KR100820247B1 - The insecticide composition comprising effective extract of pine gnarl for scuticociliates - Google Patents

Abstract

An insecticidal composition for Scuticociliates is provided to improve the Scuticociliates insecticidal effect by applying an extract of pine gnarl and treat infected fish with a simple bathing method. An insecticidal composition for Scuticociliates comprises an extract of pine gnarl. The extract is prepared by crushing pine gnarl, putting 2-4 liters of methanol in 1kg of the pine gnarl crushed material, extracting it at a temperature of 20-25 deg.C 2 times, filtering and drying the extract to prepare a methanol extract, mixing the methanol extract with water and hexane in a volumetric ratio of 1:20-50:20-50, separating a water layer using separatory funnel therefrom, obtaining the remaining hexane layer and then drying the hexane layer.

Description

Scutica insect insecticidal composition comprising the extract as an active ingredient {THE INSECTICIDE COMPOSITION COMPRISING EFFECTIVE EXTRACT OF PINE GNARL FOR SCUTICOCILIATES}

1 is a structural formula of alpha -pinene (α-pinene).

Figure 2 is a micrograph (400-fold magnification) after Scutica cultivation.

Figure 3 is a photograph showing the appearance of the cytoplasm is cavitation (lysis) when treated with a 10 mg / ℓ of alpha-pinene in Scutika insects.

Figure 4 is a flow chart obtained by extracting solvent fractions.

5 is a flow chart showing a secondary chromatography process

The present invention relates to a composition for insecticides Scutika insects containing the extract of Songjeol as an active ingredient.

Parasites of farmed fish include Ichthyophthirius multifillis, Cryptocaryon irritans, Trichodina, Chilodonella cyprini, Ambiphrya, Amphiphroma, and Epimpoma. Epistylis, Carchesium, and Scuticociliads.

Among them, Scutika is a ciliated insect of 30 to 40 μm, and frequently occurs in artificial seedlings and fish of the flounder.

Scutika insects have been reported to cause a variety of fish body surface, fins and kidneys, blood vessels and brain tissues, and most of them have been reported to kill flounder through destruction of plastic connective tissue.

In addition, unlike the parasites of other fish, it is difficult to treat during infection because the parasite is invaded through the blood vessels through the blood vessels.

Scotic worms are also detected in rotifers, the prey organisms for flounder, and parasitic from fry to sea urchins, which can be difficult to treat if not detected early, resulting in massive mortality.

Formalin was used until recently in flounder fish farms for the insecticide of Scutika.

However, due to the toxic component of formalin is prohibited, and to replace it, hydrogen peroxide, stabilized chlorine, etc., in addition to formalin, are used for in vitro parasite control, but have little effect.

Therefore, in recent years, as a way to solve this problem, a study on the insecticide Skutika insects using weak toxic components have been made.

Korean Patent Publication No. 10-0563273 (Scutica disease preventive treatment of aquaculture fish containing terpena hydrochloride as an active ingredient) is used to prevent squatic disease of cultured fish by using a drug containing terpena hydrochloride. Therapeutic effects are publicly available.

Korean Registered Patent Publication No. 10-0279281 (Fish Parasite Scutica Insecticide with Alginate Oligosaccharide Derivatives) includes low molecular weight of alginate as a polymer, preparation of oligosaccharide alginate, development of fish parasite Scutica insecticide and electric insecticide. Disclosed is a manufacturing method.

Korean Patent Laid-Open No. 10-0379026 (Method for Controlling Scutika Insects of Cultured Flounder) discloses a method for controlling Scutica insects infected inside and outside of cultured flounder using ketoconazole.

However, the above inventions are difficult to manufacture insecticides, and are not suitable in terms of economics, and the oral administration method used as a method for controlling Scutica insects is practically difficult to use.

In addition, due to the toxicity of the components used as pesticides there is a problem that the fish pass and fish toxicity occurs.

On the other hand, the present invention is an improved invention of the patent application No. 10-2005-0124329 (Skutika insecticidal composition using butyl aldehyde) that is filed by the applicant of the present invention and is unpublished.

The present invention is to solve the above problems, the object of the present invention is to apply the extract extract that was not previously used for the insecticide insecticide Skutika insects as a composition for insecticide insecticides.

In particular, there is also an object to improve the insecticidal insecticidal effect by applying the extract of the extract in the insecticidal insecticidal composition.

In addition, there is also an object to provide a composition for insecticide insecticide Scutika that can control the infected fish by a simple bathing method.

The present invention relates to a composition for insecticides Scutika insects containing the extract of Songjeol as an active ingredient.

In the insecticidal composition of the present invention, after pulverizing the pine seedling to prepare a truncated powder, methanol is added to the truncated powder per 2 kg per kg of the truncated powder, and extracted twice and filtered at 20-25 ° C. Next, after drying to prepare a methanol extract, water and hexane was added to the methanol extract, the mixture was stirred, the water layer was separated with a separatory funnel, and then removed, and the remaining hexane layer was obtained. It is an insecticidal composition.

The inventors of the present invention observed the insecticidal effect against Scutika insects using about 360 natural products in order to develop insecticidal compositions for Scutika insects.

As a result, the insecticidal effect is excellent, there is no fish toxicity, and it is used to prepare the insecticidal composition of the present invention by selecting the songjeol as an economically suitable material.

Songjeol is a node of pine tree, and it is written in the bodhisattva of the main tree and the imperial palace, "It relieves white cramps, shells, and fracture pains, and makes liquor strong when you make alcohol."

Songjeol contains rosin, where the taste of rosin is sweet and bitter, it has no poison, relaxes the five intestines, removes heat, improves flavor, heals all medicines, duyang and white poison, removes fraud, cures deafness and tooth decay, Attached to the window is said to have the effect of pain and sterilization, it is recorded in the herbal wood and imperial dongbom.

The inventors of the present invention have been doing a lot of research to find a method that exhibits a high insecticidal effect against Scutika insects at the same time to prevent fish death due to toxicity by using a songjeol with the above effects, the songjeol to methanol After extraction, it was fractionated using several organic solvents, and among them, it was found that the insecticidal effect against Scutika insects was high without being toxic in the hexane fractionation layer.

Since the organic solvent itself is toxic, since an appropriate amount must be used, in the present invention, the methanol extract, distilled water, and hexane are stirred at a volume ratio of 1:20 to 50:20 to 50, and then separated by a separatory funnel. When used as a insecticide insecticide, it was found that the highest insecticidal effect on the insect insects without killing the fish.

In addition, when using the insecticidal composition 0.2 ~ 1.0 kg per 1 ton aquaculture water, it can be usefully used as an environmentally-friendly sanitary pesticide with environmental preservation and purification as non-toxic to natural enemies, shrinkage.

Referring to the method for producing a composition for insecticide insecticide Scutika including the extract of the present invention as an active ingredient in detail.

<Method of producing Scutika insecticidal insect composition comprising pine extract as an active ingredient>

1. First Step: Methanol Extract Preparation

After pulverizing the Songjeol to prepare the Songjeol crushed product, Methanol was added to 2 ~ 4 L per 1 kg of Songjeol crushed product, extracted twice at 20 ~ 25 ℃, filtered, and dried to prepare methanol extract. do.

2. Second Step: Preparation of Scutika insecticidal insect composition

Water and hexane were added to the methanol extract, followed by stirring. After separating the water layer with a separatory funnel, the remaining hexane layer was obtained and dried to prepare a composition for the insecticidal insecticide.

At this time, it is preferable to add the methanol extract, distilled water and hexane in a volume ratio of 1:20 to 50:20 to 50 and stirred.

In addition, it is characterized by containing an alpha-pinene component (FIG. 1).

In this case, since the alpha-pinene component is insoluble in water, a mixture of alpha pinene, dimethyl sulfoxide, and a surfactant in a volume ratio of 20 to 50:50 to 80: 1 may be used.

Hereinafter, the present invention is described in detail through Examples and Experimental Examples for the insecticidal insecticidal composition comprising the extract of Songjeol as an active ingredient, but they are not intended to limit the scope of the present invention.

<Example 1> Preparation of Scutika insecticidal composition 1 containing the extract of the present invention as an active ingredient

Songjeol was commercially available and prepared.

The prepared Songjeol was pulverized with a grinder to prepare Songjeol.

Methanol extract was prepared by adding 3 L of methanol to 1.2 kg of pulverized powder, extracting twice at 20 ° C., filtering, and drying in a drier.

After distilled water 800 ml and hexane 800 ml were added to 20 g of methanol extract and stirred, the water layer was separated and removed with a separatory funnel, and the remaining hexane layer was obtained.

Example 2 Preparation of Scutika insecticidal composition 2 comprising the extract of Songjeol of the present invention as an active ingredient

Songjeol was commercially available and prepared.

The prepared Songjeol was pulverized with a grinder to prepare Songjeol.

Methanol extract was prepared by adding 3 L of methanol to 1.2 kg of pulverized powder, extracting twice at 20 ° C., filtering, and drying in a drier.

1000 g of distilled water and 1000 mL of hexane were added to 20 g of methanol extract, followed by stirring. After separating the water layer with a separatory funnel, the remaining hexane layer was obtained and dried to prepare a composition for insecticidal insecticides.

Experimental Example 1 Extraction Optimization Experiment

The extract was fractionated for each solvent according to the extraction flow chart shown in FIG.

As shown in Example 1, the Songjeol was pulverized with a grinder to prepare Songjeol, and then 1.2 liters of Songgeol was added to 3 liters of methanol, extracted twice at 20 ° C., filtered, and then dried in a drier. After preparing 20 g of methanol extract, it was dissolved in 800 ml of distilled water and 800 ml of hexane (n-Hexane) and placed in a 2 L separation filter. The cap was closed and mixed well, and then separated into a water layer and a hexane layer.

After 30 minutes, the primary hexane layer was separated using another separatory filter, 800 ml of fresh hexane was mixed with the water layer, the stopper was closed, and the mixture was shaken sufficiently to separate the water layer and the hexane layer.

800 ml of chloroform was mixed with the remaining water layer, and the stopper was sufficiently shaken to separate the water layer and the chloroform layer, and after 30 minutes, the primary chloroform layer was separated.

Again, 800 ml of fresh chloroform was mixed into the water layer, and the stopper was closed. Then, the mixture was shaken sufficiently to separate the water layer and the chloroform layer secondarily.

800 ml of ethyl acetate was poured into the remaining water layer, and the ethyl acetate layer was separated twice in the same manner as in the hexane distribution, and 800 ml of butanol was poured into the remaining water layer to separate the butanol layer.

5 fractions of hexane layer, chloroform layer, ethyl acetate layer, butanol layer, and water layer were obtained by the solvent fractionation method as described above. The water layer was concentrated by lyophilizer, and the hexane layer, chloroform layer, ethyl acetate layer and butanol layer were rotated, respectively. Concentrated in a vacuum concentrator.

The weight of each fraction was hexane fraction (6.7 g), chloroform fraction (5.5 g), ethyl acetate fraction (1.3 g), butanol fraction (4.0 g) and water soluble fraction (2.5 g).

Insecticidal capacity for Scutika insects by each fraction is shown in Table 1 below.

Table 1 Insecticidal Insecticidal Capacity

Fraction Insecticidal (100 ppm) Insecticidal (Minimum Concentration) Hexane fraction 100 100 (30 ppm) Chloroform fraction 100 100 (50 ppm) Ethyl acetate fraction Reduced mobility - Butanol fraction Reduced mobility - Water soluble fraction Reduced mobility -

As shown in Table 1, it was found that exhibited the highest insecticidal insecticidal ability in the hexane fractionation layer using the present invention Songjeol.

Experimental Example 2 Material Analysis

1) First active fractionation experiment

Material analysis was carried out by fractionation and after measuring the Scutica insecticidal activity was analyzed by HPLC and G.C-MASS for the active fraction.

10 g of hexane fractions were adsorbed onto silica gel columns (berck 70-230 mesh, 600 g, 5.5 * 70 cm) and subjected to primary chromatographic analysis with a stepwise gradient of hexane and ethyl acetate.

The gradient used was hexane: ethyl acetate ratio in the order of 90:10, 85:15, 70:30, 50:50, 0: 100, and the fraction according to each ratio was H1, H2, H3, H4, H5. Eluted.

Scutica insecticidal test was performed on the five fractions, indicating that H1 was an active fraction.

2) Second Chromatography Experiment

Silica gel chromatography was performed on the active fraction separated from the primary (FIG. 5).

The H1 fraction was further purified using high performance liquid chromatography.

H1 was carried out at a flow rate of 4.0 ml / min, hexane-ethyl acetate (9: 1), μ porasil silica (19 * 300 mm, Waters) column, absorbance 260 nm to obtain H21, H22 and H23 fractions. Through the same purification process, substance A (alpha-pinene, FIG. 1) was obtained.

The final material was identified through GC-mass.

Experimental Example 3 Investigation of the Effect on Scutika Insects

1. Preparation of experimental materials

The present invention was prepared by preparing an insecticidal insecticidal composition of Example 1 in aqueous solution (Table 2).

Based on Experimental Example 2, an alpha-pinene component, which is a component contained in the present invention, was prepared by the following method to prepare an aqueous alpha-pinene solution (Table 2).

<Preparation of alpha -pinene aqueous solution>

Alpha-pinene (α-pinene, <aldrich, USA, 98%>) is hydrophobic and is dissolved in water in small amounts, making it difficult to use the stock solution for bathing.

Therefore, alpha-pinene should be used after dissolving in soluble alcohol, dimethylsulfoxide.

Tween 80 and span 80 were used as the dispersant.

In the present invention, an aqueous solution was prepared by adding a mixture of tween 80 (junsei, japan) + span 80 ((junsei, japan) by weight 2.5: 1) as a dispersant to dimethyl sulfoxide (dimethylsulfoxide) having little toxicity.

A solution was prepared by mixing 20% by weight of alpha-pinene by weight, 74% by weight of dimethylsulfoxide, and 80% by weight of tween 80 + span.

Formaldehyde solution (36.0 ~ 38.0%; JUNSEI, Japan) was also prepared for the comparative toxicity experiments, which have been banned, but have been widely used as a control agent for Scutica.

<Table 2> Composition ratio according to concentration in preparing aqueous solution for each material (100 ℓ of seawater treatment standard)

50 ppm 75 ppm 100 ppm Aqueous solution of Example 1 Extract 5 g Ethanol 20 g Water 25 g Extract 7.5 g Ethanol 30 g Water 37.5 g Extract 10 g Ethanol 40 g Water 50 g Aqueous solution of alpha-pinene Alpha-pinene 5 g DMSO 19.75 g tween + span 0.25 g Alpha-pinene 7.5 g DMSO 29.62 g tween + span 0.38 g Alpha-pinene 10 g DMSO 39.5 g tween + span 0.5 g Formaldehyde aqueous solution 5 g of formaldehyde aqueous solution 7.5 g of formaldehyde aqueous solution 10 g of formaldehyde aqueous solution

2. Experimental method

1) Scutika insect culture

The isolates were cultured directly from flounder diseased infected Scutica ciliated caterpillars.

The tissues of the infected area were partially removed from the flounder diseased fish, placed on a slide glass, and examined under a microscope. As a result, it was confirmed that Scutica was infected.

The identified Scutica locus was inoculated with P1Y1 (1% proteose peptone, 1% yeast extract) medium in sterile seawater to perform primary culture at 15 ° C.

In the primary culture of Scutica culture, many bacteria were present. Therefore, pure cultures using fish coin cell lines were performed due to the contamination of bacteria and the limitation of the number of growth.

For pure cultivation, 10% FBS and penicillin-streptomycin were added to MEM (Modified Eagle's Medium) medium titrated with CHSE-214 (Chinook salmon embryo) with 7.5% sodium bicarbonate at pH 7.3 for pure culturing. Incubated daily.

The cultured fish-coated cell line was inoculated with the first cultured Scutica insect culture solution, and then purely cultured at 20 ° C. for 5 days.

2) Insecticide Insect Investigation

For this experiment, Scutica was centrifuged at 650 × g for 5 minutes using Scutica at the end of the logarithmic phase or at the beginning of the phase of suspension and resuspended in PBS or filtered seawater.

A pinene aqueous formaldehyde solution - hematocrit after counting to (haematocrit) 0.1 × 10 ⁴ ㎖ of Surgical urticae The composition of Table 2, the liquid present inventors carried containing charge Example 1 Surgical urticae charge insecticidal composition aqueous solution of alpha 0.9 ml of each was added to 24 wellplates, mixed and shaken at 20 ° C. incubator, and observed after 1 hour.

The aqueous solution for the insecticidal composition of the present invention Example 1, aqueous alpha-pinene solution, and aqueous formaldehyde solution were made on the basis of 100 L, and the amount was reduced to 1/100 to 1 L, and then to 1 L of filtered seawater. Dispersed.

The concentration in the final 24 wellplate was used to adjust the concentration.

After stirring, the number of Scutica insects was measured for 1 hour and 2 hours.

In the measurement, the culture solution was dropped on a slide glass, and then measured under a microscope to determine the number of normal individuals except the lysed individuals.

The exact number of individuals was counted with hematocrit.

3. Experimental Results

As a result, it is shown in Table 3 below.

<Table 3> Insecticidal insecticidal rate using the insecticidal composition of the present invention

By concentration Scutika insects Insecticide Rate (%) Remarks 0 hours 1 hours 2 hours Aqueous solution of Example 1 50 ppm 1.0 × 10 ³ 763 648 35.2 75 ppm 1.0 × 10 ³ 94 71 92.9 100 ppm 1.0 × 10 ³ 0 0 100 Aqueous solution of alpha-pinene 50 ppm 1.0 × 10 ³ 0 0 100 75 ppm 1.0 × 10 ³ 0 0 100 100 ppm 1.0 × 10 ³ 0 0 100 Aqueous solution of formaldehyde 50 ppm 1.0 × 10 ³ 1.0 × 10 ³ 1.0 × 10 ³ 0 Scutika horde progression 75 ppm 1.0 × 10 ³ 1.0 × 10 ³ 1.0 × 10 ³ 0 Scutika horde progression 100 ppm 1.0 × 10 ³ 1.0 × 10 ³ 1.0 × 10 ³ 0 Scutika horde progression

As shown in Table 3, it was found that the aqueous solution of Example 1 and the alpha-pinene component separated therefrom exhibited a high insecticidal effect against Scutica at low concentrations (50, 75, 100 ppm). , Formaldehyde aqueous solution used as a control did not show a pesticidal effect when treated at low concentrations (50, 75, 100 ppm), and the fear of Scutica insects progressed, and when treated at 150 ppm or more, it showed an insecticidal effect. .

Experimental Example 4 Investigation of Toxicity of Flounder

1. Preparation of experimental materials

The aqueous solution of the composition for insecticide insecticide Scutica of Example 1 prepared in Experimental Example 3, an aqueous solution of alpha-pinene, and an aqueous solution of formaldehyde were prepared.

As an experimental fish, flounder fry (5 cm) were prepared for each 100 tanks.

2. Experimental method

After filling 100 l of seawater in each of the five prepared round tanks, the flounder fry prepared in each tank was prepared, and the aqueous solution of the insecticidal composition of Scutica insect insecticide of Example 1, the aqueous alpha-pinene solution and the aqueous formaldehyde solution of Example 1 were prepared. 500 ml each was checked for toxicity to flounder fry.

Toxicity was confirmed after 1 and 2 hours of bathing.

3. Experimental Results

As a result, it is shown in Table 4 below.

Table 4 Safety Experiments

50 ppm 75 ppm 100 ppm  Aqueous solution of Example 1 Experimental fish-No abnormal swimming and kinetic change No death occurred after 2 hours Aqueous solution of alpha-pinene Experimental fish-No abnormal swimming and kinetic change up to 200 ppm No death occurs after 2 hours Formaldehyde aqueous solution Experimental fish-No more than 150 ppm swimming and kinetic change No death object occurs after 2 hours

That is, the commonly used aqueous formaldehyde solution was found to have a strong toxicity even at low concentrations, so that some individual fish species were changed, and one hour later, all individuals died.

However, when using the aqueous solution of Example 1 of the present invention and the alpha-pinene material separated from the present invention it can be seen that no toxicity to fish.

In particular, the alpha-pinene material isolated in the present invention was safe to 200 ppm.

<Experiment 5> Residual amount test experiment remaining on flounder fry

1. Experimental Materials

Residual amount tests were performed on the experimental words used in Experimental Example 4.

After bathing with 200 ppm of alpha-pinene aqueous solution and 200 ppm of formaldehyde aqueous solution for 2 hours, the residual amount in the flounder muscle was measured.

2. Experimental method

1) Formaldehyde residual amount measurement

10 g finely chopped muscle was placed in a centrifuge tube and homogenized well by adding 10 ml 0.5 N NaOH to homogenizer, and then homogenized again by adding 10 ml 12% ZnSO ₄ · 7H ₂ O. The mixture was allowed to stand at room temperature for 30 minutes.

After centrifugation at 800 rpm for 5 minutes, the supernatant was filtered with Whatman NO1 filter paper.

The residue was mixed with a vortex mixer, washed with 10 ml of distilled water, and then centrifuged at 8,000 rpm for 5 minutes.

The washes were combined with filtrate and this process was repeated twice.

The combined extracts were adjusted to 100 mL of final distilled water with 0.5 N HCl added for the best recovery pH 4.5.

2 ml of the final extract was taken in a test tube to determine the formaldehyde concentration according to the formaldehyde measurement method (AHMT method).

At this time, the detection limit is 0.05 μg / ml.

2) Alpha-pinene quantitative analysis

10 g of finely chopped muscle was placed in a centrifuge tube, homogenized well with a homogenizer by adding 15 ml of ethyl acetate, and then homogenized again with a homogenizer by adding 0.5 N NaOH, followed by 30 minutes. It was left at room temperature.

After centrifugation at 800 rpm for 5 minutes, the supernatant was filtered with Whatman NO1 filter paper.

The filtrate was placed in a 50 ml volumetric flask, and the residue was mixed with a vortex mixer by adding 15 ml of ethyl acetate (Ethyl acetate), and then centrifuged at 8,000 rpm for 5 minutes.

After centrifugation at 800 rpm for 5 minutes, the supernatant was filtered with Whatman NO1 filter paper.

The washes were combined with filtrate and this process was repeated twice.

The collected filtrate was placed in a 50 ml volumetric flask, and ethyl acetate was added to make 50 ml in total.

3) analysis

GC used HP GC 5890 model, and the column (column) was used HP-5MS capillary column and helium gas was flowed at 1.8 ㎖ / min.

The injector temperature was maintained at 220 ℃, the detector temperature was 320 ℃, the oven temperature was maintained at 37 ℃ for 5 minutes, and then the temperature was increased at a rate of 5 ℃ / min to 180 ℃ and then at a rate of 20 ℃ / min up to 300 ℃.

Mass was used as a detector.

3. Experimental Results

The experimental results, shown in Table 5 and Table 6 below.

Table 5 Measurement of the remaining amount of eruption-pinene and formalin remaining in halibut fry after 2-hour bathing

0 hours 24 hours 48 hours Alpha-pinene residue 0.91 0.28 0 Formaldehyde residue 1.02 0.75 0.78

Table 6 Safety of material safety data sheets for alpha-pinene and formaldehyde

Toxicity Information Environmental impact Formaldehyde 203 mg / m3 Inhalation-Rat LC 50 100 mg / kg Oral-Rat LD50 Carcinogenicity Toxicity to fish: 4,960 μg / L 96 hours LC50 (death rate) Julunivas Invertebrate Toxicity: 14,000 μg / L 48 hours EC50 (physiological) Daphnia Algae toxicity: 4,500 μg / L 48 hours (population) Alpha-pinene No carcinogenicity No environmental impact

As a result of the experiment, as shown in Tables 5 and 6, alpha-pinene, a component contained in the Songjeol extract of the present invention, was found to disappear after 48 hours as the residual amount of the flounder fry decreased.

However, it was found that formaldehyde remained its residual amount over time, and such formaldehyde had an adverse effect on the environment.

Experimental Example 6 Investigation of Disease Treatment in Disease Occurrence Conditions

An aqueous solution of alpha-pinene isolated from the present invention was treated to a fish farm in which Scutika insects were infected in the flounder fish farm, where death occurred.

Through screening, seawater was filled to a height of 30 cm in a 10 pyeong tank for bottled fish collected only from infected fish.

Sampling confirmed infection of Scutica on the foreskin and gills.

After that, the water volume reached 9.9 ton, and the α-pinene aqueous solution was evenly sprayed on the tank.

The concentration of α-pinene was 50 mg / l.

After 2 hours after treatment, the cells were collected and sampled with 5 mice, and the epidermis and gills were examined under a microscope. Scoutica was removed from the epidermis and Scutica was completely removed from the gills.

After a week of observation, no further mortality occurred and the flounder could be raised normally.

According to the present invention, the insecticidal composition with improved insecticidal insecticidal effect is provided by applying the extract of S. aeruginosa insecticide, which has not been conventionally used for the insecticidal insecticidal insecticide, as the composition for the insecticidal insecticidal insecticide.

In addition, there is provided a composition for insecticide insecticide Skutika which can control the infected fish by a simple bathing method.

Claims (3)

In the composition for insecticide insecticides, After pulverizing the Songjeol to prepare the Songjeol crushed product, Methanol was added to 2 ~ 4 L per 1 kg of Songjeol crushed product, extracted twice at 20 ~ 25 ℃, filtered, and dried to prepare methanol extract. After mixing the methanol extract with water and hexane in a volume ratio of 1:20 to 50:20 to 50, separating and removing the water layer with a separatory funnel, obtaining the remaining hexane layer, and then drying it, Scutika insecticidal composition comprising pine extract as an active ingredient. delete delete

Images