KR102610687B1 - Immunity enhancer composition for beekeeping and use thereof - Google Patents

Abstract

The immune enhancer composition for beekeeping according to the present invention has the effect of increasing the immunity of bees and improving the survival, activity, and honey production of bees. It does not harm bees, has high stability, and has strong control properties against pests such as mites. Eggplant is effective.

Description

Immunity enhancer composition for beekeeping and use thereof}

The present invention relates to an immune-enhancing composition for beekeeping and its use, and more specifically, to an immune-enhancing composition for beekeeping used to control pests parasitic on bees, such as mites.

Beekeeping refers to the farming of bees or bees to obtain honey, and is used in a variety of ways because it can produce not only honey, but also beeswax, propolis, pollen, and royal jelly. Since this beekeeping industry occupies a high share not only domestically but also overseas, it is important to protect and maintain the bee population in the beekeeping industry.

Recently, the number of bees has been rapidly decreasing due to disease, and it is known that bee diseases are caused by pesticides, herbicides, parasites, fungi, and viruses. In particular, diseases of honey bees are mainly caused by parasites or pathogens, and representative diseases of honey bees include Nosema disease, buzzard disease, and chalk disease. These bee diseases break the homeostasis of individual bees and the entire bee colony, making it difficult to protect and maintain bee populations, so it is important to strengthen immunity to prevent bee diseases.

Bees can have immunity to diseases. The higher their immunity, the better their water resistance (ability to collect honey; foraging power), and they can prevent frostbite during winter overwintering. However, it is not easy to enhance the immunity of bees, so the development of a composition to increase the immunity of bees is necessary.

Generally, the beehives used in beekeeping are made of wood, so the survival probability of bees is reduced due to pests living inside the wood. Representative pests that live inside beehives include bee mites such as Chinese thorn mite ( Tropilaelaps clareae ), honey bee mite ( Varroa jacobsoni ), and small honey bee mite ( Varroa underwoodi ). These bee mites have been distributed and expanded worldwide, and are external parasites, parasitizing bee larvae, pupae, and adults and sucking their body fluids. External parasitism as described above not only makes the normal development of honey bees difficult and reduces their activity, but if the occurrence of bee mites is severe, the power of the bee colony is greatly weakened, leading to the colony being abandoned, causing serious damage to the beekeeping industry.

To prevent damage caused by these bee mites, methods of selectively killing bee mites using bee mite control products are being adopted around the world. However, conventional bee mite control products are made of organic synthetic materials, and bee mites frequently exposed to these organic synthetic materials develop resistance, making control difficult. In addition, the organic synthetic material is toxic to the bees themselves, which also reduces the survival rate of the bees, so there is still a limit to increasing the production of bees.

Therefore, research is needed on an immune-enhancing composition that increases the immunity of honey bees and has a selective insecticidal effect on pests such as honey bee mites while being almost non-toxic to honey bees.

Korean Patent Publication No. 10-1424083 (2014.07.22)

The object of the present invention relates to an immune-enhancing composition for beekeeping that increases the immunity of bees and improves their survival, activity, and honey production.

Another object of the present invention relates to an immune-enhancing composition for beekeeping that does not harm bees, has high stability, and has a strong control effect against pests parasitic on bees, such as mites.

The immune enhancing composition for beekeeping according to the present invention includes an aqueous copper colloid solution and a diluent.

The immune enhancer composition for beekeeping according to an example of the present invention may include 1 to 10 parts by weight of the copper colloidal aqueous solution based on 100 parts by weight of the diluent, and the copper colloidal aqueous solution may include copper nanoparticles at a concentration of 1,000 to 3,000 ppm. You can.

In one example of the present invention, the diluent may include water.

In one example of the present invention, the copper colloidal aqueous solution may include copper nanoparticles and water, and the average particle diameter (D50) of the copper nanoparticles may be 2 to 10 nm.

In one example of the present invention, the copper colloidal aqueous solution is prepared by mixing and reacting copper chloride and sodium hydroxide in the water phase to produce an intermediate product including copper oxide and copper hydroxide, and mixing the intermediate product and hydrazine. and a reduction step of reacting to produce copper nanoparticles.

In one example of the present invention, in the dechlorination step, the sodium hydroxide may be mixed at 1 to 6 moles per mole of the copper chloride, and in the reduction step, the hydrazine may be mixed at an amount of 1 to 12 moles per mole of the copper chloride. It can be.

In one example of the present invention, in the desalination step and the reduction step, the reaction temperature may be 20 to 60°C.

The immune enhancer composition for beekeeping according to an example of the present invention may further include natural plant extract and natural plant fermentation liquid, and the natural plant may include sophora root and derris root.

The immune enhancer composition for beekeeping according to an example of the present invention may include 0.1 to 5 parts by weight of the natural plant extract and 0.5 to 10 parts by weight of the natural plant fermentation broth based on 100 parts by weight of the diluent.

In one example of the present invention, the natural plant extract comprises a) preparing a natural plant extract by extracting a natural plant mixture containing the Sophora ginseng root and the Derris root with an extraction solvent, and b) the natural plant extract. It can be prepared including the step of separating and filtering solid natural plants through a filter to prepare a natural plant extract.

In one example of the present invention, the natural plant fermentation broth is prepared by fermenting a yeast mixture containing the solid natural plant and yeast separated in step b) to produce a natural plant fermentation product, and d) the natural plant fermentation broth. The plant fermentation product may be prepared including the step of separating and filtering the solid natural plant through a filter to obtain a natural plant fermentation broth.

In one example of the present invention, in step a), the extraction may be ultrasonic extraction and may be performed at 20 to 60°C.

In one example of the present invention, in step a), the extraction solvent may include at least one selected from water and ethanol.

In one example of the present invention, in step a), the mixing weight ratio of the sophora root and the derris root may be 1:0.5 to 2.

In one example of the present invention, in step a), the extraction solvent may be used in an amount of 0.5 to 30 parts by weight based on 1 part by weight of the natural plant mixture.

In one example of the present invention, in step c), the yeast is a low-temperature tolerant yeast, and the fermentation may be performed at 3 to 15° C. for 3 to 15 days.

In one example of the present invention, in step c), the mixing weight ratio of the solid natural plant and the yeast may be 100:1 to 10.

In one example of the present invention, in step c), the yeast mixture may further include fucoidan and calcium-containing coral extract.

In one example of the present invention, in step c), the yeast mixture may include 0.1 to 10 parts by weight of the fucoidan and 0.1 to 10 parts by weight of the calcium-containing coral extract based on 100 parts by weight of the solid natural plant. .

The immune-enhancing composition for beekeeping according to the present invention can be used for controlling pests, including mites.

In one example of the present invention, the mites may include any one or two or more selected from Chinese spiny mites, honey bee mites, stipe mites, and varroa mites.

The immune enhancing composition for beekeeping according to the present invention has the effect of increasing the immunity of bees and improving the survival, activity, and honey production of bees.

In addition, the immune enhancer composition for beekeeping according to the present invention does not harm bees and has high stability, while having strong control properties against pests parasitic on bees, such as mites.

Hereinafter, the immune enhancing composition for beekeeping according to the present invention will be described in detail.

Unless there is another definition in the technical and scientific terms used in this specification, they have meanings commonly understood by those skilled in the art to which this invention pertains, and the gist of the present invention is not unnecessarily explained in the following description. Descriptions of known functions and configurations that may be unclear are omitted.

In some cases in this specification, in order to avoid ambiguity of the concept of the present invention, well-known structures and devices may be omitted or may be shown in block diagram form focusing on the core functions of each structure and device.

'Include' mentioned in this specification is an open description that has the same meaning as expressions such as 'comprises', 'contains', 'has', 'characterized by', etc., and includes elements that are not additionally listed, Does not exclude materials or processes.

The singular form of the terms used in this specification may be interpreted to also include the plural form unless otherwise specified.

Numerical ranges as used herein include lower and upper limits and all values within that range, increments logically derived from the shape and width of the range being defined, all doubly defined values, and upper and lower limits of numerical ranges defined in different forms. Includes all possible combinations of Additionally, unless specifically defined in the specification of the present invention, values outside the numerical range that may occur due to experimental error or rounding of values are also included in the defined numerical range.

The unit of ‘%’ used without special mention in this specification means ‘% by weight’ unless otherwise defined.

The immune enhancing composition for beekeeping according to the present invention includes an aqueous copper colloid solution and a diluent. By containing an aqueous copper colloid solution, the composition has the effect of controlling pests parasitic on bees, such as mites parasitic on bees. In addition, by further including the specific components described later, side effects on bees are minimized and the immunity of bees is improved, thereby significantly improving the activity, honey collection, and survival of bees.

The copper colloidal aqueous solution is one in which copper nanoparticles are dispersed in a colloidal state in an aqueous phase, and includes copper nanoparticles and water. Here, the copper nanoparticles may have an average particle diameter (D50) of 1 to 50 nm, preferably 2 to 10 nm. When the copper colloidal aqueous solution is used as an immune enhancer composition for beekeeping at a concentration (content) described later, it can exhibit a significant insecticidal effect on pests parasitic on bees without causing substantial harm to bees.

The content of the copper colloidal aqueous solution should be used in such a way that it can have a significant insecticidal effect on pests parasitic on bees without causing substantial harm to bees. In a preferred example, the composition has the above-mentioned amount relative to 100 parts by weight of the diluent. It contains 1 to 10 parts by weight of an aqueous copper colloidal solution, preferably 2 to 8 parts by weight, wherein the aqueous copper colloidal solution is used to contain copper nanoparticles at a concentration of 1,000 to 3,000 ppm, preferably at a concentration of 1,500 to 2,500 ppm. good night.

In a preferred example, the composition contains copper nanoparticles at a concentration such that it contains 2.5 to 5 parts by weight of the copper colloidal aqueous solution based on 100 parts by weight of the diluent when the concentration of the copper nanoparticles in the aqueous copper colloidal solution is 2,000 ppm. It may contain. If this is satisfied, it can have a great insecticidal effect on pests parasitic on bees while causing minimal harm to bees.

In this specification, when the composition ratio of the immune enhancer composition for beekeeping is described, it is described as the copper colloid aqueous solution composition and its concentration composition, but even if it is described in a different way, it should be interpreted with the same rights as long as the essence of the invention with respect to the content of each component is the same. Of course. For example, even for a composition that satisfies a specific composition ratio, which is also described as the content of copper nanoparticles among all components of the composition, if the composition ratio is substantially the same and the essence of the invention is the same, it should be interpreted as having the same scope of rights.

The diluent may be anything that can reduce the concentration of copper nanoparticles in the colloidal aqueous solution, and for example, it is preferably the same water as the colloidal aqueous solution.

The copper colloidal aqueous solution can be prepared by various methods, as long as copper nanoparticles are dispersed in colloidal form in the aqueous phase. As a specific example, the method for producing the aqueous copper colloid solution includes a desalination step of mixing and reacting copper chloride and sodium hydroxide in an aqueous phase to produce an intermediate product including copper oxide and copper hydroxide, and mixing the intermediate product and hydrazine, It may include a reduction step of reacting to produce copper nanoparticles.

The desalination step is a step in which copper chloride (CuCl ₂ ) reacts with sodium hydroxide (NaOH) in the water phase to produce copper oxide (CuO) and copper hydroxide (Cu(OH) ₂ ). Copper chloride is a precursor of copper nanoparticles. Unlike copper sulfate (CuSO ₄ ), copper chloride has an anionic functional group with relatively high electronegativity, so it produces an anion effect different from that of sulfate ions in a solution, preventing the produced nanoparticles from aggregating with each other. The phenomenon can be suppressed. Therefore, not only is it possible to manufacture finer nanoparticles within the above-described particle size range, but it also exhibits a more suitable surface shape control effect that can realize the above-mentioned effects.

The sodium hydroxide separates chlorine (Cl) from the copper atom of copper chloride to produce copper oxide and copper hydroxide. Here, the sodium hydroxide may be used in an amount of 1 to 6 moles per 1 mole of copper chloride. When sodium hydroxide is used in this molar ratio, the overall reaction can proceed smoothly by allowing the reduction reaction in the subsequent reduction step to proceed more actively, minimizing unreacted products, and minimizing the increase in impurities due to residual ions.

In the desalination step, the reaction temperature between copper chloride and sodium hydroxide may be sufficient to allow the reaction to occur smoothly, for example, 20 to 60°C, preferably 25 to 60°C. If this is satisfied, thermal stability can be improved by minimizing aggregation of intermediate products and preventing the reduction reaction in the subsequent reduction step from proceeding at an excessively high temperature.

The reduction step is a step in which hydrazine (N ₂ H ₄ ) undergoes a reduction reaction with copper oxide and copper hydroxide, which are intermediate products produced in the desalination step, to produce copper nanoparticles. Therefore, through the reduction step, an aqueous phase in which copper nanoparticles (CuO) precipitated in a non-ionic state are dispersed in a colloidal state, that is, an aqueous copper colloidal solution, can be prepared. At this time, the hydrazine can be used in an amount of 1 to 12 moles per mole of copper chloride, and when this amount is satisfied, unreacted products can be minimized to allow a reduction reaction to occur, and the agglomeration phenomenon of the produced copper nanoparticles can be minimized.

In the reduction step, the reaction temperature may be sufficient to allow the reaction to occur smoothly, for example, 20 to 60°C, preferably 35 to 60°C. If this is satisfied, the agglomeration phenomenon of the produced copper nanoparticles can be minimized and the reaction speed and reaction conversion rate can be improved.

The immune enhancing composition for beekeeping according to a preferred example of the present invention may further include natural plant extract and natural plant fermentation liquid, and the natural plant may include sophora root and derris root.

The shrubby sophora root contains alkaloids such as Matrine, Oxymatrine, Sophoranol, Anagyrine, and flavonoids such as Xanthohumol. , Isoanhydroicarin, etc. The matrine is a representative active ingredient, and when pests are exposed to the active ingredient, their nerves are paralyzed and internal proteins coagulate, inhibiting breathing, which can lead to death. In particular, when an extract is prepared and used from Sophora ginseng root using the method described below, it is effective in selectively controlling pests while minimizing side effects on bees.

The derris ( Derris eliptica ) roots contain rotenone, which is a representative active ingredient and exhibits an insecticidal effect by disrupting the mitochondrial electron transport system of pests and inhibiting ATP synthesis. In particular, when an extract from derris roots is prepared and used using the method described below, it is effective in selectively controlling pests while minimizing side effects on bees.

It is preferable that the natural plant extract is prepared by the following method in order to selectively control pests while minimizing side effects on bees. The method for producing the natural plant extract includes the steps of a) extracting a natural plant mixture containing the Sophora ginseng root and the Derris root with an extraction solvent to prepare a natural plant extract, and b) passing the natural plant extract through a filter into a solid state. It may include the step of separating and filtering the natural plant to produce a natural plant extract.

Step a) is a step of extracting a natural plant mixture containing Sophora root and Derris root with an extraction solvent. Here, the extraction is preferably ultrasonic extraction, and it is more preferable that the extraction is performed at a temperature below high temperature. Extraction can be performed, for example, in a temperature range of 20 to 60°C. When using an extraction method performed at a temperature below high temperature, it is possible to selectively control pests while minimizing side effects on bees and improve the immunity of bees.

When ultrasonic extraction is used in step a), the ultrasonic frequency and ultrasonic intensity may be sufficient to sufficiently extract the active ingredients that achieve the above-mentioned effects, for example, a frequency of 30 to 300 KHz and a frequency of 100 to 600 Watt. It could be a century. However, this is only described as a specific example and the present invention is not limited thereto.

In step a), the extraction time may be long enough to sufficiently extract the active ingredient that achieves the above-mentioned effects, and may be, for example, 0.2 to 6 hours, specifically 0.5 to 3 hours. However, this is only described as a specific example and the present invention is not limited thereto.

In step a), the extraction solvent may include one or more selected from water and ethanol, and may preferably be an ethanol aqueous solution containing both. When an aqueous ethanol solution is used, the concentration of ethanol may preferably be 50 to 95% by weight, specifically 60 to 90% by weight.

The amount of the extraction solvent used may be sufficient as long as the active ingredients that achieve the above-mentioned effects can be sufficiently extracted with the extraction solvent. For example, it can be used in an amount of 0.5 to 30 parts by weight based on 1 part by weight of the natural plant mixture. However, this is only described as a specific example and the present invention is not limited thereto.

In step a), the mixing weight ratio of the sophora root and the derris root is not greatly limited, and may be, for example, 1:0.5 to 2. If this is satisfied, the immunity of bees can be further improved and the pest control effect can be further improved.

Step b) is a step of separating and filtering solid impurities in the natural plant extract prepared in step a), for example, the extracted solid natural plant, to obtain a natural plant extract. At this time, the filtration means may be any known method. For example, it may be a method of separating and filtering solid natural plants from the natural plant extract through a filter such as a mesh net. The filter used at this time may be an example of a filter having a pore size of 100 to 300 mesh, specifically 120 to 250 mesh. Furthermore, the separated solid natural plants can be stored separately and used in the production of natural plant fermentation broth, which will be described later.

The amount of the natural plant extract used may be any amount that can achieve the above-mentioned effects. For example, it may be used in an amount of 0.1 to 5 parts by weight based on 100 parts by weight of the diluent.

The natural plant fermentation broth is preferably produced by the following method in order to selectively control pests while minimizing side effects on bees. The method for producing the natural plant fermentation broth includes the steps of c) fermenting the yeast mixture containing the solid natural plant and yeast separated in step b) to produce a natural plant fermentation product, and d) filtering the natural plant fermentation product. It may include the step of separating and filtering the solid natural plant through to obtain a natural plant fermentation broth.

Step c) is a step of fermenting natural plants with yeast to produce a natural plant fermentation product. The natural plants used here include the above-mentioned sophora root and derris root, and in this case, the natural plant is the natural plant extract of the natural plant extract. It is preferable that it is a solid natural plant isolated during manufacturing. That is, when producing a natural plant fermentation product by fermenting a natural plant from which some of the active ingredients have been extracted through the extraction in step a) in step c), a new natural plant that has not gone through the extraction step in step a) is used. Compared to the case where it is used, there is no decrease in the pest control effect and it has the effect of minimizing side effects on bees. Furthermore, it has the effect of further increasing immunity to bees.

Fermentation in step c) can be performed at any temperature that allows fermentation, but low-temperature fermentation, for example, fermentation performed in a low temperature range of 3 to 15°C, is preferable in terms of better realizing the above-mentioned effects. The relative humidity during fermentation may be sufficient as long as the yeast can facilitate metabolism, for example, it may be 60 to 90%. Additionally, the fermentation time is not greatly limited, but may be, for example, 3 to 15 days.

The yeast used in step c) may be of various types, but it is more preferable that it is a low-temperature-tolerant yeast that can perform fermentation better at low temperature fermentation. An example of the low-temperature resistant yeast is Saccharomyces. Cerevisiae includes Cheongdo ( Saccharomyces. cerevisiae Cheongdo). The amount of yeast used may be sufficient to produce a fermentation product that achieves the above-mentioned effects. For example, the mixed weight ratio of the solid natural plant and the yeast may be 100:1 to 10.

As a preferred example, in step c), the yeast mixture may further include fucoidan and calcium-containing coral extract. When the solid natural plant separated in step b) is fermented by further adding fucoidan and calcium-containing coral extract, a more favorable fermentation environment for low-temperature-tolerant yeast can be created, thereby maximizing the fermentation environment. Specifically, it is possible to better create a growth environment for yeast, maximize the content of fermentation products caused by microbial metabolism, and significantly increase the type and amount of ingredients that improve the immunity of bees. It is desirable from this point of view. In addition, the effect of enhancing the immunity of bees can be expected due to the fucoidan and calcium-containing coral extract itself. Therefore, it is possible to realize a synergistic effect of strengthening the immunity of bees by fucoidan and calcium-containing coral extract, as well as an immunity-enhancing effect by the active ingredients produced through the fermentation process of these ingredients and natural plants together.

In particular, compared to when the calcium-containing coral extract or fucoidan is used alone and fermented, when fucoidan and calcium-containing coral extract are used together, the above-mentioned effects can be further improved.

Fucoidan is a sulfuric acid polysaccharide having two or more sulfate groups. It is generally contained in large quantities in brown algae, so it can be extracted and used from it, and may be manufactured by extraction and concentration using an extraction solvent such as water. The fucoidan can be used, for example, as a fucoidan-containing seaweed extract derived from brown algae, and the fucoidan-containing seaweed extract may contain 1% by weight or more of fucoidan as a solid content, specifically 3% by weight or more. The amount of fucoidan-containing extract used can be appropriately quantified to achieve the fucoidan content described above. The average molecular weight of fucoidan is not greatly limited and may be, for example, 50,000 to 500,000 Da. However, this is only described as a specific example and the present invention is not limited thereto.

The calcium-containing coral extract contains calcium extracted from coral reefs, and can be extracted by methods such as hot water extraction. There is a known commercially available coral calcium, for example, CT-CCP50 from CaltecBio. As a specific example, calcium-containing coral extract may contain calcium, magnesium, sodium, etc. as main ingredients. More specifically, the calcium-containing coral extract may contain 20 to 70% by weight of calcium, 0.2 to 0.5% by weight of magnesium, and 0.1 to 0.4% by weight of sodium, and may further include potassium, phosphorus, iron, manganese, etc. there is. However, this is only described as a specific example and the present invention is not limited thereto.

In step c), the amount of the fucoidan and the calcium-containing coral extract used may be any amount sufficient to achieve the above-mentioned effect. For example, the yeast mixture may be used in the amount of the above-described yeast mixture relative to 100 parts by weight of the solid natural plant. It may include 0.1 to 10 parts by weight of fucoidan and 0.1 to 10 parts by weight of the calcium-containing coral extract.

Step d) is a step of separating and filtering solid phase impurities in the natural plant fermentation product prepared in step c) to obtain a natural plant fermentation broth. At this time, the filtration means may be any known method, for example, a method of separating and filtering solid impurities from the natural plant fermentation through a filter such as a mesh net. The filter used at this time may be an example of a filter having a pore size of 100 to 300 mesh, specifically 120 to 250 mesh.

The amount of the natural plant fermentation broth may be used as long as it can achieve the above-mentioned effects. For example, it can be used in an amount of 0.5 to 10 parts by weight based on 100 parts by weight of the diluent.

The immune-enhancing composition for beekeeping according to the present invention can be used for controlling pests, including mites, and the pests referred to in the present invention refer to organisms that can harm bees. Specifically, the pests may refer to parasites such as mites, and specific examples of the mites include Chinese spiny mites, honey bee mites, stipe mites, and varroa mites.

The immune-enhancing composition for beekeeping according to the present invention can control pests parasitic on bees by contacting them, and can be used in the field of beekeeping through the so-called ‘shedding treatment’ method. For example, the composition can be treated in a way that allows direct contact with the bees by spraying or flowing the composition into areas adjacent to bees, such as the bee plate, allele, and spectral plate.

Hereinafter, the present invention will be described in detail through examples. However, these are intended to illustrate the present invention in more detail, and the scope of the present invention is not limited to the following examples.

<Copper colloidal aqueous solution manufacturing process>

100 ml of a 2M copper chloride (CuCl ₂ ) aqueous solution was heated and stirred vigorously to maintain the temperature at 35°C, and then sodium hydroxide (NaOH) was added to the aqueous solution so that the amount was 6M. Afterwards, hydrazine (N ₂ H ₄ ) was added while maintaining the temperature of the aqueous solution at 45°C, and the amount added was 15M to reduce it to copper particles. Copper nanoparticles in the copper colloidal aqueous solution prepared in this way were present at a concentration of 2,000 ppm, and the average particle diameter (D50) was 3 nm.

<Manufacturing process of immune enhancer composition for beekeeping>

Three types of immune enhancer compositions for beekeeping with different composition ratios were prepared by mixing 50 g, 75 g, and 100 g of the copper colloidal aqueous solution in 2 kg of water.

An immune enhancer composition for beekeeping was prepared in the same manner as in Example 1, except that the content of the copper colloid aqueous solution was changed to 40 g in the manufacturing process of the immune enhancer composition for beekeeping in Example 1.

An immune enhancer composition for beekeeping was prepared in the same manner as in Example 1, except that the content of the copper colloid aqueous solution was changed to 110 g in the manufacturing process of the immune enhancer composition for beekeeping in Example 1.

An immune enhancer composition for beekeeping was prepared in the same manner as in Example 1, except that 20 g of natural plant extract prepared by the following process was further mixed in the manufacturing process of the immune enhancer composition for beekeeping in Example 1.

<Natural plant extract manufacturing process>

1 kg of natural plant mixture containing crushed Sophora ginseng roots and Derris roots in a 1:1 weight ratio to about 3 cm in size was added to 10 kg of ethanol aqueous solution with a concentration of 70% by weight and ultrasonic extraction was performed at 25°C for 30 minutes to obtain the natural plant mixture. An extract was prepared. The ultrasonic extraction device used at this time was an Ultrasonic cleaner (5510-DTH, Bransonic, Danbury, USA), and extraction was performed at a frequency of 100 KHz and an intensity of 300 Watt. Subsequently, the natural plant extract was left at 25°C for 2 hours so that the solid impurities were located in the lower layer to recover the supernatant, which was filtered through filter paper (Whatman No. 2, GE Healthcare Ltd., Little Chalfont, UK) to obtain the solid phase. A natural plant extract from which impurities were removed was obtained. At this time, the separated solid phase impurities, that is, the solid phase natural plants including Sophora ginseng root and Derris root that were used for extraction and some of the active ingredients were released, were stored separately.

An immune enhancer composition for beekeeping was prepared in the same manner as in Example 4, except that 30 g of natural plant fermentation broth prepared by the following process was further mixed in the manufacturing process of the immune enhancer composition for beekeeping in Example 4.

<Natural plant fermentation broth manufacturing process>

In the natural plant extract manufacturing process of Example 4, 100 g of general yeast (EC-1118, LALVIN) was inoculated into 1 kg of natural plant mixed with Sophora root and Derris root at a 1:1 weight ratio, placed in a container, and sealed. After creating an anaerobic environment, a natural plant fermentation broth was prepared by fermenting for 5 days at a temperature of 25°C and a relative humidity of 80% using a constant temperature and humidity chamber (Temi 850, Asung Tech, Gimhae, Korea).

In the natural plant fermentation liquid manufacturing process of Example 5, instead of the natural plant in which sophora root and derris root were mixed at a 1:1 weight ratio, the natural plant separately stored in the natural plant extract manufacturing process of Example 4 (the sophora root used in extraction and An immune-enhancing composition for beekeeping was prepared in the same manner as in Example 5, except that Derris root) was used.

In the natural plant fermentation broth production process of Example 6, the same process as Example 6 was performed, except that cold-tolerant yeast ( Saccharomyces. cerevisiae Cheongdo) was used instead of regular yeast, and fermentation was performed at 7°C and 10 days instead of 25°C and 5 days. An immune-enhancer composition for beekeeping was prepared using this method.

In the natural plant fermentation broth manufacturing process of Example 7, 100 g of calcium-containing coral extract (CT-CCP50, CaltecBio) and 500 g of fucoidan-containing seaweed extract (Haerim Fucoidan, Haerim) (fucoidan solids 4.7 wt%, liquid) were added to the natural plant. An immune enhancer composition for beekeeping was prepared in the same manner as in Example 7, except that it was further mixed and fermented.

[Experimental Example 1] Bee activity test

After feeding the immune enhancing composition for beekeeping prepared in the examples to bees ( Apis mellifera ), the activity of bees was measured by visually observing the number of bees per 10 cm ² area during the bee life cycle (21 days). The results are shown in Table 1 below.

[Experimental Example 2] Bee foraging ability test

After feeding the immune enhancer composition for beekeeping prepared in the examples to bees ( Apis mellifera ), the honey amount (kg) per bee group was checked on the 30th, 45th, and 60th days of feeding to determine the collecting power of the bees. Measurements were made, and the results are shown in Table 1 below.

[Experimental Example 3] Bee damage rate test

After feeding the immune enhancer composition for beekeeping prepared in the examples to bees ( Apis mellifera ), after the bee life cycle (21 days), the mite killing rate was measured through the total number of fallen mites, and the total number of fallen bees was measured. The bee damage rate was measured through the number of bees. And the results are shown in Table 1 below.

All experimental results according to the present invention were repeated three times, and through statistical analysis, the Student t-test of the GraphPad program was used to determine significance if the p value was 0.05 or less. It was determined that it was.

Claims (6)

An immune enhancing composition for beekeeping bees comprising an aqueous copper colloid solution, natural plant extract, natural plant fermentation broth, and diluent,
The natural plants include sophora root and derris root,
The natural plant extract is,
a) preparing a natural plant extract by extracting a natural plant mixture containing the sophora root and the derris root with an extraction solvent; and
b) preparing a natural plant extract by separating and filtering the solid natural plant extract through a filter;
The natural plant fermentation broth,
c) fermenting the yeast mixture containing the solid natural plant and yeast separated in step b) to produce a natural plant fermentation product, and
d) An immunity enhancer composition for beekeeping bees prepared including the step of separating and filtering the solid phase natural plant through a filter to obtain a natural plant fermentation broth. According to paragraph 1,
Containing 1 to 10 parts by weight of the copper colloidal aqueous solution based on 100 parts by weight of the diluent,
The copper colloidal aqueous solution is an immunity enhancer composition for bees for beekeeping containing copper nanoparticles at a concentration of 1,000 to 3,000 ppm. According to paragraph 1,
The copper colloidal aqueous solution contains copper nanoparticles and water,
An immunity enhancer composition for beekeeping bees wherein the average particle diameter (D50) of the copper nanoparticles is 2 to 10 nm. delete According to paragraph 1,
In step c), the yeast is a low-temperature tolerant yeast, and the fermentation is performed at 3 to 15° C. for 3 to 15 days,
In step c), the yeast mixture further includes fucoidan and calcium-containing coral extract. According to any one of claims 1 to 3 and 5,
An immunity enhancing composition for beekeeping bees used for controlling pests including mites.