KR101298097B1 - Scuticociliates Insecticide Composition Comprising Camphene as an Active Ingredient - Google Patents

Abstract

The present invention relates to a composition for controlling Skutika insects, and more particularly to a composition for controlling Skutika insects comprising camphene as an active ingredient. According to the feed additive composition of the present invention, by containing a camphene excellent in the insecticidal insecticidal effect (camphene) as an active ingredient of the feed additive for fish farming, it is possible to effectively control the Skutika insects that cause large death of fish In addition, it is possible to replace the existing toxic remedy formalin has the effect of preventing marine environmental pollution in advance.

Description

Scuticociliates Insecticide Composition Comprising Camphene as an Active Ingredient}

The present invention relates to a composition for controlling Skutika insects, and more particularly to a composition for controlling Skutika insects comprising camphene as an active ingredient.

The amount of fish produced in Jeju Island is about 340,000 tons, and the production of farmed flounder exceeds 10,000 tons, which is a significant part of the production of seafood in Jeju Island.

Scutika insects on farmed flounder are causing serious damage to fish farming by causing massive deaths of fish. The major infectious diseases of the flounder culture include viral, bacterial and parasitic diseases. Among these diseases, Scutika, a ciliary parasitic disease, is particularly from the flounder of the flounder, which is the largest producer of farmed fish in Korea. Because of the widespread parasite up to adult sex, which causes massive mortality, Scutika insects have emerged as a major threat to the productivity of aquaculture fisheries.

Scutika insects penetrate into the body surface, gills, and even the brain, causing nearly 100% of all parasitic fish to die. Recently, flounders infected with Skutika insects occur throughout the year, and the damage is increasing year by year, but there is no effective control measures until now. In the flounder farms in Korea, the occurrence of Scutika insects is mainly caused by the low temperature.

Until now, most of the countermeasures against Scutika insects are sterilization of breeding water and bath or fresh water bath using chemicals of infected individuals. Chemicals are used most in anticipation of some remedy effects, but this material is a harmful environmental pollutant and is restricted in its use. In recent years, chemicals have not been cured at all despite the use of very high concentrations of chemicals. It is true.

If chemotherapeutic agents such as chemicals used as insecticides against Scutika are banned due to environmental pollution, there is no suitable Scutika insecticide.

Therefore, there is an urgent need for the development of a remedy having excellent control against Scutika insects without causing problems of environmental pollution.

The present invention is derived from the above needs, the object of the present invention is to provide a feed additive composition excellent in the control effect against Scutika insects, without using chemicals that cause environmental pollution problems.

The present invention relates to a composition for controlling Skutika insects, and more particularly to a composition for controlling Skutika insects comprising camphene as an active ingredient.

Kampene, which is used as an active ingredient in the composition for the control of Scutica insects of the present invention, has a colorless crystal with unsaturated hydrocarbons belonging to terpenes and is widely used as a raw material for camphor synthesis or fragrance. Kampen's chemical formula is C ₁₀ H ₁₆ , a colorless camphor-like odor crystal that is insoluble in water but soluble in organic solvents such as ethanol and ether. D-type with right turn is contained in essential oils of cedar and cypress branches and leaves, roots and stems of ginger, and L-type with left turn is contained in essential oil of pine leaves. Racemic forms exist as components of various essential oils.

Kampene used as an active ingredient in the composition for the control of Skutika insects of the present invention can be extracted from a natural source (pine), ginger, etc. using a steam distillation method, a compression method or a solvent extraction method, which is a general essential oil extraction method. Specifically, the camphor can be obtained using various extraction methods known in the art, and preferably, (a) water, (b) anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (methanol, ethanol). , Propanol, butanol, etc.), (c) the mixed solvent of the lower alcohol and water, (d) acetone, (e) ethyl acetate, (f) chloroform or (g) 1,3-butylene glycol as extraction solvent. It can be obtained by. More preferably, extraction is carried out using an aqueous methanol and ethanol solution, and most preferably, extraction is performed using an ethanol aqueous solution.

On the other hand, it will be apparent to those skilled in the art that the camphor of the present invention can be obtained in addition to the above-described extraction solvents, and extracts having substantially the same effects using other extraction solvents.

In the present invention, the camphor may be prepared in a powder state by an additional process such as distillation under reduced pressure and freeze drying or spray drying.

Meanwhile, in the present invention, the camphor may be included in an amount of 0.00001 to 1.0% by weight, more preferably 0.0001 to 0.1% by weight, most preferably 0.0001 to 0.05% by weight, based on the total weight of the composition. This is because when the amount of camphor is less than 0.00001% by weight, the insecticidal efficacy against Scutica insects is weak, and when the amount of camphor exceeds 1.0% by weight, the toxicity of the substance itself may be a problem.

According to a preferred embodiment of the present invention, the composition of the present invention is a feed additive composition. Ingredients included in the feed additive composition of the present invention includes components commonly used in feed additive compositions in addition to camphor as an active ingredient, for example, a combination of various known functional feed additives such as growth promoting substances and immune enhancing substances. It may include.

The feed additive composition of the present invention may be prepared in any formulation commonly prepared in the art, and may be, for example, formulated as a solution, suspension, emulsion, oil or powder, but is not limited thereto. .

Another aspect of the invention relates to a feed for fish containing camphene (camphene), more specifically the fish feed may be in the form of a feed additive composition may be mixed with campene alone, or mixed with camphor and other functional ingredients It can also be mixed into the feed. The fish feed is, for example, raw fish (raw fish) using raw fish (frozen) such as mackerel, canary, roe, saury, sardines, shrimp, etc., fish (frozen fish), fish meal or vegetable oil and other dry feed and weight ratio Moist pellets mixed with, and vitamins and minerals are added to the dry feed to form a blended feed formed into a powder or pellet form. Common components of fish feed include protein sources, carbohydrate sources, vitamin mixtures and mineral mixtures, and the like, fish meal, meat meal, soybean meal, cottonseed meal or mixtures thereof may be used. Wheat flour, dextrin, potato starch, sucrose or a mixture thereof may be used. As the vitamin mixture, ascorbic acid, thiamine, riboflavin, pyridoxine, niacin, folic acid, vitamin A, vitamin K3, or vitamin D3 may be used. NaCl, MgSO4-7H2O, NaH2PO4-5H2O, KH2PO4, CaH4 (PO4) 2OH2O, ZnSO4-7H2O, CuCl2, AlCl3-7H2O, KIO3, MnSO4-7H2O, ferric citrate or calcium lactate ) May be used.

According to the feed additive composition of the present invention, by containing a camphene excellent in the insecticidal insecticidal effect (camphene) as an active ingredient of the feed additive for fish farming, it is possible to effectively control the Skutika insects that cause large death of fish In addition, it is possible to replace the existing toxic remedy formalin has the effect of preventing marine environmental pollution in advance.

Hereinafter, the present invention will be described in detail with reference to examples. The following examples are merely examples for describing the present invention, and the scope of the present invention is not limited thereto.

<Examples>

1. Cultivation of Scutika Insects

The culture of the Scutika caterpillar to be used in the embodiment of the present invention was directly isolated from the flounder diseased infected with Scutika ciliated larvae. The tissues of the infected area were partially removed from the flounder diseased fish, placed on a slide glass, and microscopically observed. 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 first cultured Scutica culture, many bacteria were present, and pure cultures using fish coin cell lines were performed due to the contamination of bacteria and the limitation of the number of growth. Cultured at 20 ° C by adding 10% FBS and penicillin-streptomycin to MEM (Modified Eagle's Medium) medium titrated with CHSE-214 (Chinook salmon embryo) with 7.5% sodium bicarbonate for pH 7.3 It was. Cell lines grown with monolayers were inoculated with Scutica cultivars initially cultured, and the culture was continued at 20 ° C. until the Scutica insects completely consumed the cell lines. After observing the ingestion of the cell lines completely under the microscope, the number of proliferated cells was counted by counting the number of cells of 1 mm ㅧ 1 mm using a hematocytometer. It was.

2. Sample used in experiment

The test substance for the control of the Scutica insects used in the embodiment of the present invention is camphene, and as a comparative example caffeine (Comparative Example 1), Quiacrine dihydrochloride (Comparative Example 2), Quinine sulfate (Comparative Example 3), Trimethoprim (Comparative) Example 4), Oxytetracycline (Comparative Example 5) was used, and formalin (formaline), a substance used to control Scutica insects, was used for positive control. As a negative control, only the solvent dimethylsulfoxide (DMSO) was administered without any substance (see <Table 1>).

 Sample used in the experiment Example camphene Comparative Example 1 caffeine Comparative Example 2 Quiacrine dihydrochloride Comparative Example 3 Quinine sulfate Comparative Example 4 Trimethoprim Comparative Example 5 Oxytetracycline positive control formaline negative control dimethylsulfoxide (DMSO)

3. Experimental effect of rescue on Scutika insects

① Motility Decrease and Morphology Experiment

After drying the experimental samples of 2. , the dried sample was diluted with sterile seawater at an appropriate magnification, and then the concentration was changed to 1 ppm or 5 ppm, and then inoculated into the Scutica insect culture prepared in 1. , followed by 2 hours at 20 ° C. Pesticide efficiency directly under the microscope while

 Measured. In the case of Scutika insects, the phenomena that occur when contacting the pesticidal substance go through the stages of motility, bulging morphological changes and morphological changes. Experimental results were determined by applying the method of Novotony et al. It was made.

 Test Result Criteria (Novotony, 1996) score Motility 4 If there is no loss of mobility 3 Decreased mobility by more than 50% of all Scutika insects 2 If you stop exercising near 50% of all Scutika insects One If more than 50% of all Scutika insects stop working 0 If all of the Scutika insects stop exercising score Morphological changes ( Morphology ) 4 If there is no morphological change of the Scutika insects 3 Less than 50% of all Scutica insects swell into balls or irregular shapes 2 Approximately 50% of all Scutica insects swell into balls or irregular shapes One If more than 50% of all Scutika insects swell into balls or irregular shapes 0 If most of the Scutika insects are lysed

The experimental results are as shown in Table 3 below. According to Table 3 below, when inoculated with camphene (Example), as compared to Comparative Examples 1 to 5, which were inoculated with other samples, the motility of the sukutica insects was significantly lowered over time, and 2 hours after the inoculation. At this point, it can be seen that all of the sukutica insects are stopped. In addition, looking at the morphological changes, the inoculation of the campene of the present invention has a greater degree of swelling in a ball shape compared to Comparative Examples 1 to 5 inoculated with other samples, and 1 hour 30 minutes (90 minutes) has elapsed. At this point, it can be seen that most of the Scutica insects are lysis.

When comparing the embodiment of the present invention with the inoculated camphor and the positive control inoculated with formalin, a conventional remedy, the inoculation of camphor was faster than the inoculation of formalin. It can be seen that it lowers the swelling and swells into a ball shape. Therefore, it can be seen that it is advantageous to use the camphor of the present invention rather than the conventional formalin to see a more rapid Scutica insect control effect. In addition, it is possible to prevent the environmental pollution in advance by using the environmentally friendly camphor, replacing the toxic material formalin.

Motility Decrease and Morphology Test Results of Scutica Insects
Target substance density 10 min 30 min 60 min 90 min 120 min ppm A * B ** A B A B A B A B Example
camphene One One 2 One 2 One One 0 One 0 0 5 0 0 0 0 0 0 0 0 0 0 Comparative Example 1
caffeine One 4 4 4 4 4 4 4 4 4 4 5 4 4 4 4 4 4 4 4 4 4 Comparative Example 2 Quiacrine dihydrochloride One 4 4 4 4 3 3 3 3 3 2 5 4 4 2 2 One One One One One One Comparative Example 3 Quinine sulfate One 3 3 3 3 3 3 2 2 2 2 5 3 3 3 3 3 3 2 2 2 2 Comparative Example 4 Trimethoprim One 3 3 3 3 3 3 2 2 2 2 5 2 3 2 3 2 3 2 2 One 2 Comparative Example 5 Oxytetracycline One 3 3 3 3 3 2 2 2 2 2 5 2 3 2 3 2 3 2 2 2 2 positive control formaline One 3 3 2 3 One 2 0 One 0 0 5 3 3 0 0 0 0 0 0 0 0 negative control DMSO 5 4 4 4 4 4 4 4 4 4 4

* A: morphology

** B: Motility

② Field test for Scutika insects

In the flounder fish farm, the infection of the Scutika insects was generated, and the camphor of the present invention was mixed in the feed to 1,000 ppm. The feed containing camphor was then fed twice a day (experimental). On the contrary, the control group was fed twice a day without campen.

The number of dead animals was confirmed once a day for each of the experimental and control groups. The dead flounder was dissected and the gills and epidermis were dissected. In addition, five healthy flounders were separately dissected from each of the experimental and control groups to dissect the gills and epidermis, and then, the presence of Scutika worms was confirmed under a microscope at 400 times. The results are shown in Table 4 below.

According to the following Table 4, the experimental group fed the camphor-containing feed showed less mortality and apparently a small number of Scutika insects were seen in the gills and epidermis even when the healthy flounder was dissected. In the non-feeding control group, the mortality did not decrease, and even when the apparently healthy flounder was dissected, a significant number of Scutika insects were found in the gills and epidermis, indicating that the Scutika insects are in the breeding state.

After one week of observation, no further mortality occurred in the experimental section and the flounder could be raised normally.

 Field test results for Scutika insects Date camphene treatment tank
Mortality (mari) Control mortality (mari) 0 (2010.07.01.) 32 34 One 10 31 2 9 36 3 11 28 4 10 30 5 11 27 6 8 32 7 10 35 8 9 34

As described above, those skilled in the art will understand that the present invention can be implemented in other specific forms without changing the technical spirit or essential features. It will be understood by those skilled in the art that the present invention may be embodied in many other specific forms without departing from the spirit or scope of the invention as defined by the appended claims and their equivalents. .

Claims (6)

Fish feed additive composition for the control of Skutika insects containing camphene (camphene) as an active ingredient.
The method of claim 1,
The content of the camphen (camphene) is fish feed additive composition, characterized in that 0.00001 to 1.0% by weight relative to the total weight of the composition.
delete The method of claim 1,
The composition of claim 1, wherein the composition has a formulation selected from the group consisting of solutions, suspensions, emulsions, oils and powders.
Fish feed comprising the fish feed additive composition of any one of claims 1 to 2.

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