KR100500397B1 - Bacillus strain having antibiotic activity against Salmonella gallinarum inducing fowl typhoid and a mixed microbial agent comprising the strain - Google Patents

Abstract

The present invention relates to a strain having antimicrobial activity against Salmonella gallinalium, which is a causative organism of poultry typhoid, and a composite microbial agent for poultry containing the same as an active ingredient, and a livestock manure or soil having high antimicrobial activity against Salmonella gallinalium, which is a causative organism of poultry typhoid. There is an excellent effect to provide novel Paenibacillus polymyxa Gu88 strain, Bacillus subtilis Gu24 and Lctobacillus plantarum Hans 1-19 strain isolated from. In addition, the composite microbial agent of the present invention has high antimicrobial activity against poultry typhoid causative bacteria and can be used as an antibiotic substitute for animals, and when the feed is 0.2% mixed, it reduces the number of Salmonella and E. coli in the fecal total of laying hens. The mortality can be reduced, and the egg laying rate, egg shell thickness and egg yolk are excellent.

Description

Bacillus strain having antibiotic activity against Salmonella gallinarum inducing fowl typhoid and a mixed microbial agent comprising the strain}

The present invention relates to a strain having antimicrobial activity against Salmonella gallinalium, which is a causative organism of poultry typhoid, and a complex microbial preparation for poultry containing the same as an active ingredient. More specifically, the present invention isolates and formulates microorganisms having antimicrobial activity against Salmonella gallinalium, the causative agent of poultry typhoid, from livestock manure, soil, feces, etc. to inhibit or kill the growth of poultry typhoid causative bacteria, The present invention relates to a poultry complex microorganism for improving egg production and mortality.

In late August 1992, poultry typhoid fever occurred in Korea at 150,000 chicken farms in Gimpo-gun, and spread almost nationwide. In the hot summer months, the damage appeared to be very serious. As pollution increases, the frequency of occurrence in other seasons is also increasing. Vaccines and antimicrobial agents were used to suppress the spread of the disease, but it was very difficult to control. The domestic poultry typhoid causative organism is very pathogenic, and in most cases, once a farm is almost empty until all the chickens in the farm are empty. The onset persisted, and despite the post-treatment using various means of prevention, it failed to prevent and treat effectively.

Currently, the use of antibiotics for poultry, cows, etc. is prohibited, the hormonal drugs are also prohibited, and the US Food and Drug Administration (FDA) is prohibited to use antibiotics such as penicillin or teramicin to promote animal growth. In addition, consumers' desire for safe and hygienic livestock products is increasing, and the government has also set up a policy to strengthen inspections, and concentrators are becoming more apprehensive about using probiotics instead of antibiotics and antimicrobial agents due to anxiety about residual substances. Mainly, began to be a probiotic is highlighted containing Lactobacillus genus (Lactobacillus), Bacillus (Bacillus), Claus tree dumsok (Clostridum), yuyonggyun of yeast and the like.

Control measures for poultry typhoid caused by Salmonella gallinarum include hygiene-based disinfection methods, immunological methods using vaccines and competitive removal of Salmonella by probiotic administration.

Among them, the competitive eviction method of Salmonella using probiotics is to administer various beneficial probiotic cultures to newborn chicks to induce the intestinal tract of these chicks into a beneficial bacterial flora, thereby preventing the colonization of Salmonella from the natural environment or feed in advance.

With this in mind, the present inventors have isolated microorganisms having antimicrobial activity against poultry typhoid causative bacteria from livestock wastewater, soil, feces, and formulating them to treat the microbial agent on the laying hens, thereby producing Salmonella gallinalium ( Salmonella gallinarum ) In addition to inhibiting or killing growth, there has been provided a complex microbial agent that improves egg production and mortality.

Accordingly, an object of the present invention is to provide a strain having antimicrobial activity against poultry typhoid causative bacteria isolated from livestock wastewater, soil, feces, and the like.

Another object of the present invention is a complex microbial agent containing live microorganisms or metabolites that can prevent and treat poultry typhoid, improve the intestinal microflora, improve egg laying and mortality of laying hens, and improve egg shell thickness and egg yolk color. To provide.

Still another object of the present invention is to provide a composition for poultry feed containing a microbial agent as an active ingredient effective for the prevention and treatment of poultry typhoid.

The above object of the present invention includes a strain obtained after separating and identifying a strain having excellent grain adsorption drying ability in livestock manure or soil, and having high acid, bile and heat resistance, and excellent antibacterial activity against Salmonella gallinalium. A composite microbial preparation was prepared and added to the feed and fed to laying hens to investigate the protective effect against poultry typhoid, the effects on laying rate and mortality of laying hens, and egg shell thickness and egg yellow color.

Hereinafter, the present invention will be described in detail.

The present invention is an isolation and identification step of bacteria having antimicrobial activity against Salmonella gallinalium in livestock manure or soil; Preparation of the complex microbial agent comprising a strain isolated from the above; Comprising the protective effect of poultry typhoid fever, egg laying rate and mortality of laying hens, egg thickness and egg yellow color of the laying hens.

Paenibacillus polymyxa Gu88 KCTC 10498BP strains having high antimicrobial activity against Salmonella gallinalium of the present invention are positive for gram staining, form spores, have resistance to heat, acid and bile, and have motility In the present invention, the composite microbial agent contains 6 to 10 Log cfu / g, preferably 6 Log cfu / g.

Bacillus subtilis Gu24 KCTC 10496BP strain having high antimicrobial activity against Salmonella gallinalium of the present invention has good resistance to heat, acid and bile, and produces a lot of useful enzymes. Tracin (Bacitracin), the composite microbial agent of the present invention is characterized by containing 7 to 10 Log cfu / g, preferably 7 Log cfu / g.

Lactobacillus plantarum Hans 1-19 KCTC 10497BP strain of the present invention has high antimicrobial activity, and the present invention contains 7-10 Log cfu / g, preferably 7 Log cfu / g. Unlike other lactic acid bacteria, it is easy to cultivate in the industrial field, and it is excellent in acid resistance, bile resistance, and heat resistance, so that many bacteria survive in the intestine. It is characterized by contributing to the normalization of intestinal flora.

Saccaromyces cerevisiae ( Saccaromyces cerevisiae ) CYC-809 KCTC 8910P strain included in the complex microbial agent of the present invention is a strain that was distributed in the microbial deposit center in the Korea Research Institute of Bioscience and Biotechnology 7 ~ 10 Log cfu / g is contained, preferably 7 Log cfu / g, and added to the formulation to help the growth of the strain of the present invention, the bacteria anaerobic the intestinal environment by generating CO ₂ using harmful oxygen in the intestine It contributes to the composition and has the characteristic of helping the growth of lactic acid bacteria by this action. Saccaromyces cerevisiae , which is killed in the intestine, is a polysaccharide that elutes the oligosaccharides found on the cell walls of yeast and binds them with harmful bacteria to excrete it in vitro.It also contains abundant amino acids, vitamins, minerals and unknowns. Has growth factors

Salmonella gallinalium bacteria, which are the causative organisms of poultry typhoid used in the embodiments of the present invention, are characterized in that the present inventors use them separately from the wild.

The composite microbial agent of the present invention is Paenibacillus polymyxa Gu88 KCTC 10498BP, Bacillus subtilis Gu24 KCTC 10496BP, Lactobacillus plantarum Hans 1-19 KCTC 10497BP and Sakaroma Each Saccaromyces cerevisiae CYC-809 KCTC 8910P strain was incubated alone, mixed with the excipient grains, and aged at a predetermined temperature, and then dried in a hot air dryer to 12% or less of moisture content, respectively. It is crushed according to the 20 kg unit characterized in that the packaging.

The composite microbial agent of the present invention is added to the poultry feed in an amount of 0.2%, and may be used regardless of the type or form of the feed.

In the feed containing the complex microbial agent of the present invention as an active ingredient, the complex microbial agent is characterized in that it comprises any of live microorganisms or dry microbial powder or culture products or metabolites thereof.

Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

EXAMPLE

Example 1 Screening of Microbial Strains of the Invention

In order to isolate strains having antimicrobial activity against Salmonella gallinalium , a causative fungus of the present invention, to isolate lactic acid bacteria including Bacillus spp. Species from livestock manure or soil and to measure antimicrobial activity- It was used by three subcultures if necessary while storing at 78 ℃. Three passages were inoculated in tryptic soy broth, incubated at 37 ° C for 24 hours, centrifuged, the supernatant was filtered and sterilized, and 0.5 mL was added to 4.5 mL of sterile TSB medium. 10%) was inoculated with Salmonella gallinalium 1x10 ⁵ cfu / mL incubated for 18 hours at 37 ℃ was selected for the bacteria that Salmonella gallinalium do not grow. The selected strains were identified using Gram staining, morphology observation, sporulation, motility test and Bergey's Manual of Systematic Bacteriology and API 50 CHL kit and API 50 CHL database V4.0. In addition, the unknown bacteria were combined with the analysis method of the nucleotide sequence of 16S rRNA.

As a result, it was confirmed that the strains exhibiting the strongest antimicrobial activity were Paenibacillus polymyxa (FIG. 1), Bacillus subtilis (FIG. 2), and Lactobacillus plantarum . Among the strains, the penivacillus polymysia was named Paenibacillus polymyxa Gu88 and was deposited on July 10, 2003 with the Genetic Resource Center Gene Bank of Korea Research Institute of Bioscience and Biotechnology and received accession number KCTC 10498BP. In addition, Bacillus subtilis was named Bacillus subtilis Gu24 and deposited on July 10, 2003 with the Korea Institute of Biotechnology Genetic Center Gene Bank and received accession number KCTC 10496BP. The Lactobacillus plantarium strain was named Lactobacillus plantarum Hans 1-19 and was deposited with the Korea Institute of Biotechnology Genetic Center Gene Bank on July 10, 2003 and received accession number KCTC 10497BP.

The characteristics and substrate availability of the Paenibacillus polymyxa Gu88 strain are shown in Tables 1 and 2.

division Characteristic Sporulation Forming motility has exist Gram Dyeing positivity Culture characteristics Saponification of the culture solution

temperament Usability temperament Usability temperament Usability Glycerol + Inositol - Inulin + Erythritol - Mannitol - Melechitozu - D-arabinose - Sorbitol - D-Raffinose + L-arabinose + α-methyl-D-mannoside - Amidodon + Ribose + α-methyl-D-glucoside + Glycogen + D-Xylose + N-acetyl glucosamine - Xylitol - L-Xylose - Amigdalin + β-genthiobioses + Adonitol - Arbutin + D-turanoose + β-methylxyloxide + Estulin + D-Rizozu - Galactose + Salinity + D-Tagatozu - D-glucose + Cellobios + D-fucose - D-Plactose + Maltose + L-putose - D-Mannose + Lactose + D-Arabitol - L-Soboz - Melibiose + L-Arabitol - Rhamnose - Sakaros + Gluconate - Dulcitol - Threhalose + 2 Seto Gluconate - 5 Seto Gluconate -

In the table, + is positive and-is negative (using API 50 CHL kit).

Example 2: The present invention penivacillus polymyisha ( Paenibacillus polymyxa Investigation of Antimicrobial Activity against Salmonella gallinalium by Gu88 Strains

The antimicrobial activity of Salmonella gallinalium of the present invention, Paenibacillus polymyxa Gu88 strain isolated in Example 1 was investigated according to the paper disk method. 1x10 ⁵ cfu / mL of Salmonell gallinarum ( Salmonell gallinarum ) was cultured on a Tryptic Soy Agar (TSA) plate medium to filter the supernatant of the Paenibacillus polymyxa Gu88 strain and sterilized by 8 mm paper disc. When the antimicrobial activity was measured by dropping to 40 μl. The results are shown in FIG.

Comparative Example 1 of the present invention Penivacillus polymyisha ( Paenibacillus polymyxa ) Antimicrobial Activity of Gu88 Strains (I)

The culture supernatant of the Paenibacillus polymyxa Gu88 strain was added to a test tube containing 2 mL of Tryptic Soy Broth (TSB) medium containing 10 ⁵ cfu / mL of Salmonell gallinarum . 0.3% and 0.5% were added to the incubator in which the temperature was adjusted to 25 ° C., 30 ° C. and 37 ° C., respectively, and cultured for 24 hours. The results are shown in FIG. 5.

At this time, A1 adds 0.3% of the test solution (25 ℃), A2 adds 0.5% of the test solution (25 ℃), B1 adds 0.3% of the test solution (30 ℃), B2 adds 0.5% of the test solution (30 ℃), and C1 adds the test solution. 0.3% addition (37 ℃), C2 0.5% addition of the test solution (37 ℃), D was the result of incubating TSB medium containing only 10 ⁵ cfu / mL of Salmonella gallinalium (Salmonell gallinarum ) without 37 Is shown in the photo.

Comparative Example 2: The present invention penivacillus polymyisha ( Paenibacillus polymyxa Investigation of Antimicrobial Activity of Gu88 Strains (II)

In order to investigate whether the present invention, Paenibacillus polymyxa Gu88 strain kills Salmonell gallinarum (bacteriocidal), 0.5% of the strain culture solution and Salmonella 1x10 ⁵ cfu / mL were mixed for 2 hours. After incubation, the sample was taken, diluted, and plated on a plate medium.

As a result, as shown in Figure 6, Salmonella lost its viability when in contact with the test solution, it can be seen that the present invention, the peniebacillus polymyxa Gu88 strain kills Salmonella.

Comparative Example 3: Investigation of antimicrobial activity against wild type Salmonella of the present invention strain (III)

In order to determine whether the present invention can inhibit the Salmonella gallinalium bacteria directly isolated from the field in Tryptic Soy Broth medium, Salmonella bacteria and competitive culture.

As a result, as shown in Figure 7, the present invention, Paenibacillus polymyxa Gu88 strain was shown to completely kill Salmonella gallinalium.

Comparative Example 4: The present invention penivacillus polymyisha ( Paenibacillus polymyxa ) Gu88 strain and Bacillus subtilis ( Bacillus subtilis Investigation of Antimicrobial Activity against Salmonella from Gu24

As a result of mixing and culturing the penicillin polymysia Gu88 bacteria and Bacillus subtilis Gu24 strain in a Erlenmeyer flask containing Salmonella gallinalium bacteria with 10% sterile feed, as shown in Figures 8 and 9, the two strains All were shown to kill Salmonella completely after 15 hours.

Example 3 Toxicity Test of Strains of the Invention

The present invention was tested for hepatotoxicity in the ICR mice of the strains of the Penivacillus polymysia Gu88. Two-week-old ICR mice were fed and 4 animals per breeding cage were placed for 1 week to stabilize. Breeding environment was 21 ℃ ± 3 ℃, relative humidity was 40 ~ 60%, litter in the cage was changed every 3 days. The inoculation amount of the experimental strain was divided into 1.2 g of the culture medium in pellet feed to 60 g of the feed so that the inoculum of the bacteria was 0.2% of the feed and left in the feed feed container for about 20 minutes. Toxicity test was indirectly compared to the control group daily weight gain, feed efficiency and feed requirements compared to the control group, and directly or by measuring the number of red blood cells and white blood cells of each individual to determine the toxicity. Table 3 is a data showing the specification effect of the ICR mouse by the feed of each strain, Table 4 is a data showing the number of red blood cells and white blood cells in the blood of the ICR mouse by the feed of each strain. There was no difference between the control group and the blood group, which was not toxic.

                           Specification Effect Daily weight increase Feed efficiency Feed requirement Control 0.521 0.031 1.95 Penivacillus Polymyxia Gu88 0.62 0.032 1.93 Bacillus subtilus Gu24 0.62 0.031 1.95 Lactobacillus Plantarium Hans 1-19 0.63 0.032 1.93

(Unit: g)

                             Hemococci Red blood cell count (1x10 ⁶ / mL) White blood cell count (1x10 ³ / mL) Control 8.659 ± 1.436 6.439 ± 0.879 Penivacillus Polymyxia Gu88 8.320 ± 1.089 7.126 ± 1.013 Bacillus subtilus Gu24 7.245 ± 1.096 6.665 ± 1.045 Lactobacillus Plantarium Hans 1-19 8.888 ± 0.653 7.773 ± 1.675

Example 5 Preparation Process and Medium Component Content of the Complex Microorganism Preparation Including the Strain of the Invention

In order to prepare a composite microbial agent for preventing and treating poultry typhoid fever, it was prepared by mixing the subsidiary grains with the strain selected in Example 1 and drying.

First, in the case of the Penivacillus polymysia Gu88 strain and Lactobacillus plantarium Hans 1-19 strain, strain culture medium was prepared according to the medium composition of Tables 5 to 6 below and sterilized for 30 minutes at 121 ℃, and then the penivacillus poly Misha Gu88 strain and Lactobacillus plantarium Hans 1-19 strain were inoculated at 2% in the liquid medium, respectively, and incubated at 30 ° C and 37 ° C, respectively. Thereafter, jade powder, powdered powder and degreasing steel, which were mixed in a ratio of 1: 1: 1, were added at a ratio of 4: 5, and hot-air dried at 53 ° C. for 24 hours to give a final moisture content of 12% or less. Then crushed in mesh size 32 and packed in 20 kg increments.

Culture Medium Composition of Penibacillus Polymyx Gu88 Component Content (g / liter) Anhydrous Glucose 10 Soy flour 15 LS 300 One

Culture medium composition of Lactobacillus plantarium Hans 1-19 Component Content (g / liter) Cane molasses 50 Corn Steep Liquor (CSL) 50 pH 7.0

In the case of the Bacillus subtilis Gu24 strain, the solid culture grains (mixing the powder of clay, powder, and skim in a ratio of 1: 1) were sterilized at 121 ° C. for 30 minutes, and then cultured in sterile distilled water for 18 hours. (10%) of the culture solution was mixed with the sterile grains at a ratio of 4: 5 and incubated for 48 hours at 37 ° C. Thereafter, the mixed solution was hot-air dried at 53 ° C. for 24 hours to give a final moisture content of 12% or less, which was ground at a mesh size 32 and packed in 20 kg units.

Strain powder obtained from the above was added according to Table 7 to prepare a composite microbial preparation. 10 shows a manufacturing process of the composite microbial agent of the present invention.

Component Content (g / kg) Penivacillus Polymyxia Gu88 1 x ^{10 10} Lactobacillus Plantarium Hans 1-19 1 x ^{10 9} Bacillus subtilis Gu24 1 x ^{10 10} Sakaromyces Celebes 1 x ^{10 10} Jade powder 333 g Powder 333 g Degreasing Steel 333 g

Example 6 Protective Effect of the Complex Microorganisms of the Invention Against Poultry Typhoid

The protective effect of poultry typhoid challenge inoculation was investigated by feeding the microbial preparation prepared in Example 5 to the herbivorous chicks. When the microbial agent of the present invention was added 0.4% to the feed and treated with a natural feed and a control without adding anything to the feed, when artificially inoculated 10 ⁸ cfu / grain of Salmonella gallinalium, the causative agent of poultry typhoid The protective effect of the strain selected in 1 was performed.

Microbial Supplement Microbial Supplements Starting head Lung Factor Final survival Starting head Lung Factor Final survival Primary 12 0 12 12 7 5 Secondary 12 4 8 12 8 4 3rd 12 6 6 12 7 5 gun 36 10 26 36 22 14

As shown in Table 8, 10 of 36 pigs were killed and 27.8% were killed, whereas the control group of non-paying rats was killed at 61%. The protective effect was protected in 12 of the 22 lung pulmonary system in the unpaid group, which was 55%.

Example 7 Investigation of the Effect on the Laying Rate of Laying Hens

In order to investigate the effect on the laying rate of the laying hen of the present invention composite microorganism prepared in Example 5, 1 dong of 10,000 s were selected from Samju Farm, a 70,000 hen laying hen farm located in Dokjeong-ri, Hwaseong-gun, Gyeonggi-do, Korea. This experiment was performed. The control period ranged from 51 weeks of age (July 9, 2002) to 53 weeks of age. The actual test period was about 3 months from August 16, 54 weeks, to November 11, 68 weeks. No antibiotics were used during the experiment, and the amount of probiotics used was adjusted to a final concentration of 0.2% by mixing 2 kg per tonne of feed. The farming environment of this farm was about the same as other laying farms.

As shown in FIG. 11, the laying rate of laying hens was improved by 5% on average when the present invention was administered with the composite microbial agent.

Example 8 Effect on Mortality of Laying Hens of the Inventive Complex Microorganisms

The mortality of the laying hens was investigated after feeding the inventive microbial microorganisms according to Example 7.

As a result, as shown in Figure 12, the mortality of laying hens when the present invention is fed a composite microbial agent showed a tendency to decrease.

Example 9 Investigation of the Effect of Fecal Bacteria on the Complex Microorganisms of the Present Invention

In order to investigate the effect on the fecal bacteria of the present invention the microbial agent, the number of bacteria produced in Salmonella culture medium and the number of bacteria produced in the lactic acid bacteria culture medium and TSB medium of the fecal bacteria of laying hens according to Example 7 was measured It was.

As a result, as shown in FIG. 13, the number of lactic acid bacteria and the total number of bacteria grown in TSB medium were kept constant, but the number of colonies formed in E. coli and Salmonella medium decreased significantly after feeding.

Example 10: Investigation of the effect of the present invention on the egg shell thickness and egg yolk color when laying the laying hens

The effects of the present invention on the egg yolk thickness and egg yolk color of the fed microbial preparations were investigated in Iljuk Farm, Iljuk-myeon, Anseong-si, Gyeonggi-do. The farm was fed a compound microorganism of the present invention 0.2% of the daily feed amount to a window shed containing 30,000 of them in a total scale of 120,000.

As shown in FIG. 14, the egg shell thickness and egg yolk color were remarkably improved when the composite microbial agent of the present invention was fed.

As described in the above Examples and Comparative Examples, the present invention relates to a strain having antimicrobial activity against Salmonella gallinalium, which is a causative organism of poultry typhoid, and a composite microbial agent for poultry containing the same as an active ingredient, Salmonella gallina, which is a causative organism of poultry typhoid. Novel Penibacillus polymyxa Gu88, Bacillus subtilis Gu24, and Lctobacillus plantarum Hans 1-19 strains isolated from livestock manure or soil with high antimicrobial activity against cerium Has an excellent effect. In addition, the composite microbial agent of the present invention has high antimicrobial activity against poultry typhoid causative bacteria and can be used as an antibiotic substitute for animals, and when the feed is 0.2% mixed, it reduces the number of Salmonel and E. coli bacteria in the fecal flora of laying hens. The mortality can be reduced, and the egg laying rate, egg shell thickness and egg yolk are excellent. Therefore, the present invention is a very advantageous invention in the livestock control industry.

Figure 1 is a photograph showing the present invention Penenbacillus polymyxa Gu 88 strain having antimicrobial activity against Salmonella gallinalium isolated from livestock manure or soil.

Figure 2 is a photograph showing the Bacillus subtilis Gu 24 strain of the present invention having antibacterial activity against Salmonella gallinalium isolated from livestock manure or soil.

Figure 3 is a photograph showing the Lactobacillus plantarum Hans 1-19 strain of the present invention having antimicrobial activity against Salmonella gallinalium isolated from livestock manure or soil.

Figure 4 shows the results of measuring the antimicrobial activity against Salmonella galanalium bacteria according to the paper disk method of the present invention Penibacillus polymyxa ( Paenibacillus polymyxa ) Gu 88 strain.

5 is a photograph showing the antimicrobial activity after the mixed culture of Paenibacillus polymyxa Gu 88 strain and Salmonella gallinalium (Salmonell gallinarum) of the present invention.

FIG. 6 is a photograph showing the results obtained by mixing a culture medium of Salmonella galanalium bacteria with a culture solution of the present invention Penibacillus polymyxa Gu 88 strain and plated on a plate medium.

Figure 7 is a result showing the antimicrobial activity of the wild-type Salmonella gallinalium of the present invention Penibacillus polymysia Gu 88 strain.

Figure 8 shows the results of the mixed culture of the present invention Penibacillus polymycelium Gu 88 strain and Salmonella gallinalium.

Figure 9 shows the results of the mixed culture of Bacillus subtilis Gu 24 strain and Salmonella gallinalium of the present invention.

Figure 10 shows the manufacturing process of the composite microbial agent of the present invention.

Figure 11 shows the effect on the laying rate when fed to the present invention complex microbial laying hens.

Figure 12 shows the effect on mortality when fed to the present invention complex microbial laying hens.

Figure 13 shows the effect on the fecal bacteria when fed to the present invention complex microbial laying hens.

Figure 14 shows the effect on egg shell thickness (olk high) and yolk yellow color when fed the present invention the microbial formulations laying hens.

Claims (6)

Produces metabolites that have antimicrobial activity against Salmonella gallinalium , the causative agent of poultry typhoid, produces positive reactions to gram staining, forms spores, is motility, and cultures the cultures in culture Paenibacillus polymyxa Gu88 KCTC 10498BP. Penibacillus polymyxa ( Paenibacillus polymyxa ) Gu88 KCTC 10498BP antimicrobial composition for the antimicrobial agent characterized in that it contains the strain as an active ingredient. Paenibacillus polymyxa ( Paenibacillus polymyxa ) Gu88 KCTC 10498BP strain culture broth (mixture of powdered clay, powdered and skim steel in a ratio of 1: 1: 1) in a ratio of 4: 5 Make the content 0-12% and grind it in mesh size 32 to obtain strain powder, Three strains ( Bacillus subtilis ) Gu24 KCTC 10496BP strain, Lactobacillus plantarum Hans 1-19 KCTC 10497BP strain and Saccaromyces cerevisiae CYC-809 KCTC 8910P ) To obtain the strain powder according to the method of the above step, Mixed powder obtained by mixing four strain powders obtained above (1 × 10 ¹⁰ g of Paenibacillus polymyxa ) Gu88 KCTC 10498BP strain powder, 1 × 10 ¹⁰ g Bacillus subtilis, per kg of the combined microorganism ( Bacillus subtilis ) Gu24 KCTC 10496BP strain powder, 1 × 10 ⁹ g Lactobacillus plantarum Hans 1-19 KCTC 10497BP strain powder, 1 × 10 ¹⁰ g Saccaromyces cerevisiae CYC- 809 KCTC 8910P strain powder) is added to the grain subsidiary agent (mixes of clay, powder and skim steel in a ratio of 1: 1: 1) to prepare a composite microbial preparation, However, the four microbial strains are combined microbial preparations for the prevention and treatment of poultry typhoid, characterized in that the content of 1 × 10 ⁹ ~ 1 × 10 ⁷ cfu / g, respectively. delete A composition for poultry feed comprising the complex microbial agent according to claim 3. The composition of claim 5, wherein the complex microbial agent is added in an amount of 0.2% per total weight.