JP2020162587A - Feed for aquatic animals, growth improving agents for aquatic animals, and methods for rearing cultured fishes using the same feed and agents - Google Patents

Abstract

To provide feed for aquatic animals and growth improving agents for aquatic animals which lead to healthy growth of fishes and can impart strong restoring power to fishes even at a water temperature significantly higher than ordinary growth water temperature, and can improve stress tolerance of not only fishes but also aqueous animals such as crustaceans.SOLUTION: The feed for aquatic animals and growth improving agents for aquatic animals are to be used for fishes and crustaceans growing in water and contain sanguinarine or a derivative thereof or a salt thereof. Not only sanguinarine, zerumbone, etc. are also effective.SELECTED DRAWING: Figure 1

Description

The present invention relates to aquatic animal feed and an aquatic animal growth improver that reduce various stresses on aquatic animals such as fish and crustaceans that grow in water and contribute to healthy growth.

Fish that grow in water, such as oceans and rivers, are less tolerant of temperature changes than terrestrial animals. This is because the change in water temperature is small compared to the change in air temperature, so resistance to large temperature changes is not usually required.

However, due to recent environmental changes such as abnormal weather and global warming, changes in water temperature have become larger than before. Therefore, changes in water temperature are becoming a major problem in fish farming.
In addition, at high water temperatures, the propagation of viruses and bacteria becomes remarkable, and the incidence of fish diseases increases, which is another factor that aggravates the problem.

On the other hand, the growth of many fish is more remarkable as the water temperature is higher, and it is preferable to grow at the highest possible water temperature in order to increase the catch.
Under such circumstances, feeds for improving stress such as water temperature of fish are being studied. For example, a fish feed having an ameliorating effect on stress fortified by DHA or its ester and / or phospholipid has been proposed (for example, Patent Document 1).

JP-A-8-98659

Although the fish feed disclosed in Patent Document 1 can enhance the high temperature resistance to red sea bream, it has not had the effect of significantly improving the survival rate in a high water temperature environment. For example, it has been difficult to bring about healthy growth at a temperature higher than the normal growth temperature by 10 ° C. or more.

The present invention has been made to solve the above problems, and it is possible to bring about healthy growth of fish even at a water temperature much higher than the normal growth temperature, and not only fish but also crustaceans. It provides a feed for aquatic animals that enhances the stress tolerance of aquatic animals such as crustaceans, and a growth improver for aquatic animals.
In addition, aquatic animal feed and water that improve resistance to various stresses such as light such as low temperature and ultraviolet rays, pH, osmotic pressure, pathogens, diseases, chemical substances, chemicals, water pressure, starvation, and rot as well as high temperature. It provides a growth improver for animals.

The aquatic animal feed according to the present invention is used for fish and crustaceans growing in water, and is characterized by containing sanguinarine or a derivative thereof, or a salt thereof.

The aquatic animal feed according to the present invention is used for fish and crustaceans growing in water, and contains zerumbon as a main component.

The aquatic animal feed according to the present invention is used for fish and crustaceans growing in water, and is a cis-trans isomer of salicylic acid, polyamine, geldanamycin, isothiocyanate, trimethylglycine, perillaldehyde, piperine, and pyripene. It is mainly composed of crustacean, which is the body, or citral.

The aquatic animal feed according to the present invention is used for fish and crustaceans growing in water, and is a plant of pepper, yabatsui, arnica, oguruma, inula linarii, salvia, majuram, scutellaria baicalensis, amomum villosum, or shukusha. The main component is an extract or crushed plant product.

Further, the growth enhancer for aquatic animals according to the present invention is used for fish and crustaceans growing in water, and is characterized by containing sanguinarine or a derivative thereof, or a salt thereof.

Further, the growth-improving agent for aquatic animals according to the present invention is used for fish and crustaceans growing in water, and contains zerumbon as a main component.

The aquatic animal growth-improving agent according to the present invention is used for fish and crustaceans growing in water, and is a cis-cis of salicylic acid, polyamine, geldanamycin, isothiocyanate, trimethylglycine, perillaldehyde, piperine and piperine. It is mainly composed of the trans isomer, kabisin or citral.

Further, the growth enhancer for aquatic animals according to the present invention is used for fish and crustaceans growing in water, and is used for pepper, yabatsui, arnica, oguruma, inula linarii, salvia, majuram, scutellaria baicalensis, amomum villosum, or sukusha. The main component is the plant extract of Scutellaria baicalensis or crushed plant product.

Furthermore, the method for breeding farmed fish according to the present invention is characterized by using the above-mentioned feed for aquatic animals.

The aquatic animal feed and the aquatic animal growth improver according to the present invention reduce the stress given to the aquatic animals by the environment and bring about healthy growth by giving them to aquatic animals such as fish and crustaceans growing in water. It is a thing.
In particular, the aquatic animal feed and the aquatic animal growth improver according to the present invention have a new property of being able to impart high high temperature stress resistance and resilience to aquatic animals, and are healthy for aquatic animals. It exerts a great effect on healthy growth.
That is, it imparts high-temperature stress tolerance that enables growth even in a high water temperature environment of 10 ° C. or higher than the optimum temperature at which aquatic animals normally grow.
In addition, even in the case of falling into a state showing an extreme abnormality that makes it almost impossible to swim, if the water temperature drops, the resilience of starting swimming normally again is imparted.
It also improves resistance to various stresses such as light such as low temperature and ultraviolet rays, pH, osmotic pressure, pathogens, diseases, chemical substances, chemicals, water pressure, starvation, and rot as well as high temperature.

It is a verification experiment result when the feed for aquatic animals according to the present invention and the grown zebrafish of the growth improver for aquatic animals were used, and it is a figure which shows the relationship between the water temperature and the survival rate in each test group. It is a verification experiment result when the feed for aquatic animals and the growth improver for aquatic animals zebrafish according to the present invention were used, and it is a figure which shows the relationship between the water temperature and the survival rate in each test group. It is a verification experiment result when the feed for aquatic animals according to the present invention and the growth improver for aquatic animals, Minaminuma shrimp, were used, and it is a figure which shows the relationship between the water temperature and the survival rate in each test group.

In each of the following embodiments, the feed for aquatic animals and the growth improver for aquatic animals according to the present invention, and the method for raising farmed fish using them will be described. The following description discloses a good example of the present invention, and the present invention is not limited to the embodiment.
The aquatic animal feed and the aquatic animal growth-improving agent according to the present invention consist of the substances shown in the following embodiments, and even if these substances are given to the aquatic animals as feed daily or regularly. It may be given as a growth-improving agent for a specific purpose such as growth improvement, specific stress reduction, or disease prevention. In the following, in order to avoid redundant expressions, "aquatic animal feed and aquatic animal growth improver" are expressed by the term "aquatic animal growth improver".

The growth improver for aquatic animals according to the present invention is used for fish and crustaceans growing in water, and reduces various environmental stresses, promotes growth, or improves quality. In addition, it may have the effect of improving resistance to disease.
In particular, with regard to various environmental stresses, the stress is reduced, and even if the stress causes an abnormality such as inability to swim, when the stress factor disappears, a strong resilience such as starting swimming normally is given. Has the effect of stress.

Further, by breeding aquatic animals using the growth-improving agent for aquatic animals according to the present invention, the above-mentioned effects can be brought about in general, but in particular, seawater acclimatization of farmed fish, breeding of larvae, or It will be a breeding method suitable for disease control.

Embodiment 1.
In the present embodiment, the results of a verification experiment on the effect of improving resistance to high temperature stress, which is one of the most susceptible stresses to aquatic organisms such as fish and crustaceans growing in water, will be described.
<Regarding the main component>
The growth enhancer for aquatic animals according to the present invention contains sanguinarine or a derivative thereof, or a salt thereof as a main component.
Sanguinarine is an alkaloid with a benzophenanthridine skeleton that is often found in poppy plants such as Macleaya cordata, California poppy, Greater celandine, and Bloodroot.

Any plant may be used as long as it is a poppy family plant or a plant containing a derivative thereof, but in the present embodiment, the active ingredient was extracted from Macleaya cordata containing a large amount of sanguinarine.
Macleaya cordata contains some alkaloids in addition to sanguinarine, but these may be contained. In particular, chelerythrine, which is an alkaloid having a benzophenanthridine skeleton similar to sanguinarine, has the same efficacy as sanguinarine as a sanguinarine derivative.
Other alkaloids contained in Macleaya cordata are kerylbin, sangiltin, sangilbin, keryltin, marcapine, and protopine, and these alkaloids, like sanguinarine and kerellisulin, have a predetermined effect, and therefore have a small amount of these. You may.

The active ingredient was extracted by a solvent extraction treatment in which Macleaya cordata was dried and then immersed in a solvent. As the solvent, alcohol such as ethanol, acid or the like can be used. The extraction method is not particularly limited, and any method may be used as long as it is a method for extracting the active ingredient from the plant.
As described above, in this verification experiment, the method of extracting the active ingredient from Macleaya cordata was used, but a crushed product obtained by crushing the plant into powder after drying may be used.

<Verification experiment 1>
Experiments were conducted to verify the efficacy of sanguinarine or its derivatives extracted from Macleaya cordata on fish. The experiment was carried out using zebrafish. The experiment was performed twice, the first using grown zebrafish and the second using growing zebrafish.

The procedure of the verification experiment will be described below. Table 1 shows the experimental conditions.

1. 1. Test plots 10 zebrafish were placed in each of the 4 aquariums to make each test plot. Each test plot was fed regular diet and Macleaya cordata extract daily during the acclimatization period. The amount of Macleaya cordata extract in each test group was 0 times (none), 1 time, 10 times, and 100 times. Here, the 1-fold amount means that the weight of sanguinarine contained in the Macleaya cordata extract is 125 μg per body weight (kg) of zebrafish. A 10-fold dose of the test plot was given 10-fold dose, and a 100-fold dose of the test plot was given 100-fold dose of Macleaya cordata extract daily.
The acclimatization period was 28 days in the first experiment and 6 days in the second experiment. The water temperature during the acclimatization period was around 25 ° C, which is the optimum temperature for zebrafish.

2. Preliminary temperature rise was performed over 24 hours on the day before the test day. The temperature was set to 35 ° C. in the first experiment and 30 ° C. in the second experiment.
3. 3. On the day of the test On the day of the test, the water temperature was raised under the temperature rising conditions shown in Table 1. Then, every 5 minutes, the survival rate of zebrafish in each test plot was measured.

The relationship between the time from the start of temperature rise, the water temperature, and the survival rate in each test group is shown in FIGS. 1 and 2. FIG. 1 shows the results of the first verification experiment, and FIG. 2 shows the results of the second verification experiment.
In each figure, the horizontal axis is the elapsed time from the start of temperature rise, and the vertical axis is the survival rate and the water temperature in each test group.

In the first experiment (Fig. 1), the water temperature was 43 ° C, which was 18 ° C higher than the optimum growth temperature of zebrafish, so that the overall survival rate was low. In the 0-fold and 1-fold test plots, the survival rates were 0% and 10%, respectively, when the water temperature was 43 ° C.
However, in the 10-fold test plot, the survival rate did not decrease at this point, and the survival rate was 1 for nearly 1 hour.
I was able to keep 00%.
In the 100-fold test group, a survival rate of 50% was obtained even after about 4 hours had passed after the water temperature was set to 43 ° C.
In the 1x and 10x dose test plots, even living zebrafish were in a state of imbalance or inability to swim, and eventually died. On the other hand, in the 100-fold dose test plot, all surviving zebrafish continued to swim normally.

In the second experiment (FIG. 2), the maximum water temperature was 41 ° C., so that the survival rate was high even after 5 hours from the start of temperature rise in the 10-fold and 100-fold test plots. In particular, in the 100-fold dose test plot, 100% survival rate was obtained, and all of them continued to swim normally.
In the 10-fold dose test plot, the surviving zebrafish became imbalanced or incapable of swimming, but subsequently recovered to normal swimming when the water temperature was lowered.
In addition, it was verified that the acclimatization period of 6 days was sufficient.

From the above experimental results, it was found that the Macleaya cordata extract has the effect of improving the heat stress tolerance of zebrafish. It was also verified that resistance can be further improved by increasing the dose to be administered.

In particular, in the 10-fold and 100-fold test plots, a remarkable effect was obtained that the plants could grow even at a water temperature 15 ° C. or higher higher than the optimum growth temperature of 25 ° C.
It was also found that even if the patient showed a very serious symptom of being unable to swim, lowering the water temperature would recover until he could swim normally.

It should be noted that the Macleaya cordata extract works effectively not only for changes in water temperature, but also for changes in water pressure, pH, changes in osmosis, ultraviolet irradiation, heat / low temperature, chemical substances, chemicals, and rot. It was separately confirmed that stress related to environmental factors can be reduced. Alternatively, it was confirmed that resistance to microorganisms such as viruses, bacteria, and molds also increased. It also improves resistance to hunger. That is, it was found that resistance to various stresses as well as heat is improved.

The above results were valid not only for zebrafish, but also for other fish such as medaka, guppy, rainbow trout, Japanese eel, mahaze, midorifugu, flounder, red sea bream, sardines, and tilapia.

<Verification experiment 2>
Next, a similar verification experiment was conducted on the crustacean Minaminuma shrimp. Table 2 shows the experimental conditions. In addition, the test group was set to two test groups of 0 times amount and 100 times amount.
FIG. 3 shows the relationship between the time from the start of temperature rise, the water temperature, and the survival rate in each test group.

As shown in FIG. 3, it was confirmed that the Macleaya cordata extract works effectively also on the crustacean Minaminuma shrimp.

<Verification experiment 3>
Zerumbon, which is the essential oil component of ginger plants, especially ginger (commonly known as white turmeric) of the genus Zingiberaceae, was able to impart similar stress tolerance. Zernbon is a type of annular sesquiterpene.
As shown below, administration of Zerumbon also significantly improved the survival rate against rising water temperature in aquatic animals such as fish and crustaceans, as well as the strong resilience of Macleaya cordata extract. It was confirmed that it could be granted.

A grown zebrafish was used in the experiment. The average total length of zebrafish is 34 mm and the average body weight is 370 mg.
100 zebrafish were placed in each of the four aquariums and used as each test plot. Each test plot was fed regular diet and ginger extract daily for a 6-day acclimatization period. The water temperature during the acclimatization period is around 25 ° C. The amount of ginger extract in each test group was 0 times (none), 1 time, 10 times, and 100 times. Here, the 1-fold amount is an amount in which the weight of zebrafish contained in the ginger extract is 125 μg per body weight (kg) of zebrafish.

2. Preliminary temperature rise to 30 ° C. on the day before the test day. Then, the temperature was maintained at 30 ° C. for 24 hours.
3. 3. On the day of the test On the day of the test, the temperature was raised from 30 ° C. to 2.5 ° C. per hour, and after 4 hours, the water temperature was raised to 40 ° C. Then, the water temperature of 40 ° C. was maintained for 30 minutes, and the number of normal zebrafish in each test plot was counted. A normal zebrafish is an individual who has not lost equilibrium or become unable to swim. The number of normal zebrafish in each test plot is shown below.

Normal population / 100 tails in each test group

Formulation-free (0-fold dose test plot)
4 tails

Formulation: Ginger extract Zerumbon 1x: 76 10x: 92 100x: 100

It can be seen that normal zebrafish is greatly increased by feeding ginger extract. In particular, all zebrafish were normal in the 100x plot.

Further, when the water temperature was lowered to 25 ° C. after the above measurement in the 1-fold group and the 10-fold group, the zebrafish in the unbalanced state recovered to a state in which they could swim normally.
On the other hand, in the 0-fold dose test group in which no preparation was used, even if the water temperature was lowered to 25 ° C., the zebrafish in the imbalanced state did not recover and died.

From the above experimental results, it was found that the ginger extract has an effect of improving the heat stress tolerance of zebrafish. It was also verified that resistance can be further improved by increasing the dose to be administered. Furthermore, it was found that it has the effect of recovering damage caused by high water temperature.
It was separately confirmed that the ginger extract has the effect of improving heat stress tolerance also for the crustacean Minaminuma shrimp.

<Verification experiment 4>
In addition, salicylic acid, polyamine, Benzoquinone ansamycin antibiotic geldanamycin, isothianate, which is a pungent component of radish and wasabi, trimethylglycine, which is abundant in vegetables and mushrooms, and perill, which is a type of Labiatae plant. Even when aldehyde, piperine contained in pepper fruits and its cis-trans isomer, cabisin, or citral, which is a characteristic aroma component of lemon, is administered, to aquatic animals such as fish and shellfish, It was confirmed that the survival rate was improved with respect to the rise in water temperature. It was also confirmed that the resilience was improved.

<Verification experiment 5>
Furthermore, it was confirmed that the plant extracts of the following plants were also administered to aquatic animals such as fish and crustaceans to improve the survival rate against an increase in water temperature.
Zingiberaceae, Zingiberaceae, Yabatsui, a above-ground dry medicine of Sawahiyodori, Arnica, a perennial plant of Asteraceae, Oguruma of Asteraceae, Hosobaoguruma of Asteraceae, Salvia, a herb native to Brazil belonging to the genus Sage of Labiatae, These plant extracts are also related to Majuram, which is also a perennial plant of the Labiatae family and used as an herb, Koganebana, which is also a perennial plant of the genus Asteraceae, Yoshunsha, which is a perennial plant of the Asteraceae family, and Shuksha, which is also a perennial plant of the Asteraceae family. It was effective in improving stress tolerance. At the same time, it was confirmed that the resilience to recover from serious symptoms can be imparted.
Regarding the above plants, the same effect can be obtained by using a crushed plant product instead of the plant extract.

Embodiment 2.
In the first embodiment, the verification experiment results on the resistance improving effect on high temperature stress have been described, but in the present embodiment, the verification experiment results on the resistance improving effect on various other stresses will be described.

<Verification experiment 6>
First, a verification experiment on low water temperature stress tolerance was conducted.
As the preparation, (A) sanguinarine or a derivative thereof extracted from Macleaya cordata, and (B) zerumbone, which is the essential oil main component of ginger of the genus Zingiberaceae, were used. The experiment used grown zebrafish. The average total length of zebrafish is 32 mm and the average body weight is 329 mg.

The procedure of the verification experiment will be described below.

1. 1. Test group The respective amounts of the above preparations (A) and (B) were taken as 1-fold amount, 10-fold amount, and 100-fold amount. Therefore, there are a total of 6 types of preparations. Here, the 1-fold amount means that, for the preparation (A), the weight of sanguinarine contained in the Macleaya cordata extract is 125 μg per body weight (kg) of zebrafish. On the other hand, for the preparation (B), the weight of zebrafish contained in the ginger extract is 125 μg per body weight (kg) of zebrafish.
Each of the seven aquariums was filled with 100 zebrafish and used as each test plot. In each test plot, a normal diet and one of the above six formulations were given daily during the acclimatization period. In one other test plot, only normal diet was given daily during the acclimatization period (0x dose of test plot).
The acclimatization period was 16 days. The water temperature during the acclimatization period was around 25 ° C, which is the optimum temperature for zebrafish.

2. On the day of the test On the day of the test, the water temperature in the water tank was lowered from 25 ° C. to 4 ± 1 ° C. over 1 hour. After that, this low water temperature was maintained for 13 hours. After 13 hours, the number of normal zebrafish in each test plot was counted. A normal zebrafish is an individual who has not lost equilibrium or become unable to swim. The number of normal zebrafish in each test plot is shown below.

Normal population / 100 tails in each test group

Formulation-free (0-fold dose test plot)
17 tails

Formulation (A) Macleaya cordata extract Sanguinarine 1x: 63 10x: 84 100x: 100

Formulation (B) Ginger extract Zerumbon 1x: 46 10x: 66 100x: 100

It can be seen that normal zebrafish is significantly increased by giving formulation (A) or (B). It was also verified that resistance can be further improved by increasing the dose.
In particular, in the 100-fold group of preparations (A) and (B), all zebrafish were normal.

Further, when the water temperature was raised to 21 ° C. after the above measurement in the 1-fold group and the 10-fold group of the preparations (A) and (B), the zebrafish in the imbalanced state recovered to a state where it could swim normally. ..
On the other hand, in the 0-fold dose test group in which no preparation was used, even if the water temperature was raised to 21 ° C., the zebrafish in the imbalanced state did not recover and died.

From the above experimental results, it was found that the Macleaya cordata extract and the ginger extract have the effect of improving the low water temperature stress tolerance. It was also verified that resistance can be further improved by increasing the dose to be administered. Furthermore, it was found that it has the effect of recovering damage caused by low water temperature.
It was also confirmed separately that Macleaya cordata extract and Hanashoga extract have the effect of improving low water temperature stress tolerance and the effect of recovering damage caused by low water temperature, even for the crustacean Minaminuma shrimp. did.

When aquaculture is carried out in winter, the water temperature may be artificially raised to the optimum growth temperature for fish and crustaceans for the purpose of promoting growth and preventing diseases. If this preparation is used to improve low water temperature stress tolerance, it is possible to minimize the artificial rise in water temperature and reduce energy costs. Alternatively, it is possible to breed fish in warm waters in cold regions, and the range of selection of farmed fish can be expanded.

<Verification experiment 7>
Next, a verification experiment on low pH stress tolerance was conducted.
As in the verification experiment 6, the preparations used were a total of 6 types in which the amounts of the preparations (A) and (B) were changed in 3 steps. The acclimatization period was 25 days. The average total length of the zebrafish used was 34 mm and the average body weight was 331 mg.
Dilute hydrochloric acid was used to lower the pH of the aquarium. Dilute hydrochloric acid was infused into a water tank, and the pH value was lowered from 7.0 to 3.0 over 4 hours, and then this low pH value was maintained for 30 minutes. At that time, the number of normal zebrafish in each test plot was counted. The number of normal zebrafish in each test plot is shown below.

Normal population / 100 tails in each test group

Formulation-free (0-fold dose test plot)
15 tails

Formulation (A) Macleaya cordata extract Sanguinarine 1x: 26 10x: 63 100x: 90

Formulation (B) Ginger extract Zernbon 1x: 44 10x: 82 100x: 93

It can be seen that normal zebrafish is significantly increased by giving formulation (A) or (B). In particular, it was found that the ginger extract (formulation (B)) has a greater effect.

From the above experimental results, it was found that Macleaya cordata extract and Ginger extract have the effect of improving low pH stress tolerance. In particular, ginger extract showed a remarkable effect. It was also verified that resistance can be further improved by increasing the dose to be administered.
In addition, it was separately confirmed that the Macleaya cordata extract and the Hanashoga extract have an effect of improving low pH stress tolerance, and the Hanashoga extract in particular has a remarkable effect on the crustacean Minaminuma shrimp.

In artificial captivity, the pH can be extremely high, and it is important to tolerate low pH stress in order to keep fish and shellfish healthy. In addition, at aquaculture sites, as a countermeasure against diseases, fish and shellfish are bred in a low pH environment in which the growth of bacteria and bacteria can be suppressed. By improving resistance to low pH stress, it becomes possible to breed fish and shellfish in a healthy manner in a low pH environment suitable for prevention or treatment of various diseases.
Further, in a low pH environment, the meat quality may be denatured and the quality may be deteriorated. However, by improving the resistance to low pH stress, the meat quality degeneration called "burnt meat" is prevented and the deterioration of the quality is suppressed. can do.

<Verification experiment 8>
In verification experiment 7, stress tolerance to ammonia was verified.
As with the verification experiments 6 and 7, the preparations used were a total of 6 types in which the amounts of the preparations (A) and (B) were changed in 3 steps. The acclimatization period was 6 days. The average total length of the zebrafish used was 33 mm and the average body weight was 324 mg.
Ammonia water was infused into the water tank over 4 hours to raise the ammonia concentration in the water tank to 150 ppm. At this time, the pH value increased from 7.0 to 9.5. After that, the ammonia concentration was maintained at 150 ppm for 30 minutes. At that time, the number of normal zebrafish in each test plot was counted. The number of normal zebrafish in each test plot is shown below.

Normal population / 100 tails in each test group

Formulation-free (0-fold dose test plot)
35 tails

Formulation (A) Macleaya cordata extract Sanguinarine 1-fold group: 72 fish 10-fold group: 80 fish 100-fold group: 91 fish

Formulation (B) Ginger extract Zerumbon 1x: 76 10x: 94 100x: 100

It can be seen that normal zebrafish is significantly increased by giving formulation (A) or (B). In particular, it was found that the ginger extract (formulation (B)) has a greater effect. It was also verified that resistance can be further improved by increasing the dose to be administered.
Similarly, it was confirmed that administration of Macleaya cordata extract and Hanashoga extract to the crustacean Neocaridina denticulata increases the number of normal individuals under high ammonia concentration compared to the case without administration. In particular, it was separately confirmed that the ginger extract showed a greater effect.

When food in water rots, the ammonia concentration rises significantly, damaging fish and shellfish. For example, elevated ammonia levels can cause fish to lose balance, increase excitability, increase respiratory volume, heart rate, and oxygen consumption, ultimately leading to convulsions, coma, and death. It is also said to be a factor that causes a decrease in hatching rate, a decrease in growth rate, pathological changes in gill tissue, and the like. At aquaculture sites, excessive feeding may occur, so it is important to take measures against ammonia. In addition, in artificial captivity, the ammonia concentration may become an extreme value due to other factors.
High pH also damages and damages the mucous membranes on the body surface of fish.
Therefore, by improving the stress tolerance to ammonia and high pH, it becomes possible to breed fish and shellfish in a healthy manner.

<Verification experiment 9>
Next, the resistance to nitrite stress was verified.
As with the verification experiments 6 to 8, the preparations used were a total of 6 types in which the amounts of the preparations (A) and (B) were changed in 3 steps. The acclimatization period was 6 days. The average total length of the zebrafish used was 32 mm and the average body weight was 312 mg.
An aqueous sodium nitrite solution was infused into a water tank, the sodium nitrite concentration was raised to 150 ppm over 4 hours, and this concentration was maintained for 30 minutes thereafter. At that time, the number of normal zebrafish in each test plot was counted. The number of normal zebrafish in each test plot is shown below.

Normal population / 100 tails in each test group

Formulation-free (0-fold dose test plot)
18 tails

Formulation (A) Macleaya cordata extract Sanguinarine 1-fold group: 63 fish 10-fold group: 81 fish 100-fold group: 95 fish

Formulation (B) Ginger extract Zerumbon 1x: 64 10x: 78 100x: 92

It can be seen that normal zebrafish is significantly increased by giving formulation (A) or (B). It was also verified that resistance can be further improved by increasing the dose to be administered.
In addition, it was separately confirmed that the administration of Macleaya cordata extract and Hanashoga extract to the crustacean Neocaridina denticulata increases the number of normal individuals under high sodium nitrite concentration compared to the case without administration. ..

When food in water rots, not only the ammonia concentration but also the nitrite concentration rises significantly, damaging fish and shellfish. At aquaculture sites, excessive feeding may occur, so it is important to take measures against nitrite. In addition, in artificial captivity, the nitrite concentration may become an extreme value due to other factors. Therefore, by improving the resistance to nitrite stress, it becomes possible to breed fish and shellfish in a healthy manner.

<Verification experiment 10>
Next, the tolerance to light stress was verified.
The amount of the preparation used is 100 times the amount of each of the preparations (A) and (B). The acclimatization period was 16 days. The average total length of the zebrafish used was 33 mm and the average body weight was 341 mg.
Light stress was applied by irradiation with an ultraviolet lamp. The wavelength of the ultraviolet lamp is 253.7 nm, and the output is 4.9 W. An ultraviolet lamp was installed at a position 25 cm from the water surface, and continuous irradiation was performed for 22 hours. At that time, the number of normal zebrafish in each test plot was counted. The number of normal zebrafish in each test plot is shown below.

Normal population / 100 tails in each test group

Formulation-free (0-fold dose test plot)
17 tails

Formulation (A) Macleaya cordata extract Sanguinarine 100 times group: 65 fish

Formulation (B) Ginger extract Zernbon 100 times group: 100 fish

It can be seen that normal zebrafish is significantly increased by giving formulation (A) or (B). In particular, it was found that the ginger extract (formulation (B)) has a greater effect.
Furthermore, it was separately confirmed that the effects of ultraviolet stress can be reduced by administering Macleaya cordata extract and Hanashoga extract to the crustacean Minaminuma shrimp. It was also confirmed that the ginger extract shows a more effective effect of reducing UV stress.
Under normal environment, the light that causes the greatest damage to living organisms is ultraviolet rays with large photon energy, and by improving the resistance to ultraviolet stress, it is possible to improve the resistance to various light stresses such as visible light and infrared rays. Conceivable.

Strong light stress can damage seafood and adversely affect growth and quality. In particular, ultraviolet stress causes the pigments of seafood to turn black, resulting in a decrease in commercial value.
Therefore, by improving the resistance to light stress, it is possible to breed fish and shellfish in a healthy manner, maintain the original body color of the fish species, and prevent a decrease in commercial value.

<Verification experiment 11>
Next, the resistance to drug stress was verified.
The preparations used are 100 times the amount of each of the preparations (A) and (B), and three types of mixed preparations of the preparation (A) 50 times and the preparation (B) 50 times. The acclimatization period was 6 days. In addition, a 1-year-old cherry salmon fish having an average weight of 83 g was used. Fifty cherry salmon were bred in each test plot.
As the drug, phenoxyethanol, which is used for anesthesia and euthanasia of fish, was used. A chemical water tank in which phenoxyethanol was dissolved at a ratio of 0.6 ml to 1 liter of seawater was separately prepared, and after sufficient aeration, cherry salmon was added to this chemical water tank for 5 minutes. Then, it was returned to a normal seawater tank, and the number of cherry salmon surviving in each test group was measured after 24 hours had passed. The surviving numbers of cherry salmon in each test plot are shown below.

Surviving population / 50 tails in each test group

Formulation-free (0-fold dose test plot)
41 fish (survival rate: 82%)

Formulation (A) Macleaya cordata extract Sanguinarine 100 times group: 46 fish (survival rate: 92%)

Formulation (B) Ginger extract Zernbon 100 times group: 47 fish (survival rate: 94%)

Formulation (A) 50 times amount + Formulation (B) 50 times amount
50 fish (survival rate: 100%)

Survival was improved by using formulation (A) or (B). And, by using both preparations in combination, all cherry salmon could be survived.
From the above, it was confirmed that the preparation (A) or (B) can improve the resistance to drug stress, and in particular, the effect can be enhanced by using both preparations in combination.

While various drugs are used for the prevention and treatment of diseases, they can also stress seafood, inhibit growth and degrade quality. Therefore, by improving the resistance to drug stress, it becomes possible to use various drugs with peace of mind.

<Verification experiment 12>
In this verification experiment, resistance to pathogens was verified.
The preparations used are 100 times the amount of each of the preparations (A) and (B), and three types of mixed preparations of the preparation (A) 50 times and the preparation (B) 50 times. The acclimatization period was 6 days. In addition, a 0-year-old Ranchu fish, which is a kind of goldfish, was used. 100 Ranchu were bred in each test plot.
Five ranchus infected with columnaris disease were put into each test group, and after swimming for 3 days, only infected fish were taken out, and two days later, the number of infected ranchus was counted in each test group. The number of infected Ranchu in each test plot is shown below. The population was classified into a serious population, a mild population, and a healthy population.

Number of infected individuals / 100 tails in each test group

Formulation-free (0-fold dose test plot)
Severe: 100 tails Mild: 0 tails Healthy: 0 tails

Formulation (A) Macleaya cordata extract 100 times the amount of sanguinarine
Serious: 0 tails Mild: 13 tails Healthy: 87 tails

Formulation (B) Ginger extract Zernbon 100 times
Serious: 0 tails Mild: 18 tails Healthy: 82 tails

Formulation (A) 50 times amount + Formulation (B) 50 times amount
Serious: 0 tails Mild: 0 tails Healthy: 100 tails

The infection rate could be significantly reduced by using the preparation (A) or (B). In addition, the symptoms of infected Ranchu were mild and were completely cured in about one week. By using both preparations in combination, the infection rate could be reduced to zero.
On the other hand, in the test plot where no preparation was used, all Ranchu were infected and fell into a serious condition, and all the infected Ranchu subsequently died.

From the above, it was demonstrated that the preparation (A) or (B) improved the resistance to the pathogen, and in particular, the effect was remarkably enhanced by the combined use of both preparations. Moreover, even when infected, the symptom could be suppressed to a mild state, and the patient recovered early.

<Verification experiment 13>
In addition, salicylic acid, polyamine, Benzoquinone ansamycin antibiotic geldanamycin, isothianate, which is a pungent component of radish and wasabi, trimethylglycine, which is abundant in vegetables and mushrooms, and perill, which is a type of Labiatae plant. Even when aldehyde, piperine contained in pepper fruits and its cis-trans isomer, cabisin, or citral, which is a characteristic aroma component of lemon, is administered, to aquatic animals such as fish and shellfish, It was confirmed that the resistance to each stress of low water temperature, low pH, high pH, ammonia, nitral salt, ultraviolet rays, chemicals and pathogens was improved. It was also confirmed that the resilience was improved.

<Verification experiment 14>
Furthermore, the plant extracts of the following plants can also be administered to aquatic animals such as fish and crustaceans to prevent stress from low water temperature, low pH, high pH, ammonia, nitrite, ultraviolet rays, drugs, and pathogens. It was confirmed that the resistance was improved.
Zingiberaceae, Zingiberaceae, Yabatsui, a above-ground dry medicine of Sawahiyodori, Arnica, a perennial plant of Asteraceae, Oguruma of Asteraceae, Hosobaoguruma of Asteraceae, Salvia, a herb native to Brazil belonging to the genus Sage of Labiatae, These plant extracts are also related to Majuram, which is also a perennial plant of the Labiatae family and used as an herb, Koganebana, which is also a perennial plant of the genus Asteraceae, Yoshunsha, which is a perennial plant of the Asteraceae family, and Shuksha, which is also a perennial plant of the Asteraceae family. It was effective in improving stress tolerance. At the same time, it was confirmed that the resilience to recover from serious symptoms can be imparted.
The same effect can be obtained by using a crushed plant product instead of the plant extract for the above-mentioned plants.

Embodiment 3.
In the first and second embodiments, the effect of improving resistance to various stresses was verified by a water tank experiment. In this embodiment, the results of verification experiments at the aquaculture site will be described.

<Verification experiment 15>
In this verification experiment, a field test was conducted according to the cultivation method used at the actual grouper farming site.
The formulation used is 1 times the amount of formulation (A). The period of feeding the preparation and feed was 30 days, and the fish used were grouper fry. Fry of 50 groupers were bred in each test plot.
The average body weight at the start of breeding in each test plot and the average body weight after breeding for 30 days are shown below.

Average body weight / average value of 50 fish in each test group

Formulation-free (0-fold dose test plot)
At the start of breeding: 42 g After breeding for 30 days: 53 g (Growth rate: 26.2%)

Formulation (A) Macleaya cordata extract Sanguinarine
At the start of breeding: 42 g After breeding for 30 days: 58 g (Growth rate: 38.1%)

The growth rate was improved by using the preparation (A). It is considered that this factor is that sanguinarine, which is an extract of Macleaya cordata, has a growth-promoting effect, or that it promotes healthy growth by improving resistance to various stresses.
Unlike aquariums in the laboratory, a lot of stress is generated in the actual aquaculture site. For example, high temperature or low temperature stress, or stress due to a sudden temperature change can be considered first. In addition, light stress such as ultraviolet rays is constantly generated. Furthermore, psychological stress due to high-density breeding, physical stress due to contact between individuals, and stress due to changes in seawater pH are also considered to have an adverse effect on growth. There are concerns about stress caused by an increase in ammonia concentration and nitrite concentration due to spoilage of food, and diseases caused by pathogens.

In an actual aquaculture site, it is difficult to sufficiently control the environment, and fish and shellfish are subject to a lot of environmental stress. Therefore, it is considered that the growth rate is improved by increasing the resistance to them.

<Verification experiment 16>
Next, a field test was conducted at the Yamame fish farm.
The preparations used are 100 times the amount of each of the preparations (A) and (B), and three types of mixed preparations of the preparation (A) 50 times and the preparation (B) 50 times. The period of feeding the preparation and feed was 35 days, and the fish used was a 1-year-old yamame trout with an average weight of 36 g. 100 yamame trout were bred in each test plot.
The breeding water temperature during the verification period was around 10 ° C, and the water from the mountain stream was flushed as it was, which was a condition in which bacterial gill disease was likely to occur. Flavobacterium brunchiofila, a bacillus that causes bacterial gill disease, becomes most active at water temperatures of 10 ° C to 15 ° C.
During the test period, bacterial gill disease developed and mortality occurred. The number of mortality and survival rate considered to be caused by bacterial gill disease in each test plot on the 35th day from the start of breeding are shown below.

Number of deceased individuals / 100 tails in each test area

Formulation-free (0-fold dose test plot)
26 fish (survival rate: 74%)

Formulation (A) Macleaya cordata extract 100 times the amount of sanguinarine
8 fish (survival rate: 92%)

Formulation (B) Ginger extract Zernbon 100 times
14 fish (survival rate: 86%)

Formulation (A) 50 times amount + Formulation (B) 50 times amount
5 fish (survival rate: 95%)

From the above, it was demonstrated that the preparations (A) and (B) improved the resistance to the pathogen, and in particular, the effect was remarkably enhanced by the combined use of both preparations.

<Verification experiment 17>
In this verification experiment as well, a field test was conducted at the Yamame fish farm.
The preparations used are 100 times the amount of each of the preparations (A) and (B), and three types of mixed preparations of the preparation (A) 50 times and the preparation (B) 50 times. The period of feeding the preparation and feed was 35 days, and the fish used was a 1-year-old yamame trout with an average weight of 35 g. 100 yamame trout were bred in each test plot. These yamame trout are selected after confirming that they have developed seawater resistance and have become smoltified.
Seawater acclimatization was carried out according to the following method practiced at the actual aquaculture site.
・ Put the smoltified yamame trout in a freshwater aquarium. Add seawater to the aquarium, gradually increase the concentration over 2 hours to 30% seawater (water temperature 13 ° C), and then breed for 1 day.
-Furthermore, breed in 70% seawater (water temperature 13 ° C) for 1 day.
・ After that, the concentration is gradually increased over 6 hours to change the 70% concentration seawater to 100% concentration seawater (water temperature 13 ° C → 18 ° C), and the seawater transfer is completed. Table 3 shows the survival rate in each test plot during this seawater acclimatization period.

From Table 3, it can be seen that the survival rate at the time of acclimatization to seawater is increased by using the preparations (A) and (B). In particular, it was demonstrated that the combined use of both preparations further enhances the survival rate.
It is considered that some strong stress adversely affects the health of yamame trout when acclimatizing to seawater. The first is stress due to sudden changes in salinity. This results in abrupt changes in osmotic pressure. In addition, it is probable that temperature change stress and stress when moving the water tank were also generated. It can be inferred that the survival rate was increased by improving the resistance to these stresses.

<Verification experiment 18>
Next, a field test was conducted at a seawater farm in Sakuramasu.
The preparations used are 100 times the amount of each of the preparations (A) and (B), and three types of mixed preparations of the preparation (A) 50 times and the preparation (B) 50 times. The period of feeding the preparation and feed was 28 days, and the fish used were cherry salmon 1 year old fish having an average weight of 51 g. Fifty cherry salmon were bred in each test plot.
The average body weight at the start of breeding in each test group and the average body weight after breeding for 28 days are shown below.

Average body weight / average value of 50 fish in each test group

Formulation-free (0-fold dose test plot)
At the start of breeding: 51.0 g After breeding for 28 days: 75.5 g (Growth rate: 48.0%)

Formulation (A) Macleaya cordata extract 100 times the amount of sanguinarine At the start of breeding: 51.0 g After 28 days of breeding: 82.2 g (growth rate: 61.2%)

Formulation (B) Ginger extract Zerunbon 100 times amount At the start of breeding: 51.0 g After 28 days breeding: 98.6 g (Growth rate: 93.3%)

Formulation (A) 50 times amount + Formulation (B) 50 times amount At the start of breeding: 51.0 g After 28 days breeding: 102.2 g (Growth rate: 100.4%)

The growth rate is improved by using the preparation (A) and the preparation (B). In particular, the combined use of both preparations further significantly improved the growth rate. The reason for this is that, as in the verification experiment 15, the preparation (A) and the preparation (B) have a growth promoting effect, or the resistance to various stresses is improved to promote healthy growth. Conceivable. Since this verification experiment was conducted at a seawater farm, it is considered that the resistance to stress due to changes in osmotic pressure due to changes in salinity could be improved, which made it possible to greatly improve the growth rate. In addition, temperature stress (heat / low temperature), light stress such as ultraviolet rays, psychological stress due to high-density breeding, physical stress due to contact between individuals, pH stress, stress due to increase in ammonia / nitrite concentration due to spoilage of food, etc. It is considered that various stresses such as stress caused by chemical substances and stress caused by pathogens also occurred at the farming site, and it is highly possible that the improvement of resistance to them also contributed to the improvement of the growth rate.

<Summary of the present invention>
The excellent features and social significance of the present invention are summarized below.
First, what was newly found in the present invention is that the substances such as sanguinarine disclosed above are excellent in improving the stress tolerance and resilience of aquatic animals by administering them to aquatic animals such as fish and crustaceans. To have the characteristics.

This newly discovered property is optimal as an aquatic animal feed for aquatic animals such as fish and crustaceans, and as a growth enhancer. For example, when administered to aquatic animals for several days, high temperature, low temperature, temperature change, light, low pH, high pH, increase in ammonia / nitrite concentration due to spoilage of food, osmotic pressure, stress due to drugs and chemical substances , Relieves stress such as pathogens and diseases, and enables healthy breeding.

It goes without saying that the feed for aquatic animals and the growth-improving agent for aquatic animals according to the present invention can be used for appreciation fish and the like to enable healthier breeding, but more importantly, they are cultivated. When used for fish.

As mentioned above, organisms that grow in water are more vulnerable to temperature changes than terrestrial organisms. And, with the recent environmental changes due to abnormal weather and global warming, the change in water temperature is larger than before. Therefore, changes in water temperature are becoming a major problem in fish farming.
In addition, at high water temperatures, the propagation of viruses and bacteria becomes remarkable, and the incidence of fish diseases increases, which is another factor that aggravates the problem.
On the other hand, the growth of many fish is more remarkable as the water temperature is higher, and it is preferable to grow at the highest possible water temperature in order to increase the catch.

Therefore, if the healthy breeding of fish at high water temperature is possible, the resistance to viruses and bacteria is enhanced, and diseases can be prevented, the aquaculture yield can be greatly improved. In addition, the quality of fish can be improved. In particular, since the resilience from diseases is improved, the survival rate is increased and a stable yield can be obtained even when there is a problem such as a temporary rise in water temperature due to abnormal weather or water pollution. Furthermore, because it grows faster, the time to shipment can be shortened.

In addition, in order to suppress the growth of viruses and bacteria, pH reduction, salt content and drug administration may be performed, but as a result of these stresses, side effects such as growth inhibition, quality deterioration and malformation occur. It is often difficult to control bacterial growth. The administration of the aquatic animal feed and the aquatic animal growth-improving agent according to the present invention can suppress the occurrence of such side effects, so that bacterial growth can be easily suppressed.

In order to further improve the yield, fish should be cultivated at high density. However, the concentration of ammonia, nitrite, nitric acid, etc. generated from excrement becomes high, which causes great damage to fish. Since the administration of the aquatic animal feed and the aquatic animal growth improver according to the present invention has the effect of relieving stress such as ammonia, it is possible to cultivate at a higher density than usual, and it is expected that the yield will be improved. it can.

Even cold-water fish such as salmon can be cultivated in the season when the water temperature is high, and can be shipped in the off-season, which can improve the added value. Even if it is difficult to completely shift the season, adding value can be increased by slightly shifting the shipping time.

Since cold water fish can be cultivated even in warm regions where the water temperature is high, the options for cultivated fish can be expanded. On the contrary, when aquaculture is carried out in a cold region, it is possible to cultivate fish in a warm water area.
Since the resistance to changes in water pressure can be increased, deep-sea fish and the like can be cultivated, and the options for cultivated fish can be expanded.
Further, since the resistance to osmotic pressure can be enhanced, saltwater fish can be bred in freshwater or brackish water, or freshwater fish can be bred in seawater or brackish water.
In this way, it is possible to greatly expand the selection range of fish to be cultivated.

Since resistance to low water temperature stress can be improved, fish and the like can be bred soundly in winter without artificially raising the water temperature, so that energy costs can be reduced. On the contrary, in summer, the resistance to high water temperature stress can be improved, so that it is not necessary to artificially lower the water temperature, and the energy cost can also be reduced.

Larvae are less resistant to various stresses than adult fish. For example, resistance to temperature changes and starvation stress is particularly poor. It also has low resistance to bacteria. Therefore, it is necessary to breed the larvae in a specially controlled environment. However, by using the feed for aquatic animals and the growth enhancer for aquatic animals according to the present invention, the larvae have the same environment as the adult fish. It becomes possible to cultivate in, and management becomes easy. Alternatively, complete aquaculture is possible.

When exposed to strong UV light, epidermal proteins are destroyed and melanin pigments are produced to prevent it. By using the feed for aquatic animals and the growth enhancer for aquatic animals according to the present invention, it is possible to prevent the destruction of proteins in the epidermis and suppress the production of melanin pigment. By suppressing the production of melanin pigment, it becomes possible to prevent fading of seafood.
For example, the colors of Thailand, leopard coral group, tilefish, shrimp, octopus, etc. can be maintained.

In tuna, yellowtail, etc., the stress at the time of fishing causes an increase in body temperature, an increase in lactic acid, etc., and a decrease in pH, and muscle proteins are denatured. Then, a deterioration of meat quality called "burnt meat" occurs. The aquatic animal feed and the aquatic animal growth improver according to the present invention can suppress protein denaturation and prevent the occurrence of burnt meat.

The most important taste in fish quality is determined by the amount of glutamic acid and inosinic acid. Inosinic acid is produced by decomposing adenosine triphosphate. When exposed to various stresses, proteins are denatured and physiological dysfunction occurs, and the synthesis of glutamic acid and adenosine triphosphate is inhibited. The aquatic animal feed and the aquatic animal growth enhancer according to the present invention have an action of preventing physiological dysfunction even under stress and maintaining the synthesis of glutamic acid and adenosine triphosphate. As a result, even under stress, the amount of inosinic acid, which is a decomposition product of glutamic acid and adenosine triphosphate, can be maintained, and the taste can be improved.

As a breeding method for fish and shellfish containing a large amount of functional components, exposure to light stress (irradiation of visible light, ultraviolet rays, infrared rays, and radiation of various wavelengths), pH adjustment of breeding water (pH stress), and ion concentration (Ionizing stress), mixing various substances and drugs into food (drug / substance stress), and breeding at low water temperature (temperature stress). By applying such various stresses, fish and shellfish containing a large amount of functional components can be obtained, but on the other hand, deterioration of quality and yield due to stress has become a problem. Therefore, by alleviating various stresses by using the aquatic animal feed and the aquatic animal growth improver according to the present invention, it is possible to increase the functional components without causing a decrease in quality or a decrease in yield. ..

When acclimatizing to seawater, strong osmotic stress is generated due to a large change in salinity, but resistance to osmotic stress can be improved, so that fish and the like are not damaged during acclimatization to seawater, and therefore the survival rate can be improved. Therefore, the yield is increased. It also improves quality.

For the purpose of preventing the depletion of marine resources and enriching marine resources, fish and shellfish are bred for seedlings and then released. However, immediately after the release, there is a problem that the survival rate is lowered due to exposure to various stresses. However, after the release, it was difficult to control the breeding conditions, and there were no effective measures. Therefore, by administering the feed for aquatic animals and the growth-improving agent for aquatic animals according to the present invention at the seedling breeding stage, various stress tolerances can be improved and the survival rate after release can be improved.

The substances such as sanguinarine disclosed in each embodiment have a great effect even when used alone, but the effect can be further remarkable by using a plurality of kinds of substances in combination. This is because each substance has the effect of repairing damage to cells and specific proteins that make up cells, and when multiple substances are used together, damage to more types of proteins can be repaired. Is considered to be.

As described above, in fish farming, improvement of yield, improvement of quality (taste, color, prevention of burnt meat, etc.), shortening of shipping time, expansion of aquaculture time, expansion of selection range of fish to be bred, resistance to diseases It offers many benefits such as improved strength and resilience, full aquaculture, and ease of aquaculture of functional fish.

Claims (1)

The invention described in the specification.

Images