KR101427187B1 - Composition for Eco-friendly Microorganism Formulation Comprising Microorganism Probiotics as Effective Ingredient and Production Method Thereof - Google Patents

Abstract

The present invention relates to an environmentally friendly microorganism preparation containing a microbial prodrug as an active ingredient and a method for producing the microbial prodrug. More particularly, the present invention relates to a lyophilized product of Bacillus bacteria and Streptococcus bacterium, Aspergillus bacteria A solid culture, and a liquid culture product of a yeast bacterium; and an eco-friendly microbial preparation for adding an animal feed containing the microbial live bacteria and the culture as an active ingredient. The environmentally friendly microorganism preparation of the present invention promotes the fermentation of livestock manure, eliminates the odor in the inside and outside of the house and the manure processing plant, inhibits flies and mosquitoes in the house, inhibits the activity of the enteropathogenic bacteria in the intestines and inhibits the activity of pathogenic bacteria And it has an effect of improving the digestion and absorption rate of the compound feed, so that it can be utilized as a multifunctional feed additive.

Description

TECHNICAL FIELD [0001] The present invention relates to an eco-friendly microorganism preparation containing a microbial prophylactic agent as an effective ingredient and a method for producing the same. More particularly, the present invention relates to a microbial prophylactic agent,

The present invention relates to an eco-friendly microbial agent containing a microbial probiotics as an active ingredient and a method of preparing the same, and more particularly, to an eco-friendly microbial agent containing a microbial prodrug, .

Generally, antibiotics are frequently added to feed additives to promote the growth of livestock and to prevent and treat diseases such as diarrhea. However, long-term use or abuse of antibiotics causes antibiotic resistance to livestock and also causes problems with antibiotic residues in livestock products.

Recently, consumers are increasingly pressing government to restrict the use of antibiotics in feeds or to ban them completely. In fact, the European Union has banned the use of antibiotics for four major stages of feed since July 1, 1999, and other countries are likely to be subject to a similar ban, with producers and feed manufacturers establishing some alternatives And whether it should be done. In recent years, in some countries such as Denmark and Sweden, regulations on the use of antibiotics have been intensified in livestock, such as prohibiting the use of antibiotics in raising finishing pigs. Thus, the development of feed additives to replace antibiotics, , Oligosaccharides, etc. have been actively studied and developed.

Probiotics can be defined as living microbial additives that have beneficial effects on the host by multiplying beneficial bacteria in the intestinal tract during feeding to the livestock, inhibiting the growth of harmful microorganisms, and keeping the intestinal microflora in a balanced manner.

In general, commercially been widely used as a Lactobacillus bacteria (Lactobacilli) [Republic of Korea Laid-Open Patent Publication No. 98-78353 Arc, Streptococcus (Streptococci) and Bacillus (Bacillus) [Republic of Korea Laid-Open Patent Publication No. 98-25282 Arc in the microorganism as a feed additive Has been developed and used in livestock. It has been reported that the use of probiotics for pigs improves the growth rate and feed efficiency when the probiotics are fed to pigs.

As a probiotic agent, lactic acid bacteria have various characteristics to be provided. One of the important ones is the ability to survive at a low stomach pH, that is, acid resistance. In case of normal livestock, the pH tends to be slightly increased due to the buffering effect of the ingested food. However, in case of pigs, in order to reach the small intestine of the probiotic strains, the bile bain should be strengthened against bile secreted from the pancreas to the duodenum.

Although many studies have been conducted to investigate the mechanism of action of probiotics, the mechanism of action of probiotics has not been clarified due to lack of scientific research reports on the use of probiotics. However, the mechanism of action of probiotics reported so far is summarized as follows.

In other words, lactate production reduces intestinal pH and decreases pH to change intestinal microflora and reduce E. coli. On the other hand, they cause the production of antibiotics or inhibit the production of toxic substances such as amines, ammonia and phenols. It also inhibits implantation and colony formation in the digestive tract, promotes the production of digestive enzymes and vitamins, and activates the immune response to maximize the effect of probiotics.

As described above, the probiotic agent can maintain the normal germ cell when the livestock is fed by various complex functional groups and improve the productivity of the livestock. However, in order to clarify the mechanism of action, molecular biology knowledge of bacteria in the intestinal tract is required more.

Generally, the probiotics are rich in the one or more strains in the intestines of livestock or dairy lactic acid bacteria (Lactobacilli), in that Lactobacillus acidophilus is the most commonly used, and other species such as Lactobacilli and Enterococcus faecium are used.

Also, in Bifidobacteria Bifidobacterium bifidum , B. pseudolongum and B. thermophilum have been added to the feed of livestock as a probiotic. Bacillus , Clostridium, and various yeasts have also been used as spores.

However, the linkage of the strains in these probiotic strains to the intestinal mucosa, the beneficial physiological effects of the strains and the antagonistic action of antagonistic antibiotics have yet to be clarified, and thus the selection requirements of the probiotic strains are arbitrarily determined at present .

For the selection of microbial strains (bacterium, fungus and yeast) for use as probiotics, the general requirements of the source, viability, processing and storage of the strain, resistance to in vitro and in vivo conditions, intestinal implantation, colonization and antimicrobial activity of the strain Of course, there is a need for requirements such as biological stability, production, processing, methods of administration of probiotics, and sites where microorganisms of probiotics act in the body of livestock.

As a feed additive, there are various conditions that the probiotics should have. First, they should be able to survive in stomach and bile acid. Second, they should be resistant to various enzymes secreted from the intestines. Generally, gram-positive bacteria are resistant. Third, the probiotics should reach the small intestine and inhabit and have high fixation.

(2) a non-pathogenicity and non-toxicity; (3) an antimicrobial substance against intestinal pathogenic bacteria is produced; (4) , And to be active in the intestinal environment (resistance to gastric acid and bile acid). Fifth, it is required to maintain viable bacterial counts exceeding the standard standard during the preservation period.

In addition, Chesson (1994) suggests that, as a probiotic agent, first, the strain must be isolated from the intestinal tract and can survive well in the intestinal tract. Second, The ability of the strain to produce a substance capable of inhibiting pathogens; and (5) the ability of the strain to induce cell-mediated and the ability to stimulate cell-mediated immune responses.

As mentioned above, the probiotics reach the surface of the livestock and perform various actions. One of the reasons that the effect of the probiotics is not clear, like the efficacy of the antibiotics in general, is that the probiotic strains of general probiotics have a strong antagonistic effect in the intestinal tract like antibiotics Because it has not been able to select strains that have the ability to produce the substances that make them.

Over the years, we have been using growth promoters such as copper sulfate or antibiotics to solve the digestive problems of pigs. However, recent welfare and environment problems of livestock have been raised, consumers' preferences have become more sensitive, and various countries are avoiding the use of this type of additives, and the livestock industry voluntarily participates and regulates them.

Due to such a change of environment, the present invention was intended to develop a product that can replace antibiotics and growth promoters. There is an increased risk for disease, which is the best way for livestock to overcome various diseases by improving their natural immunity.

In addition, the domestic livestock industry is becoming more and more commercialized, sequenced, and cultivated. Environmental pollution caused by livestock manure and wastes arising from group breeding is becoming a serious problem. The harmful gases (ammonia, hydrogen sulfide, monoxide / carbon dioxide, etc.) generated in animal manure are closely related to mucous membrane damage, respiratory disease induction, and productivity deterioration. For example, Table 1 below lists the damage by concentration of ammonia. There is an urgent need for eco-friendly formulations to promote the growth of livestock and to improve the environment in the farm and the efficiency of manure disposal.

Exposure damage by ammonia concentration Person pig density Degree of damage density Degree of damage 5 ppm
7 to 10 ppm
6 to 20 ppm
40 ppm
400 ppm / hour Detection function
Maximum allowed impressions
Concerns about eye sting, respiratory disease
Headache, runny nose, loss of appetite
Bleeding nose and throat 10 ppm
25 ppm
50 ppm
100 ppm
200 ppm Decreased Growth
Respiratory disease concerns
Respiratory disease soaring
Coughing, sneezing, loss of appetite
Abnormal breathing, bleeding nose and mouth

Accordingly, the present inventors have made efforts to develop a microorganism preparation that can be used for feed addition, including microbial probiotics that can ultimately contribute to safe and clean production of livestock products. Among them, Bacillus, Streptococcus, Microorganisms of Aspergillus and Yeast Microorganisms containing microorganisms containing zeolite and wheat shorts in live cells and cultures were found to be effective in fermenting livestock manure, The present invention has been accomplished by confirming the efficacy of a multifunctional feed additive, such as the elimination, activation of enteric bacterium in the intestinal tract, inhibition of pathogenic bacterium activity, and enhancement of digestion and absorption rate of compound feed.

It is an object of the present invention to provide a multifunctional microbial preparation that can be added to animal feed.

In order to achieve the above object, the present invention provides a method for producing a lysate of Bacillus bacterium and Streptococcus bacterium, 2) a solid culture of Aspergillus bacterium, and 3) A microbial microorganism including the microbial cell culture and the cultured product as an active ingredient.

The present invention also provides a method for producing a lyophilized bacterium comprising the steps of 1) a step of mixing a solid culture product of Aspergillus bacteria and a liquid culture product of yeast bacteria into a lyophilized bacterium of Bacillus and Streptococcus, ; And 2) mixing the mixed microorganism live cells and the culture with zeolite and horse meal.

Hereinafter, the present invention will be described in detail.

The present invention relates to a lyophilized product of 1) a lyophilizate of Bacillus and Streptococcus bacteria, 2) a solid culture of Aspergillus bacteria, and 3) a liquid culture of Yeast bacteria; Provided is an eco-friendly microorganism preparation for the addition of animal feeds containing microorganism live cells and cultures as active ingredients.

In the eco-friendly microorganism preparation for livestock feed addition, the Bacillus bacterium is Bacillus subtilis subtilis) and Bacillus particulate core Lance (Bacillus coagulans) is preferable, wherein the Streptococcus strain is Streptococcus faecalis (preferably the Streptococcus faecalis), and the Aspergillus seugyun is preferred that the Aspergillus duck chair (Aspergillus oryzae), and the yeast (Yeast) The bacterium is preferably Saccharomyces cerevisiae . In particular, it is more preferable that the saccharomyces cerevisiae is cultured and the starch is mixed and dried. Bacillus subtilis is B. subtilis MG401, Bacillus coaculans is B. coagulans MG520, Streptococcus faecalis is S. faecalis MG511, Aspergillus oryzae is A. oryzae It is more preferable that MG601 and Saccharomyces cerevisiae are S. cerevisiae MG111.

In addition, in the eco-friendly microorganism preparation for livestock feed addition of the present invention, it is preferable that the microorganism preparation further contains zeolite and / or wheat shorts in addition to the above components. Specifically, 5 to 20 parts by weight of bacillus subtilis freeze-dried product, 3 to 10 parts by weight of freeze-dried Bacillus coacullans, 3 to 10 parts by weight of freeze-dried Streptococcus faecalis, 10 parts by weight of yeast culture dried matter 50 More preferably 150 to 600 parts by weight of zeolite and 350 to 1,500 parts by weight of horsepowder. In the aspergillus oryzae, 12.5 parts by weight of the bacillus subtilis freeze-dried product, 100 parts by weight of Bacillus coacule 6.25 parts by weight of lyophilized lyophilized product, 6.25 parts by weight of freeze-dried Streptococcus faecalis, 100 parts by weight of yeast culture dried product, 312.5 parts by weight of light and 712.5 parts by weight of horse parts are most preferable.

In the eco-friendly microorganism preparation for livestock feed addition according to the present invention, it is preferable that the microorganism preparation contains 10 ⁸ CFU or more of each microorganism live bacteria per 1 kg as a frying-free microorganism preparation coated with a biopolymer, 0.0 > 5.0X10 < / ^RTI > ¹⁰ CFU, Bacillus Coaculant 2.5X10 ¹⁰ CFU, Streptococcus faecalis 2.5X10 ¹⁰ CFU or more is more preferred. The livestock to which the microorganism preparation of the present invention is applied is preferably a livestock selected from the group consisting of pigs, chickens, cows and ducks.

The lactic acid bacteria contained in the microorganism preparation of the present invention produce lactic acid during the cultivation process to lower the acidity (pH 4) and generate hydrogen peroxide (H ₂ O ₂ ), thereby inhibiting the growth of pathogenic microorganisms and preventing soil disease. In addition, by producing various physiologically active substances (vitamins, amino acids, nucleic acids, etc.), antimicrobial substances and various enzymes, it increases plant self defense ability in the case of the crops. In the case of livestock, stabilization of intestinal microorganism group, Effect.

Yeast is the main microorganism of heat generation during the fermentation of agricultural by-products such as rice bran, wheat bran, etc. It is excellent in decomposition ability of organic matter and excellent in fermentation ability and viability in any bad condition. Therefore, when it is included in the feed and sprayed on the stall, the odor removal effect is excellent. It also produces a large amount of amino acids, vitamins, and other essential components for livestock growth, enhancing the palatability of feed, and the most commonly used strain is Saccharomyces cerevisiae .

Bacillus subtilis Subtillis , etc. promote the absorption of feed by converting the organic matter of the polymer into small molecules that are easy to absorb. Bacillus subtilis, also called Bacillus subtilis, is a non-pathogenic aerobic bacterium widely distributed in nature and exists in the air as well as in dry grass, sewage and soil. It is very resistant to drying and high temperature, and the cells are Gram positive bacteria containing glycogen, which decompose a large number of carbohydrates to generate acids.

Bacillus coagulans is a spreadable lactic acid bacterium that has excellent stability because it forms spores. It does not die in the intestines when added to feeds, but works in conjunction with enzymes to normalize the intestinal flora and to exert its effect.

Streptococcus faecalis is a kind of lactic acid bacteria. It is a kind of enterococcus which grows well in human or animal intestines. It has various strains. It is used for the preparation of formulators and microorganisms, and ferment cheese or yogurt. . Streptococcus faecalis attaches well to the intestinal mucosa of the host, is easy to cultivate, and is non-pathogenic. It has a beneficial effect on the host such as producing enzymes and physiological end products that can be used by the host, can survive for a long time, and can survive to the intestinal tract with the bile salts of hydrochloric acid and small intestine on the host.

Aspergillus oryzae is a fungus that is widely used for the production of soybean fermentation, brewing (yakju, takju, shochu, etc.).

According to the present invention, a useful fungus such as yeast ( Saccharomyces cerevisiae) can provide essential amino acids, vitamins, trace elements and various physiologically active substances produced in cells to improve productivity and quality, In addition, the microbial activity is continued even after the manure is excreted, thereby reducing the ammonia gas and volatile fatty acid, which are the main causes of the smell of the animal manure, and thus, the odor of the housing is remarkably reduced.

By activating the digestive activity by the microorganism's own metabolism, it facilitates the complete nutrition of the feed. Also, many enzyme producing strains produce and activate the digestive enzymes that promote the fermentation of the nutrients, And improves the utilization rate of feed such as decomposition.

Zeolite, also called zeolite, is a silicate hydrate of mainly aluminum, sodium and calcium and is represented by the general formula W _m Z _n O _2n SH ₂ O. W is Na, Ca, K (Ba, Sr), Z is Si + Al (Si: Al1), and S is not constant. Has a property that it has cation exchanged easily such as Na ⁺ (or K ⁺ ) and is substituted with Ca ^{2 +} or Mg ^{2 +} in water to soften the water. Also, since it has a relatively large space in the interior, it is also used as an adsorbent.

During the milling process, the wheat flour is removed, and the by-products remaining in the sieve are pulverized again and separated again according to the particle size. Here, those having the middle size of wheat bran and flour wheat flour are referred to as minute flour.

The present invention also provides a method for producing a lyophilized bacterium comprising the steps of 1) a step of mixing a solid culture product of Aspergillus bacteria and a liquid culture product of yeast bacteria into a lyophilized bacterium of Bacillus and Streptococcus, ; And 2) mixing the mixed microorganism live cells and the culture with zeolite and horse meal.

The effects of the microbial formulation of the present invention are as follows.

First, it excellently suppresses the odor removal and flies generation of excrement. It reduces the generation of hydrogen sulfide (H ₂ S), methyl mercaptan (CH ₃ SH), and ammonia (NH ₃ ) gas in the stem, thereby excelling the odor removal effect of the excrement, inhibiting the scattering environment of the fly, .

Second, the composting of the plant is promoted. Digestion of livestock is accelerated by various decomposition enzymes, and the production of enzymes such as amylase, protease and lipase accelerates decomposition of various organic matter and compost fermentation.

Third, it inhibits the activation of enteric bacterium and the activity of pathogenic bacteria. Three kinds of probiotics are prescribed at high levels to help the proliferation of beneficial bacteria in the intestines and produce lactic acid and antimicrobial substances to inhibit the activity of the bacteria and pathogenic bacteria.

Fourth, the growth rate and feed efficiency are improved. The addition of the yeast culture enhances palatability and flavor, and nutrient supply has the effect of improving the growth rate and feed efficiency.

Fifth, Probiotics used in the microorganism preparation of the present invention are resistant to gastric acid and bile acid by coating with Biopolymer during lyophilization, and thus survival rate is excellent when stored for a long time.

The microbial preparation of the present invention as described above promotes fermentation of livestock manure, eliminates odors in the inside and outside of the house and in the manure treatment plant, inhibits flies and mosquitoes in the house, And it can be used as a multifunction feed additive because it has an effect of improving digestion and absorption rate of compound feed.

FIG. 1 is a process flow diagram of the preparation of an environmentally friendly microorganism preparation of the present invention.
FIG. 2 is a graph showing the average weight change (change in weight gain) of the experimental group and the control pig to which the environmental microorganism preparation of the present invention was added to the feed.
FIG. 3 is a graph showing the results of measurement of the average amount of ammonia produced in the experimental group and the control feces in which the environmentally-friendly microbial agent of the present invention was added to the feed.
FIG. 4 is a graph showing the results of measurement of the average amount of carbon dioxide produced in the experiment group and the control group in which the environmentally friendly microbial agent of the present invention was added to the feed.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

< Example  1 > Preparation of microorganism preparation of the present invention

The species used in the present invention is B. subtilis MG401 (50 billion cfu / g), S. faecalis Three freeze-dried fungi such as MG511 (50 billion cfu / g) and B. coagulans MG520 (1 billion cfu / g) and Aspergillus oryzae MG601 solid culture powder (1 billion cfu / g) and S. cerevisiae MG111 liquid culture (100 million cfu / g) were used.

Streptococcus spp . Contained in the microorganism preparation was plated on BCP plate count agar. Bacillus spp. Was spiked on nutrient agar medium and Saccharomyces spp. Was cultured on potatoes dextrose agar. After culturing for 48 hours at 30-37 ° C, the number of colonies formed was measured, The number of viable cells per g was calculated by multiplying the number by the dilution factor.

The composition ratios of the microbial preparations of the present invention are shown in Table 2 below.

Raw material name content Bacillus coagulans
Bacillus subtilis
Streptococcus faecalis
Aspergillus oryzae solid culture construct
Yeast liquid culture construct
Zeolite
Minute 5 g
10 g
5 g
80 g
80 g
250 g
570 g

Hereinafter, the eco-friendly microorganism preparation of the present invention prepared in the above-mentioned Examples was fed to pigs (weaning pigs) and Korean beef cattle and compared with the control group (commonly used feedstuff), the feed intakes, the growth rate, the degree of generation of noxious gas, , And the effluent was mixed with manure directly to check whether the gas production was reduced or not, and the efficacy of this product was improved by improving productivity and controlling environmental pollution. Specifically, 1 kg of the microbial preparation prepared in Example 1 was added to 1,000 kg of ordinary feed, and the feed was fed to pigs and Korean lambs.

< Experimental Example  1> Farm selection and experiment method

First, the selection of the farm was selected as the pig farm in mountain B of Cheju Island.

1) 122 weeks (4 days old) pigs (weaned pigs) (58 of the microbial feed group and 64 of the negative control feed group) Respectively.

2) The test items were: intake of feed, weight gain per week, diarrhea (daily diarrhea and number of diarrhea were recorded), harmful gas concentration (ammonia (NH ₃ ), hydrogen sulfide (H ₂ S), carbon dioxide ₂ ), methyl mercaptan (CH ₂ SH)), the concentration of carbon dioxide (CO ₂ ) in the pot and the amount of harmful gas generated by the fodder and composting.

3) Measurement of harmful gas was done by using gas detector pump (Model 400S or APS) and disposable gas detector tube.

For the test method, the manure was collected for 48 hours before inspecting, put into a 20 ℓ bucket, left in a closed space for 5 minutes, and then the toxic gas was measured at a distance of about 10 ㎝ from the top of the manure. In addition, the pigs were all exported from the money pouch, and the carbon dioxide concentration was measured at a distance of about 10 cm from the bottom.

4) Weekly feed intake, growth rate and noxious gas concentration were measured and recorded and analyzed using ANOVA (pc-SAS package).

< Experimental Example  2 > After the feeding of the environmentally friendly microorganism preparation of the present invention, Growth rate  exam

The pig weight gain test was performed as follows. The test animals were fed with the eco-friendly microorganism preparation of the present invention to piglets until 11 weeks old at about 4 weeks of age (283 days). The control group (N = 20, 16, 22) and the control group (N = 21, 22, 21) were 6.9 and 6.6 ㎏, respectively. The weight of each pig was measured at 1, 2, 4, 5, 6, and 7 weeks after feeding the microbial agent. The results are shown in Table 3 and FIG.

Mean weight change of experimental group and control pig (change in weight gain) (unit: kg) By county Salary before After salary 1 week 2 weeks 4 weeks 5 weeks 6 weeks 7 weeks A. Experimental group
(N = 58,
3 groups) 6.9 9.1 12.6 18 23.6 28.5 33.1 B. Control group
(N = 64, 3 groups) 6.6 8.5 11.6 17.1 21.9 27.4 30.2 A-B 0.3 0.6 1.0 0.9 1.5 1.1 2.9

As a result, in the case of the experimental group, there was an increase in the weight gain of about 1 kg per two weeks from the second week after the administration of the microorganism preparation of the present invention to the control group, and an average increase of 2.9 (P <0.05), respectively.

< Experimental Example  3 > After the feeding of the environmentally friendly microorganism preparation of the present invention, In the house  Harmful gas Incidence rate  exam

After feeding the microbial formulation of the present invention, the concentration of noxious gas in the shaft was measured. Specifically, four kinds of ammonia (NH ₃ ), hydrogen sulfide (H ₂ S), carbon dioxide (CO ₂ ), and methyl Mercaptan (CH ₂ SH)] was measured. The results are shown in Table 4 and FIG.

Average Ammonia Emission Measurements in the Experimental and Control Feces (Unit: ppm) By county Salary before After salary 1 week 2 weeks 4 weeks 5 weeks 6 weeks 7 weeks A. Experimental group 15.7 15 14.7 12.3 10 9.3 11 B. Control group 15 13.3 15 16.3 17 19 18.3

As a result, the ammonia produced in the shrub was not significantly different from that in the experimental group until the second week after the feeding of the microbial formulation of the present invention, but the ammonia tended to decrease remarkably from the third week after the feeding. In the control group (feed use in general use), it decreased slightly in the first week of the test and then gradually increased. At the end of the test, at 7th week of the test, the average of 11 ppm in the experimental group and 18.3 ppm in the control group showed a decrease of the ammonia gas generation rate of about 40% in the experimental group.

However, there was no significant difference between the two groups in the presence of other harmful gases such as hydrogen sulfide, carbon dioxide, and methyl mercaptan in the flour.

The feces of the experimental group was well formed on the surface of the feces compared to the control group, and the occurrence of odor was very small. In addition, feces inside the capsule was relatively soft and contained a lot of moisture in comparison with the control group.

< Experimental Example  4> After the eco-friendly microbial preparation of the present invention In a pound  Hazardous gas concentration measurement

Experimental group and control group (feeds for general use) After transferring the pig from the pig farm where the pigs are being raised to the outside, harmful gas [hydrogen sulfide (H ₂ S), carbon dioxide (CO ₂ ) (CH ₂ SH)] was measured. The results are shown in Table 5 and FIG.

Average CO 2 Emission Levels in Experimental and Control Plants (Unit: ppm) By county Salary before After salary 1 week 2 weeks 4 weeks 5 weeks 6 weeks 7 weeks A. Experimental group 0 ^* 0 136.7 173.3 376.7 373.3 420 B. Control group 0 0 193.3 240 463.3 586.7 620

* Below detection limit

As a result, no significant difference was observed in the experimental group and the control group until 1 week after the feeding of the pigs in each pig group, but the gas production in the pigs fed the control group tended to increase gradually from the 2nd week . At the end of the test, at the 7th week of the experiment, the average of 420 ppm in the experimental group and 620 ppm in the control group showed a decrease of about 32% in the experimental group compared to the control group.

However, there was no significant difference between the two groups in hydrogen sulfide and methyl mercaptan.

< Experimental Example  5> Feed intake test of pig after feeding of environmentally friendly microorganism preparation of the present invention

The feed intakes of the pigs after the microbial formulation of the present invention were compared. The experimental group (N = 58) received 2575 ㎏ of total feed and the control group (N = 64) received 2600 ㎏ total feed. Therefore, the dietary saccharide intake was 44.4 ㎏ in the case of pigs in the experimental group and 40.6 ㎏ in the control group. Thus, the feed intake of the microbial composition of the present invention showed an increase of about 3.8 ㎏ (daily average 78 g).

The results of Experimental Examples 2 to 5 are summarized as follows.

The effect of the environmentally friendly microorganism preparation of the present invention on the pigs, barnyardgrass and pig man environment was investigated.

Feed efficiency and body weight gain of weaned piglets were investigated. The body weight gain was significantly increased (p <0.05) in the experimental group compared to the control group.

As a result of measurement of harmful gas concentration in the flue, the ammonia gas began to show a difference between the two groups from the third week after feeding, and the incidence rate of the ammonia gas decreased about 40% in the experimental group at the end of the test.

As a result of measurement of the concentration of harmful gas generated from the money pouch, the difference between the two groups was very slight from the 2nd week after the feeding, and the gas generation rate of the experimental group was reduced by about 32% at the end of the test.

There was no significant difference in the production of hydrogen sulfide, carbon dioxide and methyl mercaptan in pigs, hydrogen sulfide and methyl mercaptan in pigs.

< Experimental Example  6> The use of the eco-friendly microorganism preparation of the present invention Acid resistance  exam

Microorganisms must be strong against gastric acid to be effective in the intestines. The probiotics contained in the microbial formulation of the present invention are highly acid resistant. The results are shown in Table 6 below.

pH Total probiotics number (cfu / ml) Test commencement After 10 minutes 20 minutes later 30 minutes later 3.0 8.5 X 10 ¹⁰ 8.0 X 10 ¹⁰ 7.9 X 10 ¹⁰ 7.8 X 10 ¹⁰

< Experimental Example  7 > The microbial agent of the present invention Bile tolerance  exam

During digestion of the livestock, bile acid is released as well as stomach acid. Many lactic acid bacteria are weak in bile acid and die before reaching the intestines. As a result of adding the microorganism preparation of the present invention to artificial DNA solution, probiotics contained in the microorganism preparation of the present invention were found to have strong bile resistance by confirming that lactic acid bacteria survived for 12 hours. The results are shown in Table 7 below.

Bile (Oxgall)
density Total probiotics number (cfu / ml) Test commencement After 10 minutes 20 minutes later 30 minutes later 0.25% 8.5 X 10 ¹⁰ 8.3 X 10 ¹⁰ 8.1 X 10 ¹⁰ 7.9 X 10 ¹⁰

< Experimental Example  8 > The eco-friendly microbial preparation of the present invention Thermal stability  exam

When the microbial formulation of the present invention was subjected to a thermal stability test at 80 캜, it was confirmed that the microbial formulation was stable against heat. The results are shown in Table 8 below.

Time (minutes) Total probiotics number (cfu / ml) 0 10 20 30 80 ℃ 8.5 X 10 ¹⁰ 8.2 X 10 ¹⁰ 7.5 X 10 ¹⁰ 6.2 X 10 ¹⁰

< Experimental Example  9> After the feeding of the environmentally friendly microorganism preparation of the present invention, weight gain test

Hanwoo was also tested in a manner similar to that of the porcine growth rate test in Experimental Example 2 above. The results are shown in Table 9 below.

Average weight (kg) Growth rate
(Kg / day) Early 64 days 160 days Control group 102 153 254 0.95 Experimental group 128 207 351 1.39

As a result, we found that the body weight gain was significantly increased by 174% in the experimental group and 149% in the control group (the feed used generally) on the 160th day.

< Experimental Example  10> Analysis of Volatile Substance of Feedstock after Feeding Environmentally Friendly Microbial Agent of the Present Invention

After the microbial formulation of the present invention was fed, the content of volatile substances in the shaft fraction was analyzed. The results are shown in Table 10 below.

division Control group Experimental group Propionic acid
Butyric acid
Valeric acid 0.0296 ppm
0.6521 ppm
0.0005 ppm 0.0000 ppm
0.2109 ppm
0.000 ppm

As a result, it was confirmed that the volatile substances in the experimental group were greatly reduced by the analysis of the volatile substances as the main component of the smell, and the smell reduction effect of the flour was observed.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, This is possible.

Claims (13)

To Aspergillus oryzae , 5 to 20 parts by weight of lyophilized Bacillus subtilis , 3 to 10 parts by weight of Bacillus coagulans lyophilisate based on 100 parts by weight of the solid culture product, 3 to 10 parts by weight of a lyophilized Streptococcus faecalis , 50 to 200 parts by weight of a dried culture liquid of Saccharomyces cerevisiae , 150 to 600 parts by weight of zeolite, The present invention relates to an eco-friendly microbial preparation for the addition of a livestock feed containing, as an active ingredient, a microorganism live cell and a culture product,
delete delete delete delete The method according to claim 1, wherein said Bacillus subtilis is B. subtilis MG401, Bacillus coaculans is B. coagulans MG520, Streptococcus faecalis is S. faecalis MG511, Aspergillus oryzae is A. oryzae MG601, Wherein the S. cerevisiae MG111 is S. cerevisiae MG111.
delete delete [3] The method according to claim 1, wherein the Aspergillus oryzae comprises 12.5 parts by weight of the bacillus subtilis lyophilisate, 6.25 parts by weight of the bacillus coaculant lyophilisate, 6.25 parts by weight of the lyophilized Streptococcus faecalis, 100 parts by weight of a dried culture product of Saccharomyces cerevisiae, 312.5 parts by weight of zeolite, and 712.5 parts by weight of a horse chestnut.
10. The microorganism preparation according to any one of claims 1, 6, and 9, wherein the microorganism preparation comprises 10 ⁸ cfu or more of each microorganism live cell per 1 kg as a lyophilized microorganism material coated with a biopolymer material Based microbial agent.
[Claim 11] The method according to claim 10, wherein the microorganism live cells contain 5.0 x ¹⁰ &lt; ¹⁰ &gt; of Bacillus subtilis CFU, Bacillus Coaculant 2.5X10 ¹⁰ CFU, Streptococcus faecalis 2.5X10 ¹⁰ CFU or more.
The eco-friendly microbial preparation according to claim 1, wherein the livestock to which the microbial agent is applied is a livestock selected from the group consisting of pigs, chickens, cows, and ducks.
1) 5 to 20 parts by weight of a lysate of Bacillus subtilis , 3 to 10 parts by weight of a lysate of Bacillus coagulans , 3 to 10 parts by weight of a lysate of Streptococcus faecalis , Mixing 100 parts by weight of the solid culture product with Aspergillus oryzae and 50-200 parts by weight of the dried culture product of Saccharomyces cerevisiae in the dried fungal material; And
2) mixing 150-600 parts by weight of zeolite and 350-1500 parts by weight of wheat red dogs (wheat shorts) with the mixed microorganism live bacteria and culture.

Images