KR20180119285A - Eco-friendly fish egg disinfectnat comprising water containing detoxified sulfur with inhibition effect for watermold infection as effective component and uses thereof - Google Patents

Abstract

The present invention relates to an eco-friendly fish egg disinfectant containing natural sulfur water having an effect of inhibiting an aquatic fungus infection as an active ingredient, and a method for inhibiting aquatic fungus infection by treating the fish egg with the eco-friendly fish disinfectant. The eco-friendly fish egg disinfectant of the present invention can effectively reduce the water mold infection rate of the fish egg, enhance the hatchability rate of the fry, and replace the formalin known as a carcinogen, thereby being usefully used as a disinfectant of an eco-friendly material for the entire aquaculture industry in the future.

Description

TECHNICAL FIELD The present invention relates to an eco-friendly disinfectant containing natural sulfur water as an effective ingredient and a use thereof.

The present invention relates to an environmentally friendly disinfectant containing natural sulfur water having an effect of inhibiting an aquatic bacterium infection as an active ingredient, and a method for inhibiting aquatic bacterium infection using the environmentally friendly disinfectant.

Rainbow trout (rainbow trout) is called rainbow trout because it has a red iridescent color at the spawning season. At the time of chewing, there are 812 wide horizontal lines like a salmon in the surface of the body, but it grows opaque when it grows and disappears when it is over a year. As a cold water fish, the species introduced in our country inhabits in rivers only for a lifetime and lives in the rivers upstream or in the mountains. The main feeds are aquatic insects, floating creatures, small crustaceans, crustaceans and small fish. The species of rainbow trout introduced in Korea are spawning from October to March, while wild trout in North America are spawning in spring (April to June). It grows faster than other fish, is fertile and has good taste. Rainbow trout, which live in Korea for 7 ~ 8 years, have a life span of 3 to 4 years.

When a rainbow trout egg is infected with a water mold ( Saprolegnia parasitica ), it will not hatch and will die. In order to control fungal infections in fish farms, trout eggs are mainly sterilized using formalin. Formalin has a very good effect on eliminating water fungus infested with trout eggs or inhibiting the infection of water fungi on trout eggs. Despite the excellent disinfecting effect of formalin, however, it is classified as a potent carcinogen and its use is subject to many regulations.

Accordingly, the present invention uses natural sulfuric water, which is an eco-friendly disinfectant excellent in inhibiting water fungal infection, which can replace formalin, and its effect is confirmed.

Korean Patent No. 0782500 discloses an 'aquatic fungus inhibitor using a natural plant extract', Korean Patent Laid-Open Publication No. 2013-0013965 discloses an extract of at least one selected from the group consisting of gosam, rhubarb, and bokbun as an active ingredient Discloses an antifungal and antimicrobial composition for fish microbial pathogens. However, the present invention does not disclose eco-friendly antimicrobial disinfectants containing natural sulfur water having an effect of inhibiting microbial infection of the present invention and uses thereof.

SUMMARY OF THE INVENTION The present invention has been made in view of the above needs, and the present inventors have made intensive efforts to develop an environmentally friendly disinfectant that can substitute for formalin, produce natural sulfur water, As a result of the disinfection treatment, the inventors have completed the present invention by confirming that the water fungus infection rate is remarkably reduced as the concentration of natural sulfur water is increased.

In order to solve the above-described problems, the present invention provides an environmentally friendly disinfectant containing natural sulfur water having an effect of inhibiting an infectious microbial infection as an effective ingredient.

Further, the present invention provides a method for inhibiting aquatic organism infection by treating the environmentally-friendly disinfectant in an egg white.

The eco-friendly disinfectant of the present invention can effectively reduce the infection rate of water fungus in the oyster, enhance the hatchability rate of the oyster, and replace the formalin known as a carcinogen. There will be.

Fig. 1 shows a trout squib used in the present invention.
Fig. 2 is a photograph taken 6 days after infecting trout eggs with water mold. Fig.

In order to accomplish the object of the present invention, the present invention provides an environmentally friendly disinfectant containing natural sulfur water having an effect of inhibiting an infectious microbial infection as an effective ingredient.

In the eco-friendly disinfectant according to an embodiment of the present invention, the aquatic organism may be a homogeneous water mold (Saprolegniales), preferably a water mold ( Saprolegnia parasitica ), but is not limited thereto.

In addition, in the disinfectant of the present invention, the natural sulfur water means sulfur water containing sulfur which is detoxified,

(a) fermenting a mixed medium containing green tea leaves and camphor leaves as a carbon source;

(b) adding a molasses dilution solution sterilized at high temperature to the fermented mixed medium of step (a), inoculating and cultivating a useful microorganism;

(c) adding a toxic sulfur powder to the useful microorganism culture solution of step (b), followed by primary fermentation and decomposition;

(d) disrupting the primary fermentation broth of step (c) with a wet colloid mill;

(e) solid-liquid separation of the crushed primary fermentation broth of step (d), adding a useful microorganism culture solution to the separated precipitate, and then secondary fermentation and decomposition; And

(f) separating the second fermentation broth obtained in the step (e) by solid-liquid separation, drying the separated solid to obtain a first mixture of sodium hydroxide, phosphoric acid (H ₃ PO ₃ ) And liquefying the mixture with a mixed solution of a sodium silicate solution and a Lactobacillus parafarrhinis culture medium, and more particularly,

(a) fermenting a mixed medium containing green tea leaves and camphor leaves at a weight ratio of 6.5 to 7.5: 2.5 to 3.5 as a carbon source at 23 to 35 ° C for 43 to 48 days;

(b) adding a molasses dilution prepared by high temperature sterilization to the fermented mixed medium of step (a), inoculating the useful microorganism, and culturing at 23 to 35 ° C for 92 to 140 hours;

(c) adding toxic sulfur powder to the useful microorganism culture of step (b), and then primary fermenting and decomposing at 25 to 35 ° C for 22 to 26 hours;

(d) disrupting the primary fermentation broth of step (c) with a wet colloid mill;

(e) subjecting the crushed primary fermentation broth of step (d) to solid-liquid separation, adding a useful microorganism culture solution to the separated precipitate, and then performing secondary fermentation decomposition; And

(f) 220-270 kg of the virgin sulfur powder prepared by solid-liquid separation of the secondary fermentation broth obtained in the step (e) and drying the separated solid, 230-260 kg of sodium hydroxide, and phosphorous acid (H ₃ PO ₃ ) 9 And 12 to 17 kg of rock salt is mixed with 18 to 21 L of a sodium silicate solution with a mixed solution containing 790 to 830 L of a culture solution of Lactobacillus parafarraginis to liquefy the mixture But is not limited thereto.

The useful microorganism of step (b) of the present invention may preferably be a mixed microorganism consisting of lactic acid bacteria, yeast and bacteria, more preferably Lactobacillus parafarraginis , Lactobacillus paracasei , , Lactobacillus tolerans , Lactobacillus buchneri , Lactobacillus harbinensis , Lactobacillus perolens , Lactobacillus rhamnosus , Lactobacillus buckerius , Lactobacillus spp ., Lactobacillus spp ., Lactobacillus spp . North Terre kusu (Lactobacillus vaccinostercus), acetonitrile bakteo Rover you N-Sys (Acetobacter lovaniensis), acetonitrile bakteo peroxy thiooxidans (Acetobacter peroxydans), Pichia peomen Tansu (Pichia fermentans), Candida ethanol Rica (Candida ethanolica), Saccharomyces Rommie Cobb cis Scoring ( Saccharomycopsis schoenii ), thiobacillus thioox Thiobacillus thiooxidans , and Bacillus thuringiensis . However, the present invention is not limited thereto.

The toxicity of the sulfur powder added during step (c) (primary fermentation decomposition step) of the present invention is detoxified.

The first mixture of step (f) of the present invention is most preferably prepared by mixing 250 kg of the virgin sulfur powder prepared from the secondary fermentation broth, 250 kg of sodium hydroxide, 10 kg of phosphorous acid (H ₃ PO ₃ ) and 15 kg of rock salt And liquefied at 22 to 28 DEG C for 66 to 78 hours with stirring.

Also, the Lactobacillus parafarraginis strain of step (f) of the present invention was obtained from the National Institute of Agricultural Science and Technology , National Institute of Agricultural Science and Technology, Accession No. KACC91710P, isolated from plants.

In the environmentally friendly disinfectant according to an embodiment of the present invention, the natural sulfur water may be contained at a concentration of 45 to 55 ppm, but is not limited thereto. When the natural sulfur content is less than 45 ppm, the effect of inhibiting aquatic organism infection is insufficient. If the natural sulfur content is more than 55 ppm, the toxicity of natural sulfur water is increased and the mortality rate of the natural sulfur is increased.

In the eco-friendly disinfectant of the present invention, the embers are not limited thereto, but may preferably be trout eggs.

The present invention also provides a method for inhibiting aquatic organism infection by treating the environmentally-friendly disinfectant in an egg white. The environmentally friendly disinfectant of the present invention contains natural sulfur water having an effect of inhibiting aquatic organism which is an initial cause of water fungal disease of fishes as an effective ingredient. Thus, when an environmentally friendly disinfectant is treated in an oat, .

The environmentally-friendly disinfectant of the present invention can be adjusted according to conditions such as the kind of the object to be treated and / or the style of the object to be treated.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

1. Manufacture of natural sulfur water

The mixed medium containing green tea leaves and camphor leaves as a carbon source at a weight ratio of 7: 3 was fermented at 23 to 35 ° C for 43 to 48 days. Thereafter, a hot- milled sterilized molasses dilution was added to the fermented mixed medium, and then Lactobacillus parafarrhinis , L. paracasei , L. tolerans , Lactobacillus paracasei , Lactobacillus boots Tenerife (L. buchneri), Lactobacillus Har binen system (L. harbinensis), Lactobacillus pages Lawrence (L. perolens), Lactobacillus Caucasus ramno (L. rhamnosus), Lactobacillus beksi North Hotel Syracuse (L. vaccinostercus), acetonitrile bakteo Rover you N-Sys (Acetobacter lovaniensis), acetonitrile bakteo peroxy thiooxidans (A. peroxydans), Pichia peomen Tansu (Pichia fermentans), Candida ethanol Rica (Candida ethanolica), Saccharomyces Rommie Cobb Scoring system Enigma (Saccharomycopsis schoenii ), thio Bacillus thio oxy Dunn when (Thiobacillus thiooxidans) and Bacillus Turing Gen cis inoculated with beneficial microorganisms mixture of 15 kinds consisting of (Bacillus thuringiensis), W and cultured at 23 ~ 35 ℃ for 92 ~ 140 hours. Thereafter, the toxic sulfur powder was added to the culture solution cultured by inoculating the useful microorganisms, followed by primary fermentation decomposition at 25 to 35 ° C for 22 to 26 hours. The primary fermentation broth was then disrupted with a wet colloid mill. The crushed primary fermentation broth was subjected to solid-liquid separation, and the above-mentioned useful microorganism mixed culture broth was again added to the separated precipitate, followed by secondary fermentation and decomposition. The second fermentation broth was subjected to solid-liquid separation, and the separated solid was dried to obtain 250 g of the crude powder of the extract, 250 kg of sodium hydroxide, 10 kg of phosphorous acid (H ₃ PO ₃ ) and 15 kg of rock salt, ( Lactobacillus parafarraginis , Accession No. KACC91710P), 790 ~ 830 L of a culture solution was mixed with 18 ~ 21 L of a sodium silicate solution and stirred at 22 ~ 28 C for 66 ~ 78 hours to finally liquefy with sulfur containing sulfur Water.

2. Disinfection of trout eggs in natural sulfur water

The trout eggs were used after the 6th day after fertilization with eyes (Fig. 1). The amount of water in the water tank was adjusted to 15 L, and the amount of trout egg was added to the water tank of 50 ± 10. The amount of dissolved oxygen in the water tank was adjusted to 10 to 11 mg / L and the temperature was adjusted to 18 ± 1 ° C.

On day 1, S. parasitica was submerged in trout eggs. After the water fungi infection, trout eggs were taken from each tank and treated with natural sulfur water and formalin immediately (within 1 minute) to disinfect trout eggs. The trout farm income was carried out on the 1st, 3rd, and 5th day at intervals of 2 days. Each disinfection was carried out for 20 minutes. Six days after the test, trout eggs infected with water fungus were measured to calculate the infection rate.

3. Toxicity test of natural sulfur water

90 ± 15 pieces of trout were put into the water tank, and the test method was the same as the above disinfection test. After 6 days of the test, hatching and mortality were measured and hatching and mortality rates were calculated.

Example 1. Analysis of inhibition rate of water fungal infection of natural sulfur water

As a result of sterilization of trout eggs with natural sulfur water after infecting trout eggs with water fungus, it was confirmed that the number of trout eggs infected with water fungi decreased as the concentration of natural sulfur water increased as shown in Table 1 below. In particular, it was found that the natural sulfur water was effective for water fungicidal control, since the natural sulfur water disinfection zone of 50 ppm concentration was more than 3 times more effective than the untreated negative control zone. It was also found that the initial time to confirm the water fungal infection was delayed as the concentration of natural sulfur water treated in trout eggs increased.

Disinfection test results of water fungal infections of natural sulfur water Number of test days Disinfection Water fungal infection status Natural sulfur water (ppm) Formalin (ppm) 0 50 100 200 400 800 100 1 day Water mold immersion infection 1 day 20 minutes - - - - - - - 2 days - - - - - - - 3 days 20 minutes + - - - - - - 4 days ++ - - - - - - 5 days 20 minutes +++ + + - - - + 6 days +++ + + + - - + Number of infections at the end of the test 29 8 6 3 One 0 3 Infection rate 58% 16% 12% 6% 2% 0% 6%

(Infected with water fungal infection: -, no infection; +, less than 20% infection; ++, less than 40% infection; +++, less than 60% infection)

Example 2. Toxicity analysis of natural sulfur water

Trout eggs were immersed in watery molds, and after 3 days and 5 days of infection, the trout eggs were disinfected with natural sulfur water or formalin. Finally, the hatching rate and mortality of trout eggs were analyzed.

As a result, as shown in Tables 2 and 3, the hatching hatching rate of trout eggs was increased by two times or more and decreased by three times, compared with the untreated group, in the 50 ppm sulfur water treatment group. However, when the concentration of treated natural sulfur water increased to more than 50 ppm, the hatching rate of trout eggs tended to be lower than that of the untreated group, and the mortality rate also increased compared to the untreated group.

Hatchability of trout after natural sulfur water disinfection Natural sulfur water formalin Concentration (ppm) 0 50 100 200 400 800 100 Test trout unknown 106 91 98 97 90 91 86 Hatching population 44 73 33 43 8 One 81 Hatching rate (%) 41.5 80.2 33.7 44.3 8.9 1.1 94.2

Mortality of trout after natural sulfur water disinfection Natural sulfur water formalin Concentration (ppm) 0 50 100 200 400 800 100 Test trout unknown 106 91 98 97 90 91 86 Our population 62 18 65 54 82 90 5 Mortality (%) 58.5 19.8 66.3 55.7 91.1 98.9 5.8

From the above results, it was confirmed that the water fungicide inhibitory effect of the natural sulfur water of the present invention is excellent, but the use thereof at a high concentration has adverse effects on the hatching of eggs. In order to inhibit water fungal infection, 50 ppm Concentration was preferred.

Claims (6)

An environmentally friendly disinfectant containing natural sulfur water as an effective ingredient that has an effect of inhibiting aquatic bacteria infection. The environment-friendly disinfectant according to claim 1, wherein the aquatic organism is a water mold ( Saprolegnia parasitica ). The environmentally friendly disinfectant according to claim 1, wherein the natural sulfur water is contained at a concentration of 45 to 55 ppm. The method of claim 1, wherein the natural sulfur water
(a) fermenting a mixed medium containing green tea leaves and camphor leaves as a carbon source;
(b) adding a molasses dilution solution sterilized at high temperature to the fermented mixed medium of step (a), inoculating and cultivating a useful microorganism;
(c) adding a toxic sulfur powder to the useful microorganism culture solution of step (b), followed by primary fermentation and decomposition;
(d) disrupting the primary fermentation broth of step (c) with a wet colloid mill;
(e) subjecting the crushed primary fermentation broth of step (d) to solid-liquid separation, adding a useful microorganism culture solution to the separated precipitate, and then performing secondary fermentation decomposition; And
(f) separating the second fermentation broth obtained in the step (e) by solid-liquid separation, drying the separated solid to obtain a first mixture of sodium hydroxide, phosphoric acid (H ₃ PO ₃ ) And liquefying the mixture with a mixed solution of a sodium silicate solution and a Lactobacillus parafarrhinis culture medium. The eco-friendly antiseptic agent according to claim 1, wherein the egg white is trout egg white. A method for inhibiting aquatic organism infection by treating the environmentally friendly disinfectant according to any one of claims 1 to 5 in an egg white.

Images