KR20180051023A - Manufacturing method of probiotics for feed additives and probiotics manufactured by the method - Google Patents

Abstract

Disclosed is a manufacturing method of probiotics for livestock feed additives which can suppress the activities of putrefactive bacteria and pathogenic bacteria. According to an embodiment of the present invention, the manufacturing method of probiotics for livestock feed additives comprises: a) a step of inoculating a medium composition having 2 parts by weight of a microorganism medium mixed with respect to 100 parts by weight of purified water, with yeast fungi, lactic acid bacteria, and bacillus bacteria to cultivate the yeast fungi, lactic acid bacteria, and bacillus bacteria; b) a step of fixing the liquid microorganisms cultivated in step a) on a solid medium consisting of rice bran, zeolite, and wheat bran at a water content of 40% and a temperature of 30°C, and manufacturing a microorganism culture while injecting air and agitating on a solid culture medium for three days; c) a step of agitating and transporting the microorganism culture of step b) once every three hours, and blowing hot air generated by an electric heater by a blower to dry the microorganism culture for three hours; and d) a step of grinding the microorganism culture dried in step c) into prescribed particles to package the ground microorganism culture.

Description

[0001] The present invention relates to a method for producing a probiotic feed additive,

The present invention relates to a method for producing a prodrug for the addition of an animal feed and a probiotic agent produced thereby. More specifically, the present invention relates to a method for producing a probiotic agent capable of inhibiting the activity of a microorganism and pathogenic bacteria by producing lactic acid and an antibacterial substance, .

So far, livestock industry has rapidly become intensified, commercialized, and large-scale, and various techniques have been applied to maximize productivity. However, as the livestock facilities are modernized and livestock are kept in high density, antibiotics for growth promotion and disease prevention have been abused. When animals are kept in a confined space, they are weakened by disease and poor growth, so that they can only use antibiotics for growth promotion and disease prevention.

The use of antibiotics can be satisfactory for immediate effect, but it induces microorganisms resistant to antibiotics in the intestines of livestock, induces the problem of residual antibiotics in livestock products, And the like.

For this reason, there is a growing interest in the development of safe materials that can replace antibiotics. For example, probiotics may be used as a substitute for antibiotics. The probiotics are used to maintain the intestinal microflora of the livestock normally to improve productivity, and to produce physiologically active substances such as enzymes, antimicrobials, and organic acids during fermentation .

Due to such a change of environment, the present invention was intended to develop a product that can replace antibiotics and growth promoters.

Korean Patent No. 10-0812667 Korean Patent No. 10-1613440

SUMMARY OF THE INVENTION The present invention provides a method for producing a multi-function probiotic agent which can be added to an animal feed, and a probiotic microbial agent prepared by the method.

In order to achieve the above object, the present invention provides a method for producing microorganisms, which comprises the steps of: (a) inoculating and culturing yeast cells, lactic acid bacteria and Bacillus bacteria in a medium composition comprising 2 parts by weight of microbial culture medium per 100 parts by weight of purified water; b) The liquid microorganisms cultured in the step a) are fixed in a solid medium composed of rice bran, zeolite and wheat bran at a temperature of 30 ° C with a moisture content of 40%, air is injected and stirred in a solid incubator for 3 days to prepare a microorganism culture step; c) stirring the microbial cultures of step b) for 3 hours with stirring, blowing the hot air generated by the electric heater with a blower and drying for 3 hours; and d) pulverizing the microorganism cultures dried in step c) into predetermined particles and packaging the microorganisms cultured in step c).

The yeast strain is saccharomyce cerevisiae and cultured at 25 ° C for 15 hours. The lactic acid bacterium is Lactobacillus plantarum and cultured at 35 ° C for 24 hours. The bacillus bacterium is bacillus subtilis Bacillus subtilis and it is preferable to incubate at 28 DEG C for 18 hours.

The solid medium is composed of 90% by weight of rice bran, 5% by weight of zeolite, and 5% by weight of wheat bran, and the solid incubator is preferably rotated and rotated at a rate of one rotation per 10 minutes.

Yeast is the main microorganism of heat generation during the fermentation of agricultural byproducts such as rice bran, wheat bran, etc., and 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 the feed, and the most commonly used strain is Saccharomyces cerevisiae.

Yeast strains of the present invention are obtained by inoculating yeast strains into a culture composition prepared by mixing 2 parts by weight of a microorganism culture medium with 100 parts by weight of purified water, and culturing at 25 DEG C for 15 hours.

The lactic acid bacteria contained in the probiotics of the present invention produce lactic acid during the cultivation process to lower the acidity (pH 4) and produce hydrogen peroxide (H2O2), thereby inhibiting the growth of pathogenic microorganisms and preventing soil disease. In addition, the production of various physiologically active substances (vitamins, amino acids, nucleic acids, etc.), antimicrobial substances and various enzymes increases the ability of plants to self-defense plants. In the case of livestock, stabilization of intestinal microorganisms, Increase effect.

The lactic acid bacterium of the present invention is Lactobacillus plantarum and is produced by culturing at 35 DEG C for 24 hours.

Bacillus subtilis (Bacillus subtilis) of the present invention promotes the absorption of feed by converting an organic matter of a polymer into a small molecule that is easy to absorb. Bacillus subtilis, also known as Bacillus subtilis, is a non-pathogenic aerobic bacillus that is 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.

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, 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.

In the present invention, the rice bran is separated when the rice is crushed, and refers to the rice husk excluding the white rice and the outer part thereof. Rice bran contains various active ingredients such as unsaturated fatty acid, protein, carbohydrate, vitamin E, dietary fiber and orizanol as a nutrient source, and it is known that rice bran has various physiological effects including an effect of suppressing the increase of cholesterol. The rice bran used in the present invention is a substrate used by the microorganisms contained in the probiotics, and various by-products such as other cereal by-products such as cornflakes and sesame pest can be used instead, and the culture conditions of the microbicide contained in the probiotic agent can be appropriately controlled have.

The embodiments of the present invention help the proliferation of beneficial bacteria in the intestines and produce lactic acid and antimicrobial substances, thereby inhibiting the activities of the spoilage bacteria and pathogenic bacteria.

In addition, the palatability and flavor of the feed is enhanced, and nutrient supply improves the growth rate and feed efficiency.

In addition, the generation of hydrogen sulfide (H2S) and ammonia (NH3) gas is reduced, and the deodorizing effect of the excrement is excellent.

In addition, digestion of livestock is facilitated by the inclusion of various degrading enzymes, and the production of enzymes such as amylase, protease, and lipase accelerates the decomposition of various organic materials and the compost fermentation.

In addition, it promotes the digestion of livestock and inhibits the growth of pathogenic microorganisms, so that the growth of livestock can grow normally, and various kinds of degrading enzymes, amino acids, vitamins, nucleic acids and antimicrobial substances are produced, Thereby increasing the disease resistance and treating and preventing diseases.

FIG. 1 is a view showing a manufacturing process of a probiotics for livestock feed according to an embodiment of the present invention.

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 .

Liquid seed culture

Yeast, lactic acid and Bacillus were inoculated into a culture composition prepared by mixing 2 parts by weight of a microbial culture medium with 100 parts by weight of purified water. Cultivation is done independently by different conditions of seed bacteria. Saccharomyces cerevisiae was cultivated for 15 hours at 25 ° C. Lactobacillus plantanum was cultured at 35 ° C for 24 hours. Bacillus bacteria were cultured on Bacillus subtilis Subtilits) were incubated at 28 ° C for 18 hours.

The optimum culture medium was used for the microbial culture.

Yeast culture medium contained 22.62% by weight of yeast extract, 67.87% by weight of anhydrous glucose, 4.52% by weight of potassium phosphate (K ₂ HPO ₄ (0.1%)), potassium phosphate (KH ₂ PO ₄ ), 4.52% by weight of zinc sulfate, and 0.45% by weight of zinc sulfate heptahydrate (ZnSO ₄ .7H ₂ O).

The lactic acid bacteria medium contained 4.53% by weight of peptone derived from soybean (Hydrolysed Soy peptone), 15.85% by weight of yeast extract, 56.61% by weight of anhydrous glucose, 11.32% by weight of sodium acetate, 2.26% by weight of sodium bicarbonate, 2.26% by weight of potassium phosphate (K ₂ HPO ₄ (0.1%)), 2.26% by weight of potassium phosphate monohydrate (KH ₂ PO ₄ (0.1% 0.23% by weight of whiskers (MgSO ₄ .7H ₂ O), 0.02% by weight of manganese (MnSO ₄ .H ₂ O), and 0.11% by weight of zinc sulfate heptahydrate (ZnSO ₄ .7H ₂ O) was used.

Bacillus bacteria medium is soybean-derived peptone (Hydrolysed Soy peptone) 18.18% by weight of yeast extract (yeast extract) 45.45% by weight ammonium sulfate (Ammonium sulfate) 18.18%, refined salt (Sodium Choride) 13.64%, J. potassium phosphate (K ₂ HPO ₄ (0.1%)) 4.55%.

Preparation of solid culture probiotics

The cultured liquid seeds were fixed in a solid medium composed of rice bran, zeolite, and wheat bran at a temperature of 30 ° C with 40% water content, air was injected, and the microorganism cultures were prepared by stirring in a solid incubator for 3 days.

The solid medium is mixed with 90% by weight of rice bran, 5% by weight of zeolite, and 5% by weight of wheat flour, and the medium is adjusted to a water content of 40% by using purified water. When the water content is too low, the growth rate of the microorganisms is low and the culture takes a long time. On the contrary, when the water content is too high, the growth rate of the microorganisms is high, but the fungi and the like show contamination, There is a problem that the manufacturing cost in the drying process performed in the post-process increases. To the solid fermentation medium suitably regulated in the above-mentioned range, 1 to 10% by weight was inoculated while adding the same liquid fertilizer seed bacterium prepared in the previous step. The inoculated solid fermentation broth was stirred for 3 days at a rate of 1 turn per 10 minutes to prepare a solid culture broth.

Drying of solid culture probiotic

It is stored in agitating and transporting machine, stirred once every 3 hours, and heat generated by electric heater is blown to dry during transferring. The dry heater was 30 kW and the blower was dried at 7.5 hp for 3 hours. The dried probiotics were pulverized into certain particles to complete the product.

The number of microorganisms contained in the probiotics after the completion of the probiotics is preferably not less than 1.0 × 10 ¹⁰ cfu / kg of Bacillus subtilis, not less than 1.0 × 10 ⁹ cfu / kg of Lactobacillus plantarum, It was confirmed that more than 1.0 × 10 ⁹ cfu / kg of saccharomyce cerevisiae was contained.

<Experimental Example 1> Test of growth rate of pigs after feeding of probiotics of the present invention

The pig weight gain test was performed as follows. The test animals were fed with the probiotics of the present invention for feeding the piglets of the present invention to the piglets for up to 11 weeks of age at about 4 weeks of age with a hybrid weaning pig, and the control group (N = 15, 15, 16) N = 15, 16, and 16) were 6.8 and 6.5 kg 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 1 below.

Mean weight change of experimental group and control pig (change in weight gain) (unit: kg)
Group

Salary before
After salary
1 week 2 weeks 4 weeks 5 weeks 6 weeks 7 weeks A. Experimental group
(N = 46,3
groups)
6.8
8.9
12.2
18.2
23.8
28.5
33.3 B. Control group
(N = 47, 3 groups)
6.5
8.3
11.1
17.1
22.1
26.5
30.2 A-B 0.3 0.6 1.1 1.1 1.7 2.0 3.1

As a result, in the experimental group, the growth rate of the average of about 1.6 kg per two weeks was increased from the second week after the feeding of the microorganism preparation of the present invention to the control group, and an average of 3.1 kg was increased at the 7th week after the end of the experiment Respectively.

<Experimental Example 2> Effect of the present invention on diarrhea prevention of pigs after feeding of probiotics

In addition, diarrhea could be prevented by a considerable level of prevention of diarrhea by more than 90%.

Prevention of diarrhea in experimental and control pigs
Group Diarrhea Prevention Effect Incidence rate Preventive effect Experimental Section 1.2% 91.9% Control 18.5% -

&Lt; Experimental Example 3 > Experiments on the growth effect of the poultry after the feeding of the probiotics of the present invention

For the experiment, 30,000 chickens from chicken farms in Chonbuk province were selected for about 40 days from August 31, 2016 to October 9, 2016, and 15,000 of experimental chickens and 15,000 of control chickens were treated with 0.1% I ate the chicken. The result was that the average delivery time of the control was 40 days, and that of the control was 6 days ahead of time. As a result, the weight gain effect was about 15%

Blood effect of poultry Until the time of release
Days Release
Shortening rate Out of season
weight Experimental Section 34 days 15% 1.7kg Control 40 days - 1.7kg

As a result of measuring the concentration of noxious gas in the stem after feeding the probiotics of the present invention. It was confirmed that the concentration of noxious gases such as ammonia (NH 3) and hydrogen sulfide (H 2 S) generated from the starch was markedly reduced compared with before feeding the probiotics.

This is because not only the metabolism in the intestines of the shaft body is activated but also the activity of the bacteria continues even after the manure is excreted, thereby reducing the ammonia gas and the volatile fatty acid which are the main cause of the smell of the animal manure.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit and scope of the invention. Accordingly, such modifications or variations are intended to fall within the scope of the appended claims.

Claims (5)

a) Inoculating and culturing yeast cells, lactic acid bacteria and Bacillus bacteria in a culture medium prepared by mixing 2 parts by weight of a microbial culture medium with 100 parts by weight of purified water;
b) The liquid microorganisms cultured in the step a) are fixed in a solid medium composed of rice bran, zeolite and wheat bran at a temperature of 30 ° C with a moisture content of 40%, air is injected and stirred in a solid incubator for 3 days to prepare a microorganism culture step;
c) stirring the microbial cultures of step b) for 3 hours with stirring, blowing the hot air generated by the electric heater with a blower and drying for 3 hours; And
and d) pulverizing the dried microbial cultures of step c) into predetermined particles and packaging. The method according to claim 1,
The yeast strain is saccharomyce cerevisiae and cultured at 25 ° C for 15 hours. The lactic acid bacterium is Lactobacillus plantarum and cultured at 35 ° C for 24 hours. The bacillus bacterium is bacillus subtilis (Bacillus subtilis) and cultivated at 28 ° C for 18 hours. The method according to claim 1,
Wherein the solid medium is composed of 90% by weight of rice bran, 5% by weight of zeolite and 5% by weight of wheat bran, and the solid incubator is rotated and stirred at a rate of one rotation per 10 minutes. A pharmaceutical composition comprising at least 1.0 × 10 ¹⁰ cfu / kg of Bacillus subtilis and 1.0 × 10 ⁹ cfu / kg or more of Lactobacillus plantarum, which is produced by any one of claims 1 to 4, Probiotics for the addition of animal feed containing 1.0 x 10 ⁹ cfu / kg or more of saccharomyce cerevisiae. 5. The method of claim 4,
α-Amylase 250,000Unit, β-Amylase 7,200Unit, proteolytic digestion enzymes (Protease) 20,000Unit, proteolytic digestion enzyme (Lipase), fiber decomposition digestive enzyme (Cellulase) 14,000Unit, photosynthetic bacteria (Photosynthetic bocteria) 2.0 × 10 ⁸ cfu / kg or more.

Images