KR20190088804A - Composition comprising fermented extract of pine gnarl for preventing or treating scuticociliatosis - Google Patents

Abstract

The present invention relates to a composition for preventing or treating scuticociliatosis containing a pine gnarl extract. The composition containing the pine gnarl extract has an excellent extermination effect of scuticociliates, thereby being able to be usefully used as a pharmaceutical composition or a feed composition for fish such as halibut and the like.

Description

[0001] The present invention relates to a composition for preventing or treating scuticacid infectious disease,

The present invention relates to a composition for the prevention or treatment of scuticacid infectious diseases containing fermented products of mixed extract of mulberry leaves, safflower seeds, dead leaves and camphor leaves.

The number of fish produced in Cheju Island is about 340 thousand tons, among which the production of cultured flounder is more than 20 thousand tons, which makes up a considerable portion of the production of fishery products in Jeju Island. The coastal aquaculture farms of Jeju Island have developed rapidly in a short period of time because they have the environmental conditions to produce and breed large quantities of seedlings every year regardless of the season, using underground seawater that maintains a clean and stable water temperature. However, as the number of farms increases and aquaculture grows, the disease is also increasing. Especially in recent years, the damage caused by the parasitic disease Sukutika is rapidly increasing. Scutica, a type of ciliate, is classified as Protista, Ciliophora, Oligohymenophorea, Hamenostomatia, Scuticociliatida and so far dozens of species (Evans and Corliss, 1964; Small, 1967 Corliss, 1979).

After the infection cases were reported by 乙 竹 and 松 里 (1986), Sututika spp. Infected with cultured flounder was cultured and propagated at a temperature (Yoshinaga and Nakazoe, 1993), the onset trend of Sututika spp., Culture, drug susceptibility, , Et al., 1993), pathological histology of flounder infected with Sutuca cordata (Lee et al., 1994; Lee et al., 2001). Jee et al., 2002; Jee et al., 2003b), infection (Jin et al., 2003b) and remedies by compounds (Choi et al., 1997; and Jo, 2002). (Kang et al., 2003a; Kang et al., 2003b).

The major infectious diseases that occur in the flounder culture process are viral, bacterial and parasitic diseases. Among these diseases, Sucutica spp., A disease of the ciliated parasite, is the most abundant among the cultured fishes of Korea, , Which leads to massive deaths, Sukutika is a major threat to the productivity of 'fisheries'.

Recently, the number of flounder infected with Sututika infestation has been growing throughout the year, and the amount of damage has been increasing year by year, but until now there is no effective control measure. In the domestic flounder farm, outbreaks of Sutica spp. Occurred mainly in the low temperature season, but in recent years, Sutica spp.

It has been known that formalin bathing is effective as a therapeutic agent for the infection of Suktica, (Former, 1992; former, 2000; Jee and Jo, 2002; There is a lot of treatment by hydrogen peroxide bath. In addition, some compounds known to be parasiticidal agents have been shown to have an insecticidal effect against the scuticacea (Iglesias et al., 2002) but have not yet been put to practical use. In addition, when chemotherapy drugs such as chemical drugs used as insecticidal substances against Sututika insecticides are prohibited from being used as environmental pollution problems, there is no Sututika insecticide that is still deserved, and there is a strong demand for safe food for consumers It is true.

Therefore, it is necessary to develop safe medicines for relieving sutuchikae which cause mass damage in fish farms.

Songjeon is a place where Songjin (松 津) in the stem and branch of a pine tree, an evergreen tree, is settled. The pine tree can be taken from any pine tree growing in various places, but the time of harvest is not particularly limited. Songjeol cuts the branches of the branches of the pine tree, dips them in water, and cuts them again. The flavor is sweet, spicy, poisonous, makes the five days easier, eliminates heat, improves flavor, and is temperate. Songjeol acts on the heart and the menopause, eliminates customs, calms the spasms, passes through the meridian, and is used for joint pain, cramps, legs, bruises. When taking, 12 ~ 20g of the day is made in the form of ganjeja or liquor. In the main hall gangmok and the royal palace gyeongbokgwa, it heals the gangbang, the angular ridge, the fracture barrel, and strengthens the leg when it is drunk and heals all medicines, It is said that it has the effect of pain and sterilization when attached to the window.

Sendal tree ( Machilus japonica ) is an evergreen tree of the dicotyledonous plants of the family Ranunculaceae, with a height of 10 m, and the bark is grayish brown or tan. The new branch has no hairs with winter eyes. Leaves are alternate, basal or inverted, with narrow ends, no hairs, and flat edges. The leaf surface is bluish green and the back side is blue white. Flowers bloom in May, light yellow-green color, and hang on cones of long peduncle. There is a small peduncle, the horns are arranged in 2 lines of 3, and the stamen is arranged in 3 lines of 3, and the pistil is 1. The fruit is round as a berry, about 1 cm in diameter, ripened in rust black around September of the following year, and a piece of falconry remains on the bottom. It is distributed in Korea, Japan, and Taiwan.

Macleaya cordata , also known as macroraya , is a perennial plant that grows about 1.5 to 2 meters in length. Its stem has a diameter of about 2 centimeters. Its stem is hollow and has sparse leaves. Leaves are horny, broad ovate, 20 ~ 40㎝ in length and 15 ~ 30㎝ in width. The back side of leaf is white with white petiole. The flowering period is from July to August, and a large number of flowers are densely bloomed and form a large cone flower. Origin is distributed in the temperate zone of the northern hemisphere.

The camphor ( Cinnamomum camphora ) is an evergreen large tree growing at the foot of a mountain with a height of 20-50m. The bark is brown or dark gray. Leaves are alternate phyllotaxis, 5-9cm long, 3-6cm wide, broad ovate, pointed with narrowed leaf tips. The leaf vein has three branches and the back side is green and shiny. Flowers bloom in May-June, hanging on the end of a sprig from the axilla's branch, hanging in a cone, and light yellow. The horns are wide elliptical, and they are arranged in 2 lines of 3 pieces. The fruit is berry, round and ripe black. It grows in Jeju Island in Korea. It is distributed in Japan, China and Taiwan (Taiwan). Wood is planted as a work of art, and trees are planted as ornamental.

On the other hand, the inventors of the present invention conducted research on the infection diseases of fishes and methods of preventing or treating them, and found that fermented products of mixed extracts of mullet, It was confirmed that the effect of treating the infectious disease was excellent, and the present invention was completed.

Korean Patent No. 10-0820247 (Title of the invention: composition for insecticidal action against schizophrenia comprising zucchini extract as an active ingredient, applicant: Creation Biotech Co., Ltd., registered on Apr. 1, 2008) Korean Patent Publication No. 10-2012-0070015 (Title of the invention: composition for relieving scuticacid containing alpha-pinene as an active ingredient, applicant: Creation Biotech Co., Ltd., publication date: June 29, 2012) Korean Patent No. 10-1158364 (entitled: Pest insecticidal composition isolated from aquatic plants, pest control agent containing the pest insecticidal composition and method for controlling the same, applicant: Kyung Hee University Industry & Academy Collaboration Foundation) Month) Korean Patent No. 10-1762762 (the name of the invention: a method for producing a composition capable of simultaneously controlling pest insect pests containing an extract of camphor tree or pictorial tree as an active ingredient, and a composition prepared by the method, applicant: agricultural company corporation Nature and future , Posted on July 24, 2017)

It is an object of the present invention to provide a composition for the prevention or treatment of scuticacid infectious disease containing fermented products of mixed extract of mulberry leaves, safflower seeds, dead leaves and camphor leaves.

The present invention relates to a composition for preventing or treating scuticacid infectious disease, which comprises a fermented product of a mixed extract of mulberry leaves, Machilus japonica fruit, Macleaya cordata leaf and Cinnamomum camphora leaf .

The mixed extract may contain 100 to 200 parts by weight of the seeds of Sendal nuts, 10 to 50 parts by weight of dead leaves, and 50 to 100 parts by weight of camphor leaves on the basis of 100 parts by weight of the mint.

The mixed extract may be a hot-water extract, which is extracted by adding 1000 to 2000 parts by weight of water on the basis of 100 parts by weight of melon.

The fermented product may be a fermented product obtained by co-culturing a mixed extract of mulberry jelly, safflower seeds, mulberry leaf, and camphor leaf with a mixed strain of lactic acid bacteria and yeast. At this time, molasses may be added when co-culturing with a mixed strain of the above-mentioned lactic acid bacteria and yeast, and 0.1 to 0.2 weight of the molasses may be added to the extracted extract by adding water. The mixed strain may be added in an amount of 0.0001 to 0.001 part by weight based on 1 part by weight of the extract. In this case, the fermentation time is preferably 5 to 10 days, and the fermentation temperature is preferably 25 to 40 ° C. When the fermentation is completed, it is preferable to react at 80 to 90 ° C for 1 to 3 hours to inactivate the activity of the mixed strain.

The lactic acid bacterium may be selected from Lactobacillus plantarum and Lactobacillus casei , and those distributed on the market for food production may be used. Preferably, those produced for yogurt production can be used .

The yeast may be Saccharomyces cerevisiae , and any of those produced in the form of wine, beer or bread-making seeds may be used.

The mixed strains may be mixed with lactic acid bacteria and yeast in a weight ratio of 99: 1 to 90: 1 on the basis of the number of cells.

The present invention also provides a feed additive composition for fishes comprising a fermented product of a mixed extract of Macaday japonica fruit, Macleaya cordata leaf and Cinnamomum camphora leaf. The composition is characterized in that it has an effect of preventing or ameliorating scuticacid infectious disease. The fish may be preferably flounder.

Accordingly, the present invention also provides a fish feed comprising the composition.

The mixed extract may be added in an amount of 0.001 to 30% by weight to the feed composition for fish rearing. The feed additive composition may be added to feeds that prevent or ameliorate infectious diseases of fish according to a conventional method, and may be formulated into various types of feed compositions.

In addition, the present invention provides a pharmaceutical composition for fishes that prevents or treats infectious diseases of Sukhtuka japonica containing a fermented product of a mixed extract of Sendal nuts, Safflower larvae and camphor leaves. The gangrene extract may be added in an amount of 0.001 to 30% by weight to the pharmaceutical composition of the present invention. The pharmaceutical composition may be formulated into various forms used for infectious disease of fish according to a conventional method.

The present invention also relates to a method for producing a hot-water extract, comprising the steps of (1) adding water to a mixture of linden , Machilus japonica fruit, Macleaya cordata leaf and Cinnamomum camphora leaf, step; And

(Step 2) Mixing molasses in the hot-water extract solution and adding a mixed strain of lactic acid bacteria and yeast to co-culture to obtain a culture product. Thereby producing a fermented product.

The fermentation product may be provided as a feed additive composition of fish.

In the first step, 100 to 200 parts by weight of the seeds of deciduous trees, 10 to 50 parts by weight of dead leaves, and 50 to 50 parts by weight of camphor tree leaves, 100 parts by weight may be mixed.

In the first step, the water may be added in an amount of 1000 to 2000 parts by weight based on 100 parts by weight of the gypsum.

In the first step, it is appropriate that the heating extraction conditions are 70 to 90 占 폚 for 12 to 48 hours.

In the second step, the molasses is preferably added in an amount of 0.1 to 0.2 wt% based on the weight of the hot-water extract.

The present invention relates to a fermented product of a mixed extract of mulberry jelly, safflower nuts, mulberry leaf, and camphor leaf. The composition containing the fermented product is excellent in the remedial effect of Sucutika spp., And is useful as a pharmaceutical composition for fish or a feed additive Can be easily used as a composition.

Fig. 1 shows a process of isolating and cultivating Sukutika, wherein A is a flounder infested with Sukutika, B is a microscope photograph of Sukitika, C is a microscope photograph of a coin cell subjected to an infection test of Sukikaca, This indicates a flask in which the Tikapugi is being cultured.
2 is a photomicrograph showing a state (A) of a normal state of the scutica and a state (B) in which a scutica is killed.
Fig. 3 shows the results of measurement of flounder mortality, which confirmed the insecticidal effect of the sututa crab on the fermented product of the present invention.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the concept of the invention to those skilled in the art.

<Example 1> Preparation of fermented product of mixed extract of mulberry, safflower,

The mulberry leaves, safflower seeds, mulberry leaves and camphor leaves were dried and mixed with water under the conditions shown in Table 1, and then extracted at 80 ° C for 12 hours. Only the liquid phase was collected. When the weight of the collected liquid phase was 1, molasses equivalent to 0.1 weight was added, and 0.0001 weight of seed bacterium mixed with lactobacillus plantarum and saccharomyces cerevisiae at a weight ratio of 99: 1 was added. The Lactobacillus plantarum was purchased as a yoghurt preparation, and the Saccharomyces cerevisiae was purchased from a beer brewer.

One week after the addition of the mixed strains, the fermentation broth was reacted at 90 ° C. for 1 hour to inactivate the activity of the strain, and the fermentation broth was prepared as a powder by lyophilization.

Condition The gods (g) Sendal nut (g) Dead leaves (g) Camphor leaf (g) Fermentation Example 1-1 100 150 25 75 ○ Examples 1-2 100 100 50 100 ○ Example 1-3 100 190 10 50 ○ Examples 1-4 100 100 50 100 ○

&Lt; Comparative Example 1 > Preparation of Various Extracts to be Comparative and Fermented Product &gt;

On the condition of Table 2, fermented water or extract was prepared from mulberry jelly, Sendal nuts, mulberry leaves and camphor leaves. The fermented product was prepared in the same manner as in Example 1 except that only the molasses was added and the fermentation process was not performed at all.

Condition The gods (g) Sendal nut (g) Dead leaves (g) Camphor leaf (g) Fermentation Comparative Example 1-1 350 0 0 0 ○ Comparative Example 1-2 0 350 0 0 ○ Comparative Example 1-3 0 0 350 0 ○ Comparative Example 1-4 0 0 0 350 ○ Comparative Example 1-5 200 0 150 0 ○ Comparative Example 1-6 0 200 0 150 ○ Comparative Example 1-7 100 100 150 0 ○ Comparative Example 1-8 100 40 190 20 ○ Comparative Example 1-9 100 150 25 75 x

<Experimental Example 1: Isolation and culture of Sutuca crab>

Sucutica spp. Was directly isolated from flounder, which was infected with Sututika spp., And cultured. The tissues of the infected area were partially excised from the flounder and placed on a slide glass and observed under a microscope. As a result, it was confirmed that they were infected with Sukutika (Fig. 1. A-B). Confirmed Sukutika worms were inoculated with sterilized seawater in P1Y1 (1% proteose peptone, 1% yeast extract) medium to conduct primary culture at 15 ° C. There are many bacteria in the primary cultured Sututika crab culture, and pure culture using a fish coin cell line was carried out with the limitation of bacterial contamination and proliferation (Fig. 1C). Next, for the pure culture of scutica, fish CHSE-214 (Chinook salmon embryo) was supplemented with 10% FBS and penicillin-streptomycin in a MEM medium (Modified Eagle's Medium) titrated to pH 7.3 with 7.5% sodium bicarbonate And cultured at 20 ° C. The monolayer-grown cell line was inoculated with the cultured scuticacid fungus and cultivation was continued at 20 ° C until the scuticha crab completely consumed the cell line (Fig. 1D). After observing under the microscope that the scutica larvae were completely consumed, count the number of scutica cells in 1 mm x 1 mm using a hematocytometer (blood count plate) to determine the number of cells that were proliferated. Respectively.

<Experimental Example 2: Determination of insecticidal ability against Sutuca puffer I>

To determine the insecticidal activity, Sukutika pupae at the end of the logarithmic growth period or at the early stage of the pause were centrifuged at 650 ㅧ g for 5 minutes and then resuspended in PBS or filtered seawater. After counting with a hematocytometer, 0.9 ml of each fermentation product or extract was added to a 24-well plate in 0.1 ml of a solution containing 104 sutucca cells, mixed and incubated at 20 ° C in an incubator, shaking for 1 hour, . At this time, the final concentration of each fermentation product or extract was adjusted to be 10 ppm, and the insecticidal ability was confirmed based on the appearance of the sututa crab (see Figs. 2A and 2B).

Table 3 shows the insecticidal activity of Sucutika on the fermented products or extracts of Example 1 and Comparative Example 1 through these criteria.

Conditions of fermented water or extract Insecticide (%) Example 1-1 100 Examples 1-2 100 Example 1-3 100 Examples 1-4 100 Comparative Example 1-1 16 Comparative Example 1-2 12 Comparative Example 1-3 15 Comparative Example 1-4 7 Comparative Example 1-5 3 Comparative Example 1-6 12 Comparative Example 1-7 15 Comparative Example 1-8 26 Comparative Example 1-9 25

As a result, it was confirmed that all of the fermented product of Example 1 was killed by 100% of the Sututika crab, but the fermentation product or extract of Comparative Example 1 showed almost no insecticidal effect.

<Experimental Example 3: Confirmation of insecticidal activity against Sutica spp .- II>

The insecticidal power of the fermented product of Example 1-1 against Sukutika was directly confirmed in the flounder. A 10 ppm solution of the fermented product of Example 1-1 was sprayed with 5 g based on 100 g of the flounder feed and sufficiently dried. Experimental flounder was artificially used for 30 days (for each group) infected with Sukutika infestation for 3 days.

The mortality rate of the flounder was shown in FIG. 3 when the feed was supplied until the 26th day. The final mortality rate of the control was 80%. In the fermented water treated group of Example 1-1 of the present invention, the mortality rate was 60% 20%, and it was confirmed that the fermented product of the present invention has an effect of inhibiting the infectivity of Sucutica in flounder.

Claims (7)

A composition for preventing or treating scuticacid infectious disease, which comprises a fermented product of a mulberry , a Machilus japonica fruit, a Macleaya cordata leaf and a Cinnamomum camphora leaf mixture. The method according to claim 1,
Wherein said mixed extract is extracted by containing 100 to 200 parts by weight of safflower seeds, 10 to 50 parts by weight of safflower leaf, and 50 to 100 parts by weight of camphor leaf on the basis of 100 parts by weight of gypsum. Or &lt; / RTI &gt; The method according to claim 1,
Wherein the fermented product is a fermented product obtained by co-culturing a mixed extract of mulberry jelly, safflower seeds, mulberry leaf, and camphor leaf with a mixed strain of lactic acid bacteria and yeast. Wherein the composition comprises a fermentation product of a mixed extract of Macalaya japonica fruit, Macleaya cordata leaf and Cinnamomum camphora leaf. A fish feed comprising the composition of claim 5. (Step 1) Water is added to a mixture of jellyfish , Machilus japonica , Macleaya cordata leaf, and Cinnamomum camphora leaf, followed by heating and extracting, and collecting only the liquid phase to obtain a hot water extract; And
(Step 2) mixing molasses in the hot-water-extracted liquid and adding a mixed strain of lactic acid bacteria and yeast to co-culture to obtain a culture; &Lt; / RTI &gt; A feed additive composition for fish characterized by containing a fermented product produced by the method of claim 6.

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