KR101584918B1 - Composition for controlling and preventing fish diseases using red clay slurry and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a method for controlling and preventing fish diseases by using red clay slurry. The method comprises the steps of: 1) inserting and mixing 10 to 40 wt% of sodium hydroxide (NaOH) with respect to 60 to 90 wt% of red clay; 2) melting the mixture at the temperature of 1100 to 1200°C in a melting furnace installed with a graphite crucible; 3) cooling the molten material; 4) crushing the cooled material by using a crusher; 5) manufacturing red clay slurry by secondarily crushing after adding water to the crushed material; 6) neutralizing by adding sulfuric acid to the red clay slurry; and 7) adding the neutralized red clay slurry to breeding water in a fish farm.

Description

Technical Field [0001] The present invention relates to a yellow clay slurry composition for controlling fish diseases, a method for controlling fish diseases using the same,

The present invention relates to a method of producing a loess slurry-fed trout-style feed for antibiotic substitution and a trout culture method.

Korea, which is the sea on three sides, has the highest per capita consumption per capita in the world, along with Japan, and this trend is likely to continue for the time being. Korea accounts for 5 million tons of seafood, about 10% of total food (including imported beverages) of about 50 million tons a year (about 30%, 15 million tons of feed grains). Also, the consumption of aquatic products is gradually increasing due to the improvement of income level and the increase of interest in health. Korea has a geographical advantage that it can easily obtain aquatic products, but recently the fishery amount is decreasing due to changes in fish species due to abnormal temperature, decrease in population due to overfishing and illegal fishing, and pollution of the coast. In order to increase the incomes of fishermen and fisheries in this situation, aquaculture has been extensively used, and the range of aquaculture has diversified into seawater fishes, freshwater fishes and shellfishes.

When a fish is cultivated (sea-water fish, freshwater fish), unlike the natural state, it is raised in a closed environment, and various diseases depending on the breeding environment are caused. In other words, various diseases are caused by water pollution, aging of aquaculture farm, tempering of breed, high density breeding, etc., and the amount of dead fish fish is 20 ~ 30% of the total production amount. Is coming. Diseases that occur in aquaculture fish are largely classified into bacterial, parasitic, and viral diseases. Examples of bacterial diseases include Vibrio spp., Slug bacterium, Streptococcus, and Edwards spp., Which are caused by single infection or mixed infections. Examples of parasitic diseases include diseases caused by scurvy, Sexually transmitted diseases vary from species to species. One way to solve this problem is to sterilize the aquaculture. Because the number of aquaculture used in the aquaculture process is used as the habitat and infection route of the pathogens, the aquatic organisms are treated with ozone, ultraviolet rays, and chemical disinfectants to prevent contact between various pathogens and aquaculture fish. The use of antibiotics improves nutrient utilization, prevents disease, cures and promotes growth, making large-scale population rearing possible today, and has been used as the most economically effective feed additive. In the aquaculture industry, large amounts of antibiotics have been used for growth promotion, disease prevention and treatment. However, due to the overuse of these antibiotics, many pathogens have recently become resistant to antibiotics. In particular, resistant bacteria that have the same resistance to many antibiotics have emerged. In addition, when antibiotics were mixed in feed, they often remained in aquatic products, resulting in unstable water environment due to antibiotics that harm human health and flow into the natural environment. In recent years, the European Union (EU) has banned the use of antibiotics in feed due to these effects, and most countries are following it.

Since 2005, Korea has also reduced the number of antibiotics for feed additive to 44 from 2005, and since July 2009, it has banned the addition of 7 additional feeds. In July 2011, enameysin, tyrosine, , Antibiotics such as bacitracin methylenedisilisylate, chymotrypsin, thiamyrin, apramycin and uvilamycin, and antibiotics such as sulfathiazole 1, which prohibit the addition of antibiotics, It is only allowed to use the prescription according to the treatment purpose of the manager. Therefore, these antibiotic substitutes are very necessary in aquaculture.

Rainbow Trout ( Oncorhynchus mykiss ) is a cold-water fish belonging to the Salmoniformes Salmonidae. It is the dominant species in the western Pacific, and is now cultivated all over the world except the tropical regions. The beginning of rainbow trout farming in Korea began in 1965 when Henrycline schmidt, California, USA, donated 10000 bottles to Kangwon province after being donated by Jung Seok Cho. In 1982, the first batch of feed was developed in Korea, which led to a rapid increase in production volume. However, due to the limited number of fishery sites, the reduction of fish farms, the weakening of competitiveness due to import and export of foreign marine products through the WTO system, Soybeans and other social issues, the volume of production decreased due to the strengthening of water quality regulations. In 2005, it suffered the worst decline since suffering malachite green wave. Since then, the government has been expanding investment in safe food production by adopting eco-friendly fisheries systems such as non-antibiotic farming.

Accordingly, the inventors of the present invention completed the present invention by confirming that it is possible to control and prevent fish diseases by using a yellow loess slurry composition prepared by processing yellow loess while studying a method for controlling and preventing fish diseases occurring in farm fishes without using antibiotics .

1. R.C. Cipriano, J. Bertolini, Selection for virulence in the fish pathogen Aeromonas salmonicida, using Coomassie Brilliant Blue agar, J Wildl Dis. 24, 672-678 (1988) 2. S.S. Choi, J.W. Lee, B.Y. Kang, N.J. Ha, Antimicrobial resistance patterns of vancomycin-resistant Streptococcus equinus isolated from animal foods and epidemiological typing of resistant S. equinus by microbial uniprimer kit, Arch Pharm Res. 26, 638-643 (2003) 3. S. Ulitzur, Vibrio parahaemolyticus and Vibrio alginolyticus: Short generation-time marine bacteria, Microb Ecol. 1, 127-135 (1974) 4. Song Ji-Young et al., Survey on Residues of Penicillin Veterinary Drugs Among Domestic Distribution Shafts and Aquatic Products. Analytical Science & Technology, 25 (4), 257-264, 2012. 5. Detection of fluoroquinolone antibiotics by Bukhan et al. J. Korean Electrochem. Soc., 13 (1), 63-69, 2010. 6. Health and welfare policy forum, 2003, 77: 72-82; Journal of the Korean Veterinary Medical Association, 38: 305-316, 2002. 7. Huh Gang-joon, et al., Measures to Prevent Aquatic Animal Diseases and Aquatic Animal Products. Kor. J. Food Hygiene, 7 (2,3), S7-S19 (1992). 8. Actual Status Report on Participation Solidarity, Shrimp, and Aquatic Antibiotics Ⅱ (Problems and Improvement of Residual Substance Inspection System in Meat). The Korean Veterinary Medical Association, 3, 2006. 9. An Myeong-ok, Animal Antibiotic Residues in Foods Serious. Health and Welfare (Grand National Party) Press Release, October 22, 2005.

It is an object of the present invention to provide a method for preventing and preventing fish diseases using yellow clay slurry.

In order to achieve the above object,

1) mixing and mixing 10 to 40% by weight of sodium hydroxide (NaOH) with respect to 60 to 90% by weight of loess;

2) melting the mixture in a melting furnace equipped with a graphite crucible at a temperature of 1100 ° C to 1200 ° C;

3) cooling the melt;

4) pulverizing the cooled material using a pulverizer;

5) adding water to the pulverized product and then pulverizing the pulverized product to prepare a yellow loam slurry; And

6) neutralizing the yellow clay slurry by adding sulfuric acid;

7) adding the neutralized loess slurry to aquaculture water;

And a method for preventing and preventing fish diseases using the yellow loess slurry.

According to one embodiment of the present invention, the loess comprises SiO ₂ 40 to 80 parts by weight 10 to 30 parts by weight of the Al ₂ O ₃ with respect to, Fe ₂ O ₃ as an essential component parts of 0.5 to 20 parts by weight, the additional component And 0.1 to 5 parts by weight of at least one component selected from the group consisting of CaO, MgO, K ₂ O, Na ₂ O, TiO ₂ and P ₂ O ₅ . The loess can be collected from the natural world by a conventional method in the art, or can be obtained by purchasing a commercially available product.

Among the essential components of the loess, the SiO ₂ component has a strong adsorption power, and has functions such as antibacterial and deodorization, mineral elution, and increased dissolved oxygen amount. However, since SiO ₂ is substantially insoluble, it is difficult to expect such an effect when a fine powder of loess having a main component of SiO ₂ is fertilized. However, if after mixing a predetermined amount of caustic soda to the yellow soil of SiO ₂ is a main component according to the invention is heated at a high temperature, melting the tissue of SiO ₂ kkaejimyeonseo SiO ₃ ^{2 -} is the present in the form of ions can be dissolved upon contact with water have.

The step 1) is a step of mixing 10 to 40% by weight of sodium hydroxide (NaOH) with 60 to 90% by weight of loess to obtain a mixed composition, wherein sodium hydroxide (NaOH) (NaOH), or 30 wt% of sodium hydroxide (NaOH) relative to 70 wt% of the loess. As the amount of caustic soda in the mixed composition increases, the ionization yield of SiO ₂ increases. However, when caustic soda is added in an excessively large amount, sodium hydroxide (NaOH) is used in an amount of 70% by weight based on 70% by weight of the cerium oxide in the embodiment of the present invention.

Step 2) is the step 1) was heated to melt for 1 to 4 hours a mixed composition obtained in the 1100 to 1300 ℃ to obtain a melt of the liquid at, the heating, melting process via the main component of SiO ₂ of ocher SiO ₂ + NaOH → Na ₂ SiO ₃ + H ₂ O ↑ and dissolves in water.

According to the present invention, when the yellow mortar and caustic soda are blended and melted in the above-mentioned proportions, the melting temperature and the melting time are lowered to about 1100 to 1300 ° C for about 1 to 4 hours, thereby reducing the manufacturing cost of the product. The melting temperature may be 1100 to 1290 ° C, 1100 to 1280 ° C, 1100 to 1270 ° C, 1100 to 1260 ° C, or 1100 to 1250 ° C. The melting time may be from 1 to 4 hours, from 2 to 4 hours or from 2 to 3 hours. Hwangto is very weak in strength compared to pure silica (SiO ₂ ) or elvan stone used in conventional processes and has a porous structure so that it can be melted at low temperature. Especially, when caustic soda is used as alkali substance, Thereby further lowering the melting temperature of the loess. In this case, when the melting is performed at a temperature lower than 1100 ° C, the yellow earth does not melt, so that the reaction with caustic soda does not occur, so that water-soluble SiO ₃ ^{2 -} ions are hardly produced. , The same result as in the case where the yield is performed at 1200 DEG C, and the energy cost is large, which may cause economic loss.

Step 3) is a step of cooling the melt obtained in step 2), and the cooling can be carried out by a conventional method in the art. For example, but not exclusively, the melt can be rapidly cooled or cryogenically cooled using a cooling facility and naturally cooled at ambient temperature. If the cooling facility is used, it is preferable to cool the room at room temperature because additional costs are incurred due to the installation of the cooling facility. However, in the case of mass production, rapid or low-temperature cooling system may be required since many products must be produced quickly. According to one embodiment of the present invention, the melt can be spontaneously cooled at room temperature for 24 to 48 hours.

According to one embodiment of the present invention, it may further comprise the step of grinding the cooled cooling product in step 3) into microparticles or nanoparticle size. The cooled product may be pulverized into microparticles or nanoparticles and then may be prepared into a liquid, colloidal or solid product depending on the intended application. For example, microparticles or microparticles of 1 to 1000 mu m in size, Nanoparticles having a size of 1000 nm. The pulverization can be carried out by a conventional method in the art.

The crusher is preferably a hammer mill or a ball mill, and it is more preferable that the crusher is crushed using a hammer mill and then ground into a powder of nanoparticles using a ball mill, but the present invention is not limited thereto. The particle size of the crushed loess slurry by the method of step 5) may preferably be 300 to 700 nm.

By the above method, SiO ₂ 10 to 30 parts by weight of Al ₂ O ₃ and 0.5 to 20 parts by weight of Fe ₂ O ₃ are contained in an amount of 40 to 80 parts by weight and CaO, MgO, K ₂ O, Na ₂ O, TiO ₂ and P ₂ O and contains one or more components selected from the group consisting of ₅ parts of 0.1 to 5 parts by weight, the SiO ₂ is SiO ₃ ^{2 -} can be prepared ocher slurry, characterized in that is present in the form of ions.

In step 6), sulfuric acid is preferably added to neutralize the solution in the range of pH 6 to pH 8, and it is more preferable to neutralize the solution in the range of pH 6.5 to pH 7.5, but the present invention is not limited thereto.

According to an embodiment of the present invention, the particle size of the loess slurry in step 6) may be 300 to 700 nm, and the solid content may be about 20%.

The present invention can also prevent and prevent fish diseases by adding the neutralized yellow loess slurry prepared by the above method to the aquaculture water.

In the embodiment of the present invention, the addition concentration of the loess slurry in the step 7) may be added at 5 ppm to 250 ppm. Outside of the above range, the effect of preventing and preventing fish diseases may be significantly deteriorated.

In an embodiment of the present invention, the fish fish disease may be any one or more selected from the group consisting of infectious disease, gill bleeding, streptococcal disease, Aeromonas disease and Vibrio disease, but not limited to, You can include both.

Preferably, said fish disease is selected from the group consisting of Aeromonas , Streptococcus 0.0 > and / or < / RTI > Vibrio , and most preferably the fish is Aeromonas salmonicida, Streptococcus equinus and Vibrio alginolyticus , < / RTI > and the like.

In the embodiment of the present invention, by adding the loess slurry prepared in the above 1) to 6) step of the present invention to the breeding water of the aquaculture farm, Aeromonas salmonicida subsp . salmonicida (KCCM 41055), Streptococcus which causes sudden massive death such as bleeding of trout and eel culture gill equinus (KCCM 11930), Vibrio Vibrio that cause illness that causes sepsis and muscle necrosis and hemorrhage alginolyticus (KCCM 40513), it was confirmed that the method using the yellow clay slurry of the present invention has an excellent effect for the control and prevention of fish diseases.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is well known and commonly used in the art.

The addition of the loess slurry of the present invention to the water in aquaculture can prevent the antibiotic resistance due to the unreasonable abuse of antibiotics and prevent the instability of the aqueous environment. It also contributes to the development of eco-friendly aquaculture by using non-antibiotics without antibiotics.

1 is a flow chart of a method for controlling and preventing fish diseases using a loess slurry according to the present invention.
2 is a graph showing the average particle size of the yellow clay slurry according to the present invention.
FIG. 3 is a view showing a yellow clay slurry according to the present invention. FIG.
FIG. 4 shows the relative antimicrobial activity over time in the culture medium of the pathogen A. salmonicida , S. equinus , and V. alginolyticus to the untreated group (Con), the loess slurry concentration group, and the antibiotic (OTC) The graph is analyzed. ((A): A. salmonicida Experimental results on culture medium; (B): Experimental results on S. equinus culture medium; (C): Experimental results in V. alginolyticus culture medium)
FIG. 5 is a graph showing the results of a comparison between the untreated group (Con) according to the present invention, the treated group according to the concentration of the loess slurry, the pathogens A. salmonicida , S. equinus , V. alginolyticus This graph is a graph showing the relative antibacterial activity of the trout in the trout collected from a trout aquaculture tank. ((A): A. salmonicida In in the number of breeding vitro Experiment result; (B): in the number of breeding in S. equinus vitro Experiment result; (C): in V. alginolyticus in a number of breeding vitro Experiment result)
6 is a non-treatment group (Con), concentrations of the yellow soil slurry treatment group, antibiotics (OTC) pathogens for the group treated with S. equinus, V. trout mortality in a number of breeding alginolyticus processing trout form the tank according to the present invention ( (Above) and pathogen viability (below). ((A): S. equinus Processing in Experimental results in vivo feeding; (B): V. alginolyticus treatment in Results of vivo experiments)

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

< Example  1> Yellow soil Slurry  Antibacterial experiment to confirm possibility of substitution of antibiotics

<1-1> Yellow soil slurry  Ready

70% by weight of yellow loess (Boryung Hwangto Co, Korea) and 30% by weight of caustic soda were mixed in a vessel and mixed in a 50 L capacity graphite crucible in an electric furnace (STI Supreme Thermal Instrument Co., Or more. The melt was cooled at room temperature for 1 day and then pulverized to a size of 3 mm or less using a hammer mill (Wonder Blender, Sanplatech Corp., Japan), and further prepared into a loess slurry having an average particle size of 500 nm using a bead mill. Sulfuric acid was added to neutralize it to pH 7.0 ± 0.5, and then it was used for antibacterial and trout breeding experiments.

<1-2> Selection of strains and activation of strains

Aeromonas causing trout breeding salmonicida subsp . salmonicida (KCCM 41055), trout and Streptococcus equinus (KCCM 11930), Vibrio alginolyticus (KCCM 40513) which causes Vibrio illness that causes necrosis and bleeding, sepsis and muscle that causes a sudden mass mortality eel form during the gills bleeding Were tested for antimicrobial activity.

<1-3> Experimental Method

Each strain present as glycerol stock in a cryogenic freezer (-80 ° C) is activated by incubating under the conditions shown in Table 1 below. The pathogen culture was activated according to the culture conditions. 0.5 (1.5x108 CFU / mL), and inoculated in a 50 mL cornical tube filled with sterilized liquid medium and live (untreated) farms. After the inoculation, the loess slurry was treated at various concentrations (0, 5, 25, 50, 100, 250 ppm) applicable to farms and incubated at an appropriate temperature for growth of each strain at 50 rpm The culture is carried out according to the culture conditions of the transferred bacteria. As a control, oxytetracycline is used and the number of viable cells is measured at specific time by starting the culture.

Culture conditions of target strain division Test strain badge Incubation time Temperature Gram negative A. salmonicida Trypticase soy broth 18 hours 20 ℃ Gram positive S. equinus Brain heart infusion broth 72 hours 37 ℃ Gram negative V. alginolyticus LB broth + 2.5% NaCl 18 hours 30 ℃

<1-4> Evaluation of antimicrobial activity of loess material

Each strain was treated with a slurry of loess soil under the conditions of the above Example <1-3> to cultivate the strain, and then the number of viable cells was analyzed by time.

As a result, when the slurry for processing the loess material was treated as shown in Figs. 4 and 5, the growth of the strain was inhibited in a concentration-dependent manner. The results in FIG. 4 suggest that the optimal inhibition time for the pathogens of the loess slurry in the liquid culture medium is 4 hours. Based on these results, in the experiment of FIG. 5, the result of analyzing the number of live bacteria in the breeding water after treating the slurry for 4 hours as the optimal inhibition time. As a result, it was analyzed that the cultivation of the actual fish also inhibited the growth of the strain by the slurry.

The result of this embodiment is that in suggest that the yellow soil slurry is optimized culture conditions in a liquid culture medium, as well as actual fish within the growing number of breeding pathogen-inhibiting effect of each strain and on vitro.

< Example  2> Yellow soil Slurry  Experiment on breeding trout and strain for confirmation of antibiotic substitution possibility

<2-1> Experimental language  And breeding management

Rainbow trout with mean weight of 30 ± 1.6 g (mean ± SD) of experimental fishes was randomly placed in 250L rectangular water tanks in each experimental group. The average water temperature of each experimental tank was maintained at 15 ± 0.5 ℃, After the treatment, green clay slurry was treated with 25ppm for 5 days.

<2-2> Experimental design

Based on the experimental results of Example 1, a plot of trout and strain breeding experiment was prepared as shown in the following table. Control (C), loess 25ppm antibiotic additive (Tet)

Group 1: S. equinus 1.1
Oxytetracycline 1.2
Control 1.3
Yellow Loess 25ppm Group 2: V. alginolyticus 2.1
Oxytetracycline 2.2
Control 2.3
Yellow Loess 25ppm Group 3: No additives 3.1
Oxytetracycline 3.2
Control 3.3
Yellow Loess 25ppm

<2-3> Analysis Items

The amount of dead fish in trout,

<2-4> Experimental results

The trout was treated with strain and loess slurry under the conditions of the above Example <2-2>, and the amount of dead trout and the number of live bacteria in the breeding stock were analyzed by time.

As a result, the survival rate of the trout and the growth inhibition rate of the strain were maintained to be higher than those in the case where the slurry of the loess material was treated as in Fig.

The result of this embodiment is that in In vivo, loess slurry also has a pathogenic inhibitory effect.

Claims (6)

1) mixing and mixing 10 to 40% by weight of sodium hydroxide (NaOH) with respect to 60 to 90% by weight of loess;
2) melting the mixture in a melting furnace equipped with a graphite crucible at a temperature of 1100 ° C to 1200 ° C;
3) cooling the melt;
4) pulverizing the cooled material using a pulverizer;
5) adding water to the pulverized product and then pulverizing the pulverized product to prepare a yellow loam slurry; And
6) neutralizing the yellow clay slurry by adding sulfuric acid;
7) adding the neutralized loess slurry to aquaculture water;
And a method for preventing and preventing fish diseases using the yellow clay slurry. The method according to claim 1, wherein the concentration of the yellow clay slurry in step 7) is 5 ppm to 250 ppm. The method for preventing and preventing fish diseases according to claim 1, wherein the fish is at least one selected from the group consisting of poultry, gill bleeding, streptococcal disease, aeromonas disease, and vibriosis. 3. The method of claim 1, wherein the fish is Aeromonas , Streptococcus And Vibrio spp. The present invention also relates to a method for preventing and preventing fish diseases using yellow clay slurry. The method of claim 1, wherein the fish is Aeromonas salmonicida , Streptococcus equinus, and Vibrio alginolyticus . The method for preventing and preventing fish diseases using the yellow loess slurry according to claim 1, The method according to claim 1, wherein the particle size of the loess slurry in step 5) is 300 to 700 nm.

Images