TWI406635B - An additive for lowering cholesterol contents in aquatic animals - Google Patents

Abstract

This invention is to provide an additive for lowering cholesterol contents in aquatic animal. The additive comprises a probiotic complex formula and a Chinese herbal compound composition. Using one of the additives alone or using both at the same time can all effectively lower cholesterol contents in aquatic animal. The additive utilizes the microorganism in the probiotic complex colony containing blocker (Monacolin K) capable of lowering cholesterol and antioxidants, and the mechanism of the Chinese herbal compound composition capable of lowering cholesterol, to achieve the efficacy of lowering cholesterol contents in aquatic animal. The result of this invention also shows that lowering cholesterol contents does not generate significant change in the nutritional value and nature of the aquatic animal gonad.

Description

An additive for reducing cholesterol content in aquatic animals

The invention relates to an additive which can reduce the cholesterol content in an aquatic animal, which comprises a combination of a compound probiotic formula and a Chinese herbal medicine combination, which can reduce the cholesterol content of the aquatic animal, or reduce the serum of the aquatic animal, either alone or in combination. Cholesterol content in the middle, liver or gonads.

With the improvement of people's living standards, the number of hyperlipidemia patients has increased significantly in recent years, and the cholesterol in the blood is too high, which may accumulate on the inner wall of blood vessels, making the blood vessels gradually narrow, and may even block the blood vessels, if This happens when the coronary arteries that supply blood to the heart muscle cause coronary heart disease; if the blocked blood vessels are in the brain, it will cause a stroke. In the current disease killer rankings, the proportion of stroke and heart disease is very heavy. Has become an important issue in global health care. Cholesterol levels in the blood have been shown to be associated with coronary heart disease (CHD) because it causes atherosclerosis. Clinical trials have shown that for every 1% reduction in blood cholesterol levels, the incidence of coronary heart disease will be reduced by 2%. Therefore, the American Heart Association recommends that each person's daily cholesterol intake should be less than 300mg. Otherwise, they should take more omega-3 fatty acids to reduce the impact of heart disease. In order to meet market demand, there is no "low cholesterol" food. Research and development.

Many common aquatic products are high-cholesterol foods. For example, every 100 grams of fresh white shrimp contains 54 mg of cholesterol, but the dry shrimp has a cholesterol content of 434 mg, and the watery salmon has a cholesterol content of 308 mg. In addition, crab yellow cholesterol is as high as 466 mg. , 蚬 up to 454 mg; people with high blood lipid problems must be modest.

Cholesterol is only found in animals. It is synthesized by Acetyl-CoA through a series of reactions. It contains 27 carbons and has a molecular weight of 386. It is mainly present in free cholesterol and cholesterol (cholesterolester). The proportion of the two varies from organization to organization, and the main sites of synthesis in the human body are liver (50%), small intestine (15%), and other tissues (the most skin, accounting for 35%); the most important in laying hens The location is the liver, while the ovaries have little or no ability to synthesize lipoproteins. Both humans and animals need some cholesterol to maintain life and normal physiological functions. The physiological function of cholesterol in animals is (1) it is an essential component of the cell membrane, regulating the fluidity of the cell membrane, (2) cholesterol is the precursor of vitamin D, cholic acid and steroid endocrine, (3) embryo development The required material, under the natural selection pressure, maintains the nutrient environment for embryo development, so genetic selection does not significantly reduce cholesterol levels in animals (Holland et al., 1980).

Researchers have used genetic breeding techniques, feed nutrition and drugs to control the yolk cholesterol content of animals. It has been found that feed cholesterol and fat increase yolk cholesterol content, while feed fiber content or certain drugs can reduce yolk cholesterol content, but the decline is limited. The use of genetic breeding techniques has not yielded results, or only reduced 5-7% cholesterol, and the performance of laying eggs has also been affected. Therefore, it is still difficult to improve the genital gland or yolk by traditional genetic breeding techniques. Reduced cholesterol levels. The regulation of feed nutrition or drugs has become the focus of animal nutrition research.

Since the cholesterol-lowering substance (HMG-CoA reducatase inhibitor) was isolated from Penicillium brevicompactum and P. citrinum in 1976 , many similar substances have been separated from Aspergillus and Monascus. In 1979, monacolin k was isolated from the culture of Monascus ruber , and American scholars also isolated the same chemical structure from Aspergillus terreus , but named mevinolin. In 1987, Merck developed Lovatatin and was the first to market its drug. Lovastatin is the active ingredient of the fermentation product of Monascus purourous , so monacolin K, mevinolin and lovastatin are all the same.

The antioxidants and bioactive ingredients produced by the complex probiotics can reduce the cholesterol content of the eggs. The study found that 30-week-old roman hens were fed a diet fed with a combination of probiotics. As a result, the total protein of the whole egg increased by 10-13%, fat decreased by 11.7%, and cholesterol decreased by 15-91%. Since the compound probiotics group has high selectivity to provide a culture medium for beneficial bacteria such as bifidobacteria in the intestinal tract of the animal, and stimulates the rapid proliferation of the beneficial bacteria, the beneficial bacteria such as bifidobacteria and lactobacilli are in the animal body. It can synthesize a variety of vitamins, such as folic acid, niacin, vitamins B1, B2 and B6, which can promote the absorption of nutrients such as protein, calcium, zinc and iron, and significantly improve animal performance.

Because Chinese herbal medicine additives have the characteristics of promoting animal growth, enhancing their resistance, low toxicity and side effects, no drug residues, and low resistance to drug resistance, they have received much attention in the feed industry in recent years.

The study found that a compound preparation consisting of Chinese herbal medicines such as red peony, honeysuckle, eucommia, silybum, hawthorn, etc., was added to the diet of 32-week-old Nickel hens at doses of 500 mg/kg and 250 mg/kg, respectively. The concentration of cholesterol in egg yolk (mg/g egg yolk) decreased by 25.4% (P<0.05) and 17.4% (P<0.05), respectively. The plasma cholesterol, LDL-C and VLDL concentrations decreased significantly (P<0.01). And increased the HDL concentration; improved the composition of egg fat, the content of linoleic acid increased by 24.2%. 17.1%; the content of linolenic acid increased by 44% and 36%. Another study found that in the 280-day-old Roman chicken laying hen diet, 1% of Pinggan powder consisting of Chinese herbal medicines such as Gynostemma pentaphyllum and 1.5% of flavored red and white powder consisting of pepper, gypsum and Gynostemma pentaphyllum were added. Compared with the control group, the cholesterol content of eggs decreased by 15.45% and 14.81%.

In summary, although there are techniques for reducing cholesterol in eggs by using a combination of probiotics or Chinese herbal medicines, there have been no technical discoveries for reducing cholesterol in aquatic animals, and the production and application techniques of poultry and aquatic products vary greatly. Many aquatic animals, such as fish, shrimps, crabs, shellfish, etc., which have high cholesterol content, are the favorite of many hackers. In order to balance the taste and health and improve the quality of the aquaculture industry, the inventor of this case is After thinking and improving, and after many years of painstaking research, I finally succeeded in research and development of this kind of additive to reduce the cholesterol content of aquatic animals.

It is an object of the present invention to provide an additive which reduces cholesterol levels in aquatic animals, including a compound probiotic formulation which can be used to reduce cholesterol levels in aquatic animals.

Another object of the present invention is to provide an additive for reducing the cholesterol content of aquatic animals, which comprises a combination of Chinese herbal medicines, which can be used to reduce the cholesterol content in aquatic animals.

The object of the present invention is to provide an additive for reducing the cholesterol content in aquatic animals, which comprises the above-mentioned compound probiotic formula and a combination of Chinese herbal medicines, which can be used to reduce the cholesterol content in aquatic animals.

Another object of the present invention is to provide a method of adding aquafeed: including

Two kinds of feeding methods of compound probiotics added to aquatic feed: (1) Artificial feed

The above composite probiotic formula is uniformly sprayed on the artificial aquatic feed at a weight ratio of 0.5% or more (additive/feed, W/W), allowed to stand for more than 5 minutes to be completely absorbed, and then placed in a dark place for fermentation, and then cast. Feed to aquatic animals.

(2) Feeding bait

Take the weight ratio of the above composite probiotics formula to the raw bait feed by 0.5% or more (additive/feed, W/W), add to the raw bait and mix evenly, let stand for more than 10 minutes, let the raw bait adhere to the additive and then cast. Feed; the raw bait is fresh fish and minced meat.

Two kinds of feeding methods of Chinese herbal compound combination added to aquatic feed: (1) Artificial feed

Take the above combination of Chinese herbal medicine, add 0.5% or more by weight (additive/feed, W/W) to the artificial aquatic feed and mix well. Use this feed throughout the feeding.

(2) Feeding bait

Take the above combination of Chinese herbal medicine, add 0.5% or more by weight (additive/feed, W/W), mix it into the raw bait feed, and let it stand for more than 10 minutes to let the raw bait adhere to the additive and then feed it; The bait is fresh fish and shrimp.

Compound probiotic formula and Chinese herbal compound combination are added to the aquatic feed to feed: (1) Artificial feed

Take more than 0.5% by weight of the compound probiotic formula (additive/feed, W/W), spray it evenly on the artificial diet, let it stand for more than 5 minutes to completely absorb it, and then put it in the dark place for fermentation; The compound combination is added to the artificial feed at a ratio of 0.5% by weight (additive/feed, W/W), and the mixture is uniformly mixed. The special feed is used throughout the feeding.

(2) Feeding bait

The composite probiotic formula and the above-mentioned herbal compound combination powder are added to the raw bait uniformly in a ratio of 0.5% by weight (additive/feed, W/W), and allowed to stand for more than 10 minutes to completely adhere the raw bait to the additive. After feeding.

To achieve the above object, an additive for reducing cholesterol content in an aquatic animal according to the present invention comprises a composite probiotic formulation, wherein the composite probiotic formulation is based on Complex Beneficial Microorganisms (CBM). The invention comprises one or more microorganisms such as photosynthetic bacteria, lactic acid bacteria, yeast, actinomycetes and special bacteria containing cholesterol-lowering substances, wherein the special bacterial species containing the cholesterol-lowering substances are selected from one or more of the following species: P. brevicompac tu m, P. citrinum, A. terreus, M. purourous and M. ru ber, fermented by commensal culture techniques. The compound probiotic formula has several advantages: (1) because the single strain is difficult to preserve, the compound probiotic formula can be used for long-term preservation and amplification of the special strain of the cholesterol-lowering substance Monacolin K; (2) by each probiotic Antioxidant substances and bioactive compounds produced by bacteria can reduce the content of cholesterol in aquatic animals, such as hydroxymethylglutaric acid coenzyme A reductase produced by lactic acid bacteria, orotic acid, calcium, lactose, etc. Niacin, etc., plays an important role in reducing cholesterol in animals; (3) it has a high selectivity to provide a culture medium for beneficial bacteria such as bifidobacteria in the intestinal tract of animals, and stimulates the rapid proliferation of the beneficial bacteria. Bifidobacteria and Lactobacillus can produce a variety of vitamins, such as folic acid, niacin, vitamins B1, B2 and B6, which can promote the absorption of protein, calcium, zinc and iron. To improve animal performance; (4) After using the compound probiotic formula to reduce cholesterol in aquatic animals, the nutritional value and properties of the gonads of the animal did not appear obvious. Variety.

Human and aquatic physiological structures are very different. Therefore, although Chinese herbal medicines are known to reduce blood lipids and cholesterol in humans, it has not been confirmed whether it can also be applied to aquatic animals. Therefore, the present invention will prove that the Chinese herbal compound combination can lower cholesterol in aquatic animals. content.

Chinese herbal medicine can reduce the body's blood lipids and cholesterol into: (1) promote intestinal cholesterol excretion, inhibit the absorption of exogenous cholesterol; (2) competitive inhibition of intestinal cholesterol absorption; (3) inhibition of cholesterol, three In vivo synthesis of glycerides; (4) affecting the distribution, functioning and clearance of blood lipids; and (5) promoting the decomposition and excretion of cholesterol. According to the above-mentioned herbal medicine to reduce blood fat and cholesterol, the inventors formulated a combination of Chinese herbal medicine, including Codonopsis pilosula, Atractylodes, Acanthopanax senticosus, Curculigo, Polygonum, Angelica, Ai, Hawthorn, Divine Comedy, Malt and Pinus fine. The herbal combination of the Chinese herbal medicine and the feed are uniformly mixed at a ratio of 0.5% by weight or more, and then the aquatic animal is fed.

The present invention is exemplified by the following examples, but the present invention is not limited by the following examples.

Embodiment 1 A method for preparing an additive for reducing cholesterol content in an aquatic animal 1. Compound probiotic formula

The compound probiotic formula is based on Complex Beneficial Microorganisms (CBM), which contains one or more microorganisms such as photosynthetic bacteria, lactic acid bacteria, yeasts, actinomycetes, and special strains containing cholesterol-lowering substances. The special strain containing cholesterol-lowering substances is selected from one or more of the following strains: P. brevicompac tum, P. citrinum , A. terreus , M. purourous and M. ru ber, fermented by commensal culture techniques to make.

2. Chinese herbal compound combination

Humans and aquatic structures are very different, so it is known that Chinese herbal medicines can lower blood lipids and cholesterol in humans (as shown in the table below, The representative has efficacy), but it has not been confirmed whether it can also be applied to aquatic animals, so the present invention will prove that the Chinese herbal compound combination can reduce the cholesterol content in aquatic animals:

Chinese herbal medicine can reduce the body's blood lipids and cholesterol: (1) Promote the excretion of intestinal cholesterol and inhibit the absorption of exogenous cholesterol:

(2) Competitively inhibit intestinal cholesterol absorption:

(3) Inhibition of in vivo synthesis of cholesterol and triglyceride:

(4) affecting the distribution, operation and elimination of blood lipids:

(5) Promote the decomposition and excretion of cholesterol:

According to the above-mentioned Chinese herbal medicine to reduce blood fat and cholesterol, the inventors formulated a Chinese herbal compound combination comprising 50 to 150 grams of the party participating in Atractylodes, Acanthopanax senticosus, Curculigo, Polygonum multiflorum, Angelica and Ai Ye each 30 to 150 grams, Hawthorn, Divine Comedy 20 to 150 grams of malt and 100 to 300 grams of pine needles are mixed and added to the feed, and the added amount is 0.5% or more of the total weight of the feed.

Example 2: Method of using additive Two kinds of feeding methods of compound probiotics added to aquatic feed: (1) Artificial feed

Take more than 0.5% by weight of the compound probiotic formula (additive/feed, W/W), spray it evenly on the artificial aquatic feed, let it stand for 5 minutes, completely absorb it, and put it in a black plastic bag or a covered plastic bucket. Fermentation for 1 to 2 days, followed by feeding to aquatic animals.

(2) Feeding bait

The weight ratio of the compound probiotics to the raw bait feed is 0.5% or more (additive/feed, W/W), added to the raw bait and uniformly mixed, and allowed to stand for 15 minutes to allow the raw bait to adhere to the additive and then feed. Where the raw bait is fresh fish and minced meat.

Two kinds of feeding methods of Chinese herbal compound combination added to aquatic feed: (1) Artificial feed

Take the above combination of Chinese herbal medicine, add 0.5% or more by weight (additive/feed, W/W) to the artificial aquatic feed and mix well. Use this feed throughout the feeding.

(2) Feeding bait

Take the above combination of Chinese herbal medicine, add 0.5% or more by weight (additive/feed, W/W), mix it into the raw bait feed, and let it stand for 15 minutes to let the raw bait adhere to the additive and then feed it; Minced meat for fresh fish and shrimp.

Compound probiotic formula and Chinese herbal compound combination are added to the aquatic feed to feed: (1) Artificial feed

Take the compound probiotic formula 0.5% or more by weight (additive/feed, W/W), spray it evenly on the artificial feed, let it stand for 5 minutes to completely absorb it, and put it in a black plastic bag or a covered plastic bucket for 1~ 2 days of fermentation; the above combination of Chinese herbal medicine is added to the artificial feed with 0.5% by weight (additive/feed, W/W), and the mixture is evenly mixed, and the special feed is used throughout the feeding.

(2) Feeding bait

The composite probiotic formula and the above-mentioned herbal combination are added to the raw bait uniformly in a ratio of 0.5% by weight (additive/feed, W/W), and allowed to stand for 15 minutes to allow the raw bait to adhere to the additive and then fed. .

After the aquatic animals were fed with the above additive-containing feed for 1 to 2 months, the cholesterol content of the aquatic animal was measured.

After the aquatic animals were fed with the above additive-containing feed for 1 to 2 months, the cholesterol content of the aquatic animal was measured.

Example 3, cholesterol detection method

The following two methods are used to determine the cholesterol content of animal serum, muscle, fish eggs or ovary (crab cream), testis (crab); immature animals use hepatopancreas to determine total cholesterol.

Principles and methods for determination of total cholesterol

Cholesterol ester is hydrolyzed to free cholesterol and fatty acid by cholesterol esterase (CHE), and cholesterol is produced by cholesterol oxidase (COD) to form Δ4-cholestenone (cholest-4- En-3-one) and hydrogen peroxide (H ₂ O ₂ ), while H ₂ O _{2 is} subjected to the action of peroxidase (POD) in the presence of 4-aminoantipyrine (4-AAP) and phenol. A red quinone imine pigment is formed. The absorbance of the solution was measured at a wavelength of 500 nm, and the absorbance of the quinone imine pigment was directly proportional to the cholesterol content in the sample.

First, the method of determination of total cholesterol in tissues

0.5 g of tissue was added to 9 ml of cooling extract (cholorform: methanol = 2:1; v / v), homogenized, centrifuged at 3,500 rpm for 10 minutes, and the supernatant was aspirated and stored at 4 ° C; After 500 ul of tissue extract, the organic solvent was evaporated, an equal volume of DMSO was added, and the cholesterol content was determined using a cholesterol test kit (Fortress Diagnostics, UK).

Second, the blood total cholesterol determination method

The blood of the animal was extracted with an anticoagulant-washed syringe for about 5 cc. After centrifugation at 5,000 rpm and 4 ° C for 25 minutes, the supernatant was aspirated, and 10 ul of the sample was added to 1000 ul of the reagent, and the solution was applied at 37 ° C for 5 minutes. The absorbance was measured at a wavelength of 500 nm, and the total cholesterol content was converted by standard concentration; wherein the reagent composition was Good's buffer, pH 6.7, 50 mmol/L; phenol, 5 mmol/L; 4-aminoantipyrine , 0.3 mmol/L; cholesterol esterase (CHE), 200U/L; cholesterol oxidase (CHO), 50 U/L; peroxidase (POD), 3000U/L.

test results: First, reduce the cholesterol content of fish egg yolk

The zebrafish were divided into two groups. The control group was fed with normal feed, while the other group was fed with a combination of Chinese herbal medicine. After one month of feeding, the yolk of 50 zebrafish was collected to measure the cholesterol content. The experimental results showed that the artificial diet was added to the Chinese herbal medicine formula, and the cholesterol content in the fish eggs was reduced compared with the control group (as shown in Table 1).

The zebrafish were divided into four groups, the control group was fed with general feed, while the experimental group was fed only with the combination of the Chinese herbal compound, the feed group with the probiotics, and the compound and Chinese herbal compound group, each feeding for two weeks. The yolk of 50 zebrafish was collected to measure the cholesterol content.

The experimental results showed that the control group of only the general feed had a cholesterol content of 2.133 mg / dl; the zebrafish was fed with the Chinese herbal compound feed group, the cholesterol content of the fish eggs was 2.091 mg / dl; feeding with compound probiotics After the feed, the cholesterol content in the fish eggs was found to be 1.739mg/dl; and the feed group added with the compound probiotics and the Chinese herbal compound compound, the cholesterol content of the fish eggs decreased the most, and its content was 1.723mg/dl (as shown in Figure 3). ).

Second, reduce shrimp serum and liver and pancreas cholesterol content

The South American white shrimps were divided into three groups, wherein the control group was fed with general feed; the treatment group T1 was fed with a combination of a combination of a probiotic formula and a Chinese herbal compound; and the treatment group T2 was fed with a combination of a Chinese herbal compound, after one month. Measuring the body's cholesterol content, the results show that not only can reduce the serum cholesterol content of South American white shrimp ( L. vannamei ) about 15% or more (as shown in Table 2); it can also reduce the liver and pancreas cholesterol content of about 20% or more (such as Three shows).

Treatment group T1: compound probiotic formula and Chinese herbal compound combination

Treatment site T2: Chinese herbal compound combination

Treatment group T1: compound probiotic formula and Chinese herbal compound combination

Treatment site T2: Chinese herbal compound combination

Third, reduce the cholesterol content of crab gonads

The crabs were divided into two groups, the control group was fed with general feed; the other group was fed with feed supplemented with probiotic formula. After one month of culture, each group randomly collected 3~5 male crabs and measured cholesterol in the body. content. The experimental results showed that after feeding with the artificial feed of the above additives, the cholesterol content of hairy crabs was significantly reduced by more than 40% (Table 4).

The crabs were divided into two groups, of which the control group was fed with general feed; the other group was fed with a combination of a combination of probiotics and a combination of Chinese herbal medicines. After one month of culture, each group randomly collected 3 to 5 crabs. , measuring the body's cholesterol content. The experimental results showed that after feeding with the artificial feed of the above additives, the cholesterol content in the ovary of the female crab was significantly reduced by about 20% (Table 5).

The test results of the present invention also show that the additive for lowering the cholesterol content in aquatic animals can significantly reduce the cholesterol content in aquatic animals, and there is no significant change in the nutritional value and properties of the gonads (sperm eggs) of aquatic animals.

The detailed description of the above is a detailed description of a possible embodiment of the present invention, and the embodiment is not intended to limit the scope of the invention, and the equivalents or modifications, such as the additive, are not departing from the spirit of the invention. Equivalent examples of changes in cholesterol in other aquatic animals such as fish, shrimps, crabs, etc., should be included in the patent scope of this case.

In summary, this case is not only innovative in terms of method and form, but also can enhance the above-mentioned multiple functions compared with conventional articles. It should fully comply with the statutory invention patent requirements of novelty and progressiveness, and apply for it according to law. This invention patent application, in order to invent invention, to the sense of virtue.

Figure 1. An additive that reduces the cholesterol content of aquatic animals can reduce the serum cholesterol level of L. vannamei . The X axis is the group, and the Y axis is the cholesterol content, and the unit is mg/dg. The treatment group T1 is a feed supplemented with a compound probiotic formula and a Chinese herbal compound combination; the treatment T2 is a feed supplemented with a Chinese herbal compound combination.

Figure 2. An additive that reduces the cholesterol content of aquatic animals reduces the cholesterol content of the liver and pancreas of L. vannamei . The X axis is the group, and the Y axis is the cholesterol content, and the unit is mg/dg. The treatment group T1 is a feed supplemented with a compound probiotic formula and a Chinese herbal compound combination; the treatment T2 is a feed supplemented with a Chinese herbal compound combination.

Figure 3. An additive that reduces the cholesterol content of aquatic animals reduces the cholesterol content of zebrafish eggs. The X axis is the group, and the Y axis is the cholesterol content, and the unit is mg/dl.

Figure 4. An additive that reduces the cholesterol content of aquatic animals can reduce the cholesterol content of irritans in hairy crabs. The X axis is the group, and the Y axis is the cholesterol content, and the unit is mg/dg.

Figure 5. An additive that reduces the cholesterol content of aquatic animals can reduce the cholesterol content of crabs. The X axis is the group, and the Y axis is the cholesterol content, and the unit is mg/dg.

Claims (7)

An additive which can reduce the cholesterol content of aquatic animals, including a combination of Chinese herbal medicines, the combination of Chinese herbal medicines for the party participating in Atractylodes 50-150 grams, Acanthopanax senticosus, Curculigo, Polygonum, Angelica and Ai Ye each 30 to 150 grams, Hawthorn, Divine Comedy and Malt are 20~150g each and pine needles are 100~300g. An additive for reducing the cholesterol content of an aquatic animal, as described in claim 1, wherein the Chinese herbal compound combination is a feed additive for artificial feed of aquatic animals. An additive for reducing the cholesterol content of an aquatic animal, as described in claim 1, wherein the herbal combination is a feed additive for aquatic animal feed. An additive for reducing the cholesterol content in an aquatic animal according to claim 1, wherein the Chinese herbal compound combination is used in an amount of 0.5% by weight or more (additive/feed, W/W). The invention relates to a method for adding aquaculture feed, which comprises adding an additive capable of reducing the cholesterol content of an aquatic animal as described in claim 1 to a ratio of 0.5% by weight (additive/feed, W/W) to artificial aquatic product. Premixed in the feed evenly. A method for adding aquaculture feed according to claim 5, wherein the artificial aquatic feed is liquid or solid. A method for adding aquaculture feed, which comprises, in an amount of 0.5% by weight or more (additive/feed, W/W), which is an additive which can reduce the cholesterol content of an aquatic animal as described in claim 1 of the patent application. Add to the raw bait and mix evenly, let stand for more than 10 minutes to allow the raw bait to adhere to the additive completely.