KR20190142564A - Feed composition comprising melberry leaf for preventing or improving infection of Allomyrina dichotoma Nudivirus - Google Patents

Abstract

The present invention relates to a feed composition containing mulberry leaves for preventing or reducing nudivirus infection of Allomyrina dichotoma. By supplying the composition to Allomyrina dichotoma larvae and adjusting the feeding time, it is possible to breed Allomyrina dichotoma larvae well, which were mostly killed before the pupa stage due to nudivirus infection, until the adult stage.

Description

Feed composition comprising melberry leaf for preventing or improving infection of Allomyrina dichotoma Nudivirus}

The present invention relates to a feed composition for preventing or improving long beetle beetle Nudivirus infection containing mulberry leaves.

Allomyrina dichotoma is an insect belonging to Coleoptera and Family Scarabaeidae. Long-tailed beetles are not sensitive to humidity or temperature and are difficult to feed, making them easy to breed, making them popular as pet insects. The tendency to use is increasing. Slugs, silkworms, cordyceps, centipedes, etc., which have been frequently used as herbal medicines since ancient times, are used as useful insect resources. Recently, research on the function of these natural resources has been actively conducted in the development of new drugs. Like this, the beetle is widely used as a pet and is one of the useful insect resources.

Nudieviruses infected with Korean beetles are the most common infections of the beetle beetles. These viral infections cause mass mortality of the beetle beetles, causing severe economic losses. Research has been carried out to the extent that there are 95-99% homology with the genes of Orictes nudyvirus that infects the beetle ( Oryctes rhinoceros ) and [Oryctes rhinoceros virus isolate Ma07 from Malaysia, complete genome] (Genbank number: EU747721.1). As a method for diagnosing this problem is being studied (Korean Patent Publication No. 10-2018-0024813), research on a composition for preventing or improving an infection caused by such a virus is almost insignificant.

Accordingly, the present inventors have completed the present invention by confirming that the mulberry leaf powder is fed to the beetle beetle larvae infected with the virus, and the beetle beetle larvae are grown from the infectious disease to adulthood through the pupal phase while maintaining a healthy state.

Republic of Korea Patent No. 10-1672174 (Invention name: Insect feeding thing loading gel and its manufacturing method, Applicant: Ether Co., Ltd., registered date: October 28, 2016) Republic of Korea Patent No. 10-0786501 (Invention: Mixed feed composition of scarabs and larvae, Applicant: Yoon Ju-sam, Registered Date: December 10, 2007) Republic of Korea Patent Publication No. 10-2018-0002307 (Name of the invention: white spotted flower plain white leukemia control composition, Applicant: Republic of Korea (Rural Development Administration), Publication Date: January 08, 2018) Republic of Korea Patent Publication No. 10-2018-0024813 (Invention name: primer set for isothermal nucleic acid chain polymerization method for diagnosing infection of enteric beetle pathogenic virus or orictes nudievirus and infection with enteric beetle pathogenic virus or orictes nudyvirus Diagnostic Method, Applicant: Korea (Rural Development Administration), Publication Date: March 08, 2018)

An object of the present invention is to provide a feed composition for the prevention or improvement of the beetle beetle Nudivirus infection containing mulberry leaves.

The present invention relates to a feed composition for preventing or improving long beetle beetle Nudivirus infection containing mulberry leaves.

The feed composition may contain 1 to 5% by weight of mulberry leaves.

The feed composition may be added to fermented sawdust fermented with a mixture containing oak sawdust, bran, sugars and EM fermentation broth.

Fermentation conditions of the mixture is preferably 20 to 30 days at 20 ~ 35 ℃.

As the saccharide, one or more selected from the group consisting of fructose, sugar, honey and oligosaccharides may be used.

The EM fermentation broth is characterized in that the EM stock solution or EM active liquid.

The EM active liquid is 0.1 to 2 parts by weight of sugars mixed with 100 parts by weight of water, 5 to 20 parts by weight of EM stock solution mixed with the sugar is mixed and then incubated in anaerobic conditions for 1 to 5 days at 29 ~ 35 ℃ Can be.

Thus, the present invention, mulberry leaves; And, fermented sawdust fermented a mixture containing oak sawdust, bran, sugars and EM fermentation broth; provides a method of producing a feed composition of the beetle larvae by mixing. The mulberry leaf may be included in a powder state. For this reason, the present invention can provide a feed composition for preventing or improving nudyvirus infection of the beetle larvae prepared by the above method.

The present invention also provides a method for breeding the beetle by feeding the feed composition to the beetle larvae.

Preferably (step 1) egg eggs of the beetle, and obtain the hatching larvae from the eggs;

(Step 2) while breeding the hatching larvae obtained in step 1 (stage 1) while feeding the feed composition of the present invention to the larvae of the 2nd stage of breeding to obtain the species (stage 3) larvae;

(Step 3) solubilizing the seed larva obtained in step 2 to obtain a pupa;

(Step 4) to obtain the adult by allegorizing the pupa obtained in step 3; And,

(Step 5) to induce the fable adult adults obtained in step 4 to mate and spawn; may include.

In another aspect, the present invention relates to a composition for preventing or treating long beetle beetle nudyvirus infection containing mulberry leaves.

Hereinafter, the present invention will be described in more detail.

Among the terms used in the present invention, the first larvae are larvae until about 14 days after hatching, the head diameter is 2 to 3 mm, and the second larvae start at about 15 days and the head diameter is about 4 to 6 mm. The age of 3 larvae starts from about 35 days and the head diameter increases from 3mm to 1.2cm, and this period is maintained for 3-4 months. The pupa period lasts 20 to 25 days.

In the feed composition of the present invention, the mulberry leaf may be included in 1 to 5% by weight.

When the mulberry leaves in less than 1% by weight in the feed composition may not inhibit the infection of nudievirus, if the mulberry leaves in excess of 5% by weight may cause a hindrance to the growth of larva rather reduced survival rate.

The feed composition may be mixed and used in the feed contained for breeding beetle larvae. Preferably, the feed composition may be added fermented sawdust fermented with a mixture containing oak sawdust, bran, sugar and EM fermentation broth. Can be. The mixture may include 88 to 96% by weight of oak sawdust, 1 to 5% by weight of bran, 1 to 5% by weight of sugar and 0.5 to 2% by weight of EM fermentation broth.

The feed composition is preferably supplied from the beginning of the second age to the age of the larva growth process. The feeding of mulberry leaves to larvae during the 1st stage (just after hatching to just before 2nd stage) may lower survival rates.

The term 'yonghua' means that the larvae escape from the pupae, and the term 'allegory' means that the pupae become adults. 'Hatching larvae' means larvae that have just awakened from their eggs.

As the oak sawdust used in the present invention, any of sawdust such as oak, gingko, pruning oak, and oyster oak may be used. The oak sawdust is preferably in the size range from 250 mesh to 350 mesh. If the size of oak sawdust is less than 250 mesh, the larvae feed poorly, and if the size of the oak sawdust exceeds 350 mesh, the particles are small and are not preferable for maintaining water. Oak sawdust may be used to produce dried wood as oak sawdust, but preferably using sawdust (raw sawdust) of raw wood.

Mulberry leaves used in the present invention may be used both undried raw leaves and dried leaves, but preferably dry and powdered.

Wheat bran used in the present invention is leftover after the manufacture of flour, and the inner skin of the wheat is mixed about 12 to 15%. Due to its unique texture, it is also widely used as a food ingredient. It is also used as a raw material for miso and yeast. It is also well digested and used as a feed for livestock. Low calories, but contains a lot of minerals.

EM fermentation broth used in the present invention can be used EM stock solution or EM active solution. EM (Effective Microorganism) is an abbreviation of useful microbial group and is a complex microbial preparation combining a combination of 10 and 80 microorganisms beneficial to humans among many microorganisms existing in nature. Probiotics that make up EM bacteria generally include yeast, lactic acid bacteria, yeast bacteria, photosynthetic bacteria, actinomyces, and mold fungi. It is a group of microorganisms symbiosis with each other, leading to the direction of regenerating nature by producing antioxidant activity or antioxidants and inhibiting decay. Professor Higaderuo of Ryukyu University, Japan, the first developer of EM bacteria, called these fermentation microorganisms "utility microorganisms". The EM (Effective-Microorgarnisms) fermentation broth used in the present invention can be any of commercially available EM stock solutions (starters) and EM active solutions. The EM active liquid is mixed with 0.1 to 2 parts by weight of sugars (one or more selected from fructose, sugar, honey and oligosaccharides) in 100 parts by weight of water, and 5 to 20 parts by weight of EM stock solution in the water in which the sugars are mixed. After mixing, it can be made by incubating in an anaerobic state for 1 to 5 days at a temperature of 29 ~ 35 ℃, more preferably, a mixture of 100 parts by weight of water, 1 part by weight of sugar, 20 parts by weight of EM stock solution at a temperature of 32 ℃ It can be prepared by incubating for 3 days.

The present invention provides a feed composition of fermented sawdust beetle larvae added to the mulberry leaves, fermented sawdust fermented with a mixture containing oak sawdust, bran, sugar and EM fermentation broth, the fermentation conditions of the mixture is 20 ~ at 20 ~ 35 ℃ It is preferable that it is 30 days.

At this time, the mixture may include 88 to 96% by weight of oak sawdust, 1 to 5% by weight of bran, 1 to 5% by weight of sugar and 0.5 to 2% by weight of EM fermentation broth. Even if the oak sawdust is less than 88% by weight of the mixture may be slow growth of the beetle that is supplied with the growth source from the wood material. The growth of beetles can be slow even when the bran is less than 1% by weight or the sugars are less than 1% by weight. When the oak sawdust exceeds 96% by weight, the bran exceeds 5% by weight or the sugars exceeds 5% by weight, a nutritional imbalance occurs due to a decrease in the supply of relatively different components, respectively. Growth may be inhibited. If the EM fermentation broth is less than 0.5% by weight, fermentation may not be performed well or various germs may be generated and the feed composition may be corrupted. It is also undesirable because the fermentation time may be long. If the EM fermentation broth exceeds 2% by weight, the feed composition may be overfermented, the pH may change, and the larva may be deteriorated. May adversely affect growth.

When fermenting the mixture for the feed composition of the present invention, the conditions for fermenting the mixture containing oak sawdust, bran, sugars and EM fermentation broth are preferably maintained for 20 to 30 days at a temperature of 20 ~ 35 ℃. If the temperature is less than 20 ℃ microorganisms in the EM fermentation broth may not be activated fermentation is not performed well, the fermentation may not be well off the conditions suitable for the growth of microorganisms outside 35 ℃. In addition, even if the fermentation period of the mixture is less than 20 days, fermentation may not proceed well and over fermentation for more than 30 days may be difficult to use the feed composition.

The humidity during fermentation is preferably maintained in the range of 35 to 85%, more preferably 40 to 70%, and most preferably 40 to 60%. Under these conditions, the mixture containing non-baked, oak sawdust, bran, sugar and EM fermentation broth is suitable for use as a feed composition suitable for breeding beetle larvae by effectively controlling nudievirus through fermentation of EM fermentation broth. To change.

The present invention provides a method for preparing a feed composition of the beetle larvae, preferably,

Prepare the mulberry leaves,

Preparing a mixture comprising oak sawdust, bran, saccharides and a mixture comprising an EM fermentation broth and then maintaining the mixture at a temperature of 20 to 35 ° C. for 20 to 30 days to produce fermented sawdust; And,

Mixing with mulberry leaves and the fermented sawdust; may be prepared.

In the present invention, the nudie virus causing the infection with the beetle beetle larva is the same as the beetle beetle virus (Allomyrina dichotoma Virus: AdV) published in the Republic of Korea Patent Publication No. 10-2018-0024813 95% to 99% homology with Orytes nunovirus virus isolate Ma07 from Malaysia, complete genome (Genbank number: EU747721.1).

The present invention provides a method for breeding the beetle by feeding the feed composition to the beetle larvae. The breeding conditions of the beetle is not particularly limited except for feeding the feed composition of the present invention, as a specific example, (step 1) egg eggs of the beetle, to obtain the hatching larvae from the eggs;

(Step 2) obtaining the seed larvae by feeding and feeding the feed composition of the present invention to the larvae reached at the end of the second stage while breeding the hatched larvae obtained in step 1;

(Step 3) solubilizing the seed larva obtained in step 2 to obtain a pupa;

(Step 4) to obtain the adult by allegorizing the pupa obtained in step 3; And,

(Step 5) to induce the fable adult adults obtained in step 4 to mate and spawn; may include.

Feed composition of the beetle larvae of the present invention may further include a micronutrient such as nutrients, preservatives, preservatives, antibiotics, vitamins and the like commonly used in insect feed. The nutrients, preservatives, preservatives, antibiotics, micronutrients, and the like can be used arbitrarily selected by the person of ordinary skill in the art as a conventionally added content to the feed.

When the feed composition of the present invention is supplied to the beetle larvae, the size and weight are increased compared to the beetle larvae grown in the natural state, and the growth of the larvae is excellent and the growth is good while the survival rate of the larvae is greatly increased. There is an effect that can supply a high quality beetle larvae or adults significantly increased in size and weight.

In the present invention, the breeding environment of the beetle larvae temperature is preferably 23 ~ 30 ℃, photoperiod is preferably 15 ~ 17L (Light) and 9 ~ 7D (Dark; memorization).

Breeding box breeding beetle larvae in the present invention to maintain cleanliness by cleaning, disinfecting or replacing for a certain period of time so that the larvae do not catch the disease, put a tissue moistened with sterilized water in the container to maintain a constant humidity It is desirable to. At this time, the humidity is preferably maintained at 60 ~ 70% R.H. The breeding box can be used as long as it is used for the breeding of insect larvae or adults, but has a hole in the lid (preferably with a hole of 3 to 5 mm intervals), and a glass or plastic box. It is good to use. The size of the box can also be any size, for example, 270 (height) x 400 (width) x 270 (height) (mm) can be used.

When the beetle larvae are soluble in the pupa, it is desirable to replenish moisture at any time using a water spray so that the pupa does not dry out, and it is desirable to be careful not to touch a human's hand with a pupa. Dissolved chrysalis can be allegorized to obtain an adult. The allegorical adult can be mated and spawn to breed into the next generation.

The feed composition of the present invention can be used for feeding both beetle larvae and adults, but the feeding of the adult may further feed the composition for adult mixed feed containing jelly (jelly) and edible protein. The composition for adult mixed feed may be fed two to three times a week. The adult mixed feed composition may contain 4 to 6 parts by weight of an edible protein based on 100 parts by weight of jelly. The jelly is a food coagulated with gelatinous ingredients such as pectin, gelatin, agar, and alginic acid, and is prepared in a gel state by hardening sugar or sugar such as juice or sugar in gelatin and agar. have. As the edible protein, any of foods such as casein, powdered milk, soybean protein, soybean meal, soybean meal, rice bran, rice protein and the like, or foods of protein used as feed composition can be used.

The present invention provides a composition for the prevention or treatment of long-lived beetle nudyvirus infection, characterized by containing mulberry leaves, can be included in other food sources such as sawdust to treat the infection.

The present invention relates to a feed composition for preventing or improving a beetle beetle Nudivirus infection containing mulberry leaves, by supplying the composition to the beetle beetle larvae and controlling the feeding time, before the pupa period due to the existing Nudivirus infection Most of the dead beetle larvae became healthy until the adulthood.

Figure 1 is a photograph showing a comparison of the size of the beetle larvae survived after feeding the feed composition containing mulberry leaf powder 5% by weight from the second age of virus infection for 9 weeks.

Hereinafter, a preferred embodiment of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, it is provided to fully convey the spirit of the present invention to those skilled in the art so that the contents introduced herein are thoroughly and completely.

Example 1 Preparation of Feed Composition

Fermented sawdust was prepared by mixing oak sawdust 93g, bran 3g, sugar 3g, EM stock solution 1g and fermentation at 25 ℃ for 25 days. The fermented sawdust and mulberry leaf powder were mixed to prepare a feed composition, but the mulberry leaf powder was prepared to be 1% by weight and 5% by weight, respectively.

<Example 2. Feeding, survival status and weight confirmation of mulberry leaf-containing feed composition for the second larvae of beetle beetle>

Two-stage early beetle larvae infected with the virus were selected and prepared, and 30 or 15 larvae were prepared in each experimental group and the control group. For the experiment, larvae were placed in insect breeding buckets one by one and then fed.The experimental group contained 1 wt% or 5 wt% of mulberry leaf powder (1g and 5g of mulberry leaf powder in 100g feed, respectively, and the rest was fermented sawdust. ), And the control group was fed only the fermented sawdust without adding anything else.

Experiments were conducted from 28 days (4 weeks) up to 63 days (9 weeks) and weighed the surviving population or larvae. In addition, during the experiment, 5 ml of tertiary distilled water was sprinkled on the sawdust once a week to give adequate moisture to the sawdust.

-Breeding condition: Temperature 26 ~ 28 ℃, Humidity 50 ~ 60%

-Check the larvae weight and mortality rate: check every 7 days

As a result of the experiment, as shown in Table 1 and Table 2 below, in the control group provided with only fermented sawdust, as time passed, the food was not fed well in the state of viral infection, but the weight loss phenomenon occurred (week 1), Most of the larvae were killed while remaining in poor condition.

 Control start date 1 week 4 Weeks 9 Weeks Live population 30 heads 10 heads 1 head 1 head Average weight of living subjects 0.939 g 0.601 g 2.630 g 6.770 g Mulberry leaf powder 5 wt% treatment group start date 1 week 4 Weeks 9 Weeks Live population 30 heads 30 heads 25 heads 25 heads Average weight of living subjects 0.895 g 2.467 g 4.426 g 9.481 g

Control start date 4 Weeks Live population 15 heads 0 heads Average weight of living subjects 1.024 g (Not measurable) Mulberry leaf powder 1 wt% treatment group start date 4 Weeks Live population 15 heads 13 heads Average weight of living subjects 1.001 g 3.932 g

On the other hand, the experimental group fed the feed composition containing 1 to 5% by weight of mulberry leaf powder and fermented sawdust was confirmed to show a high survival rate, it is confirmed that the body weight is significantly increased, the appearance of the larva also larvae uninfected with the virus He recovered to a state of health similar to that of.

In addition, it can be seen that the weight of the 5% by weight of the mulberry leaf powder treated group was increased more at 4 weeks compared to the 1% by weight treatment group, the weight of the 5% by weight of the mulberry leaf powder treated larvae, once in the virus It was almost similar to other larvae that grew healthy without being infected.

Similar results were obtained by repeating the experiment again. In the case of virus-infected larvae fed fermented sawdust only, they were almost killed at 4 weeks, and even though they lived until 9 weeks, the size of Fig. 1 (left: fermented sawdust treatment) Group, the right: fermented sawdust + 5% by weight mulberry leaf treatment group) because it is significantly different, it can prove that the supply of mulberry leaves is very important in the breeding of larvae of beetle.

On the other hand, in the present invention, whether the beetle larvae viral infection was confirmed through visual verification. In the case of visual inspection, the healthy beetle larvae are black in the intestinal tract due to sawdust and remain solid. When infected with the virus, the larva's feeding activity was slowed down, the body fluid was excessively increased, and the lower abdomen was swollen. This is a clear contrast to fungal or bacterial infections in which blackish larvae develop. This is the reason for the diagnosis of viral diseases.

In addition, when the virus-infected larvae were killed by visual inspection, it was determined by detecting genes for the virus by the method of Korean Patent Application Publication No. 10-2018-0024813 to confirm that it died due to nudievirus infection, not other beetle disease.

Claims (5)

Feeding composition for the prevention or improvement of beetle beetle Nudivirus infection, comprising mulberry leaves. The method of claim 1,
The feed composition is a feed composition for the prevention or improvement of the beetle beetle nudivirus infection, characterized in that the fermented sawdust fermented with a mixture containing oak sawdust, bran, sugars and EM fermentation broth is added. Mulberry leaves; And fermented sawdust fermented with a mixture containing oak sawdust, bran, sugar and EM fermentation broth. (Step 1) egg eggs of the beetle, and hatching larvae from the eggs;
(Step 2) breeding the hatching larvae obtained in the step 1 while feeding and feeding the feed composition of claim 1 to the larvae of the second stage to obtain the seed larvae;
(Step 3) solubilizing the seed larva obtained in step 2 to obtain a pupa;
(Step 4) to obtain the adult by allegorizing the pupa obtained in step 3; And,
(Step 5) to induce the fable adult adults obtained in step 4 to mate and spawn; breeding method of the beetle, characterized in that it comprises a. A composition for the prevention or treatment of enteric beetle Nudivirus infection, comprising mulberry leaves.

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