KR101907194B1 - Feed additive manufactured by using complex microbe spawn and method for manufacturing the same - Google Patents

Abstract

(Bacillus subtilis) , Lactobacillus plantarum (Lactobacillus plantarum ) , Lactobacillus plantarum (Lactobacillus plantarum ) , Lactobacillus plantarum (Lactobacillus plantarum ) , and Lactobacillus plantarum using a culture medium in which lysine, propolis, molasses, plantarum and saccharomyces cerevisiae are cultured, and the feed additive is completed by filtering the liquid material whose pH value of the cultured liquid material is measured within a predetermined range do.

Description

[0001] The present invention relates to a feed additive prepared by using a complex microbial strain and a preparation method thereof,

The present invention relates to a method for preparing a feed additive using a seed microorganism having a multiplicity of microorganisms and a feed additive prepared by the method.

In recent years, there has been a growing interest in environmental pollution, and there has been a problem of livestock waste as one of environmental pollution sources. Among them, livestock wastewater collectively referred to as livestock manure and livestock wastewater discharge facility cleaned groundwater pollution and odor Residents near the house are being damaged. Therefore, studies on the removal of odor have been actively conducted along with the decrease of nitrogen and phosphorus contents in livestock manure.

Antibiotics, which are essential for the growth of livestock, are used for the purpose of promoting the growth of livestock and for the prevention and treatment of diarrhea and various diseases. However, when antibiotics are used frequently, the pathogens develop their own strength to resist antibiotics As a result of strong bacterial resistance against strong antibiotics due to strong tolerance, it has been putting strong sanctions to prevent some antibiotics from being added to feeds worldwide.

Therefore, a substance that can replace antibiotics became necessary, and the most active research now is a microbial prodrug. Probiotics are live lactic acid bacteria fed to livestock in order to improve the productivity of livestock through resistance to harmful microorganisms. It increases immunity by increasing beneficial bacteria in the intestines when fed to livestock. In addition, beneficial microorganisms contained in such a probiotic agent have an effect of reducing odor, inhibiting hydrogen sulfide (H ₂ S) and ammonia (NH ₃ ) generation, and can solve the conventional problems.

On the other hand, in the prior art disclosed in Korean Patent No. 10-0949903, microorganisms cultured through separate culture of Bacillus, lactic acid bacteria and yeast are cultured in a culture medium containing 30-70% by weight of Bacillus subtilis, 20-40% by weight of lactic acid bacteria, Manganese chloride (MnCl _x ), citric acid (C ₆ H ₈ O ₇ ), and potassium sorbate (C ₆ H ₇ O ₂ K) were mixed with 0.5 wt% manganese chloride, citric acid 0.2% by weight and 0.04% by weight of potassium sorbate are mixed together to prepare a feed additive. This conventional technology has a disadvantage in that the manufacturing process is complicated and the manufacturing time is long because the microorganisms are cultured differently under the culture conditions considering the culture time, culture temperature, initial pH, carbon source effect, nitrogen source effect and influence of inorganic salts exist.

As described above, the conventional feed additive manufactured using microorganisms such as the prior art disclosed in the above-mentioned patent is complicated and requires a long production time. However, since the number of beneficial bacteria contained in the feed additive is insufficient, When administered to a ruminant such as a cow, the beneficial bacteria contained in the feed additive were killed before reaching the ruminant. In addition, since the above-mentioned prior art uses chemical substances such as manganese chloride, citric acid and potassium sorbate in the process of preparing the feed additive, it may cause additional problems.

Friendly feed additive which can increase the number of beneficial bacteria in a short time through a simple manufacturing process. The present invention also provides a feed additive prepared by the method.

Method of manufacturing a feed additive by using a complex microbial inoculum according to one aspect of the present invention is Bacillus subtilis (Bacillus subtilis), Lactobacillus plan tareom (Lactobacillus plantarum), and a saccharide in which input to the plurality of the lysine with the composite seed microorganism, propolis, molasses, and water culture vessels containing the MRS celebrity busy (Saccharomyces cerevisiae) The pH value of the liquid material cultured in the culture step and the culture step in which the complex microorganism seed introduced into the culture vessel is cultured using the culture medium formed by mixing the lysine, propolis, molasses, and water injected into the culture vessel, And a filtration step of completing the feed additive by filtering the liquid material whose pH value is measured within a predetermined range in the measurement step and the measurement step.

The step of introducing may include 0.7 to 0.9 parts by weight of a complex microbial strain, 0.1 to 0.3 parts by weight of lysine, 0.1 to 0.3 parts by weight of propolis and 4.5 to 6.5 parts by weight of molasses in relation to 100 parts by weight of water, Wherein the culture medium is prepared by mixing 0.1 to 0.3 parts by weight of lysine, 0.1 to 0.3 parts by weight of propolis and 4.5 to 6.5 parts by weight of molasses per 100 parts by weight of the combined microorganism strain Can be cultured.

The culture step is carried out by aeration while simultaneously intermittently stirring 0.7 to 0.9 part by weight of the complex microbial species, 0.1 to 0.3 part by weight of lysine, 0.1 to 0.3 part by weight of propolis and 4.5 to 6.5 parts by weight of molasses, The complex microorganism seeds can be cultured.

The culturing step may be performed by repeating the stirring and the aeration for a second time longer than the first time after the stirring and the aeration for the first time are repeated to perform the intermittent stirring and aeration. In the culturing step, the intermittent stirring and aerating may be performed while the temperature of the culture liquid is maintained within a range of 35 to 45 ° C.

The composite seed microorganism is Lactobacillus casei (Lactobacillus casei), Lactobacillus Ecija FIG pillar's (Lactobacillus acidophilus ) , Lactobacillus flow Stock Kono (Lactobacillus leuconostoc ) , Lactobacillus Brevis (Lactobacillus brevis), Streptococcus faecalis (Streptococcus faecalis), Bacillus Pew tree blood carcass (Bacillus putrificus), Bacillus cereus (Bacillus cereus), Pseudomonas fluoro lesson's (Pseudomonas fluorescens), and Oh Spanish way Bruce duck claim (Aspergillus oryzae) As shown in FIG.

The filtration step may complete the feed additive by filtering the liquid material measured in the pH range of 3 to 4.5 in the measurement step. The liquid phase material corresponding to the feed additive may be discharged from the filter by performing filtration at least once using a filter having a pore size of 200 mesh in the measurement step, have.

A feed additive according to another aspect of the present invention is prepared by a method for preparing a feed additive using the complex microorganism.

Lysine, propolis, molasses, and water were mixed to prepare a bacillus Subtilis (Bacillus subtilis), Lactobacillus tareom Plan (Lactobacillus plantarum), and to Mrs Saka A feed additive having a greatly increased number of microorganisms can be prepared by culturing a seed microorganism containing a plurality of microorganisms including Saccharomyces cerevisiae .

A mixed microorganism having 0.7 to 0.9 part by weight based on 100 parts by weight of water is mixed with 100 parts by weight of water using 0.1 to 0.3 parts by weight of lysine, 0.1 to 0.3 parts by weight of propolis and 4.5 to 6.5 parts by weight of molasses. By culturing the seed culture, a feed additive having a greatly increased number of microorganisms can be produced in a short time.

The feed additive according to the present invention can be produced by a simple process consisting of the purification step 10, the addition step 20, the cultivation step 30, the measurement step 40, and the filtration step 50 and the like. In addition, any process in the feed additive manufacturing process does not require any chemicals, so it is possible to provide eco-friendly feed additives.

1 is a flow chart of a method of manufacturing a feed additive according to an embodiment of the present invention.
FIG. 2 is a graph comparing the daily gain in the whole breeding period of Example 1 prepared according to the manufacturing method shown in FIG. 1 with another example.
FIG. 3 is a graph comparing feed efficiency with other examples during the entire breeding period of Example 1 produced according to the production method shown in FIG. 1; FIG.

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

The present invention relates to a method for producing Bacillus subtilis, Lactobacillus plantarum, and Sakae using a culture medium containing lysine, propolis, molasses, and water. The present invention relates to a feed additive prepared by culturing a seed microorganism having a multiplicity of microorganisms, including Saccharomyces cerevisiae, and filtering the cultured liquid material.

The feed additive according to the present invention is prepared by mixing 0.7 to 0.9 part by weight of a complex microorganism strain, 0.1 to 0.3 part by weight of lysine, 0.1 to 0.3 part by weight of propolis, and 4.5 to 6.5 parts by weight of molasses, relative to 100 parts by weight of water, Is incubated with intermittent agitation and aeration while maintaining the temperature in the range of 35 to 45 ° C.

Bacillus used in the manufacture of feed additives Subtilis (Bacillus Subtilis), Lactobacillus tareom Plan (Lactobacillus plantarum), the MRS celebrity busy as Saccharomyces (Saccharomyces cerevisiae) a description of a number of microorganisms, lysine, propolis, molasses, water and the nature and purpose of the microorganism is a composite containing silver will be described later.

Bacillus subtilis (Bacillus subtilis) is a type of bacteria belonging to the genus Bacillus, widely distributed in the natural world such as soil, hay, and dust, and is called aerobic microorganism . Bacillus subtilis produces proteases, increases the number of beneficial bacteria in the intestines, and reduces odor and insect pests. It is also excellent in feed efficiency and growth effect, so that healthy growth of livestock can be achieved.

Lactobacillus plantarum is lactic acid bacteria (bacteria that produce lactic acid by decomposing saccharides such as glucose, lactic acid bacteria), and corresponds to fluid anaerobic. Lactobacillus plantarum is a byproduct that produces organic acids, prevents bacterial enteritis, inhibits the growth of harmful bacteria in the body, inhibits the growth of other bacteria from outside through harmful bacteria and foods in the intestines, Keeping it clean and preventing diarrhea and constipation increases feed efficiency and digestibility. It also suppresses the generation of ammonia and hydrogen sulfide present in the manure of livestock, thereby eliminating odors.

Saccharomyces cerevisiae (Saccharomyces cerevisiae) is a typical yeast belonging to the carrot fungus (bacteria that form adenocarcinoma by the sexual reproduction in the fungus and produces the ascospores), and most of the yeast used for brewing Can be said to be close to this species. Sakaromisses Saccharomyces cerevisiae supplies useful physiologically active substances such as amino acids, vitamins, minerals, and digestive enzymes, and improves livestock productivity by promoting the metabolism of yeast itself. In addition, it accelerates fermentation, promotes decomposition of organic matter using microorganisms, and promotes palatability. The above-mentioned palatability refers to a characteristic that an animal can perceive and ingest a feed as a result of factors such as appearance, taste, odor, texture, and temperature of the feed.

Composite microbial inoculum of the present invention can be obtained from bean water, molasses, rice, etc. and the above-described Bacillus Subtilis (Bacillus Subtilis), Lactobacillus tareom Plan (Lactobacillus plantarum), Lactobacillus casei (Lactobacillus casei) in addition to the MRS celebrity busy as Saccharomyces (Saccharomyces cerevisiae). Lactobacillus < RTI ID = 0.0 & gt; acidophilus), Lactobacillus flow Stock Kono (Lactobacillus leuconostoc ), Lactobacillus Brevis (Lactobacillus such as Streptococcus faecalis , Bacillus putrificus, Bacillus cereus, Pseudomonas fluorescens, Aspergillus oryzae, and the like, It contains various kinds of seeds. Generally, each microorganism strain is mixed with 1 to 20% by weight of each microorganism strain, and the number of each microorganism strain is about 10 ² to 10 ³ cfu / g.

The combined microorganism strain of the present invention is 0.7 to 0.9 parts by weight based on 100 parts by weight of water. When the amount is less than 0.7 part by weight, microorganisms necessary for cultivation are not sufficiently supplied. Therefore, the growth of the microorganism population through the cultivation step is not effectively performed, The proportion of microorganisms is higher than the ratio of lysine, propolis, molasses, and water used as a food for microorganisms, so that it is difficult for microorganisms to propagate through sufficient feeding activity And excessive mixing of the medium contained in the complex microbial strain may cause contamination of the culture liquid depending on the by-products.

Lysine is a basic amino acid that acts as a growth promoter that promotes growth and development and is also called lysine. In the present invention, lysine is produced by Bacillus subtilis Subtilis ), Lactobacillus Plan tareom (Lactobacillus plantarum ), Saccharomyces Serenity busy (Saccharomyces cerevisiae , etc., which is used as a culture food of complex microbial species, thereby promoting the growth of the mixed microbial species. Some of the remaining lysine, which is used as a feed for the culture of complex microorganisms, remains as a feed additive and acts as a growth promoter for livestock. Lysine is a kind of amino acid that promotes the growth and development of livestock. It is involved in growth hormone and promotes the growth of livestock. In addition to helping microbes breed and promoting the growth of livestock, lysine also helps to reduce the nitrogen content of livestock feces and prevent environmental pollution caused by ammonia.

When the amount of lysine used in the present invention is 0.1 to 0.3 part by weight based on 100 parts by weight of water, the amount of the lysine used in the present invention is less than 0.1 part by weight, the microbial proliferation efficiency of the microorganism is not sufficient, The feed additive due to the culture to be obtained in the present invention may not be produced due to difficulties and if the amount of lysine is more than 0.3 part by weight, the microorganisms in the culture liquid may lack in the habitat and breeding space, , Livestock that consume excessive amounts of lysine, which acts as a growth promoter for livestock, may cause non-ideal growth.

Propolis is a substance made by mixing bees with their own needles and enzymes in the same material as resin (resin) extracted from various plants for their survival and propagation. Propolis is composed of organic matter and minerals Many minerals, vitamins, amino acids, fats, organic acids and flavonoids play an important role in cell metabolism and terpenes have anticancer activities. In the present invention, it is used in the step of culturing a microorganism of the composite food microorganisms Bacillus Subtilis (Bacillus Subtilis), Lactobacillus tareom Plan (Lactobacillus plantarum), helps to increase the yuikgyun populations, such as the MRS celebrity busy as Saccharomyces (Saccharomyces cerevisiae). In addition, some of the propolis remains intact and fed to livestock to enhance anti-inflammatory, antioxidant, and immunity.

When the propolis used in the present invention is 0.1 to 0.3 parts by weight per 100 parts by weight of water, the propolis is not sufficiently supplied to the food of the complex microorganism strain during the culturing step, so that the propagation efficiency of the microorganisms may be lowered, The feed additive due to the culture to be obtained in the present invention may not be produced. If the amount of propolis is more than 0.3 part by weight, the culture and microbial growth space in the culture liquid may be insufficient and the proliferation efficiency of the microorganisms may be decreased. , Excessive use of the remaining propolis, which is used as food for microorganisms, may cause side effects such as fever and heart rate increase due to excessive antibiotic ingredient addition, and problems with livestock growth.

Molasses is a black syrup-like liquid that is extracted from sugars during the process of sugar production through sugarcane, sugar beet, etc., through the processes of dissolution, purification and crystallization. It contains 20 ~ 30% moisture The main ingredient is sugar. The molasses is divided into sugar cane molasses, sugar beet molasses, cornstarch molasses, citrus molasses, molasses, and molasses, depending on the raw material, and about 5 to 10% is used for livestock feed. Molasses is a substance having various nutrients such as dietary fiber, folic acid, protein, vitamin, and potassium. In the present invention, the molasses is used as a culture food for a complex microorganism strain.

The molasses is a liquid viscous material having a viscosity of 500 Pa · s and a viscosity higher than 1 mPa · S (20 ° C) of water. Therefore, in the present invention, molasses is melted separately at 40 ° C and put into a culture vessel. In addition, although glucose and brown sugar can be used as the fermented food of the complex microorganism strain instead of molasses, the cost saving effect is obtained by using molasses which is relatively easy to obtain and low in cost.

If the molasses used in the present invention is 4.5 to 6.5 parts by weight based on 100 parts by weight of water and less than 4.5 parts by weight, the multiplication efficiency of the microorganisms may be lowered due to insufficient feeding into the culture food of the complex microorganism strain during the culturing step, If the weight of the molasses is excessively high, the microbial growth and the propagation space of the microorganism in the culture liquid may be insufficient, and the efficiency of proliferation of the microorganisms may decrease. If the molasses having high viscosity is used excessively, the viscosity of the cultured liquid material becomes excessively high, have.

In the present invention, all components other than the complex microbial strain, lysine, propolis, and molasses are all composed of water, and water is used as a solvent for the raw materials. Water has a specific heat that is large enough to control the body temperature of a thermophilic animal, so it is not sensitive to changes in temperature, such as temperature, and does not cause a chemical reaction with other substances, so it is suitable for use as a solvent.

The feed additive according to the present invention is prepared by mixing 0.7 to 0.9 parts by weight of a complex microbial strain, 0.1 to 0.3 parts by weight of lysine, 0.1 to 0.3 parts by weight of propolis, and 4.5 to 6.5 parts by weight of molasses, based on 100 parts by weight of water, The mixing ratio of one ingredient is an optimal condition for culturing the multiple microorganism strain, and a preferred embodiment will be described below.

Hereinafter, a method for preparing a feed additive using a complex microbial strain according to an embodiment of the present invention will be described in detail with reference to the drawings.

1 is a flow chart of a method of manufacturing a feed additive according to an embodiment of the present invention. Referring to FIG. 1, the method for preparing a feed additive according to the present embodiment comprises a purification step 10, an input step 20, a cultivation step 30, a measurement step 40, and a filtration step 50 .

In the water purification step (10), tap water is purified to remove by-products such as chlorine from tap water. More specifically, in the water purification step 10, tap water can be poured into a heating container provided separately from the culture container and purified by heating for 3 to 24 hours in a temperature range of 60 to 80 ° C. Tap water contains chlorine (Cl), fluorine (F), calcium (Ca), magnesium (Mg), iron (Fe), other byproducts and mixed germs other than certain bacilli. In the water purification step (10), tap water is purified by water suitable for the growth of microorganisms by removing such chlorine, bacteria, and other by-products from tap water.

In the injecting step 20 , a seed microorganism having a multiplicity of microorganisms including Bacillus subtilis, Lactobacillus plantarum and Saccharomyces cerevisiae is mixed with lysine, propolis, , Molasses, and water are put into a culture container. According to the present embodiment, 0.7 to 0.9 parts by weight of complex microbial species, 0.1 to 0.3 parts by weight of lysine, 0.1 to 0.3 parts by weight of propolis and 4.5 to 6.5 parts by weight of molasses are added to 100 parts by weight of water . This input step 20 follows the following series of steps. First, water purified in the water purification step (10) is cooled and water corresponding to about 80% of the total amount of water to be added to the culture vessel is introduced into the culture vessel when the temperature of the water reaches about 40 ° C which is the optimum culture temperature of the microorganism .

Molasses dissolved at about 40 占 폚 is then introduced into a culture vessel, and then the complex microorganism strain, lysine, and propolis are put into a culture vessel. The reason why the molasses is first added to the mixed microorganism seedlings, lysine, and propolis is that the molasses is highly viscous, and when it is added later than the complex microbial strain, lysine, and propolis, This is because mixing can be disturbed. Lastly, about 20% of the total amount of water to be fed into the culture container is put into the culture container until the space corresponding to about 85% of the culture container is filled, and then the lid of the culture container is closed. To ensure a smooth supply of oxygen to the culture medium containing lysine, propolis, molasses, and water, approximately 15% of the culture space should be empty. Water is injected into the culture vessel in two portions as described above in order to precisely empty space corresponding to about 15% of the culture container.

In the incubation step 30, Bacillus subtilis , Lactobacillus plantarum , and Saccharomyces cerevisiae , which are introduced into a culture container using a culture medium containing lysine, propolis, molasses, and water, Saccharomyces cerevisiae is cultivated in the seed microbes. In the prior art, microorganisms are cultured mainly by feeding molasses. As shown in Test Example 2 to be described later, when the lysine, propolis and molasses are added together, the number of microorganisms is greatly increased compared with the case where only the molasses is injected. According to this example, only by incubating the complex microorganism strain as described above for about 72 hours, the population of the microorganism was sharply increased as shown in Test Example 1 to be described later.

In the incubation step 30, 0.7 to 0.9 part by weight of complex microorganism seeds, 0.1 to 0.3 part by weight of lysine, 0.1 to 0.3 part by weight of propolis, and 4.5 to 4.5 parts by weight of molasses are added to 100 parts by weight of water, And 6.5 parts by weight of water are simultaneously stirred while being aerated to cultivate the complex microorganism strain. As shown in Test Example 1 to be described later, when the complex microorganism strain is cultured in the composition ratio as described above, the microbial population is explosively increased in a short time. In particular, for explosive growth of these microbial populations, agitation and aeration must be carried out at appropriate intervals. Agitation of the culture liquid may be carried out by means of a device for wetting the culture liquid, and the aeration of the culture liquid may be carried out by an air pump or the like provided inside the culture vessel.

If the agitation time of the culture medium is too long, the microorganisms can not take a sufficient rest and the increase in the number of microorganisms is slow. If the agitation time of the culture medium is too short, the food is concentrated in the culture medium and the increase in the number of microorganisms becomes slow. If the aeration time of the culture medium is excessively long, anaerobic microorganisms that do not require oxygen are killed and the growth of the microbial population is slow. If the aeration time of the culture medium is too short, the aerobic microorganisms requiring oxygen are killed and the increase of microbial population is delayed.

In the incubation step 30 according to the present embodiment, in order to explosively increase the number of microorganisms for the reasons described above, the culturing step is a step of stirring and aeration for about 2 hours and then stopping stirring and aeration for about 4 hours Intermittent stirring and aeration are carried out by repeating. The reason why the time to stop stirring and aeration is twice longer than the time to stir and aeration is because the microbes need more time to rest after metabolic activity than the time they eat food, .

In the measuring step 40, the pH of the liquid material cultured in the culturing step 30 is measured using a pH meter. The pH of the liquid material may be measured while the pH meter is immersed in the liquid material cultured in the culturing step 30. Alternatively, a small amount of the liquid material cultured in the culturing step 30 may be collected and measured by a pH meter. When the measured pH value is between 3 and 4.5, the cultured liquid material is filtered and used as a feed additive. The cultured liquid material is characterized by having a herbaceous and sour taste. On the other hand, if the measured pH value of the liquid material cultured in the cultivation step 30 is not between 3 and 4.5, the cultured liquid material is discarded.

The filtration step (50) completes the feed additive by filtering the liquid material measured in the predetermined range of the pH value in the measuring step (40). Here, the predetermined range is preferably 3 to 4.5. For example, the filtration step 50 may filter the liquid material measured at a pH value of 3-4.5 in the measuring step 40 with a filter having a porosity of 200 mesh size. A filter with a mesh size of 200 mesh can be made by drilling several holes of 200 mesh size on the bottom of a container made of acrylic material. When a liquid substance whose pH value is measured within 3 to 4.5 is put into such a container type filter, the liquid substance discharged from the filter becomes a feed additive.

In the measurement step 40, the liquid substance measured at a pH value of 3 to 4.5 includes Bacillus subtilis , Lactobacillus plantarum , Saccharomyces cerevisiae, Lysine and propolis, as well as molasses and residual by-products remaining in the seeds. The molasses in the liquid material has an easily rotting property, so that when a large amount of molasses is contained in the liquid material, the liquid material is deteriorated during long-term storage of the liquid material. Likewise, residual by-products in the liquid material alter the liquid material during the long-term storage of the liquid material.

Therefore, the molasses and remaining by-products in the liquid material whose pH value is measured within 3 to 4.5 should be removed as they alter the feed additive. The aforementioned complex microbial species, lysine, and propolis have the property of passing through a hole of 200 mesh size, while molasses and the remaining byproduct do not pass through a hole of 200 mesh size. Thus, molasses and remaining by-products in the liquid material can be removed by filtering the liquid material measured at a pH value of 3 to 4.5 by the above-described method. Thus, since the feed additive having been subjected to the filtration step 50 does not contain molasses and residual by-products, alteration of the feed additive can be prevented, and the state of the feed additive can be preserved for a long period of time. For example, when the feed additive was stored for 6 months, not only did not rot, but also the number of beneficial bacteria was almost unchanged.

On the other hand, the filtration process as described above can be repeated several times in order to reliably remove unnecessary components and by-products from the liquid substance whose pH value is measured within 3 to 4.5. For example, four filters with pores of 200 mesh size can be prepared and the liquid material measured at pH values between 3 and 4.5 can be passed through the four filters in turn. That is, the liquid material filtered by any one of the filters is passed to the next filter to be filtered. The liquid material passing through the fourth filter is equivalent to a feed additive and is put in a container and released to the market.

The feed additive according to this embodiment is a simple process comprising the purification step 10, the feeding step 20, the cultivation step 30, the measuring step 40, and the filtration step 50 according to the above- . ≪ / RTI > In addition, any process in the feed additive manufacturing process does not require any chemicals, so it is possible to provide eco-friendly feed additives. On the other hand, the use of feed additive can be divided into two. When added to the feed, 0.5% of the feed amount can be fed to the livestock by spraying on the feed, and when it is used in the negative water, 0.5% of the minced feed is added to the negative vessel, To the livestock.

Hereinafter, feed additives using the complex microorganism strain of the present invention will be described in detail through examples, comparative examples, and test examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

≪ Example 1 >

According to the above-described embodiment of the present invention, feed additives were prepared as follows. The contents of the embodiments described above are followed even if omitted. 1000 Kg of tap water was put in a heating container, and the mixture was heated to 80 캜 and maintained for 12 hours. Subsequently, while the water was slowly cooled, when the water temperature reached 40 DEG C, 800 Kg of water was added to the culture container. Then, 50 kg of molasses, 7.5 Kg of complex seed, 1.5 Kg of lysine and 1.5 Kg of propolis were added to the culture container. Subsequently, 200 kg of water remaining in the culture vessel was added. Subsequently, the culture medium in the culture vessel was stirred and aerated for 2 hours, followed by stirring and aeration for 4 hours. The culture was repeated for a total of 72 hours, and the temperature of the culture medium was maintained at 40 占 폚. Subsequently, the liquid material having a pH value of 4 was filtered four times using a filter having a pore size of 200 mesh, and the liquid material having passed through the fourth filter was placed in a storage container to complete the preparation of the feed additive.

≪ Comparative Example 1 &

The feed additive according to Comparative Example 1 was prepared according to the above-described Example 1 by changing the conditions as follows. 1000 Kg of tap water was put in a heating container, and the mixture was heated to 80 캜 and maintained for 12 hours. Subsequently, while the water was slowly cooled, when the water temperature reached 40 DEG C, 800 Kg of water was added to the culture container. Then, 50 kg of molasses and 7.5 kg of complex seed were added to the culture container. Subsequently, 200 kg of water remaining in the culture vessel was added. Subsequently, the culture medium in the culture vessel was stirred and aerated for 2 hours, followed by stirring and aeration for 4 hours. The culture was repeated for a total of 72 hours, and the temperature of the culture medium was maintained at 40 占 폚. Subsequently, the liquid material having a pH value of 4 was filtered four times using a filter having a pore size of 200 mesh, and the liquid material having passed through the fourth filter was placed in a storage container to complete the preparation of the feed additive.

≪ Comparative Example 2 &

The feed additive according to Comparative Example 1 was prepared according to the above-described Example 1 by changing the conditions as follows. 1000 Kg of tap water was put in a heating container, and the mixture was heated to 80 캜 and maintained for 12 hours. Subsequently, while the water was slowly cooled, when the water temperature reached 40 DEG C, 800 Kg of water was added to the culture container. Then, 50 kg of molasses, 7.5 kg of complex seed, 3.5 kg of lysine and 4 kg of propolis were added to the culture container. Subsequently, 200 kg of water remaining in the culture vessel was added. Subsequently, the culture medium in the culture vessel was stirred and aerated for 2 hours, followed by stirring and aeration for 4 hours. The culture was repeated for a total of 72 hours, and the temperature of the culture medium was maintained at 40 占 폚. Subsequently, the liquid material having a pH value of 4 was filtered four times using a filter having a pore size of 200 mesh, and the liquid material having passed through the fourth filter was placed in a storage container to complete the preparation of the feed additive.

≪ Test Example 1 >

The results shown in Table 1 below were tested by Chungnam National University Agricultural Science and Technology Center located in Daejeon City, Chungcheongnam-do, Korea. The result is to confirm the superiority of the feed additive through the microorganism test according to the embodiment of the present invention.

Inspection items unit test results Bacillus subtilis cfu / g 3.0 × 10 ⁹ Lactobacillus plantarum cfu / g 2.0 × 10 ¹⁰ Saccharomyces cerevisiae cfu / g 7.0 × 10 ⁹

Table 1 Bacillus subtilis of a feed additive prepared according to Example 1 subtilis (Bacillus subtilis ), Lactobacillus Plan tareom (Lactobacillus plantarum ) and This is a test report on the number of microbial populations of Saccharomyces cerevisiae .

When Bacillus subtilis, Lactobacillus plantarum, and Lactobacillus plantarum were cultured in the presence of lysine, propolis, and molasses at the same time as the feed of the complex microorganism strain according to Example 1 of the present invention, At least 3.0 x 10 ⁹ microorganisms were detected in Saccharomyces cerevisiae .

≪ Test Example 2 &

Inspection items unit test results Bacillus subtilis cfu / g 5.0 × 10 ⁶ Lactobacillus plantarum cfu / g 4.0 × 10 ⁶ Saccharomyces cerevisiae cfu / g 1.5 x 10 ⁷

Table 2 Comparative Example Bacillus of the feed additive made in accordance with the first subtilis (Bacillus subtilis ), Lactobacillus Plan tareom (Lactobacillus plantarum ) and This is a test report on the number of microbial populations of Saccharomyces cerevisiae .

In Comparative Example 1, molasses was cultured as a microbial feed, and the complex microbial strain was cultured as in the prior art. Comparison of Table 1 and Table 2 showed that the number of microorganisms was greatly increased when lysine, propolis and molasses were added together, compared to the case where only molasses was added. Therefore, it can be seen that the addition of lysine, propolis, and molasses at the same time to the culture feed of the microorganism is effective for increasing the number of microorganisms.

≪ Test Example 3 >

Inspection items unit test results Bacillus subtilis cfu / g 4.0 × 10 ⁷ Lactobacillus plantarum cfu / g 6.0 × 10 ⁸ Saccharomyces cerevisiae cfu / g 4.0 x 10 ⁸

Table 3 shows test results for Bacillus subtilis, Lactobacillus plantarum and Saccharomyces cerevisiae microbial populations of feed additives prepared according to Comparative Example 2.

In the case of Comparative Example 2, the complex microorganism strain was cultured by feeding lysine, propolis and molasses at the same time as the food of the complex microorganism strain. However, the amount of lysine and propolis was excessively added Lt; / RTI > As a result, it can be seen that the numbers of microorganisms of Bacillus subtilis, Lactobacillus plantarum and Saccharomyces cerevisiae were decreased in comparison with those of Table 1.

Therefore, the feed additive is prepared in a mixing ratio of 0.7 to 0.9 part by weight of the complex microbial species, 0.1 to 0.3 part by weight of lysine, 0.1 to 0.3 part by weight of propolis and 4.5 to 6.5 parts by weight of molasses, relative to 100 parts by weight of water according to the present invention The feed additives can be prepared in which the number of microorganisms is greatly increased within the same time.

<Test Example 4>

FIG. 2 is a graph comparing the daily gain (Kg) of the case of Example 1, commercially available probiotics and general diets fed to livestock, wherein the amount of gain refers to the body weight increased over a certain period of time. 2, a feed additive prepared according to Example 1 of the present invention and having a large number of microbial populations of Bacillus subtilis, Lactobacillus plantarum and Saccharomyces cerevisiae And it is more effective in increasing the amount of livestock gain than that of ordinary feed and in the market.

&Lt; Test Example 5 >

FIG. 3 is a graph comparing the feed efficiency (%) of the above-described Example 1 with the feed efficiency of commercially available probiotics and general feeds, wherein the feed efficiency is the weight gain And feed intake. 3, when the same amount of feed was consumed, Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae were produced according to Example 1, Feed additives with high numbers of individuals show excellent feed efficiency compared with those of ordinary probiotics.

When all the results of the examples, comparative examples and test examples of the present invention are taken into consideration, when lysine, propolis, and molasses are simultaneously used as the culture food of the complex microorganism strain, it is effective to increase the population of the complex microorganism strain. It is estimated that the effect of increasing the population is higher when the proposed mixing ratio is followed.

The present invention has been described with reference to preferred embodiments, comparative examples and test examples. Although the present invention has been described in detail by way of examples, comparative examples and test examples, it is not intended to limit the present invention but merely to illustrate the present invention. Therefore, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these embodiments. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

10: Integer steps
20: input step
30: Culture step
40: Measuring step
50: filtration step

Claims (9)

A method for preparing a feed additive using a combined microorganism strain,
Propolis, molasses, and water (together with seeds of a complex of microorganisms, including Bacillus subtilis , Lactobacillus plantarum , and Saccharomyces cerevisiae) Into a culture vessel;
Culturing the complex microorganism seed introduced into the culture vessel using a culture solution formed by mixing lysine, propolis, molasses and water introduced into the culture vessel;
A measuring step of measuring a pH value of the liquid material cultured in the culturing step; And
And a filtration step of completing the feed additive by filtering the liquid material whose pH value is measured within a predetermined range in the measurement step,
Wherein the step of introducing is carried out by adding 0.7 to 0.9 parts by weight of a complex microbial strain, 0.1 to 0.3 parts by weight of lysine, 0.1 to 0.3 parts by weight of propolis and 4.5 to 6.5 parts by weight of molasses to 100 parts by weight of water,
The culturing step is carried out by using a culture medium in which 0.1 to 0.3 parts by weight of lysine, 0.1 to 0.3 parts by weight of propolis and 4.5 to 6.5 parts by weight of molasses are mixed with 100 parts by weight of water, Wherein the method comprises culturing a complex microorganism strain having a part of a microorganism. delete The method according to claim 1,
The culture step is carried out by aeration while simultaneously intermittently stirring 0.7 to 0.9 part by weight of the complex microbial species, 0.1 to 0.3 part by weight of lysine, 0.1 to 0.3 part by weight of propolis and 4.5 to 6.5 parts by weight of molasses, And culturing the complex microbial strain. The method of claim 3,
Wherein the culturing step is performed by repeating the steps of stirring and aeration for a second time longer than the first time after performing the stirring and the aeration for a first time period so as to perform the intermittent stirring and aeration A method for producing a feed additive. The method of claim 3,
Wherein the culturing step is carried out with intermittent stirring and aeration while maintaining the temperature of the culture liquid within a range of 35 to 45 占 폚. The method according to claim 1,
The composite seed microorganism is Lactobacillus casei (Lactobacillus casei), Lactobacillus Ecija FIG filler's (Lactobacillus acidophilus ) , Lactobacillus flow Stock Kono (Lactobacillus leuconostoc ) , Lactobacillus Brevis (Lactobacillus brevis), Streptococcus faecalis (Streptococcus faecalis), Bacillus Pew tree blood carcass (Bacillus putrificus), Bacillus cereus (Bacillus cereus), Pseudomonas fluoro lesson's (Pseudomonas fluorescens), and Oh Spanish way Bruce duck claim (Aspergillus oryzae) &Lt; RTI ID = 0.0 &gt; 1, &lt; / RTI &gt; The method according to claim 1,
Wherein the filtration step comprises filtering the liquid material measured in the pH range of 3 to 4.5 in the measuring step to complete the feed additive. The method according to claim 1,
Wherein the filtration is performed by filtering the liquid material having a pH value measured within a predetermined range in the measuring step at least once using a filter having a pore size of 200 mesh so that the liquid material corresponding to the feed additive is discharged from the filter &Lt; / RTI &gt; A feed additive prepared by the method of claim 1.

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