KR20200060799A - A composition for the eel leptocephalus diets and manufacturing method - Google Patents

Abstract

The present invention relates to a method for manufacturing a nano-BIOFLOC colloid feed for eel leptocephalus, which comprises: (A) a step of preparing BIOFLOC feed; (B) a first processing step of using a functional group to the BIOFLOC feed to chemically process the BIOFLOC feed; and (C) a second processing step of mechanically pulverizing the BIOFLOC feed passing through the step (B). According to the present invention, the feed is dispersed in cultivation water to be prevented from being dropped on the bottom, such that the feed does not rot for a long time, thereby feeding the eel leptocephalus without a change in water quality even if the water is not exchanged. Moreover, the feed with a size suitable for the eel leptocephalus and the like, to which feed is sufficiently provided due to a small mouthpart, is fed to increase an initial survival rate of the eel leptocephalus, thereby increasing farming efficiency.

Description

A composition for the eel leptocephalus diets and manufacturing method

The present invention is a nanoparticle that can be used as an initial feed for hatchery larvae, which have limited initial feeding activity due to small goji, by nanoparticles useful organic matter of breeding water used in biofloc farming to the size of colloidal dispersion and modify the particle surface. It relates to a method of producing a bio-flock colloidal hatchery feed.

The production of marine products in Korea exceeds 50%. This is also the result of a lot of research and investment in fish farming technology over the past 20 years in order to take advantage of Korea's geographic advantage surrounded by the sea on three sides in line with the needs of the era of environmental pollution and depletion of natural resources.

At the present time, shifting from catching fisheries to cultivating fisheries, research is underway to enter each aquaculture method from seedling production to intermediate breeding and harvesting. In general, artificial propagation technology and larval management at the beginning of hatching are known as the most important stages of cultivation. In particular, increasing the survival rate of hatched larvae is directly related to productivity, and research on prey organisms in this period is actively conducted.

In the seawater, there are many microscopic zooplanktons, for example, larvae of sponges, tonics and invertebrates, reptiles of reptiles, and noplius larvae of copepods, which play an important role in the natural food chain. However, few of them have conditions as prey for the production of fish seedlings.

Until now, most of the hatchery larvae are rotifers such as rotifers and altemia. Altemia is usually imported in egg form and hatched to feed, and rotifers, etc., have been supplied with algae cultured in outdoor ponds using fertilizer, or commercially available chlorella, yeast, etc. have been used.

However, in recent early stages of larvae, much effort has been devoted worldwide to produce seedlings of fish with a small mouth size, mainly the nopleus larvae and protozoa of copepods as candidate organisms. However, in order to produce them, copepods Noplaus has to cultivate copepods at all stages and has extremely low culture density. Protozoa are promising such as Euphrates sp and Fabra salina. Although species have been cultivated, they are not widely used for seedling production because they are pathogenic or parasitic protozoa.

Also, the prey organisms that are absolutely used in Korean farming, Brachionus plicatilis (large larvae, 130~140 ㎛) and medium-sized species B. rotundiformis, small larvae, 100 ~120㎛) is also known as an unsuitable prey for neung fish, which are classified as classy fish species with small mouth and throat. In order to produce seedlings of fish with a small mouth and throat in this way, there is an urgent need to research small-sized and easily mass-cultivated fish.

On the other hand, BioFloc Technology (BioFloc Technology) grows useful microorganisms and cultured species of heterotrophic bacteria and autotrophic bacteria, while bacteria grow in aquaculture, such as ammonia, etc. It refers to a farming method that decomposes organic by-products harmful to farmed fish and converts it into food that can be consumed by farming fish, and also cleans up the breeding water.

During the bio-floc farming, bio-floc sludge is naturally produced in the rearing water, and the unused feed residue, photosynthetic bacteria, attached algae, tagatrophic bacteria, phytoplankton, animal plankton, copepods, nematodes, etc. are entangled together. It is a lump that is said to be composed of over 80% of organic matter. Biofloc sludge degrades over time by some biofloc microorganisms and is reduced to breeding water, while some accumulate on top of the breeding water to form thickly stacked cakes, adversely affecting the aquaculture process.

That is, the amount of oxygen input is increased by reducing the surface area of the livestock that comes into contact with the atmosphere, and it is prevented that the farmed fish and independent nutrients are exposed to light of appropriate illumination. Therefore, in the biofloc farm, the biofloc sludge has been taken out and removed, and in this process, the management cost has to be increased.

As a result of analyzing the components of the recent biofloc sludge, the crude protein contains 25~41%, crude fat 2~3%, and 20~30% of lookup powder. In particular, highly unsaturated fatty acid, carotenoid, phosphorus, methionine, lysine and arginine, etc. It has been shown that even the amino acids of fish contain a lot of ingredients that are very useful for fish farming, and these useful ingredients increase the possibility that biofloc sludge is provided as a feed composition for farmed fish or as a feed additive that can increase growth and immunity.

However, the preceding studies have provided the steps of preparing the BIOFLOC feed of the present invention (A), the first process step (B) of chemical treatment using a functional group (fuctional group) on the BIOFLOC feed, and the BIOFLOC which has undergone the step (B). It shows a difference in its composition and effect from the manufacturing method of the nano-bio-flock colloidal hatchery feed, which is composed of the second process step (C) of mechanically crushing the feed.

In Korean Patent Registration No. 10-0908928, Bacillus subtilus KS-3, Bacillus amyloliquefaciens KS-4, Bacillus agaradhaerens KS-5, Paeni Bacillus lentimobus KS-6, Bacillus laevolacticus KS-7, Leuconostoc paramesenteroides KS-9, Cursia sibirica KS- Aquaculture fish containing at least one microorganism selected from the group consisting of 13 and Sphingobacterium spiritivorum GC subgroup B (Flavobacterium) KS-18. It discloses a feed additive or a feed composition. In Korean Patent Registration No. 10-1420392, after mixing a mixture of mushrooms containing two or more of ink mushroom extract and shiitake mushroom, oyster mushroom, and mandarin mushroom, after adding a fat and oil mixture containing skim milk and skim milk powder, mushrooms containing fermented lactic acid bacteria It characterized in that the production of fermented milk containing lactic acid bacteria, wherein the mushroom-containing lactic acid bacteria fermented milk is 0.05 to 20 parts by weight of mushroom mixture, 0.05 to 20 parts by weight of mushroom mixture, including two or more of shiitake mushrooms, oyster mushrooms and ganoderma mushrooms, shiyu And 1.0 to 30 parts by weight of a fat-and-oil mixture mixed with skim milk powder, and characterized by being prepared by mixing a lactic acid bacteria fermentation broth, wherein the mushroom-containing lactic acid bacteria fermented milk is used as an additive for fermented milk itself or freeze-dried as an eel-type feed. Disclosed is a feed for eel and feed additives for energy savings in a farm with a feature. In Korean Patent Registration No. 10-0860111, the new microorganism Traustocatrium sp. Thraustochytrium sp.KJS-1, Bacillus polyfermenticus KJS-2, and the two new microorganisms and Bacillus licheniformis. Disclosed are feed additives and feeds. In the Republic of Korea Patent Registration No. 10-0692378, the length of the intestinal larvae in the range of 50㎛~60㎛ and the width of the chest is 44㎛~46㎛ in the optimal culture conditions of 29 ~ 30 ℃ temperature and 32 ~ 34‰ salinity 4 Isochrysis galbana, which has a cell size in the range of µm to 5 µm, is proliferated remarkably excellent as a prey, thereby producing a fertilizer for hatchery cultivation of tortoises and seedling of tortoises that were impossible to grow. Disclosed is a southern larvae that can be used as a hatchery larvae having an effect of enabling production and a method for culturing the same.

The present invention has an object to provide a nano-bio-floc colloidal-enriched sage feed and a method for manufacturing the bio-floc organic material to be nano-particled and dispersed in a water layer so that it can be fed to eels hatching eel with small goji. .

The present invention is a step of preparing a BIOFLOC feed (A), a first process step (B) of chemical treatment using a functional group (fuctional group) on the BIOFLOC feed, and mechanically crushing the BIOFLOC feed that has undergone the step (B) It provides a method for producing a nano-bio-flock colloidal hatchery feed consisting of the second process step (C).

The (B) step is characterized in that the surface modification of the BIOFLOC feed with ??OH or ??COOH functional groups, the ??OH functional group modification is characterized by using ethanol or glycerol, the ??COOH functional group modification is It provides a method for producing a nano-bio-flock colloidal hatchery feed characterized by using acetic acid.

In addition, it provides a nano-bio-flock colloidal hatchery feed prepared by the method of manufacturing the nano-bio-flock colloidal hatchery feed.

By using the biofloc colloidal hatchery feed according to the present invention and a method for manufacturing the same, the food is distributed in the breeding water so that it does not settle on the floor, so that the food does not rot for a long time and evenly distributed in a large amount in the water layer without hatching without changing water quality. The eel can be fed, and it can increase the efficiency of aquaculture by increasing the initial survival rate by supplying an appropriately sized food to an eel hatching eel or the like, which is unable to provide sufficient feeding due to small goji.

1 is a particle size distribution diagram of the feed for BIOFLOC.
2 is a structural formula of acetic acid, glycerol, ethanol.
Figure 3 is a schematic diagram showing the process of nano-dispersion of the feed for BIOFLOC.
Figure 4 is a particle size distribution (10min process) of the feed for Ethanol-treated BIOFLOC.
Figure 5 is a particle size distribution (20min process) of the feed for Ethanol-treated BIOFLOC.
6 is a particle size distribution (40min process) of the feed for Ethanol-treated BIOFLOC.
7 is a particle size distribution (10min process) of Glycerol-treated BIOFLOC feed.
Figure 8 is a particle size distribution (20min process) of the feed for Glycerol-treated BIOFLOC.
Figure 9 is a particle size distribution of the feed for Glycerol-treated BIOFLOC (40min process).
Figure 10 is a particle size distribution (10min process) of the feed for acetic acid-treated BIOFLOC.
11 is a particle size distribution chart (20min process) of the feed for BIOFLOC treated with acetic acid.
12 is a particle size distribution chart (40min process) of the feed for acetic acid-treated BIOFLOC.
13 is a particle size distribution chart after 3 days of the feed for acetic acid-treated BIOFLOC.
14 is a photograph showing dispersion over time of the second process step of the feed for BIOFLOC.

Biofloc useful organic materials are very diverse and a large number of living microorganisms, such as plant plankton-animal plankton-protozoa, organisms and feed surplus, and incubated larvae such as amino acids, unsaturated fatty acids, and low molecular proteins degraded by microorganisms can be used as food. There are a variety of food sizes, large and small.

However, the size of the floc present in the biofloc breeding water is generally too large to feed the small goji, and when it is supplied to the hatchery, it sinks and accumulates on the bottom after a certain time. In this case, it decays from the bottom and deteriorates the livestock environment, threatening the survival of hatchery fish. The same problem occurs even if the biofloc useful organics are collected and dried and crushed and then fed to farmed fish.

The inventors of the present invention are nanoparticles to a size sufficient for a hatched stool to provide a biofloc useful organic material capable of supplying sufficient nutrients to a hatchery shark, which is rich in nutrients. It is to devise a method of manufacturing a nano-bio-flock colloidal hatchery chow feed that allows the hatchery shark to continue to feed by preventing it from sinking to the floor.

Through this technology, the bio-flock organic matter can be evenly distributed in the water layer for a long period of time without being rotten. In particular, it is possible to maintain good water quality without frequent return to the hatchery fish, whose survival rate rapidly decreases in the process of water quality management for breeding water. It was able to supply food. Hereinafter, the present invention will be described in detail with specific examples.

1. BIOFLOC analysis before experiment

Natural organic matter (NOM) is divided into hydrophobic (hydrophobic) and hydrophilic (Hydrophilic). These particles are chemically attracted and have a nature of aggregation with each other. The sample feed for BIOFLOC is a hydrophobic material composed of aliphatic carbon, protein, and carbohydrate, which ensures stability when producing nanoparticles of particles, and requires pretreatment for interaction between particles.

The present inventors investigated the particle size of the BIOFLOC feed using dynamic light scattering (DLS). 1 is a particle size distribution diagram of the feed for BIOFLOC. Table 1 summarizes the particle sizes shown in FIG. 1.

Initial particle size of feed for BIOFLOC D ₁₀ 464 nm D ₅₀ 8.23 μm D ₉₀ 20.4 μm

As shown in Figure 1, Table 1, the particle size of the feed for BIOFLOC was observed in three parts: D ₁₀ , D ₅₀ , and D ₉₀ , and D ₁₀ 464 nm, D ₅₀ 8.23 μm, D ₉₀ 20.4 μm, and the particle distribution was very high. I could see a wide variety. In addition, it can be found that the particles become larger and precipitate due to the aggregation phenomenon between the particles, and it was found that a technique is needed to prevent mechanically and chemically reagglomerating each of these particles.

2. Nano dispersion process of feed for BIOFLOC

In order to use the pre-treatment material having an eco-friendly functional group, chemical pre-treatment was performed using acetic acid having a structure of COOH and glycerol and ethanol having an OH structure.

2 is a structural formula of acetic acid and glycerol and ethanol. In order to increase the water dispersion capacity of the feed for BIOFLOC, the particles of the BIOFLOC feed were nanonized and the particle surface was modified to have a functional group. At this time, pre-chemical pretreatment was performed for the particles of the BIOFLOC feed using Acetic acid having a structure of -COOH and Glycerol and Ethanol having a functional group of -OH.

Figure 3 is a schematic diagram showing the process of nano-dispersion of the feed for BIOFLOC. Preparing the BIOFLOC feed of the present invention (A), the first processing step (B) chemically treated with a functional group (fuctional group) on the BIOFLOC feed, the second step of mechanically pulverizing the BIOFLOC feed that has undergone the step (B) It provides a method for producing a nano-bio-flock colloidal hatchery edible feed consisting of the step (D) of packaging the BIOFLOC feed that has been processed (C) and (C). Through the above process, the BIOFLOC feed prevents re-aggregation by interacting between particles, and improves the stability of the dispersion by increasing the charge stability of the organic matter in the breeding water.

(A). Steps to prepare BIOFLOC feed

In the BIOFLOC farming process, biofloc sludge is naturally produced in the breeding water, and the unused feed residue, photosynthetic bacteria, attached algae, tagatrophic bacteria, phytoplankton, animal plankton, copepods, nematodes, etc. are entangled together. It is formed into a loaf. The biofloc sludge floats on the upper part of the stock with air bubbles generated during the aeration process. If these are not removed in time, the light transmittance is lowered, and the photosynthesis of phytoplankton is inhibited and the biofloc efficiency can be lowered and must be collected continuously. .

The biofloc sludge is periodically collected by a collection net and dried, and then pulverized in a grinder so that the reaction takes place at a constant rate, thereby preparing a BIOFLOC feed as shown in FIG. 3.

(B). 1st processing step

The BIOFLOC feed is chemically treated with a functional group to modify the particle surface. The functional group may be a carboxyl group (-COOH) or hydroxyl group (-OH).

In the case of a chemical having a carboxyl group, acetic acid having high biological safety can be selected and treated, but is not limited thereto. Glycerol and Ethanol can be selected and treated for chemical substances having hydroxyl groups, but are not limited thereto.

Through this process, as the nano-sized BIOFLOC feed is dispersed in the water layer, it prevents the particle size from growing due to the reaggregation process between particles, so that farmed fish such as small eels and hatched eel can be easily fed.

(C). 2nd processing step

After completing the first processing step, the BIOFLOC feed is subjected to a second processing step of mechanically pulverizing to nanoparticle size. Generally, colloidal particles are larger than molecules or ions, and fine particles having a diameter of about 1 nm to 1000 nm can be dispersed in a gas or liquid.

The process may use ball milling, bead milling, or the like, and determines the time until particles are crushed in nano units through dynamic light scattering. At this time, the grinding time and thus the size of the particles may vary depending on the treatment chemical of the first processing step, which is a chemical pretreatment process.

The speed at which the centrifugation is performed in the above process may range from 5000 rpm to 20000 rpm, or may range from 6000 rpm to 10000 rpm. The time for performing the centrifugation may be, for example, in the range of 1 second to 24 hours, and more specifically, in the range of 1 minute to 30 minutes.

The size of the ball or bead used for ball milling or bead milling may have a diameter of 0.1 mm to 0.3 mm, and an ultrasonic device may be used.

3. BIOFLOC analysis after the experiment

3.1 BIOFLOC Analysis Using Ethanol

After the primary processing was performed using Ethanol having a -OH functional group, the particle size of the BIOFLOC feed was analyzed by varying the processing time of the secondary processing. 4 to 6 is a particle size distribution of the ethanol-treated BIOFLOC feed (10 minutes, 20 minutes, 40 minutes process), it is shown in Tables 2 to 4.

Particle size of Ethanol-treated feed for BIOFLOC (10min process) D ₁₀ 283nm D ₅₀ 562nm D ₉₀ 1.76um

Particle size of ethanolol-treated feed for BIOFLOC (20min process) D ₁₀ 227nm D ₅₀ 451nm D ₉₀ 1.81um

Particle size of ethanolol-treated feed for BIOFLOC (40min process) D ₁₀ 239nm D ₅₀ 426nm D ₉₀ 759nm

As seen from the above, the particle size of the feed for BIOFLOC primarily treated with Ethanol was observed to be about 420 nm to 450 nm at D ₅₀ . When the second process step (C) was performed for 40 minutes, it showed a distribution similar to that of the feed for BIOFLOC, which performed the first process step (B) for 10 minutes, but the particle size was reduced by about 100 nm (D ₁₀ 303 nm -> 239, D ₅₀ 539nm -> 426nm, D ₉₀ 962nm -> 759nm), and thus it was found that it was formed in a particle size capable of stably forming a colloid in a water layer and maintaining a dispersion state.

3.2 Nano dispersion of feed for BIOLOC using glycerol

After the first processing was performed using Glycerol having a -OH functional group, the particle size of the BIOFLOC feed was analyzed by varying the processing time of the second processing. 7 to 9 is a particle size distribution of the feed for Glycerol-treated BIOFLOC (10 minutes, 20 minutes, 40 minutes process), it is shown in Tables 5 to 7.

Glycerol-treated particle size of feed for BIOFLOC (10min process) D ₁₀ 296nm D ₅₀ 528nm D ₉₀ 941nm

Glycerol-treated BIOFLOC feed particle size (20min process) D ₁₀ 254nm D ₅₀ 461nm D ₉₀ 916um

Glycerol-treated particle size of feed for BIOFLOC (40min process) D ₁₀ 163nm D ₅₀ 368nm D ₉₀ 1.17um

As seen from the above, the particle size of the feed for BIOFLOC primary treated with Glycerol was observed from D ₅₀ to about 360 nm to 430 nm. The particle size distribution was characterized by the appearance of two peaks, such as the shape of a camel. Unlike the Ethanol-treated feed (D ₁₀ 239 nm D ₅₀ 759 nm), the values of D ₁₀ and D ₅₀ of the Glycerol-treated feed were 163 nm, 1.17. um showed the largest difference.

3.3 Nano dispersion of feed for BIOLOC using acetic acid

-After the primary processing was performed using acetic acid having a COOH group, the particle size of the BIOFLOC feed was analyzed by varying the processing time of the secondary processing. 10 to 12 are particle size distribution diagrams (10 minutes, 20 minutes, 40 minutes) of the feed for acetic acid-treated BIOFLOC, which are shown in Tables 8 to 10.

Particle size of feed for BIOFLOC treated with acetic acid (10min process) D ₁₀ 283nm D ₅₀ 505nm D ₉₀ 901nm

Particle size of feed for BIOFLOC treated with acetic acid (20min process) D ₁₀ 243nm D ₅₀ 433nm D ₉₀ 785nm

Particle size of feed for BIOFLOC treated with acetic acid (40min process) D ₁₀ 189nm D ₅₀ 336nm D ₉₀ 599nm

The particle size of the feed for BIOFLOC that was first treated with acetic acid was observed from D ₅₀ to 330 nm to 380 nm after the second process. Unlike the particle size distribution of glycerol-treated BIOFLOC feed, it was characterized by showing one peak with the sharpest shape. In addition, the BIOFLOC feed treated with Acetic acid, which was subjected to the second process step (C) for 40 minutes, was compared to the Ethanol and Glycerol treated feed, and the difference between the values of D ₁₀ and D _{50 in} the diet treated with Acetic acid was 189 nm and 599 nm. It was showing the smallest difference. This means that the BIOFLOC feed particles can be distributed to a fairly uniform size by pre-treatment with acetic acid, and can be regarded as advantageous for forming a stable dispersion in the breeding water.

After dispersing the feed for BIOFLOC pretreated with acetic acid in water, the particle size was confirmed after 72 hours. FIG. 13 is a particle size distribution chart after 3 days of acetic acid-treated BIOFLOC feed, which are shown in Table 11. The feed for BIOFLOC treated with acetic acid tended to increase in particle size overall after 72 hours after dispersion, but it was confirmed that the difference was not large and maintained within a certain size by showing the level within the error range.

 Comparison of particle size after 3 days of feed for acetic acid-treated BIOFLOC D ₁₀ D ₅₀ D ₉₀ Early 3 days elapsed Early 3 days elapsed Early 3 days elapsed 189nm 205nm 336nm 365nm 599nm 672nm

4. Result of dispersion

For the nano-dispersion of the BIOFLOC feed, it was first treated in various ways, and then dispersed. 14 is a photograph showing dispersion over time of the second process step of the feed for BIOFLOC. A certain amount of sedimentation was observed in the feed that did not undergo the first process step (B) after a period of time (not shown). As shown in FIG. 14, it was confirmed that the BIOFLOC feed can maintain a more stable colloidal state as the time of the second process step (C) is increased. In addition, compared to the feed that was first treated with ethanol and acetic acid, the feed with glycerol was precipitated at the earliest time, and precipitation occurred in the order of ethanol and acetic acid.

As a result of this, by chemically modifying the surface of BIOFLOC feed and crushing it to nano size, it is stably dispersed in the breeding water and maintains the colloidal state, so that it can be used as the initial food for small farmed fish such as eel hatching eel. Confirmed to complete the present invention.

By using the biofloc colloidal hatchery chow feed and its manufacturing method in the present invention, it is possible to supply foods of an appropriate size to eel hatchery sharks, etc., which do not have sufficient feeding due to small goji, and the food is dispersed in the breeding water. Since it does not settle on the bottom and the food does not rot for a long period of time and is distributed evenly in a large amount of water, the hatching fish can rapidly feed without changing water quality without returning water, which can greatly increase aquaculture efficiency, and thus has industrial applicability.

Claims (5)

Preparing a BIOFLOC feed (A);
A first process step (B) of chemical treatment using a functional group in the BIOFLOC feed; And
A second process step (C) of mechanically pulverizing the BIOFLOC feed subjected to step (B); Method of producing a nano-bio-flock colloidal hatchery feed consisting of.
The method of claim 1, wherein the step (B) is a method for producing a nano-bio-flock colloidal hatchery roe feed, characterized in that the surface of the BIOFLOC feed is modified with -OH or -COOH functional groups.
3. The method of claim 2, wherein the -OH functional group is modified using ethanol or glycerol.
3. The method of claim 2, wherein the -COOH functional group modification uses acetic acid.
According to any one of claims 1 to 4, wherein the nano-bio-flock colloidal hatchery feed prepared by the method of manufacturing the nano-bio-flock colloidal hatchery feed.

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