KR20210109301A - Animal feed additives for substituting antibiotics and preparation method thereof - Google Patents

Abstract

The present invention relates to an animal feed additive for substituting antibiotics and a manufacturing method thereof. According to the present invention, the manufacturing method comprises: a mealworm powdering process (S1); a mealworm degreasing and chitin removing process (S2); an antioxidant mixing process (S3); an inoculation and fermentation process (S4); and a mixing process (S5) of adding and mixing complex L. pamtarum and P. acidlactici lactic acid bacteria probiotics to a fermented product, or comprises: a primary seaweed powdering process (S11); a secondary seaweed powdering process (S12); an antioxidant mixing process (S13); an inoculation and fermentation process (S14) of obtaining a fermented product of inoculating and fermenting prebiotics together with an emulsifier and yeast on a solid medium; and a mixing process (S15) of adding and mixing complex L. pamtarum and P. acidlactici lactic acid bacteria probiotics to the fermented product. Accordingly, a degreased and chitin-removed mealworm extract or an alginic acid-removed seaweed by-product powder is used as prebiotics, chitin-degradable Bacillus thuringiensis T3 and yeast are inoculated and fermented together on a solid medium, and complex L. pamtarum and P. acidlactici lactic acid bacteria probiotics are mixed so that the animal additive is manufactured, thereby providing an effect of substituting antibiotics for feed as well as treatment. When the animal additive is added to feed, physical improvement, immunity improvement, constitution improvement, and a feed demand rate are significantly increased, thereby providing an advantage of providing significant economic benefits.

Description

Animal feed additives for substituting antibiotics and their manufacturing method {Animal feed additives for substituting antibiotics and preparation method thereof}

The present invention relates to an animal feed additive for replacing antibiotics and a method for producing the same, and more particularly, using an insect brown mealworm extract treated with defatting and talchitin or a seaweed by-product powder treated with alginic acid as prebiotics, and chitin A novel livestock feed additive for pig poultry and cows produced by inoculating and fermenting decomposable Bacillus thuringiensis T3 and yeast together on a solid medium, then adding L. pamtarum and P.acidlactici complex lactic acid bacteria probiotics, and manufacturing method thereof is about

The level of technological development of immune-enhancing or anti-military functional feed additives in the domestic feed additive market is estimated to be 70 to 80% of that of advanced countries such as the United States, Europe, and Japan, and after the complete ban on antibiotics in 2011, a lot of R&D investment for technology discovery has been made. However, the infrastructure is still lacking.

In addition, although various physiological, metabolic and immunological basic and applied studies are being conducted on bioactive substances for the purpose of replacing antibiotics so far, these are the results of simple antibiotic replacement studies, and they are Research on the development of feed additives is still insufficient.

On the other hand, in general, feed additives are manufactured by mixing technology, processing technology, quality assurance and evaluation technology similar to compounded feed. In July 2011, the use of antibiotics added to feed for domestic growth was completely banned (Ministry of Agriculture, Food and Rural Affairs, 'National Antibiotic Resistance Safety Management Project', from May 2005, 28 out of 44 antibiotics were banned, from January 2009 to 7 Additional species ban, full ban in July 2011) was imposed. The Ministry of Agriculture, Food and Rural Affairs presented a roadmap to significantly increase the supply and demand of insects in the “Second Five-Year Plan for Insect Industry Promotion” from 2016 to 2020, and the use of feed in feed is expected to increase significantly.

However, developed countries such as the EU have been thinking about the ban on antibiotics 10 to 20 years ago, and as a result, they have technical flexibility and excellent systemic infrastructure to cope with livestock diseases. Since the use of antibiotics is prohibited, there is an urgent need to develop and apply feed additives to replace them.

However, although technical attempts are being made on various forms of feeding such as pellets, powders, and injections for the replacement of antibiotics, including basic research on physiological and metabolic and immunological responses, these feed additives are efficiently recycled using their own raw materials. However, as it is difficult to apply directly in Korea, there is an urgent need to discover useful natural by-products, to efficiently convert them into resources, and to develop functional feed additives.

In addition to silkworms and bees, as part of a feed resource recycling of insect protein raw materials that have natural antibiotic effects, feed additives containing the larvae or excrement of recent brown mealworm (T. It is disclosed (refer patent document 1). However, there is no disclosure on the antibiotic function in the above public document, only the meat quality improvement effect by changing the composition ratio of unsaturated fatty acids on chicken meat is disclosed.

Document 1: Patent Publication No. 2011-0054692 Patent Publication,

Therefore, the present invention uses degreasing and dechitin-treated insect brown mealworm extract or dealginic acid-treated seaweed by-product powder as a raw material for prebiotics, and inoculates a solid medium with chitin-decomposable Bacillus thuringiensis T3 and yeast together. The purpose of this is to provide an animal feed additive for antibiotic replacement that has an effect of replacing antibiotics for feed use by adding, mixing, and manufacturing the mixed lactic acid bacteria L. pamtarum and P.acidlactici composite lactic acid bacteria probiotics after fermentation and a method for manufacturing the same.

Another object of the present invention is to provide an animal feed additive for antibiotic replacement, which can enjoy significant economic benefits by not only improving constitution, improving immunity, improving constitution, but also significantly improving the feed requirement when used in addition to feed, and a method for manufacturing the same. There is a purpose.

The present invention provides a method for producing an animal feed additive for an alternative to antibiotics of the present invention for achieving the above object, comprising: a brown mealworm powdering process (step S1) of drying and powdering brown mealworm; A brown mealworm degreasing process (step S2) to obtain a brown mealworm powder obtained through the brown mealworm powdering process (step S1) by defatting talchitin treatment; Antioxidant mixing process (step S3) of mixing antioxidant or Vitamin C 0.05 to 0.5% to the defatted dechitinized brown mealworm to obtain prebiotics and sealing; An inoculation fermentation process (step S4) of obtaining a fermented product of inoculating and fermenting the prebiotics with chitin-decomposable Bacillus thuringiensis T3 and yeast together in a solid medium; It is characterized in that it consists of a mixing process (step S5) of adding and mixing L. pamtarum and P.acidlactici complex lactic acid bacteria probiotics to the fermented product.

In addition, the present invention provides a method for preparing an animal feed additive for antibiotics, comprising: a primary powdering process of seaweed (step S11) of drying the collected seaweed in a concrete trench silo, drying it in sunlight, and then pulverizing; Secondary powdering process of seaweed (step S12), in which the pulverized seaweed raw material is put in a rotary kiln (cylindrical hot air dryer) and dried, then put into a hammer mill and pulverized into fine grains; an antioxidant mixing process (step S13) of mixing 0.05 to 0.5% of antioxidant or Vitamin C with powdered seaweed that has undergone the secondary powdering process of seaweed (step S11) to obtain prebiotics and sealing; An inoculation fermentation process (step S14) of obtaining a fermented product of inoculating and fermenting the prebiotics in a solid medium with 0.9 to 1.6% emulsifier and 0.08 to 0.7% yeast; It characterized in that it consists of a mixing process (step S15) of adding and mixing L. pamtarum and P.acidlactici complex lactic acid bacteria probiotics to the fermented product.

The present invention uses degreasing and dechitin-treated insect brown mealworm extract or dealginic acid-treated seaweed by-product powder as prebiotics, and inoculates and ferments chitin-decomposable Bacillus thuringiensis T3 and yeast together on a solid medium, followed by complex By mixing and manufacturing lactic acid bacteria L.pamtarum and P.acidlactici complex lactic acid bacteria probiotic, it has the effect of replacing antibiotics for feed as well as treatment. There is a special advantage that can be improved and enjoy significant economic benefits.

1a to 1c is a graph showing the growth curve (CFU count) of lactic acid bacteria according to the addition of Mealworm brown;
2a to 3c are graphs showing changes in the pH of the medium according to the addition of Mealworm brown;
Figure 3 is a view showing the chitinase gene confirmation results in Basillus, (a) B. licheniformis T2, (b) B. thuringiensis T3, (c) b. subtilis T4
Figure 4 is a view showing the results of identification of protease and chitinase genes in Bacillus;
5 is a graph showing the chitin decomposition ability of the bacillus strain as a result of sugar reduction assay;
Figure 6 is a graph showing the growth change of B. thuringiensis T3 according to the addition of NAG,
(a) OD, (b) CFU
7 is a graph showing the PH change of B. thuringiensis T3 according to the addition of NAG,
8 is a view showing the antibacterial ability evaluation (first, second) of WMW and DFDC;
Figure 9 is a view showing the pH and growth curve according to the addition of brown mealworm of Basillus,
(a) T2, (b) T3, (c) T4
10 is a view showing an antimicrobial activity test against E. coli K99 of a complex microbial probiotic including L. plantarum and P. acididilactici;
11 is a process flow chart for executing the method for producing an animal feed additive for the replacement of antibiotics of the present invention using brown mealworm;
Figure 12 is a process flow chart of executing the method for producing an animal feed additive for the replacement of antibiotics of the present invention using seaweed by-products;
13 is a security photograph of the final formulation of an animal feed additive using brown mealworm of the present invention;
14 is a security photograph of the final formulation of the animal feed additive using the seaweed by-product of the present invention;
15 is a photograph showing the final formulation powder type (15a), crumble type (15b) and pellet type (15c) of the seaweed by-product of the present invention.

Hereinafter, with reference to the accompanying drawings, a preferred embodiment of the present invention antibiotic substitute for animal feed additive and its manufacturing method will be described in detail.

The method for producing an animal feed additive for antibiotic replacement of the present invention comprises: a brown mealworm powdering process (step S1) of dry and powdering brown mealworm; A brown mealworm degreasing process (step S2) to obtain a brown mealworm powder obtained through the brown mealworm powdering process (step S1) by defatting talchitin treatment; An antioxidant mixing process (step S3) of mixing 0.05 to 0.5% of antioxidants or Vitamin C with the defatted dechitined brown mealworm to obtain prebiotics and sealing; An inoculation fermentation process (step S4) of obtaining a fermented product of inoculating and fermenting the prebiotics with chitin-decomposable Bacillus thuringiensis T3 and yeast together on a solid medium; It consists of a mixing process (step S5) of adding and mixing L. pamtarum and P.acidlactici complex lactic acid bacteria probiotics to the fermented product.

In the brown mealworm powdering process (step S1), it is preferable to irradiate the brown mealworm with microwaves so that the moisture content is 8% or less.

It is preferable to degrease the brown mealworm to a fat content of 6% or less in the talchitin step (step S2).

In the mixing process (step S5), it is preferable to use any one or a mixture of two or more of lactic acid bacteria, Bacillus, and yeast as the composite lactic acid bacteria probiotic.

In addition, the method for producing an animal feed additive for antibiotic replacement includes: a seaweed primary powdering process (step S11) of drying the collected seaweed by-products in a concrete trench silo, drying them in sunlight, and then pulverizing them; Secondary powdering process of seaweed (step S12), in which the pulverized seaweed raw material is put in a rotary kiln (cylindrical hot air dryer) and dried, then put into a hammer mill and pulverized into fine grains; an antioxidant mixing process (step S13) of mixing 0.05 to 0.5% of antioxidant or Vitamin C with powdered seaweed that has undergone the secondary powdering process of seaweed (step S11) to obtain prebiotics and sealing; An inoculation fermentation process (step S14) of obtaining a fermented product of inoculating and fermenting the prebiotics in a solid medium with 0.9 to 1.6% emulsifier and 0.08 to 0.7% yeast; It consists of a mixing process (step S15) of adding and mixing L. pamtarum and P.acidlactici complex lactic acid bacteria probiotics to the fermented product.

In the seaweed primary powdering process (step S11), the seaweed raw material is spread out in a concrete trench silo so that it does not overlap and dried in sunlight for 48 hours, then put into a pin mill and pulverized. It is preferably 0.5 cm in length.

The raw material pulverized in the secondary powdering process (step S12) is dried in a rotary kiln (cylindrical hot air dryer) with a diameter of 1.2 m and a length of 12 m for 4 hours, then put into a Hammer Mill and powdered so that the moisture content of the powder is 12% or less. desirable.

In the mixing process (step S15), it is preferable to use any one or a mixture of two or more of lactic acid bacteria, Bacillus, and yeast as the composite lactic acid bacteria probiotic.

Example 1

(Selection of raw material)

Insects currently being mass-cultured in East Asia including Korea were analyzed for nutrients of mealworm (hereinafter referred to as 'mealworm'), soleer fly, and king cricket.

Among them, mealworms are recognized as having the greatest value as feed when considering both economic and nutritional aspects. This is because the nutrients are balanced compared to insects (see Table 1).

Nutritional composition of insect candidate species for feed Sample name Analysis result by component (%) moisture crude protein george crude fiber views brown mealworm
(Mealworm) 0.76 46.42 39.42 5.13 3.26 in a fraternity 1.74 41.54 36.45 7.09 8.24 king cricket 4.97 65.92 12.10 6.78 5.75

※ National Academy of Agricultural Science. 2015. p36

Example 2

(mealworm skim, talchitin)

First, in the farm, the moisture in the mealworm is extremely dried by irradiating it with microwaves at the same time as the harvest of the mealworm (you can also use the dried mealworm purchased from the shopping mall “Mealworm Nara”). By maintaining it, it prevents spoilage caused by moisture in advance.

On the other hand, mealworms contain abundant fat and a large amount of chitin, an effective substance. In Korea, there are no factories that extract these fats and chitin, whereas there are extraction and processing plants in China. In this processing plant, fat and chitin are extracted from mealworms and sold at a high price, and the remaining extract is converted into feed and sold to feed companies and farms.

Due to the fat extraction process, the fat content of skim talchitin mealworms is lowered to 6%, so the problem of rancidity caused by fat is not highlighted. Nevertheless, the present inventors prevented rancidity of the mealworms by mixing antioxidants with defatted talchitin mealworms and sealingly packaging them to block contact with air in case rancidity problems occur.

The present inventors purchased the raw mealworm raw material and analyzed the components, and then analyzed the general components of the defatted/talchitinized mealworm and the raw wheat source. Tables 2 and 3 show the results.

General component analysis division
(unit: %) Defatted talchitin mealworms Mealworm raw material Primary Secondary average Primary Secondary average moisture 7.88 8.24 8.06 4.39 6.36 5.38 crude protein 68.96 68.90 68.93 52.68 51.96 52.32 george 6.86 - 6.86 28.40 - 28.40 crude fiber 2.11 2.45 2.28 6.13 6.82 6.48 views 6.94 - 6.94 3.97 4.26 4.12 chitin 0.38 - 0.38 4.28 - 4.28

※ Chungnam National University Agricultural Science Research Center

Amino Acid Analysis division
(unit: %) Defatted talchitin mealworms Mealworm raw material Primary Secondary average Primary Secondary average aspartic acid 4.76 4.69 4.73 4.55 4.24 4.40 threonine 2.66 2.54 2.60 2.28 2.13 2.21 serine 4.33 3.60 3.97 2.79 2.58 2.69 glutamic acid 9.51 8.86 9.19 6.91 6.34 6.63 proline 4.78 4.12 4.45 3.83 3.33 3.58 glycine 5.97 4.70 5.34 2.99 2.65 2.82 alanine 4.09 3.89 3.99 3.82 3.38 3.60 valine 3.00 2.98 2.99 2.86 3.26 3.06 isoleucine 1.99 1.91 1.95 2.14 1.92 2.03 leucine 5.29 5.05 5.17 3.99 3.58 3.79 tyrosine 1.85 1.64 1.75 3.91 3.47 3.69 phenylalanine 2.66 2.53 2.60 2.05 1.86 1.96 histidine 1.07 1.14 1.11 1.63 1.46 1.55 lysine 4.28 4.09 4.19 2.92 2.95 2.94 arginine 4.60 4.14 4.37 2.87 2.70 2.79 Sistine 2.55 1.87 2.21 0.81 0.86 0.84 methionine 1.65 1.89 1.77 0.77 0.83 0.80 tryptophan 0.40 0.43 0.42 0.58 0.53 0.56 Sum 65.44 60.07 62.76 51.7 48.07 49.89

※ Chungnam National University Agricultural Science Research Center

Comparison of amino acids between mealworms and fishmeal Defatted talchitin mealworms foreign fish meal domestic fish meal income aginine 4.37 - - 3.44 glycine 5.34 4.29 1.76 5.67 lysine 4.19 - - 5.19 methionine 1.77 2.43 1.60 2.00 phenylalanine 2.60 2.16 2.17 2.40 isoleucine 1.95 3.47 2.32 2.82 leucine 5.17 4.38 3.10 5.25 threonine 2.60 0.04 0.07 2.64 tryptophan 0.42 - - - valine 2.99 4.19 3.69 1.90

※ Feed Resources Handbook 4th Edition. 2011.

<Experimental Example 1> Microbial material selection

1-1. Selection of lactic acid bacteria material

With respect to the candidate probiotic lactic acid bacteria is Lactobacillus 3 jong (Lactobacillus reuteri KLR 3004, Lactobaillus KLW001 salivarius, Lactobacillus plantarum GS-1) of the evaluate check that the number of bacteria such as prebiotics function enhancing effect when milwom added.

To confirm this, each lactic acid bacteria was incubated in De Man, Rogosa and Sharpe (MRS, later MRS) Broth (mealworm (0.1% w/v)) mixed with powdered mealworms at 0.1% w/v for 36 hours. Sampling the culture medium for each 0, 2, 4, 6, 8, 12, 24, and 36 time period, spreading it on MRS agar medium, counting the number of colony forming units (CFU, hereinafter CFU) generated after 24 hours and showing the result 1, and after measuring the pH at the incubation time, it was compared with MRS broth (No mealworm) to which no mealworm was added, and it is shown in FIG. 2 .

1 and 2, it was confirmed that when the mealworm was treated at 0.1% w / v, it did not affect the pH change through the increase in the number of bacteria or the production of lactic acid for three types of lactic acid bacteria, and there was no special negative effect. there was.

<Experimental Example 2> Bacillus material selection

Among the probiotic candidates, the material selection process was carried out for three types of Bacillus (Bacillus licheniformis T2, Bacillus thuringiensis T3, Bacillus subtilis T4, hereinafter referred to as T2, T3, and T4, respectively). The strains were each deposited with the deposit number 0000 on 00/00 on 0000 in the microorganism depository institution 0000.

In milwom may expect a synergistic effect of the probiotic and that can be used to break down chitin high chitin content, ginseng relative to them (called chi, since chi) genes, chitinase to decompose chitin is the presence or absence of the gene. This was confirmed through extraction and PCR of each bacterial DNA using an existing reference primer, and the results are shown in FIG. 3 .

In addition, since there is a content that the secretion of protease plays an important role when polysaccharides such as chitin are isolated from shrimp, etc., it was judged that it is highly likely to act in the same way in the decomposition of chitin in mealworms. The presence of the gene was also confirmed through PCR, and the results are shown in FIG. 4 .

Finally, through a sugar assay, it was confirmed whether there was a change in the sugar content according to the decomposition of chitin by the corresponding gene, and the results are shown in FIG. 5 .

On the other hand, N- acetyl glucosamine (NAG, hereinafter referred to as NAG), a monomer form that can be produced by decomposition of chitin contained in mealworms, was added to Lysogeny broth (LB, hereinafter referred to as LB) broth at 1.5% w/v. By giving, the operation of checking whether the monosaccharide can be used as a nutrient necessary for the growth of Bacillus bacteria is confirmed by optical density (OD, hereinafter referred to as OD) measurement and CFU count and pH measurement, and the results are shown in FIGS. 6 and 7 showed

<Experimental Example 3-1> Check the presence of chi gene

As a result of the experiment, it was confirmed from FIG. 3 that the chi gene was present in the case of T2 and T3 among the three Bacillus strains. there was. That is, the possibility that T2 and T3 can use mealworms as prebiotics was confirmed.

<Experimental Example 3-2> Confirmation of presence of protease gene

In addition, as a result of the experiment, in the case of T2 and T3 among the Bacillus candidate groups from FIG. 4, it was confirmed that amplification was performed when PCR was performed using a primer for protease detection.

It was confirmed that T2 and T3, which were previously confirmed to have chi genes, also have protease genes. means you can

<Experimental Example 3-3> Confirmation of presence of protease gene

As a result of the experiment, using T2 and T3, which confirmed the possibility of decomposing chitin, a sugar reduction assay was performed to see if there was a change in sugar content due to chitin decomposition, and the results are shown in FIG. 5 .

From FIG. 5, it was confirmed that sugar was produced in T2 and T3, and in particular, in the case of T3, it was confirmed that the sugar content increased according to the addition of chitin. This means that chitin is actually degraded by T3. In other words, it was confirmed that T3 is highly likely to use mealworms as prebiotics.

<Experimental Example 3-4> Confirmation of T3 growth change according to the addition of NAG

As a result of the experiment, when NAG was added at 1.5% w/v to the existing LB medium through the confirmation of the number of bacteria and the change in pH through the OD and CFU counts from FIGS. . This means that T3 does not prefer NAG, the monomer form of chitin, as a nutrient source.

<Experimental Example 4> Degreasing insect material in vitro Functional evaluation

Because it is known that the antimicrobial peptide represented by tenecin, an antibacterial material, is abundant in mealworms, raw mealworms (whole mealworm, WMW in this section) and defatted talchitin mealworms (hereinafter, "de-fatted de-chitin mealworm, antibacterial for DFDC "referred to) each representative of pathogens by using a medium haejun added with 0.5% or 1/0% w / v in LB medium Escherichia coli K99 (named E.coli K99, E.coli K99 hereinafter) The effect was confirmed through the CFU count of E. coli K99, and the results are shown in FIG. 8 .

In addition, in order to check whether there is a bacterial growth enhancing effect according to the addition of DFDC using the Bacillus candidates that are used as candidates, the growth curve of the bacteria when mixed with LB at 0.5% w/v with basic LB medium and 0.5% w/v mealworm Bacterial growth curves and comparison time zones in the added LB medium were shown in FIG. 9 after sampling the culture solution for each time zone 0, 6, 12, 24, 36, 48, 60, 72, 84.

After spreading this on an LB agar plate, the generated CFU was counted and the pH of the culture medium was measured to confirm the effect of the addition effect on the bacterial growth.

<Experimental Example 4-1> Confirmation of antibacterial effect of Mealworm (WMW) and DFDC on E.coli K99

As a result of the second experiment, it was confirmed that there was no difference in the number of CFU counts of E. coli K99 according to the addition of WMW or DFDC as a result of the second trial, and is shown in FIG. 10 . This means that mealworm itself and DFDC degreasing it have no antibacterial effect on the pathogen E.coli K99.

<Experimental Example 4-2> CFU count (0.5% w/v DFDC added LB, 0.5% w/v mealworm LB, basic LB medium)

As a result of the experiment, in terms of bacterial growth in T2 and T3, it was confirmed from FIG. 1 that mealworm (WMW) supplemented medium rapidly promoted growth in the first 6 hours to 12 hours. It was found to be somewhat helpful for the promotion of bacteria.

In particular, it was confirmed that when WMW was added in T3, the growth rate increased significantly during the first 24 hours, and in T4, it was confirmed that WMW did not significantly affect the growth.

Also, as shown in FIG. 9, it was confirmed that the pH increase rate was generally slow in the WMW-added medium, and through this, it was confirmed that neither WMW nor DFDC had a negative effect on bacterial growth. It was confirmed that further growth was promoted and the number of bacteria in the final equilibrium state could be increased by the addition of DFDC.

In particular, in the case of DFDC, it was confirmed that there was no difference from the basal LB medium in pH, and it was shown that it did not negatively affect the bacterial physiology.

<Experimental Example 5> of the prototype in vitro Functional evaluation

As a prototype of the present invention, Mealworm and B. thuringiensis T3 and yeast were solid-fermented together.

The reason for choosing solid fermentation rather than conventional liquid or dry powder form is that during solid fermentation, the spore-forming ability is improved, which is advantageous for the survival of the strain. This is because it contains effective metabolites and degradation products produced by the strain.

In addition to the Bacillus strain and mealworm, active ingredients and yeast in the industrial medium were included together, making it easy to use as a complex probiotic.

Considering the applicability of livestock feed additives, the form of a complex microbial probiotic using multiple strains in combination is more suitable than a single strain additive.

Considering the formulation as a complex microbial probiotic, a combination of a strain with excellent antibacterial activity was required because the combination of mealworm and Bacillus cannot expect antibacterial activity against pathogenic microorganisms.

A product produced through solid fermentation and a lactic acid strain (Lactobacillus plantarum , Pediococcus acidilactici ), which is currently widely used as a probiotic and is known to have antibacterial activity against pathogenic microorganisms (E. coli K99), was mixed to establish a formulation in the form of a complex microbial probiotic. .

<Experimental Example 5-1> Complex microbial probiotic E. coli Confirmation of antibacterial activity against K99

E. coli K99 is a pathogen that causes Escherichia coli diarrheal disease, and it causes diarrhea in piglets, especially in piglets during weaning.

Two lactic acid bacteria, L. plantarum and P. acidilactici , known to have antibacterial activity against E. coli K99 in complex microbial preparations (including medium components, mealworms, B. thuringien sis T3, yeast and metabolites) produced through solid fermentation Each strain was mixed to test the antimicrobial activity of the complex microbial probiotic.

Both L. plantarum and P. acidilactici strains showed an inhibition zone diameter of 15 mm or more, confirming the antibacterial activity against E. coli K99, a pathogenic microorganism.

In particular, in the case of L. plantarum among lactic acid bacteria selected as a candidate material for the complex microbial probiotic in the present invention, the average diameter of the inhibition zone was 18.4 mm, demonstrating excellent antibacterial activity.

Inhibition zone diameter (Unit: mm) L. plantarum P. acidilactici Sample 1 17.01 15.28 Sample 2 17.02 14.95 Sample 3 17.26 15.98 Sample 4 17.71 16.06 Sample 5 19.26 15.37 Sample 6 19.80 16.50 Sample 7 16.50 15.20 Sample 8 17.12 15.64 Sample 9 21.42 19.06 Sample 10 21.09 18.57 average 18.419 16.243

As a result of the experiment, when lactic acid bacteria, Bacillus, and yeast were used as a complex microbial probiotic, the antibacterial activity of lactic acid bacteria, the enzyme activity of Bacillus and the improvement of the livestock environment, the protein and mineral supply effect of yeast, and the effect of non-specific immunity promotion could be expected together.

In addition, it can be said that useful metabolites derived from microorganisms included through solid fermentation can be used together, so it can be said that it is a form with excellent usability as a feed additive.

<Specification Experiment Example 1>

In order to confirm the functionality of the brown mealworm product of the present invention, the following breeding experiment was conducted on pigs (weaned piglets) from June to July 2019.

A feeding test was conducted to confirm the safety and functionality of the prototype by feeding the mealworm prototype to 22 weaned pigs at 24 days of age. After mixing 2% of the compounded feed and feeding it for 21 days, the daily weight gain, feed requirement, and mortality were checked.

Although the weight gain did not reach the target, the normal weaning piglet experiment was 4 weeks, but the experiment period was limited to 3 weeks due to the circumstances of the farm where this experiment was conducted, so the effect of increasing the weight gain in the late weaning period was not enjoyed. In addition, the daily weight gain varied according to the environment, feed feed, and climatic conditions of individual farms.

However, during the experiment period, the feed requirement ratio of the experimental group fed the mealworm prototype was 1.25, which was significantly better than the feed requirement ratio of the control group that was not fed with 1.69.

It was possible to confirm the functionality of the present invention by showing that the piglets fed the prototype mealworm and the defatted/talchitin mealworm of the present invention grew faster even though randomly placed piglets were fed the same feed in the same environment.

To analyze the economic feasibility, the feed for weaned pigs used in the experiment was 1,025 won per kg, and when 2% of skim/talchitin mealworms were added, only 100 won per kg was increased. Therefore, it was judged to be able to enjoy considerable economic profits by feeding mealworms at actual farms.

According to the qualitative evaluation of the workers of Yangdong Farm, who conducted the experiment, there were no weak pigs/dead pigs in the experimental group during the experiment period, and the physique was enlarged and the luster of the coat could be maintained. This is a result that is expected to improve physique, improve immunity, and improve constitution according to mealworm feeding.

During the period of the experiment, health/immunity-related problems of the experimental animals did not appear without using antibiotics at all, so it was judged that there was an alternative effect of antibiotics for feed as well as treatment.

Certificate of Analysis using mealworms (Certificate of Analysis) Unit: kg division control 1 treatment group (mealworm + probiotics)
2% added) 1 treatment area (mealworm only
2% added) number of tests 35 heads 35 heads 35 heads starting total weight
Gaechi Sejung (two parties) 270
7.71 261
7.46 245
7.00 end total weight
End weight (per head) 415
11.36 441
12.60 416
11.39 Total feed amount
Feed amount (per head) 245
7.00 225
6.43 225
6.43 total weight gain
Weight gain (per head) 145
4.14 180
5.14 171
4.89 Daily weight gain (g)
Feed demand rate 197.28
1.69 244.90
1.25 232.65
1.32

Note) Experiment name: Weaning pig breeding experiment using wheat source

Experimental location: Jeongil Farm, Hongseong-gun, Chungcheongnam-do

Experiment period June 7 - 28, 2019 (21 days)

Actual animals: 105 piglets (35 pigs × 3), 23-day-old weaned piglets

Weighing method: Weighing using electronic scales for each group of pigs

Example 3

(Manufacture of seaweed by-products extracted from alginic acid)

After the seaweed raw material was spread out in a concrete trench silo so as not to overlap, it was dried in sunlight for 48 hours, and then samples of the raw material were taken from 3 places at random and moisture was measured.

Sun-dried raw materials were put into a pin mill with a crushing capacity of 500 kg per hour and pulverized. After pulverization, the size of the particles was 0.5 cm in width and 0.5 cm in length, but the shape and size of the particles were not uniform.

After drying the pulverized raw material in a rotary kiln (cylindrical hot air dryer) having a diameter of 1.2 m and a length of 12 m for 4 hours, a raw material sample was collected and moisture was measured. After hot air drying, the raw material was put into a Hammer Mill with a grinding capacity of 5 tons per hour and pulverized into fine particles. After drying in the sun and hot air, the moisture of the raw material was lowered to 12%, which became a level usable as a raw material for feed.

Through the grinding process twice, it was possible to produce a product with a uniform particle size that can be used as a raw material for feed. It succeeded in formulating the particle size into a fine powder. The formulated powder was judged to be suitable for use as a raw material for feed and feed additives.

However, it was found that excessive manpower is required during the drying process, and the raw material is adsorbed to the machine due to the adhesive force of alginic acid, which significantly lowers productivity.

Example 4

(Formulation of alginic acid extract seaweed by-product)

After extracting alginic acid from the alginic acid processing plant in Gamwi County, Jiangsu Province, China, the remaining seaweed by-product solution is collected and moved to the by-product treatment area of the plant. was used.

Through this process, the semi-dried seaweed by-product raw materials were collected, moved to the drying plant in the same area and the plant of the governing organization, and the raw materials were randomly collected from three places and moisture was measured.

The raw materials were divided into two groups, and the first group applied the raw materials thinly to a thickness of 3 to 5 cm on the ground of the concrete trench silo using a 1-ton payloader, and then turned the raw materials over at 11 am and 15 pm, After that, solar drying was performed for 72 hours.

The second group put raw materials into a rotary kiln (cylindrical hot air dryer) with a diameter of 1.2 m and a length of 12 m and dried for 4 hours. , The finely pulverized raw material was molded to a diameter of 8 mm using a 200 horsepower pellet molding machine.

change in moisture Raw material (semi-dry solution) 72 hours sun drying Hot air drying 4 hours 46% 14% 13%

The moisture content of the first group that was sun-dried and the second group that was dried with hot air was lowered to 14% or less, so that it could be used as a raw material for feed.

Particle size change trend Raw material (semi-dry solution) dry Hammer mill grinding thick porridge A mixture of lumps and powders with a maximum diameter of 7 cm 0.1cm wide, 0.1cm long

During the drying process, the raw materials agglomerated by themselves due to the algin contained in the seaweed to form large lumps. In order to use this as a raw material for feed, it was subjected to a grinding process to produce a product with a uniform particle size that can be used as a raw material for feed.

During the solar drying process, if the top and bottom were not continuously turned upside down, the drying of the lower raw material was sluggish. In further experiments, a significant part could be solved by adding 1.5% emulsifier.

In this experiment, it was possible to save drying time and cost by using the raw material in a semi-dry state, and since most of the active substances including alginic acid remain after alginic acid extraction, it is an advantageous formulation compared to processing raw seaweed in consideration of process convenience and economic feasibility was judged by the method.

First, in the case of raw seaweed, it was difficult to pulverize due to the thick and tough fiber peculiar to seaweed insoluble parts. In addition, when hot air drying using a rotary kiln was performed after pulverizing the raw seaweed, the raw material was adsorbed on the wall of the rotary kiln due to the alginic acid component of seaweed, which caused a decrease in productivity.

On the other hand, during the drying process, seaweed turns black and almost disappears at the same time flavor, which is expected to decrease the preference of final consumers. However, when exposed to air, the moisture gradually increased.

To solve this problem, steam was added to the dry powder and formed into pellets. As a result, the addition of steam showed the effect of reproducing the color and flavor of the raw seaweed, and the hard outer wall of the pellet prevents moisture penetration and maintains the dry state for a long time. was able to confirm

Through this, it was determined that alginic acid extraction seaweed by-product as a raw material was sun-dried and finely pulverized and then pelleted satisfies all of the qualitative preference, economic feasibility, and distribution stability of the product (see FIGS. 15a, 15b, 15c).

<Specification Experiment 2>

Contents and results of cow breeding experiment using seaweed by-products

One). Experimental Design

Thirty-four Holstein cows were assigned to two treatment groups of 17 cows each, and divided into an experimental group (with 2% of experimental feed) and a control group (with no addition), and the test was conducted at Hwani Ranch in Hwaseong, Gyeonggi-do for 4 weeks.

All milking cows were raised in a free stall and fed Suwon Livestock Cooperative TMR feed. The feed was divided into 2 times a day, designed to be a free vegetarian diet, and water was freely ingested.

The experimental group was fed by adding 2% level (400 g) of the daily feed to the TMR feed and mixing it. Milk production and somatic cell levels were measured daily for each milking cow, and health status, intake status, and mortality were frequently checked during the experiment period.

2).Experiment result

Holstein dairy cows are one of the livestock most vulnerable to high temperature stress due to the characteristics of their breed. Diarrhea and decreased milk production occurred in cows, so the research team temporarily stopped the experiment.

As this phenomenon occurred in the experimental group to which the seaweed product was fed as well as the control group that did not feed it, it was judged that the problem was not caused by the seaweed product.

Results of monitoring the health of dairy farms due to high temperature stress (estimated) on the 4th day of the experiment Reduced rumination activity flow reduction somatic increase experimental group 5 4 One control 3 4 One

Flow rate and somatic cell warning due to high temperature stress (estimated) on the 4th day of the experiment somatic increase reduced flow experimental group 2 2 control 4 4

On the other hand, the experimental period is too short to judge the result according to the wetness of the prototype of the present invention, but as shown in the table below, the amount of oil produced during the experiment exceeded 30 kg of oil production per day, the goal of the present invention for both the experimental group and the control group, as shown in the table below. A slight high was seen.

The amount of milk produced by cows during the experiment Oil production (kg/en/day) Day 1 Day 3 Day 6 average experimental group 33.8 34.5 34.2 34.2 control 33.0 32.9 33.1 33.0

Claims (8)

A brown mealworm powdering process (step S1) of drying and powdering brown mealworm; A brown mealworm degreasing process (step S2) to obtain a brown mealworm powder obtained through the brown mealworm powdering process (step S1) by defatting talchitin treatment; An antioxidant mixing process (step S3) of mixing 0.05 to 0.5% of antioxidant Vitamin C with the defatted dechitinized brown mealworm to obtain prebiotics and sealing; An inoculation fermentation process (step S4) of obtaining a fermented product by inoculating and fermenting the prebiotics with chitin-decomposable Bacillus thuringien sis T3 and yeast together in a solid medium; A method for producing an animal feed additive for antibiotic replacement, comprising a mixing process (step S5) of adding and mixing L. pamtarum and P.acidlactici complex lactic acid bacteria to the fermented product. The method according to claim 1, wherein the brown mealworm powdering process (step S1) is irradiated with microwaves to dry the brown mealworm to a moisture content of 8% or less. . The method according to claim 1, wherein the fat content is degreased to 6% or less in the degreasing process (step S2) of the brown mealworm. a primary powdering process of seaweed (step S11) of drying the collected seaweed by-products in a concrete trench silo, drying them in sunlight, and then pulverizing them; Secondary powdering process of seaweed (step S12), in which the pulverized seaweed raw material is put in a rotary kiln (cylindrical hot air dryer) and dried, then put into a hammer mill and pulverized into fine grains; an antioxidant mixing process (step S13) of mixing 0.1% of antioxidant Vitamin C with the powdered seaweed that has undergone the secondary powdering process (step S11) to obtain prebiotics and sealing; An inoculation fermentation process (step S14) of obtaining a fermented product of inoculating and fermenting the prebiotics in a solid medium with 0.9 to 1.6% emulsifier and 0.08 to 0.7% yeast; A method for producing an animal feed additive for antibiotic replacement, comprising a mixing process (step S15) of adding and mixing L. pamtarum and P.acidlactici complex lactic acid bacteria to the fermented product. [Claim 5] The method according to claim 4, wherein, in the seaweed primary powdering process (step S11), the seaweed raw material is spread out in a concrete trench silo so as not to overlap, dried in sunlight for 48 hours, and then put into a pin mill to pulverize and pulverize the particles. A method for producing an animal feed additive for antibiotics, characterized in that the size is 0.5 cm in width and 0.5 cm in length. According to claim 4, wherein the raw material pulverized in the secondary powdering process (S12 step) of the seaweed is dried in a rotary kiln (cylindrical hot air dryer) having a diameter of 1.2 m and a length of 12 m for 4 hours, then put into a Hammer Mill and powdered. A method for producing an animal feed additive for antibiotic replacement, characterized in that the moisture content of the water content is 12% or less. According to claim 1 or 4, wherein in the mixing process (step S5) or the mixing process (step S15), the complex lactic acid bacteria probiotic is any one or a mixture of lactic acid bacteria, bacillus, yeast (yeast), characterized in that A method for manufacturing an animal feed additive for an alternative to antibiotics. An animal feed additive for antibiotic replacement prepared by the method according to any one of claims 1 to 8.

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