since the kujin extract is bitter, this bitter taste may cause a decrease in food intake. In this test, feeding cucumber extract-added feed to red sea bream with a weight of 50 g or more to see how the appetite decreases, and whether the appetite is improved by coating this extract so that the bitter taste component of the kudin extract does not elute. investigated.
A feeding test was conducted using 13 fish of red sea bream (average fish weight 71.7 g) in which no disease was recognized. The test group was fed with feed containing 1.0% of kujin hot water and ethanol extract as raw powder (1.0% of kudin extract added), and 1.0% of coated kudin extract was added as raw powder. A total of 3 groups were divided into a group fed with feed (coating cucumber extract 1.0% added group) and a group fed with additive-free feed (control group).
The coating of the kudin extract was performed by mixing casein, carrageenan, and kudin extract (hot water extract: Et extract = 1: 1) at a ratio of 10: 1: 15. The method is shown below. The water extract 100g obtained by the preparation method 1 of a kujin extract and the ethanol extract 50g were mixed, and 100g of casein was added in 100 degreeC hot water, and was mixed sufficiently. The pH was adjusted to 7 with an aqueous sodium hydroxide solution, and 10 g of carrageenan was added and stirred well until smooth. After the mixture was cooled to room temperature, the mixture was dried with a freeze dryer. The dried product was pulverized with a mixer to obtain a powder having a 300 μm mesh and a low particle size. This powder was used as a kujin extraction coating. Production of a feed containing a kujin extract coating was performed as follows. White meal, krill meal, squid meal, dextrin, sardine oil, vitamin mixture, mineral mixture, monobasic calcium phosphate, calcium carbonate, gluten, guar gum are mixed according to a conventional method, and a kujin extract coating is 1.0 per feed weight in terms of bulk. Add to the amount and mix well. Water was added to the mixture, pelleted with a granulator, and dried at 40 ° C. for 3 hours. This feed was used as a kujin extract-coated feed. In addition, pellets prepared without adding the kudin extract coating were used for the feed in the control group, and 1.0% kujin hot water and ethanol extract were sprayed on the pellets in the control group to the control group pellets, respectively. .
The feeding test was conducted by storing each fish in a 100 liter water tank and feeding each test feed until they were satiety. The number of feedings was twice a day, and the times were 9:00 am and 16:00 pm. The daily feed for each ward was recorded. The test period was 13 days, and the water temperature during the test period was 16 to 17 ° C. Water exchange during the test period was 13 revolutions / day.
The results of the daily feeding rate are shown in FIG. The feeding rate per fish body weight in the control group, the 1.0% addition of the cucumber extract, and the 1.0% addition of the coated kudin extract was 0.9 ± 0.2% / day, 0.4 ± 0.2% / day, and 0.6 ± 0.2% / day, respectively. It was a day. The feeding rate in the 1.0% addition group of the kujin extract and the 1.0% addition of the coated kudin extract was significantly lower than that in the control group (P <0.01). However, the feeding rate of the coated cucumber extract 1.0% group was significantly higher than that of the kudin extract 1.0% group (P <0.01). Therefore, although a kujin extract addition feed may reduce the appetite of a red sea bream, it was shown that the appetite is improved by coating a kujin extract so that a bitterness ingredient may not elute.

<Improvement of the effect of cucumber extract-added diet on red sea bream feeding-Effect of improving appetite by moist diet>
Moist pellets are feed obtained by mixing raw mash and horse mackerel. Fish have a high preference for moist pellets, and moist pellets induce fish feeding behavior. This study examined whether adding appetite extract to moist pellets would improve appetite of red sea bream.
A feeding test was conducted using 10 fish of red sea bream (average fish weight 148.8 g) in which no disease was recognized. The test plots were fed with moist pellet feed supplemented with 0.4% kujin hot water and ethanol extract each in terms of bulk (Cudin 0.4% moist), and each kujin hot water and ethanol extract was 0.7% in bulk. % Of the moist pellet feed added (% kujin 0.7% moist group) and the group fed the non-added moist pellet feed (control group).
The kujin extract was obtained in the same manner as in Preparation Method 1. Moist pellets were produced by the following method. In the mincher, raw bait (mash): mash = 1: 1 was added and mixed, and then the kujin extract was added to 0.4% or 0.7% in terms of bulk powder and further mixed. After mixing, it was pelletized with a granulator. The control group fed only moist pellets without adding kudin extract.
The feeding test was conducted by storing each fish in a 100 liter water tank and feeding each test feed until they were satiety. The number of feedings was twice a day, and the times were 9:00 am and 16:00 pm. The daily feed for each ward was recorded. The test period was 14 days, and the water temperature during the test period was 19 ± 1 ° C. Water change during the test period was 22 revolutions / day.
The results of daily feeding rate are shown in FIG. Feeding rates in the control group, 0.4% moist group, and 0.7% moist group are 2.3 ± 0.5%, 2.2 ± 0.5%, and 2.4 ± 0.5%, respectively. It was. Therefore, it was shown that the kujin extract can be administered without reducing the appetite of the fish by feeding the kujin extract to the moist pellet and feeding.

<Anti-white spotted insect effect of kudin extract-added feed for fish species other than red sea bream -Effects of flounder on white spotted bug>
The effect of feed addition of cucumber extract against white spotworms was examined using 17 fish in each section of flounder (average fish weight 11.5 g) in which no disease was recognized. The test plots consisted of a total of three plots, one fed with feed containing 0.5 or 2.0% of kujin 20% ethanol extract in terms of bulk powder (0.5% kudin, 2.0% kudin), and the control fed with additive-free feed. It was set as a ward. The amount of feeding was 3% / day with respect to the fish weight of each ward. The kudin extract was obtained in the same manner as in Example 2. A kujin-containing feed was obtained in the same manner as in Example 4.
In the attack test, each fish is placed in a 100 liter tank and fed with each test feed. Two weeks after the start of feeding, 85,000 white spotted larvae are placed in each tank and infected for 2 hours with water stopped. Was done. Thereafter, the animals were bred continuously on the test feed for 2 days. The water temperature during the test period was 24-25 ° C. Water exchange during the test period was 24 rotations / day.
The evaluation was performed by sampling all test fish 3 days after the start of the attack and comparing the number of parasites of the worms parasitizing the moth on one side and the correction value.
As a result of the test, the number of white spotted insects in the control group, the 0.5% kudin group, and the 2.0% kudin group is 7.0 ± 6.1 individuals / tail (mean ± standard deviation), 1.5 ± 1.8 individuals / tail, 0.3 ± 0.5, respectively. Individual / tail. The correction values for these values are shown in FIG. There was no significant difference in the number of parasites of the worms in the 0.5% kujin group compared with the control group, but there was a tendency to decrease (P = 0.26). The number of parasites of worms in the 2.0% kujin group was significantly less than that in the control group (P <0.01). It was shown that the cucumber extract-added diet also prevented the infection of flounder white spotted caterpillars.
From the results of this example, it is considered that the kudin extract-added feed prevents infection of white spotworms of various fishes.

<Effects of kudin extract and goat root extract against flounder white spotted insects>
In Examples 14 to 16 using red sea bream, one of the anti-white spot worm active ingredients in the kudin extract is matrine, and the white bean extract, which is a herbal medicine containing matrine, is the same as the kujin extract. The feed addition effect on insect infection was observed. Then, the control effect with respect to white spotted insects of the diet with added soybean root extract was examined using Japanese flounder.
The effect of dietary addition of goat root extract and cucumber extract against white spotted worms was examined using 17 flounder (average fish body weight 11.4g) in each section with no disease. The test plots are fed with feed containing 0.5% or 1.0% of yam-root 20% ethanol extract in terms of raw powder (0.5% of goat root, 1.0% of bean-root), 20% ethanol extract of kujin A total of 5 districts were fed: 0.5% or 1.0% of feed added to the feed (0.5% kudin, 1.0% kudin) and no control feed (control). Feeding was carried out twice a day, and satiety. A 20% ethanolic extract of goat root was obtained in the same manner as in Example 2. The bean root-containing feed was obtained in the same manner as in Production Example 4.
In the attack test, each fish is placed in a 100 liter aquarium and fed with each test feed. Two months after the start of feeding, 58,000 individuals of white spotted larvae are placed in each aquarium and infected for 3 hours with water stopped. Was done. Thereafter, the animals were continuously raised on the test feed for 2 days. The water temperature during the test period was 24 ± 1 ° C. Water exchange during the test period was 24 rotations / day.
The evaluation was performed by sampling all test fish 3 days after the start of the attack and comparing the number of parasites of the worms parasitizing the moth on one side and the correction value.
The number of white spotted insects in the control group, 0.5% of the bean root, 1.0% of the bean root, 0.5% of the kujin, and 1.0% of the kujin is 111.7 ± 88.4 individuals / tail (mean ± standard deviation), 56.9 They were ± 30.5 individuals / tail, 65.9 ± 95.8 individuals / tail, 41.7 ± 25.8 individuals / tail, and 70.9 ± 65.2 individuals / tail. The correction values of these values are shown in FIG. 16 at a ratio where the value of the control group is 1. The number of parasites on the moths in the 0.5% kudin group was significantly less than that in the control group (P <0.05). There was no significant difference in the number of parasites on the moths in the 0.5% and 1.0% areas of the bean root extract, although there was no significant difference compared to the control area (P values in the 0.5% area were 0.069 and 1.0). The P value of% ward is 0.212). It has been found that the diet supplemented with the bean root extract inhibits the infection of white spotted flounder. Moreover, it was reproduced that the cucumber extract-added feed also prevents the infection of flounder white spotted insects.
Therefore, it was shown that adding a herbal extract containing matrine to the feed is effective against white spotworm infection of various fishes.

  The present invention can provide a composition that has a high antiparasitic activity against parasites such as white spotworms, and that is extremely less affected by side effects of the drug and residual drug on fish. That is, it is possible to provide an orally administrable composition having an antiparasitic action that can be used even when a bath salt cannot be used.

It is explanatory drawing which showed the anti-white spot insect effect of a kujin extract addition feed (* P <0.01). It is explanatory drawing which showed the anti-white spot insect effect (short-term administration) of a kujin extract addition feed (* P <0.01). It is explanatory drawing which showed the white spotworm anthelmintic effect of the kujin extract addition feed. It is explanatory drawing which showed the treatment and survival improvement effect (water temperature transition during a test period) with respect to the white spot insect infection of a kujin extract addition feed. It is explanatory drawing which showed the treatment and survival improvement effect (comparison of the number of parasites 20 days after an attack) (P <0.01) with respect to the white spot insect infection of the feed which added a kujin extract. It is explanatory drawing which showed the treatment and survival improvement effect (comparison of the number of parasites 30 days after an attack) with respect to the white spot insect infection of the feed containing a kujin extract (P <0.01). It is explanatory drawing which showed the treatment and survival improvement effect (comparison of a survival rate) with respect to the white spot insect infection of a kujin extract addition feed (P <0.01). It is explanatory drawing which showed the comparison of the anti-white spot insect effect of a hot water extract and an ethanol extract, and the synergistic effect presence (* P <0.01) by both extract addition. It is explanatory drawing which showed the anti-white spot insect synergistic effect presence / absence (additional test) (* P <0.01) by hot water extract and ethanol extract addition. It is explanatory drawing which showed the anti-white spot insect effect (* P <0.01) of a matrine addition feed. It is explanatory drawing which showed the anti-white spot insect effect (additional test) (* P <0.01) of a diet supplemented with matrine. It is explanatory drawing which showed the anti-white spot insect effect (Yamazu-root extract) (* P <0.01) of the crude drug extract containing matrine. It is explanatory drawing which showed the appetite improvement by the coating of a kujin extract. It is explanatory drawing which showed the appetite improvement by adding a kujin extract to the moist feed. It is explanatory drawing which showed the anti-white spot insect effect (* P <0.01) of the cucumber extract addition feed in Japanese flounder. It is explanatory drawing which showed the anti-white spot insect effect (* P <0.05) of the cucumber extract and goat root addition feed in Japanese flounder.

Claims (20)

An orally administrable composition for preventing and / or treating parasitic diseases for fish containing matrine. The composition for prevention and / or treatment of parasitic diseases according to claim 1, wherein the composition containing matrine is kudin, ground bean root extract or extract. The composition for prevention and / or treatment of parasitic diseases according to claim 2, wherein the extract of kujin is water and / or an organic solvent extract. The composition for prevention and / or treatment of parasitic diseases according to claim 1, 2, or 3, wherein the fish is marine fish. The composition for prevention and / or treatment of parasitic diseases according to claim 1, 2 or 3, wherein the fish is classified as a fish species belonging to the order of perch, puffer, flounder, salmon, catfish, carp. The composition for the prevention and / or treatment of parasitic diseases according to any one of claims 1 to 5, wherein the parasitic diseases are caused by protozoan ciliates or parasites belonging to the flat animal monopods. White spotted insects belonging to the protozoan ciliate, blue clinella, kilodonella, epistilis, sukutka, trichodina, neobenedenia, or neobenedenia belonging to the flatlife monophyte, daktyrogillus, pseudodactyrogillus, microcotile, vivagina, hexaxine The composition for the prevention and / or treatment of parasitic diseases according to any one of claims 1 to 5, wherein the composition is a heterobotulinum or neoheterobotulinum. A method for preventing and / or treating fish parasitic diseases by orally administering a composition containing matrine. Oral administration of 1 to 50 mg / day of fish weight per kg of matrine, 10 to 2000 mg / day of fish weight of 1 kg of fish weight, or 10 to 2000 mg / kg of fish weight of yam root A method for preventing and / or treating a parasitic disease according to claim 8. A method for preventing and / or treating fish parasitic diseases by feeding a feed containing a composition containing matrine. The feed contains a composition containing matrine as a feed for fish farming in an amount of 0.002 to 0.1% in terms of matrine, 0.1 to 5% in terms of raw material of kudin, or 0.1 to 5 in terms of yam root powder The method for preventing and / or treating parasitic diseases according to claim 10, wherein the parasitic diseases are added. The method for preventing and / or treating a parasitic disease according to claim 10 or 11, wherein the matrine-containing composition is processed to make it difficult to elute bitter components. The parasitic disease according to claim 12, wherein the processing for making the bitter taste component of the composition containing matrine difficult to dissolve is encapsulated, microencapsulated, coated tableted, or powdered with a water-insoluble component. A method for preventing and / or treating The method for preventing and / or treating a parasitic disease according to any one of claims 10 to 13, wherein the composition containing matrine is kujin, ground bean root extract or extract. 15. The method for preventing and / or treating parasitic diseases according to claim 14, wherein the extract is a water and / or organic solvent extract. The method for preventing and / or treating a parasitic disease according to any one of claims 10 to 15 , wherein the feed is a moist pellet, EP (Extruded Pellet). The method for preventing and / or treating parasitic diseases according to any one of claims 10 to 16, wherein the fish is marine fish. The method for preventing and / or treating a parasitic disease according to any one of claims 10 to 17, wherein the fish is classified as a fish species belonging to the order of perch, puffer, flounder, salmon, catfish, carp. The method for preventing and / or treating a parasitic disease according to any one of claims 10 to 18, wherein the parasitic disease is caused by a protozoan ciliate or a parasite belonging to a flat animal monopod. White spotted insects belonging to protozoan ciliates, blue clinella, kilodonella, epistilis, sukutka, trichodina, neobenedenia, or neobenedenia belonging to the flatlife monopoda, dactyrogillus, pseudodactyrogillus, microcotile, vivagina, heteraxine The method for preventing and / or treating parasitic diseases according to any one of claims 10 to 18, wherein the method is caused by heterobotulinum or neoheterobotulinum.

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