KR19990040962A - Fish farming and fry farming methods for improving the immunity of fry - Google Patents

Abstract

The present invention relates to the development of nutritional fish feed for fry in the field of fisheries, and more specifically, to find immunity enhancing substances from natural products and add them to the existing feed formulation for fry for production of immunity enhanced fry. It is to develop and lower fish feed for fry.

In other words, according to the present invention, by first placing the ganban stone known as a water purifier and infrared emitter in the fish farm to increase the cleanliness of the water to prevent the growth of microorganisms, extracts of garlic, coriander, ginseng, perilla, etc. To provide a cultured fry for fry made by adding each of chitosan or organic minerals added with vitamin B1 and B2 to the existing fry feed, can improve the immunity of fry and at the same time normal fish feed By administering antibiotics to fish meal, pupa, etc., it reduces the pathogenicity of fry and reduces the lethality of fry, which has threatened human health by transferring antibiotics to the human body as well as water pollution caused by the antibiotics. It is possible to solve the problem in the conventional fish feed for.

Description

Fish farming and fry farming methods for improving the immunity of fry

The present invention relates to a fish feed method for fry and aquaculture method for fry for enhancing the immunity of fry that can significantly reduce the lethality by increasing the immunity of fry.

In general, it has been reported that 40% of the fry are dead, mainly due to the lack of nutrition or contamination of the water due to the breeding of microorganisms caused by the decay of the feed is not eaten. . In particular, deficiency of pathogenic immunity was known to be the leading cause of death of fry. Therefore, in order to reduce the mortality rate of the 40% of the fry to develop a special fish feed for the development of the fish, the most representative of the conventional fish feed, fishmeal, pupa, etc., the antibiotics have been used a lot now. However, this feed contains antibiotics, so the fry that consumed it could not grow healthy because of its lack of immunity to pathogenicity, as well as contamination of water quality by antibiotics and transfer of antibiotics to the human body, threatening human health. Has been given.

The present invention is to solve the above problems, without the use of artificial antibiotics by searching for immune-enhancing substances from natural products and adding to the existing fry feed formulations to enhance the immunity of the fry to significantly reduce the mortality of the fry The aim is to develop a fish farm for juvenile fish that can produce large quantities of fish.

1 is a perspective view of a fish farm showing the form and structure of a fish tank for farming fry by feeding the feed according to the present invention.

<Description of Symbols for Main Parts of Drawings>

A: Groundwater outlet B1: PVC pipe inside the fish tank

B2: PVC pipe outside the fishbowl C: Groundwater outlet

D: oxygen drop E: oxygen generator

Features of the present invention are premixed by adding 0.005 to 0.015 wt% of vitamin B1 and B2 to 1-5% of the chitosan solution, and then adding 0.5-2 wt% of the mixture to the existing fry feed for fish farming. To provide feed.

Another feature of the present invention is to mix the garlic, Astragalus, ginseng and perilla in the ratio of 1 to 2: 1 to 2: 1 to 3: 1 to 2 (w / w) ratio of the extract extracted from the mixture to the existing fry feed It is to provide a feed for fish farming made by adding 0.5 ~ 1wt%.

In addition, the present invention is characterized in that it provides a fish feed for fish farming made by adding 5 to 8wt% of organic mineral solution, which is a mineral in a form combined with an in vivo material, to an existing fry feed.

The present invention feeds the garlic, Astragalus ginseng, the extract of perilla, chitosan with the addition of vitamins B1 and B2, or organic mineral additives while feeding the ganban stone to raise the cleanliness of the water and breeding of microorganisms It is also characterized by farming cheerfish to prevent it.

When explaining the method of preparing a feed for improving the immunity of the fry according to the present invention and the use of the feed prepared according to the present invention and the measurement of the immune strength.

Fish farming for enhancing the immunity of the fry according to the present invention is pre-mixed with each added vitamin B1 and B2 to chitosan solution; Garlic, Astragalus, Ginseng and Perilla extracts are extracted by mixing a certain ratio; the organic mineral solution; is prepared by simply adding to each of the existing fry feed.

Chitosan is widely distributed in crustaceans such as crabs and shrimps, as well as in insects, mushrooms, and cell membranes of bacteria. It can obtain by high heat processing (deaselification). The characteristics of chitosan produced in this way are anti-aging, which activates senescent cells to become vital cells, prevents the invasion and proliferation of bacteria, viruses, cancer cells, etc., strengthens immune cells, and weakens the cause of illness. By giving vitality to the immune cells of somatic cells has the function of total control of life activities, such as preventing diseases.

In addition, if vitamin B1 is insufficient as a vitamin necessary for carbohydrate metabolism, the sugar is not completely decomposed, and it becomes pyruvate or lactic acid, which is an intermediate substance, causing nerves to circulate in the circulation, and vitamin B2 carries hydrogen in the body. Calorie metabolism is inhibited, and it is known to affect inflammation, dermatitis, and growth and is recognized as a major influencer.

Garlic, Astragalus, Ginseng, Perilla, etc. are widely known as immune enhancing substances. In the mid-yellow season, 2 ', 4'-dihydroxy-5', 6-dimethoxyisoflavan, 5,4'-dioxy -3,7-dimethoxyflavones, flavonoids of molecular formula C ₁₆ H ₁₂ O ₅ , cumategenin, β-cyrrosterol, fructose, glucose, mucus and alkaloids, saponin-reactive substances, choline, betaine, linoleic acid, linol Lean and Amino Acids with Leucine (18mg /%), Glycine (24mg /%), Serine (28mg /%), Alanine (55mg /%), Glutamic Acid (77mg /%), Arginine (88mg /%), γ- Amino Butyric Acid (24 ~ 66mg /%) is included, and Ginseng contains amino acids such as choline (0.1 ~ 0.3%), arginine (1%), glutamic acid, cysteine, tyrosine, α-aminobutyric acid and more than 7 alkyloids , Ash (8.7%) and other ingredients are known to act on the nervous system.

Organic minerals are minerals that are combined with in vivo materials and can be obtained through alkali treatment and hydrolysis of shark bones and bones.

The immune-enhanced feed and aquaculture method according to the present invention as described above will be described in detail through the following examples. In the present embodiment,% refers to a weight percentage, and in this embodiment, trout fry is added, but the same result is obtained even when other fry are added.

EXAMPLE

○ Fish farms

As a fish farm for fry, as shown in FIG. 1, the amount of water coming in on one side and the amount of outgoing water are the same so that groundwater can continue to flow for 24 hours in a fishbowl having a size of 900 x 300 x 450 (mm) in size. A hole was made and PVC pipes were connected to the inside and outside of the tank to adjust the height of the groundwater in the tank.The temperature of the water was about 17 ℃, and five sub-tanks were purchased to purchase oxygen cylinders. Later in the present embodiment, 30 of the trout fry in each of the fry in each fish farm.

Group (C group) fed to the trout fry added to the fish farm according to the present invention, 5% organic mineral solution was added to the existing fry feed (group C); (D) group fed with a premixer added with 0.005% to 0.015% of non-minerals B1 and B2 in chitosan 1% solution to a conventional fry feed; Garlic: Astragalus: Ginseng: Perilla perilla 1: 1: 2: 1 (w / w) ratio of extract extracted by adding 0.5% to the existing fry feed to feed (group E); A group of conventional feed (group A) as a control; Group B that fed the existing feed in fish farms in which elvan was introduced; Each group was fed three times a day to feed each group, and the weight and survival rate were measured at 10-day intervals and fish farmed for 60 days.

In addition, each of the above-mentioned fish tank of the present invention was put into the ganban stone known as a water purifier and infrared emitter and set to C group using organic minerals, D group using chitosan, E group using garlic, Astragalus, ginseng and perilla extract, respectively. The results were measured.

○ Dissection and Treatment of Cheer

In order to avoid stress for 60 days of farmed fry, 7 fish per group (because only 7 fry from the control group survived) were placed in a large plastic bag and brought to the laboratory and immediately dissected by a skilled expert. After the liver, spleen and tissue were extracted and measured, and then cryopreserved, lipid peroxidation activity was measured by the following reagents and procedures to determine immuno-enhancing activity.

● Reagent

1.8.1% sodium dodecyl sulphate (SDS) → heat dissolution

2. 20% acetic acid

3. 0.8% tuovaltbric acid (TBA) → heating

4. 1.15% KCl buffer

● Course

200 ml of 10% homogenate

Vortexing 30 seconds

Heating 60 minutes (95 ℃)

Cool to room temperature in ice or running water

Centrifuge 10 minutes (3000 rpm)

Absorbance Measurement (λ = 532 nm)

Using a spectrophotometer according to the above-described treatment, the Tuvalvrt acid (TBA) value was obtained and lipid peroxide activity was measured.

○ Fatty acid analysis of total lipid of liver, spleen and tissue of fry

Tissue 0.5g

↓

15 ml methanol

↓

↓ ← Homogenization

Chloroform 30ml

↓

Heat extraction (shake warmed for 30 minutes at 40 ° C)

↓

Cooling in ice

↓

Add 50 ml of chloroform: methanol (2: 1, V / V)

↓

Whatman fillter paperNo. Filtration with two

↓

Distilled water addition

↓ ← Mix

It is left at 4 ℃ for 24 hours

↓

Remove water-methanol (MeOH) layer

↓

Vacuum drying

↓

Add 24ml Hexane

↓

Trans methylation with 2 ml of sulfuric acid-methanol (MeOH)

↓

Argon gas injection

↓

1 hour heating to 80 ° C

↓

Hexane gas removal

↓

2 ml of hexane and 2 ml of saturated sodium chloride (NaCl)

↓

Shaking for 5 minutes

↓

Centrifuge (at 3000 rpm for 30 seconds)

↓

Hexane layer recovery

↓

Gas Liquid Chromatograph (GLC) Analysis

(10μℓ injection)

Fatty acid content was analyzed using gas liquid chromatograph (GLC) according to the above procedure.

○ Assessment of immune activity of fry by histamine assay

Sodium Chloride 0.75g-0.78g

↓

Sample (1g) was added to Tyrode solution (1ml)-2ml sample

1N NaCH 0.5ml

B-C solution (butanol: chloroform) → 3: 2 5 ml

↓

2 minutes mixed

↓

B-C (butanol-chloroform) layer extraction (upper layer)

↓

Centrifuge (3.000 rpm, 5 minutes)

↓

4 ml of B-C layers

2 ml of n-heptane

0.1N HCl 1.5ml

↓

Mixed, 2 minutes

↓

Centrifuge 3,000 rpm, 5 minutes

Fluorometer → Measured in 2-3 hours

<Result of Example>

○ Measurement of survival rate of fry

The survival rate of the larvae was visually observed for 60 days at 10-day intervals. The survival rate of the control group was 92.3%, 88.55, 61.5%, 42.3%, 30.8%, and 26.9%.

As shown in Table 1, in group D with chitosan, one mortality rate was not seen during the 60-day experiment. Even in the extract group E, which received 40 g of extracts such as ginseng for 40 days, the mortality rate was zero. However, after 50 days, the mortality rate was reduced to 80.8%. % Was kept as it is. Group C with minerals maintained 100% survival for 30 days, but then decreased to 96.2%, 84.6%, and 80.8%, showing a similar trend to the extract. Survival rate was higher in group B> C> E> D. Table 1

Table 1 Comparison of survival rate of fry by each treatment group

○ Changes in weight and size of fry

In the control group, the mortality rate of the fry was very high, which made it impossible to make an accurate comparison for each group. However, the weight of the 7 fry of the last 60 days was measured. Showed an increase in weight.

The average size change after 60 days of fish farming was 10.9 cm, 4.0 cm above the original mean size. In order of size, group A <B <D <E <E (see Tables 2A and B).

(A)

(B)

Table 2 Histogram of weight and size change of fry of each treatment group

○ Weighing Cheer Spleen

There was a slight drop in group B and C, but the weight of the remaining group A and each group was heavier than the group. Table 3

Table 3 Bar graph of weight change of larva spleen by each treatment group

※ of the bar graph Represents the effective deviation

This result suggests that 60 days of feeding of the developed diet is more likely to involve Gut-Associated Lymphoid Tissues rather than spleen immunity.

○ Effect on Triacylalycerol (TG) in Cheerleaders

As can be seen in Table 4, the triacylglycerol content of each treatment was significantly reduced compared to the control. In particular, the chitosan-treated group D showed a significant decrease.

The results of TG conversion are thought to contribute to the enhancement of immunity by converting the physicochemical and nutritional properties of muscles and by participating in polyunsaturated fatty acid content changes such as EPA (see Table 6).

Table 4 Changes in Triacylglycerol Contents of Young Fishes in Different Treatment Groups

○ Antioxidant activity of fry

As can be seen in Table 5, the antioxidant activity was strong in each treatment group, especially D and E groups. This is thought to reduce the lethality by enhancing the in vivo immune activity by inhibiting the production of peroxides from fry.

Table 5 Comparison of Antioxidant Activity in Young Fishes by Treatment Group

○ Changes in fatty acids of fry

Table 6 shows fatty acid changes according to various additive treatments. In each treatment group, all, especially the group D containing chitosan, the control group significantly increased (10-fold) EPA content compared to A.

In group D, the ratio of 20: 3 + 20: 4/18: 2 as an index of immune activity was 0.20 ± 0.03, which was significantly higher than group A 0.15 ± 0.01. Therefore, chitosan of group D is considered to be related as an immune enhancing substance.

Table 6 Comparison of fatty acid composition by treatment group

* Values are mean and ± standard deviation of three animals

* Significance difference (P) is 0.05 or more in comparison with subscript a-c value.

○ Histamine content of fry

Table 7 shows the histamine content accumulated in the body of the fry. Feeding chitosan-added feed (group D) resulted in very low histamine content in the body. This proves that we are healthy without growing infected. The high histamine content is known to increase the amount of histamine for self-defense by increasing foreign substances or infectious agents in the body. Therefore, the low histamine content indicates that the risks from the outside, ie, pathogens are growing normally without being invaded, are consistent with the highest survival rate.

Table 7 Comparison of accumulated histamine content of fry of different treatment groups

* Values are mean and ± standard deviation of three animals

* Significance difference (P) is 0.05 or more in comparison with subscript a-c value.

By developing and manufacturing a functional feed for fry that can enhance the immunity of the fry of the present invention and feed it to the fry, it is possible to produce healthy fry without administering artificial antibiotics harmful to the human body, and thus does not harm the human body. At the same time, it is possible to increase the income of the younger farmers.

In addition, it is possible to increase the cleanliness of the water and prevent the growth of microorganisms by drastically reducing the lethality of the fry by installing ganban rock known as a water purifier and an infrared emitter in the fish farm, thereby obtaining a large harvest in the fry farming.

Claims (4)

To improve immunity of the fry, which is achieved by adding 0.005 ~ 0.015wt% of vitamin B1 and B2 to chitosan solution 1 ~ 5% wt, respectively, and then adding the mixture 0.5 ~ 2wt% to the existing feed for fry. Feed for fish. Garlic: Astragalus: Ginseng: Persimmons made by adding 0.5-1wt% extract of perilla extract in 1-2: 1: 1-2: 1-3: 1-2 (w / w) ratio to existing feed Feed for fish farming to enhance immunity. Fish feed for improving the immunity of the fry, characterized in that the organic mineral 5 ~ 8wt% solution is added to the existing fry feed. Conventional fish farming, fish farming garlic feed with the chitosan solution and vitamins B1 and B2; Astragalus; A method of farming larvae characterized in that the fry are farmed while feeding a fish feed with the extract extracted from ginseng and perilla, or a fish feed with the addition of organic minerals to the fry farms in which elvan is added.