KR20090078065A - The composition of improved lactic acid bacteria feed additive and method for manufacturing thereof - Google Patents

Abstract

A method for manufacturing the lactobacillus preparation composition for a feed additive, and the lactobacillus preparation composition are provided to improve antibacterial effect. A method for manufacturing the lactobacillus preparation composition for a feed additive comprises the steps of adding betaine and yucca extract to the culture solution of lactobacillus at a constant temperature for a certain time to make the culture solution of killed lactobacillus; and spraying the culture solution of killed lactobacillus on an adsorbent to adsorb it, and mixing it with an excipient.

Description

The composition of improved Lactic acid bacteria feed additive and Method for manufacturing et al.

The present invention relates to a feed lactic acid bacterium for enhancing the antimicrobial effect and a method for manufacturing the same, and in particular, the conventional lactic acid bacteria are cultured for a predetermined time, followed by heat treatment by addition of bitine, followed by two steps of reculturing by adding a yucca extract. The present invention relates to a lactic acid bactericidal agent having an enhanced antimicrobial effect, comprising adsorbing a bactericidal culture solution to a conventional pharmaceutically acceptable adsorbent, and then mixing the same with an excipient and repeating the same. .

Probiotic is derived from the Greek word for life and, in contrast to antibiotic, refers to a beneficial relationship between living organisms and in the ecological term probiosis, meaning symbiosis. It was first used by Lilly and Stillwell (1965). Since then, many researchers have defined probiotics. Parker (1974) is a microbial and metabolite that has beneficial effects on the host through control of the intestinal microflora. It was. Mitsuoka (1975) and Crawford (1979) are probiotics that are microorganisms or cultures with moderate physiological activity, and antagonistically act against antagonistic bacteria in their intestines by administering them. It should be possible to guarantee that the gun is returned to normal. Fuller (1989) later defined probiotics as living microorganisms that benefit the host by improving the balance of intestinal microflora. In Korea, Probiotics are preparations containing live beneficial microorganisms in Korea, and they are settled in the intestines of animals and compete with other pathogenic microorganisms to inhibit the growth of harmful microorganisms and to help digestion and absorption of ingested feed. It is defined as a microbial agent that promotes growth and improves feed efficiency.

Probiotics have been recognized for their effects on abnormal fermentation, diarrhea, indigestion, and constipation in the intestine, and have been used for humans. They are widely used in various livestock species for the purpose of improving the intestinal environment, improving productivity, and preventing diarrhea. It has been used. Microorganisms currently used in probiotics include lactic acid bacteria, subtilis bacteria, molds and yeasts, and lactic acid bacteria isolated from the gut of healthy livestock are generally widely used.

In 1973, Tortuero reported that dietary supplementation of 100 chickens with Lactobacillus acidophilus for 11 days resulted in increased weight gain and improved feed efficiency (Poultry Sci., 52, 197). In 1982, Watkins et al. Found that Lactobacillus acidophilus was significantly reduced in E. coli-infected chickens compared to non-administered bacteria (Poultry Sci., 61, 1298). -1308). In 1989, Myung-Won Jae et al reported that when lactic acid bacteria were fed to pigs, the gain in body weight was improved and diarrhea in piglets was prevented (Korea J. Anim. Sci ,. 31, 318-323). In 1984, Reddy G.V et al. Reported that Lactobacillus acidophilus and Lactobacillus bulgaricus exerted antimicrobial activity as natural antibiotics (Cultured Dairy Product J. 7-11). Binek M et al. Then demonstrated that lactic acid bacteria isolated from the digestive tract of chickens had antibacterial effects against Salmonella, Escherichia coli and Clostridium bacteria (Poultry J. Microbiol. 2005; 54 (4) 287-294). Belfiore et al. Reported that the growth of Escherichia coli was inhibited by bacteriocin, lactocin and nisin, which are antimicrobial peptides produced by lactic acid bacteria in the culture of lactic acid bacteria (Food microbiol, 2007 May; 24 (3)). 223-229), Stern NJ et al. Reported that bacteriocin, an antimicrobial peptide produced by Lactobacillus salivarius, significantly inhibited Clostridium bacteria (Antimicrob. Agents chemother., 2006 Sep; 50 (9) 3111-3116).

Recently, the problem of resistance due to misuse and abuse of antibiotics and antimicrobial agents and the problem of residues in livestock products due to the addition of antibiotic feed have led to trade friction between countries. According to the 2005 Food and Drug Administration data, food and livestock products are shocking to find antibiotic resistance in more than 80% of E. coli and enterococci. In order to prevent antibiotic and antimicrobial residues in livestock and food products, the US Code of Federal Regulation has created a Food Additive Compendium for antibiotics and antimicrobials and strictly restricts their use. The Annex I of council regulation strictly controls the use of antibiotics and antibiotics by species. In Korea, in accordance with the criteria for the use of veterinary medicine for formulated feed under Article 72 of the Pharmaceutical Affairs Act and Article 46 of the Handling Rules of Animal Medicine, the use of feed additive antibiotics was reduced from 53 to 25 in July 2005. Regulates the use of materials. In addition, the Ministry of Agriculture and Forestry announced comprehensive measures for the hygiene and stability of livestock products and measures to reduce the use of antimicrobial substances, including antibiotics, to support the development and supply of antibiotic substitutes such as probiotics, enzymes, organic acids, medicinal plant preparations, and natural ingredients. Probiotics make up the bulk of antibiotics used. However, because probiotics differ from the test tubes of animals, there is a problem that the evidence indicating permanent settling in the intestine is unclear and should be continuously administered if a continuous effect is needed (Fuller, 1992, Livestock Research Vol. 46, No. 1). , 44).

In addition, depending on the environmental conditions, the expected effect is insufficient and there is a problem in stability (stability), so the processing process of the distribution process or feed, especially in the case of heat-treated feed such as pellet feed, crumble feed and flake feed during the manufacturing process The high heat and steam of 120 lead to the loss of viability and deterioration of vitality (Lee, In-Ho, Technology Management Approach to Direct-fed Microbials Theory, 2000, 17-19). Republic of Korea Patent No. 10-0261352 (Clean feed additive composition and its manufacturing method), 10-0396022 (Manufacturing method for livestock probiotics and livestock probiotics), 10-0489859 (Lactobacillus for treating and preventing poultry typhoid and poultry using the same Typhoid treatment method), 10-0688441 (feed additive using citrus gourd and its manufacturing method) and US Patent No. 6,503,544 (Animal feed additive) are all patents related to animal feed additives using lactic acid bacteria, and are fundamentally stable. I have a problem with In order to solve the problem of stability of heat of probiotics, a lactic acid bacterium that has been treated with tyldallization has been developed and marketed. J. Fourniat et al. Found that mortality was significantly reduced by oral administration of Lactobacillus acidophilus treated with E. coli (Swiss OF1). It was reported that the antimicrobial peptides produced by lactic acid bacteria were greatly increased during the heat treatment (Microecology and therapy, Vol. 14, 1984, 263-264). Currently, commercially available tindal lactobacillus is listed as the Lactobacillus asidophilus tindal freeze-dried product in the 2nd Amendment of Standards and Test Methods of the Korea Food and Drug Administration. It is difficult to feed chickens, pigs and cows, which are expensive to use when applied to economical animals, and there is a disadvantage that the antimicrobial effect is insufficient as an antibiotic substitute.

An object of the present invention is to repeat the process of culturing the conventional lactic acid bacteria in order to solve the above problems for a certain period of time after the heat treatment by the addition of betaine (betaine) and adding the yucca extract (yucca extract) twice After the sterilization process by adsorbing to a suitable adsorbent and then mixed with an excipient, to provide a lactic acid bacteria agent and a method for producing the enhanced antibacterial effect.

The present invention is incubated for a predetermined time at 34 ℃ to 37 ℃ conventional lactic acid bacteria and then 0.005 to 0.2 parts by weight of betaine is added to 1 part by weight of the culture and heat treatment at 60 ℃ to 70 ℃ 10 minutes to 30 minutes and then 34 ℃ To 0.001 to 0.05 parts by weight of yucca extract per 1 part by weight of the culture medium and incubated at 23 ° C. to 37 ° C. for a certain time, and then 0.005 to 0.2 parts by weight of betaine was added again to 1 part by weight of the culture solution. Heat-treated at 70 to 80 ℃ for 10 to 30 minutes, then lowered to 34 ℃ to 37 ℃, and then added 0.001 to 0.05 parts by weight of yucca extract per 1 part by weight of the culture medium and incubated for a certain time at 34 ℃ to 37 ℃ 121 1 to 4 parts by weight of the adsorbent to 1 part by weight of the sterilized lactic acid bacteria culture medium sterilized at 15 ° C. for 30 minutes, and then to 1 part by weight of the sterilized lactic acid bacteria culture medium adsorbate. Conventional excipients 4 to 19 parts by weight of lactic acid bacteria for the antibacterial effect is enhanced by the addition to the feed mixture preparation composition and to a method of manufacturing the same.

As described above, the lactic acid bacteria were incubated for a predetermined time, followed by heat treatment with the addition of betaine, followed by re-cultivation with the addition of the yucca extract, followed by sterile treatment, adsorption and mixing with the adsorbent and the excipient to feed the lactic acid bacteria. By preparing the formulation, it is possible to achieve stability problems against heat with enhancement of the antimicrobial effect.

Hereinafter, the present invention will be described in detail.

The present invention is largely composed of (a) lactic acid bacteria culture and heat treatment step and (b) the adsorption and mixing step of the lactic acid bacteria culture solution is completed the heat treatment.

Lactic acid bacteria culture and heat treatment step of step (a) can be carried out divided into three steps,

As a first step, conventional lactic acid bacteria (Lactobacillus acidophilus, L.brevis, L.casei, L.fermentum, L.lactis, L.plantarum, L.bulgaricus, L.reuteri, etc.) approved by the US National Feed Ingredients Association After 6 to 12 hours of incubation at 34 ° C. to 37 ° C. using a culture medium, betaine was added thereto, followed by heat treatment at 60 ° C. to 70 ° C. for 10 to 30 minutes.

At this time, betaine used in the present invention is a kind of alkaloids extracted from sugar beet or sugar cane, and acts as an osmolyte to protect the lactic acid bacteria during heat treatment and activate the vitality of the lactic acid bacteria in the subsequent culture. Therefore, by adding a certain amount of betaine during heat treatment, it is estimated that the antimicrobial effect of the antimicrobial peptide material is increased by lactic acid bacteria, thereby enhancing the antimicrobial effect. The amount of the bitine added is preferably about 0.005 to 0.2 parts by weight with respect to 1 part by weight of the culture medium, when less than 0.005 parts by weight can not fully exert the effect of the lactic acid bacteria as an osmotic pressure protection agent during heat treatment, if exceeding 0.2 parts by weight Overuse is undesirable.

 In the second step, the appropriate amount of yucca extract was added to the culture solution, which was heat-treated as in the first step, incubated again for 6 to 12 hours at 34 ° C. to 37 ° C., and then, again, betaine was added, and this time at 70 ° C. to 80 ° C. Heat-treat for 10-30 minutes. Yucca extract (yucca extract) used in the present invention is a natural substance extracted from a plant called Yucca schidigera, which mainly lives in the Americas, and should contain at least 5% of sarsaponin. Yucca extract is used for animal odor removal, animal growth promotion, meat quality improvement and feed efficiency improvement in the livestock field, and in the present invention, it was used to prevent the aggregation of cells during heat treatment and to promote the growth of surviving lactic acid bacteria. The addition amount of the yucca extract is preferably about 0.01 to 0.05 parts by weight based on 1 part by weight of the culture medium, when less than 0.001 parts by weight is insufficient to prevent the aggregation of lactic acid bacteria, and when the excess exceeds 0.05 parts by weight is preferable to use excessively. I can't. The amount of bitine added is preferably 0.005 to 0.2 parts by weight based on 1 part by weight of the culture as in the first step.

As a third step, add the appropriate amount of Yucca extract to the culture solution after the second heat treatment as in the second step, incubated for 6 to 12 hours at 34 ℃ to 37 ℃ and sterilized for 15 minutes to 30 minutes at 121 ℃ (a Prepare the sterile lactic acid bacteria culture medium, which is the final product of step). The amount of the yucca extract is preferably 0.001 to 0.05 parts by weight based on 1 part by weight of the culture as described in the paragraph.

The step (b) adsorption and mixing can be carried out divided into two stages,

As the first step, the bactericidal lactic acid culture medium obtained in the third step (a) is adsorbed while being sprayed onto a common adsorbent (for example, zeolite, silicon dioxide, methyl cellulose, etc.). The amount of the adsorbent is preferably about 4 to 9 parts by weight based on 1 part by weight of the bactericidal lactic acid bacteria culture medium. When the amount of the adsorbent is less than 4 parts by weight, the moisture content is high, and when the amount of the adsorbent exceeds 9 parts by weight, the amount of the adsorbent is required. Many are not desirable.

In the second step, the adsorbed sterilized lactic acid culture medium obtained in the first step and the usual excipients (e.g. chaff powder, powder, starch, lactose, etc.) were added to the mixer and stirred at 35 RPM for 10 to 20 minutes. Lactobacillus preparations can be prepared for enhanced antimicrobial effect. The amount of the excipient added is preferably about 4 to 19 parts by weight based on 1 part by weight of the bactericidal lactic acid bacteria adsorbate. When the amount of the excipient is less than 4 parts by weight, the amount of the excipient added to the feed may cause problems in mixing. If it exceeds the weight part, it is not preferable because the amount used when added to the feed may cause inconvenience when mixing.

Hereinafter, the present invention will be described in more detail based on Examples and Comparative Examples, but the present invention is not limited thereto.

[Examples and Comparative Examples]

<Example 1>

After aseptically inoculating 2% (v / v) of Lactobacillus acidophilus KCTC 3164 starter in MRS medium and incubating for 8 hours at 37, sterilized bitine was added to 0.01 part by weight of the culture medium. After adding the parts by weight, the first heat treatment at 65 ℃ for 10 minutes and lowered to 37 ℃, and then added 0.01 parts by weight of sterilized yucca extract to 1 part by weight of the culture and incubated for 8 hours at 37 ℃ and then again sterilized Phosphorus was added 0.01 part by weight based on 1 part by weight of the culture solution, followed by secondary heat treatment at 75 ° C. for 20 minutes. Next, 0.01 parts by weight of sterilized yucca extract was added to 1 part by weight of the culture medium, followed by incubation at 37 ° C. for 8 hours, and sterilization at 121 ° C. for 15 minutes to prepare a bactericidal lactic acid bacteria culture medium.

<Example 1-1>

7 parts by weight of zeolite was added to 1 part by weight of the bactericidal lactic acid bacteria culture medium prepared in <Example 1>, and the bactericidal lactic acid culture medium adsorbed was prepared by spraying the bactericidal lactic acid culture medium while stirring at 30 RPM. 12 parts by weight of rice hull powder was added into the mixer with respect to 1 part by weight of the lactic acid bacteria culture medium adsorbate, and mixed for 15 minutes at a speed of 35 RPM to prepare a lactic acid bacteria preparation for feed addition.

<Comparative Example 1>

Lactobacillus acidophilus KCTC 3164 starter was inoculated 2% (v / v) aseptically into MRS medium and incubated at 37 ° C for 8 hours, followed by primary heat treatment at 65 ° C for 10 minutes. Next, the culture solution was incubated at 37 ° C. for 8 hours and then subjected to secondary heat treatment at 75 ° C. for 20 minutes. Next, the culture was incubated for 8 hours at 37 ℃ sterilized for 15 minutes at 121 ℃ to prepare a bactericidal lactic acid bacteria culture.

<Comparative Example 1-1>

Lactobacillus lactic acid bacteria culture solution prepared in <Comparative Example 1> was prepared in the same manner as in <Example 1-1> to prepare a lactic acid bacteria preparation for feed addition.

<Comparative Example 2>

Lactobacillus acidophilus KCTC 3164 starter was inoculated 2% (v / v) aseptically into the MRS medium and then cultured at 37 ℃ for 24 hours to prepare a lactic acid bacteria culture solution without the bactericidal treatment.

<Comparative Example 2-1>

The lactic acid bacteria culture solution that had not been sterilized in <Comparative Example 2> was prepared in the same manner as in <Example 1-1> to prepare a lactic acid bacteria preparation for feed addition.

Experimental Example 1

Comparison of Antimicrobial Effects of Lactic Acid Bacteria Cultures

1. Test Method

Kirby-Bauer disk diffusion method was used to measure and compare the antimicrobial activity of each lactic acid bacteria culture medium prepared in Example 1, Comparative Example 1 and Comparative Example 2 of the present invention. E. coli and Salmonella gallinarum were used as test strains. Dilute the cultured test solution so that the concentration of bacteria is 1 × 10 ⁸ CFU / ml, drop 100-150 μl onto each plate agar, and spread it evenly with a sterile triangular spreader. Leave for 3-5 minutes to allow the surface to absorb moisture, and within 15 minutes, put a 6.35mm diameter 1.5mm paper disk on the medium, and then culture the lactic acid bacteria in each of Examples 1, 1 and 2. Was inoculated 50 times on each original plate and left for 30 minutes, and the plate lid was placed in the incubator at 37 ° C. under the culture for 18 hours, and the diameter of the bacterial growth inhibitor around the plate was measured using a ruler.

2. Test result

Example 1, Comparative Example 1 and Comparative Example 2 all showed antimicrobial activity against test strains E. coli and Salmonella. However, Comparative Example 1 was excellent in antimicrobial activity compared to Comparative Example 2, Example 1 was excellent in antimicrobial activity compared to Comparative Example 1, the antibacterial effect was the most excellent. The experimental results are shown in Table 1.

Size of Antimicrobial Activation (Unit: nm) Escherichia coli Salmonella Galinarum Example 1 11.1 10.3 Comparative Example 1 10.4 9.5 Comparative Example 3 9.8 8.3

Experimental Example 2

Disease Prevention Effect of Lactic Acid Bacteria for Feed Additives in Chickens

1. Experimental method

Five-day-old broiler chickens (Cobbs) were divided into four groups of 24 each of 24 animals, and the control group was not added to broiler electric feed, and the test group 1 added 0.2% of the lactic acid bactericide for feed addition prepared in Example 1-1 0.2% of the feed additive lactic acid bacteria prepared in Comparative Example 1-1 was added, and test 3 group was fed up to 25 days of age by adding 0.2% of the lactic acid bacteria for feed additive prepared in Comparative Example 2-1. In order to measure the disease prevention effect of each feed supplement lactobacillus against Salmonella, the number of mortality was measured until the end of the test (25 days old) after artificially infecting Salmonella gallinarum bacteria by 10 ⁸ CFU per horse to all chicks at 15 days of age. It was.

2. Test result

In the control group that did not receive the feed supplement lactobacillus, a total of 10 animals died and showed a mortality rate of 41.7%. A total of 5 animals died in the group, resulting in 20.8% mortality. On the other hand, in Test 2 group, which was fed and fed Lactobacillus lactic acid bactericide, which had been added to the feed, was fed 4 rats, resulting in 16.7% mortality. The total number of mortality in test 1 group fed Example 1-1 to feed was 3, showing 12.5% mortality. Therefore, lactobacillus for general feed supplementation without mortality of feed group lactobacillus supplemented with bactericidal treatment by adding betaine and yucca extract (Example 1-1) (Comparative Example 2-1) The mortality rate was 8.3% lower than that of the broilers fed the diet, and the mortality was 4.2% lower than those fed the lactobacillus supplements treated with sterile feed (Comparative Example 1-1). The experimental results are shown in [Table 2].

The number of trials Our number Mortality Control 24 10 41.7% Test group 1 24 3  12.5% Test 2 group 24 4 16.7% Test Group 3 24 5 20.8%

Experimental Example 3

Disease Prevention Effect of Lactic Acid Bacteria for Feed Additives in Pigs

1. Experimental method

After weaning, we randomly harvest 180 heads of weaned pigs (Landrace × Yorkshire × Duroc) of A pig farm in Gyeonggi-do, which is known to be associated with various viral and bacterial digestive diseases and respiratory diseases. The control group was added to the piglet feed, divided into four groups of 45 animals each, and the test group 1 was 0.2% of the lactic acid bacteria preparation for feed addition prepared in Example 1-1, and the test group 2 was added to the feed prepared in Comparative Example 1-1. Lactobacillus agent 0.2%, test 3 groups were fed up to 68 days of age by adding 0.2% each of the lactic acid bacteria agent for feed additive prepared in Comparative Example 2-1. The weight change, feed intake, feed demand, atrophy and mortality of piglets were measured during the entire test period. The cause of mortality was necropsy and the tissues with lesions were identified by microbiological and pathological examination. The clinical symptom index was 0: normal, 1: mild to moderate depression, 2: severe depression, refusal to exercise, neurological symptoms and atrophy; 20: mortality was observed at regular times daily for clinical symptoms score × Number of heads = clinical symptom cumulative index. All piglets were tested under the same breeding conditions and control of the pig farm during the pre-test period. Feeding was in accordance with the farm feeding program and negative drinking was allowed.

2. Test Results

1) Feed efficiency improvement effect

The mean weight gain per head during the trial period (from 21 to 68 days of age) was 17.9 kg from the control group fed the feed without the addition of lactobacillus supplements, and the lactobacillus supplements without the sterilization treatment (Comparative Example 2-1). Test 3 group fed with feed to feed was 19.3kg, Lactobacillus feed supplemented with sterilized feed (Comparative Example 1-1) was added to feed and tested 2 group fed 19.6kg, Feed added with sterilized feed The test 1 group fed with a lactic acid bacteria preparation (Example 1-1) to feed was 20.3 kg. On the other hand, the average feed intake per head during the test period (from 21 to 68 days of age) was 39.9 kg in the control group fed the feed without the addition of the lactic acid bacterium for the feed, and the lactic acid bacterium for the feed addition without the sterilization treatment (Comparative Example 2 Test 3 group fed with -1) to feed was 40.7kg, and test 2 group fed and supplemented with lactic acid bactericide for feed sterilization (Comparative Example 1-1) was 40.6kg and sterilized. The test 1 group fed with a feed supplemented lactic acid bacterium (Example 1-1) to the feed was 40.2 kg, and the control group was 2.23, the test group 3 was 2.11, the test 2 Group 2.07, Test 1 group 1.98, pig group fed feed lactic acid bacteria was improved as compared to the control group as a whole Example 1-1 Test 1 group fed the lactic acid bacteria for feed additives improved the most.

The experimental results are shown in [Table 3].

Control Test group 1 Test 2 group Test Group 3 Test head (two) 45 45 45 45 Starting weight per head (kg) 5.8 5.7 5.7 5.9 Average head weight per head (kg) 23.7 26.0 25.3 25.2 Average head glucose gain (kg) 17.9 20.3 19.6 19.3 Soy sugar intake (kg) 39.9 40.2 40.6 40.7 Feed demand rate (FCR) 2.23 1.98 2.07 2.11 % Improvement over control 11.2 7.2 5.4

2) Clinical symptoms index and mortality reduction effect

The cumulative index of clinical symptoms for each group was 202, the control group fed the feed without the addition of lactic acid bacteria for feed, and the feed supplemented with the addition of the lactic acid bacteria for feed addition (Comparative Example 2-1). Group 139, Lactobacillus for Feed Addition (Comparative Example 1-1), added to the feed and fed to Group 2, Group 117, Lactobacillus for Feed Additive Treated (Example 1-1) The test 1 group fed to the diet was 62. The mortality rate of each group was 16.7% in the control group fed the feed without the addition of lactic acid bacteria for feed, and the test 3 group supplemented with the addition of the lactic acid bacteria for the feed without the sterilization (Comparative Example 2-1). 11.1%, Lactobacillus for feed supplementation (Comparative Example 1-1) supplemented with feed was added to the feed, and Test 2 group fed 8.9%, Lactobacillus for feed supplementation (Example 1-1). In the test group, 4.4% were added to the diet. Overall, the experimental group showed lower clinical cumulative index and mortality rate by PMWS than the control group. The results are shown in [Table 4].

Test head Cumulative Index of Clinical Symptoms * Our head Mortality (%) Cause of death Pathogen 0 One 2 3 20 system Control 45 0 11 16 15 160 202 8 16.7 PMWS Glasser's disease PCV-2, PRRSV, H. parasuis , Sal . typhimurium B. dysenteria M. hyopneumoniae P. multocida Test group 1 45 0 6 10 6 40 62 2 4.4 PMWS PCV-2, PRRSV, H. parasuis , M. hyopneumoniae P. multocida Test 2 group 45 0 7 18 12 80 117 4 8.9 PMWS PCV-2, PRRSV, H. parasuis , M. hyopneumoniae P. multocida Test Group 3 45 0 10 11 18 100 139 5 11.1 PMWS PCV-2, PRRSV, H. parasuis , M. hyopneumoniae P. multocida

*: Two heads with clinical symptoms × clinical symptoms index,

**: 0; Normal, 1; Anorexia, 2; Moderate to moderate depression,

    3; Severe depression, motor refusal, neurological symptoms and atrophy, 20; Our company.

As described above, the lactic acid bacteria were incubated for a predetermined time, followed by heat treatment with the addition of betaine, followed by re-cultivation with the addition of the yucca extract, followed by sterile treatment, adsorption and mixing with the adsorbent and the excipient to feed the lactic acid bacteria. By preparing the formulation, the antimicrobial effect can be enhanced and the stability against heat can be overcome.

Figure 1 shows the manufacturing process of the lactic acid bacteria preparation for the enhanced feed antimicrobial effect according to the present invention.

2 is a structural formula showing the chemical structure of bitine according to the present invention.

Claims (5)

In lactic acid bacteria culture which cultured normal lactic acid bacteria, Lactic acid bacteria culturing and heat treatment step of producing a bactericidal lactic acid bacteria culture medium by adding a constant bitine and yucca extract at a constant temperature and a predetermined time to the culture medium cultured normal lactic acid bacteria; The method of producing a lactic acid bacteria composition for feed supplements with enhanced antibacterial effect, characterized in that the adsorbed by spraying the adsorbent to the cultured sterilized lactic acid bacteria culture medium is completed, the adsorbing and mixing step of the excipient. The method of claim 1, Lactobacillus culture and heat treatment step A first culture solution in which ordinary lactic acid bacteria are cultured; After incubating the first culture solution at 34 ° C. to 37 ° C. for a predetermined time, 0.005 to 0.2 parts by weight of betaine was added to 1 part by weight of the culture solution, followed by heat treatment at 60 ° C. to 70 ° C. for 10 minutes to 30 minutes, and then 34 ° C. to 37 ° C. A second culture solution lowered to ℃; A third culture solution added with 0.001 to 0.05 parts by weight of yucca extract based on 1 part by weight of the second culture solution and incubated at 23 ° C. to 37 ° C. for a predetermined time; A fourth culture solution lowered from 34 ° C. to 37 ° C. after heat treatment at 70 ° C. to 80 ° C. for 10 minutes to 30 minutes, followed by the addition of 0.005 to 0.2 parts by weight of betaine based on 1 part by weight of the third culture solution; Addition of 0.001 to 0.05 parts by weight of yucca extract per 1 part by weight of the fourth culture solution and incubated for a predetermined time at 34 ℃ to 37 ℃ and sterilized lactic acid bacteria culture medium which is a fifth culture sterilized 15 minutes to 30 minutes at 121 ℃ Lactobacillus formulations for feed supplements, characterized in that the antimicrobial effect is enhanced and a manufacturing method thereof. The method of claim 1, Adsorption and mixing steps An adsorption step of adsorbing 4 to 9 parts by weight of the adsorbent based on 1 part by weight of the bactericidal lactic acid bacteria culture medium; The antimicrobial effect-enhanced feed additive lactic acid composition and its preparation method characterized in that it is mixed with 4 to 19 parts by weight of the usual excipient with respect to 1 part by weight of the bactericidal lactic acid bacteria culture medium adsorbate. After aseptically inoculating 2% (v / v) of Lactobacillus acidophilus KCTC 3164 starter in MRS medium and incubating for 8 hours at 37, sterilized bitine was added to 0.01 part by weight of the culture medium. After adding the parts by weight, the first heat treatment at 65 ℃ for 10 minutes and lowered to 37 ℃, and then added 0.01 parts by weight of sterilized yucca extract to 1 part by weight of the culture and incubated for 8 hours at 37 ℃ and then again sterilized Phosphorus was added 0.01 part by weight based on 1 part by weight of the culture medium, followed by secondary heat treatment at 75 ° C. for 20 minutes. Next, 0.01 parts by weight of the sterilized yucca extract was added to 1 part by weight of the culture medium, incubated for 8 hours at 37 ° C., and sterilized for 15 minutes at 121 ° C. to enhance the antibacterial effect of the cultured lactic acid bacteria. Lactic acid bacteria composition for feed addition. The method of claim 4, wherein 7 parts by weight of zeolite per 1 part by weight of the bactericidal lactic acid bacteria culture medium was added to the mixer, and the mixture of the bactericidal Lactobacillus culture medium was prepared by spraying the bactericidal Lactobacillus culture medium while stirring at 30 RPM, and then 1 part by weight of the bactericidal Lactobacillus culture medium adsorbate. Addition of 12 parts by weight of rice hull powder into the mixer for mixing for 15 minutes at 35 RPM speed was added to the lactic acid bacteria formulation for enhanced feed antimicrobial effect, characterized in that the composition was added.

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