KR101665334B1 - Rhodobacter sphaeroides CB 8521 strain, having the effect of reducing malodor and immune activity in livestock industry, and microbial agent using it - Google Patents

Abstract

The present invention relates to a process for the isolation of Rhodobacter CB 8521, which is isolated from soil and has reduced livestock odor generation, pathogenic bacterial inhibition and immunological activity, sphaeroides CB 8521) (accession number: KACC92014P) and microorganisms containing the same, which reduce the odor substances generated in the livestock field, exhibit antimicrobial effect against various pathogenic bacteria, enhance immunity of livestock The productivity of livestock can be improved.

Description

(Rhodobacter sphaeroides CB 8521 strain, having the effect of reducing the malodor and immune activity in the livestock industry, and microbial agent using it)

The present invention is also to separate from the soil and the jinyeo suppression and immune activity enhancement effect of reducing odor of livestock, intestinal pathogenic bacteria to enhance the productivity of livestock bakteo sphaeroides (Rhodobacter sphaeroides CB 8521 strain and a microbial composition comprising the same.

Photosynthetic bacteria are living microorganisms that use light, minerals, organic matter, etc. as sources of energy and carbon in streams, soils and oceans. These photosynthetic bacteria have an environmental purification ability to remove harmful carbon, nitrogen, and sulfur compounds, and produce microbial materials that have very high content of proteins, trace elements, nucleic acids and vitamins of the cells themselves and inhibit harmful bacteria, And microorganisms that are highly utilized for the removal of offensive odors in housing and sewage treatment plants.

Photosynthetic microorganisms that can utilize light energy include eukaryotic cells (algae), prokaryotic cells cyanobacteria, and photosynthetic bacteria. The photosynthesis mechanism of algae and cyanobacteria is similar to the photosynthesis mechanism of plants. On the other hand, photosynthetic bacteria (PSB), which are photosynthetic bacteria only in anaerobic state, are classified into purple bacteria and green bacteria according to the produced photosynthetic pigment and electron donor (hydrogen sulfide availability) Are divided into purple nonsulfur bacteria and purple sulfur bacteria. Green bacteria are subdivided into green sulfur bacteira and green nonsulfur bacteria.

The red bacterium is a gram-negative bacterium composed of about 30 species. It is unicellular. It is generally propagated by dichotomous method, but some of it is propagated by the method of propagation. Most are monocotyled with mobility, but some are non-athletic. The red-sulfur bacteria are absolute photosynthetic bacteria that can not be photosynthetically produced under aerobic conditions, whereas most of the non-sulfur non-sulfur bacteria are chemoorganotrophs that can aerobically grow in the cow. Microorganisms that use organic matter as an energy source and carbon source )to be. Red non-sulfur bacteria generally live in freshwater lakes, ponds and soils where organic matter is present but not sulfides or are present in low concentrations. Red-sulfur bacteria and green-sulfur bacteria are only present in limited habitats, so their contribution from photosynthetic productivity in the ecosystem is negligible. Therefore, Rhodobacter sphaeroides and Rhodobacter capsulatus belonging to the Rhodospirillaceae family are widely used in industry.

Photosynthetic bacteria are used in various industrial fields such as agriculture, animal husbandry, fish culture, environmental improvement, cosmetics, and therapeutic agents. In the field of agriculture, the soil is fertile due to the nitrogen fixation by photosynthetic bacteria, and the effect of improving the rhizosphere environment and improvement of root development by the large amount of protein, trace element, nucleic acid and physiologically active substance secreted by photosynthetic bacteria . Especially, it is helped to prevent the damage of soil acidification caused by the series. In the livestock sector, it can be used to remove fatty acids such as acetic acid and propionic acid in farms, amines and hydrogen sulfide-producing odor-producing substances. It provides abundant nutrition, improves livestock immune activity and suppresses intestinal pathogenic bacteria. And increase the productivity of livestock. In addition, carotenoids produced by photosynthetic bacteria can improve the color of meat and eggs and improve meat quality. In the field of fish culture, the photosynthetic bacteria become the protein source and increase the animal plactone, thereby increasing the food of the fish, so that the hatching rate and survival rate of the fish can be improved. In the field of environmental improvement, it is suitable for treatment of high concentration wastewater and is applied to purification of fresh water, seawater and aquaculture water. In recent years, attempts have been made to use cosmetics such as skin moisturizers and treatments for skin diseases and respiratory diseases by utilizing photosynthetic bacterial cells and culture media.

Patent No. 10-1200235 relates to Rhodobacter sphaeroides, a photosynthetic bacterium having crop growth promoting activity, a culture thereof, and a microorganism including the strain and the culture, which promote the growth of crops and improve productivity Friendly microbial fertilizer that can increase the bioavailability of the microorganism. Japanese Patent Registration No. 10-0913611 discloses a feed additive and a feed having improved fish growth promotion effect, immune enhancement effect, and water quality improvement effect using Rhodopseudomonas lutira suitable for fish culture among photosynthetic bacteria. Japanese Patent Application Nos. 10-1176560 and 10-1220922 disclose culture methods and conditions capable of maximizing bio-oil or hydrogen production capacity using Rhodobacter sphaeroides, a photosynthetic bacterium. Japanese Patent Registration No. 10-1320045 discloses a cosmetic composition for alleviating skin irritation or skin moisturizing comprising an extract or a bacterium of bacterial chlorophyll-containing cells obtained from Rhodobacter sphaeroides as an active ingredient, 0973176 is directed to a method for eliminating odors that occur when photosynthetic bacteria are added to a food composition. 10-0143954, 10-0261747, 10-0381371, 10-0430298, and 10-1029346 disclose improvements in the growth and feed efficiency of livestock, reduction of odor of livestock manure, and reduction of pathogenic bacteria But the present invention is not limited to the use of photosynthetic bacterium alone but may be used as an antagonistic lactic acid bacterium such as Aspergillus which can inhibit fungi and pathogenic bacteria, Bacillus, yeast, and other microorganisms.

There are more than 30 photosynthetic bacteria in the natural world, and they have different characteristics such as the ability to produce germplasm, physiologically active substances, odor removal ability, and proliferation ability. Therefore, it is necessary to improve agricultural microorganisms, animal husbandry, And microorganisms suitable for each field are different.

Therefore, there is a demand for photosynthetic bacteria and microorganisms containing the photosynthetic bacteria suitable for selection of a microorganism for domestic animal.

It is an object of the present invention to provide a photosynthetic bacterial strain suitable for animal husbandry and a microorganism containing the same, which are effective in suppressing and reducing the production of animal offensive odors, inhibiting pathogenic bacteria, and improving productivity of livestock due to immunity enhancement.

In order to achieve the above object, the present invention provides Rhodobacter sphaeroides CB 8521 (accession number: KACC92014P), which is isolated from soil and has reduced livestock odor generating substance, pathogenic bacteria inhibition and immunological activity, do.

In addition, the present invention provides a microbial composition comprising Rhodobacter sphaeroides CB 8521 (Accession No. KACC92014P).

The composition preferably contains Rhodobacter sphaeroides CB 8521 in the range of 1.0 × 10 ⁶ cfu / g to 1.0 × 10 ¹¹ cfu / g.

The present invention also relates to a method for producing a recombinant vector comprising the steps of liquid culturing Rhodobacter sphaeroides CB 8521 (Accession No .: KACC92014P) for 40 to 48 hours;

The cultured Rhodobacter sphaeroides CB 8521 (Accession No .: KACC92014P) was added to a culture medium containing 0.1 to 2.0% casein peptone or vegetable protein, 0.5 to 3.0% yeast extract, 1.0 to 3.0% sodium acetate, 0.1 to 2.0% ammonium sulfate _{{(NH 4) 2 SO 4} }, 0.1 ~ 2.0% glutamic acid, 0.1% to 2.0% sodium succinate, 0.1% to 1.0% J. potassium phosphate _{(K 2 HPO 4), 0.001} ~ 0.05% of minerals (iron, magnesium, manganese, etc. ) And the rest of distilled water at 25 to 37 ° C for 40 to 48 hours under a humid condition, and then centrifuging the cells to recover the cells; And

Mixing the recovered cells with a cryoprotectant at a weight ratio of 1: 1 to 1: 2, followed by lyophilization to obtain a final volume of 5.0 × 10 ¹⁰ cfu / g to 3.0 × 10 ¹² cfu / g of the microorganism And a manufacturing method thereof.

In the method of the present invention, it is preferable that the cryoprotectant is at least one selected from skim milk powder, maltodextrin, lactose, wheat flour, trehalose and mannitol.

The Rhodobacter sphaeroides CB 8521 of the present invention and the microorganisms containing the same reduce the odor substances such as ammonia, hydrogen sulfide, methyl mercaptan and trimethylamine, which are frequently generated in the field of livestock farming, and reduce the odor substances such as Bacillus cereus, The antimicrobial effect against various pathogenic bacteria including NOx, TNF-α and IL, which shows antibiotics against various pathogenic bacteria including Zens, E. coli, Listeria monocytogenes, Listeria innocia, Salmonella typhimurium, Salmonella entericidis, Staphylococcus aureus, -1β activity, thereby improving the productivity of livestock by enhancing the immunity of the livestock.

1 is Rhodobacter sphaeroides (Rhodobacter sphaeroides ) CB 8521 strain by electron microscope.

Hereinafter, the present invention will be described in detail.

One. Rhodobacter Spelloides CB  8521

The Rhodobacter sphaeroides CB 8521 of the present invention is a novel strain belonging to the scarlet non-sulfur photosynthetic bacteria. It is a Gram-negative region and has an ability to reduce malodor generating materials suitable for livestock production, antimicrobial efficacy against intestinal pathogenic bacteria, It has immune activation ability. The Rhodobacter sphaeroides CB 8521 strain of the present invention was deposited with Accession No. KACC92014P on Jan. 2, 2015 at the National Institute of Agricultural Science and Technology.

The Rhodobacter sphaeroides CB 8521 of the present invention has an effect of reducing malodorous substances such as ammonia, hydrogen sulfide, methyl mercaptan and trimethylamine, which are frequently generated in the livestock farming sites, by 90% or more.

In addition, the Rhodobacter sphaeroides CB 8521 of the present invention is useful as an effective ingredient for the treatment and / or prevention of diseases such as Bacillus cereus, Clostridium perfringens, E. coli, Listeria monocytogenenis, Listeria monocytogenes, Salmonella typhimurium, Salmonella entericidis, Staphylococcus aureus And antibiotics against various pathogenic bacteria including viruses, viruses, and the like, and have high immunological activities such as NO, TNF-α and IL-1β activity.

2. Microbial agent  Composition

The microbial composition of the present invention comprises the Rhodobacter sphaeroides CB 8521 described above. The microbial composition of the present invention preferably contains Rhodobacter sphaeroides CB 8521 in the range of 1.0 × 10 ⁶ cfu / g to 1.0 × 10 ¹¹ cfu / g, preferably 1.0 × 10 ⁸ cfu / g to 1.0 × 10 ¹⁰ cfu / / g. < / RTI >

The microbial composition of the present invention is an excellent microbial agent having a reduced odor of animal offspring, an antimicrobial effect against enteric pathogenic bacteria, and an immunity activity improving effect, and can be used as a substitute for a conventional probiotic agent, and is particularly excellent as a microbial agent for animal husbandry.

The microbial composition of the present invention may further contain other microorganisms, yeast or other nutritional components depending on the specific use. In addition, the microbial composition of the present invention may further include known additives such as excipients, flavor enhancers, and the like.

3. Manufacturing method of microbial agent

The method for producing the Rhodobacter sphaeroides CB 8521 strain of the present invention with a microorganism is as follows.

First, the Rhodobacter sphaeroides CB 8521 of the present invention is liquid cultured for 40 to 48 hours and used as a seed bacterium. It is preferable to use the culture medium that has been cultured overnight in 10 ml of LB liquid medium as a seed culture for mass production. For mass production of the medium, based on the weight of 0.1% to 2.0% casein peptone or vegetable protein, 0.5 ~ 3.0% yeast extract, 1.0 ~ 3.0% sodium acetate, 0.1% to 2.0% of ammonium sulfate _{{(NH 4) 2 SO 4} }, 0.1 ~ Using a liquid medium containing 2.0% glutamic acid, 0.1-2.0% sodium succinate, 0.1-1.0% potassium phosphate (K ₂ HPO ₄ ), 0.001-0.05% of minerals (iron, magnesium, manganese etc.) , And it is preferable that the culture is carried out at a temperature of 25 to 37 DEG C for 40 to 48 hours under a slight condition.

After the cultivation is completed, the cells are recovered by centrifugation at a rapid rate and then mixed with the cryoprotectant at a weight ratio of 1: 1 to 1: 2 , more preferably 1: 1, followed by lyophilization. The above-mentioned cryoprotectant is preferably at least one selected from skim milk powder, maltodextrin, lactose, wheat flour, trehalose and mannitol. More preferably, the cryoprotectant contains at least 4 kinds of these components in the range of 0.5 to 30%.

The solid microbial cell prepared by freeze-drying is in the range of 5.0 × 10 ¹⁰ to 3.0 × 10 ¹² cfu / g. It is preferable to include this source at a level of preferably 1.0 × 10 ⁶ cfu / g to 1.0 × 10 ¹¹ cfu / g, more preferably at a level of 1.0 × 10 ⁸ cfu / g to 1.0 × 10 ¹⁰ cfu / A microbicide, an excipient, a nutrient or a common additive are added to prepare the microbicide composition of the present invention.

The microorganism of the present invention uses Rhodobacter sphaeroides CB 8521, which is a strain isolated from soil, and can be safely used as a microorganism for livestock.

[ Example ]

Hereinafter, the present invention will be described in more detail with reference to specific examples. However, the scope of the present invention is not limited by the following embodiments, but may be embodied in various other forms.

< Example  1>

Isolation of photosynthetic bacteria

To isolate photosynthetic bacteria, samples were taken from the soil and river water, transferred to the laboratory, and stored at 4 ° C. The samples were weighed 1 g or 1 ml each, and then placed in 9 ml of sterile physiological saline and sequentially diluted. 0.2 ml of an appropriately diluted sample was plated on a photosynthetic bacterial separation medium (ATCC medium 650) and incubated at 5,000 lux for 3 to 5 days at 30 ° C. In the colony grown on the medium, And photosynthetic strains of Gram - negative strains were isolated at the time of microscopic observation.

< Example  2>

Identification of microorganisms

The final photosynthetic bacteria were selected for the microorganisms selected in Example 1, considering deodorizing effect on odor substances and antimicrobial ability against pathogenic bacteria. The selected bacteria were analyzed by 16S rRNA sequence, which is morphological, biochemical and molecular genetic method, and Rhodobacter sphaeroides ) CB 8521.

< Example  3>

Antimicrobial range investigation

A spot-on-lawn method was used to determine the antimicrobial coverage against pathogenic bacteria of Rhodobacter sphaeroides CB 8521 strain.

Namely, Rhodobacter sphaeroides CB 8521 strain was inoculated into LB liquid medium and cultured at 30 ° C for 2 days. Then, the culture broth was dropped on a soft cloth containing indicator bacteria (1.0 × 10 ⁷ cfu / ml) Were cultured overnight at the optimum growth temperature. The results are shown in Table 1 below.

Indicator strains Inhibition Bacillus cereus KCTC 1012 + Clostridium perfringens KCTC 3269 &lt; RTI ID = 0.0 &gt; ++ Clostridium perfringens KCTC 5100 ++ Escherichia coli KCTC 1682 + E. coli KCTC 1467 ++ Lactobacillus casei KCTC 3109 - Lactobacillus plantarum KCTC 3108 - Lactobacillus sakei KCCM 40264 - Leuconostoc mesenteroides KCTC 3505 &lt; RTI ID = 0.0 &gt; - Listeria monocytogenes KCTC 3569 ++ Listeria monocytogenes KCTC 3710 ++ Listeria innocua KCTC 3586 ++ Pseudomonas aeruginosa KCTC 1750 - Salmonella typhimurium KCTC 2515 + Salmonella enteritidis KCCM 12021 + Staphylococcus aureus KCTC 1621 +

( +: Positive, -: negative)

As shown in the results of the above Table 1, the Rhodobacter sphaeroides CB 8521 strain of the present invention is useful for the treatment of Bacillus cereus, Clostridium perfringens, E. coli, Listeria monocytogenes, Listeria monocytogenes, Salmonella typhimurium, It has the ability to inhibit intestinal pathogenic bacteria such as rheumatoid, staphylococcus aureus.

< Example  4>

In vitro  Deodorization test

20 ml of a culture solution of Rhodobacter sphaeroides CB 8521 strain was placed in a 5 L reactor and sealed. Then, ammonia, methyl mercaptan and trimethylamine as test gases were injected at an initial concentration of 50 μmol / mol, and the concentration of the test solution was measured And the concentration was determined as a sample concentration. At this time, the concentration of the test gas was measured by the detector tube gas measuring instrument of KS I 2218, the temperature was maintained at 23 ± 5 ° C and the relative humidity was maintained at 50 ± 10%. For the purpose of comparison, the blank concentration was determined in the same manner without any sample.

The deodorization rate (%) was determined by the following formula, and the results are shown in Table 2 below.

[Equation 1]

Deodorization rate (%) = (A - B) / A × 100

      (Wherein, A is a blank test value and B is a sample test value)

Test Items Test result Blank test concentration Sample concentration Deodorization rate (%) ammonia
(NH ₃₎ 0 minutes 50 50 0.0 60 minutes 49 2 95.9 120 minutes 48 One 97.9 Methyl mercaptan
(CH ₃ SH) 0 minutes 50 50 0.0 60 minutes 49 40 18.4 120 minutes 48 32 33.3 Trimethylamine
{(CH _{3) 3} N}
0 minutes 50 50 0.0 60 minutes 49 4 91.8 120 minutes 48 3 93.8

As can be seen from the results of the deodorization test shown in Table 2, the Rhodobacter sphaeroides CB 8521 strain of the present invention decreased the concentration of ammonia and trimetalamine by 90% or more after 120 minutes of reaction, and the concentration of methyl mercaptan 33% or more.

< Example  5>

Immunological activity experiment

One. Heat killing  Photosynthetic bacteria Heat - killed PSB )

The Rhodobacter sphaeroides CB 8521 strain was inoculated into LB liquid medium, cultured overnight at 30 ° C with shaking, centrifuged at 6000 rpm, and the collected cells were washed twice with RPMI medium. Heat-killed photosynthetic bacteria were prepared by heating at 100 ° C for 10 minutes, and adjusted to 1.0 × 10 ⁸ cells / ml.

2. RAW  264.7 Cell culture and sample treatment

To measure the immunological activity, RAW 264.7 cells (murine macrophage, 5.0 x 10 ⁵ cells / ml) were dispensed in a 12-well flat bottomed 12-well plate Respectively. 400 μl of each heat-killed photosynthetic bacteria was added, and 600 μl of RPMI medium was injected to make a total of 2 ml. The plate was incubated at 37 ° C. for 24 hours in a 5% CO ₂ incubator. After culturing, the supernatant was collected by centrifugation and used as a sample for measurement of NO, TNF-α and IL-1β.

3. NO ( nitrogen oxide ), TNF -α and IL -1β Generation  Measure

NO is a substance produced from macrophages, which has an anticancer effect against cancer cells and has a function of removing them in response to foreign pathogenic substances such as bacteria, viruses and parasites in the immune response. NO was measured by adding 50 쨉 l of the supernatant sample to a 98-well plate, adding 50 쨉 l of each of griess reagent I and grease reagent II, reacting for 10 minutes, and measuring the absorbance at 540 nm. TNF-α and IL-1β production were assayed by enzyme-linked immunoabsorbent assay (ELISA) (Mouse TNF-α or IL-1β ELISA set, BD OptEIA). As a control, LPS (lipopolysaccharide) was treated and compared.

The results of investigating the immunological activity as described above are shown in Table 3 below.

Item NO (μM) TNF-a (pg / ml) IL-1? (Pg / ml) CB 8521 17.8 180.1 294.5 Bacillus 5.6 64.8 260.1 LPS 24.1 480.5 283.5

As can be seen from the results in Table 3, Rhodobacter sphaeroides CB 8521 exhibited relatively higher activities of NO, TNF-α and IL-1β than those of other Bacillus strains.

< Example  6>

Rhodobacter Spelloides CB  8521 &lt; / RTI &gt; Microbial agent  Produce

Rhodobacter sphaeroides CB 8521 was mass-produced in a 300 L fermenter using a culture medium containing a carbon source, a nitrogen source and trace elements as described below, and culture conditions (culture time, temperature, shaking speed, etc.).

(NH ₄ ) ₂ SO ₄ }, 0.5 g of potassium phosphate dibasic potassium phosphate (K ₂ HPO ₄ ), 0.5 g of sodium glutamate, 0.5 g of sodium acetate, 0.5 g of sodium succinate, and 0.05 g of magnesium sulfate were sterilized at 121 DEG C for 20 minutes, and the seeds cultured in the medium were inoculated at 2% by weight. The incubation temperature was 30 캜, and oxygen was supplied at 0.1 vvm level. When the agitation speed was maintained at 80 rpm for 40 to 44 hours, viable cell counts of 6.0 × 10 ⁹ cfu / ml or more were obtained. In order to obtain a high number of viable cells, the medium was concentrated at 20 hours and 30 hours, and when cultured by aseptically adding the medium, it was grown at 2.0 × 10 ¹⁰ cfu / ml or more.

The culture broth was centrifuged to recover the cells, followed by lyophilization after mixing with a cryoprotectant (5% skim milk powder, 3% lactose, 5% wheat flour, 5% maltodextrin and 2% trehalose) at a weight ratio of 1: 1 . The viable cell count of the lyophilized Bacillus sp. Source was measured and found to be 2.0 x 10 ¹² cfu / g.

< Example  7>

Specification Test

To test odor reduction efficacy of Rhodobacter sphaeroides CB 8521, 600 scaled finishing pigs were selected and divided into control and treated groups, respectively. The form of the pilgrimage company was a kind of pilgrimage and slurry. A 3-member hybrid with a body weight of 56.1 ± 1.6 kg at the time of initiation of the test was disclosed and a field test was conducted for 4 weeks. As a test diet, commercially available diets were fed at a fixed interval, and a certain amount of diets were fed for free diet. In the treatment area, solid or liquid microorganism containing Rhodobacter sphaeroides CB 8521 (1.0 × 10 ⁷ cfu / g or ㎖) as the main ingredient was fed at 1 kg per ton of feed or at a concentration of 500 ml per 100 sq. Sprinkled evenly on the floor, and no action was taken on the control.

One. Rhodobacter Spelloides CB  8521 confirmed odor reduction effect

The odor reduction effect by Rhodobacter sphaeroides CB 8521 was confirmed as follows.

The concentrations of hydrogen sulfide (H ₂ S) and methyl mercaptan (CH ₃ SH), which are odor compounds of sulfur compounds, were measured using a gas detection method (Kitagawa, Japan) Using a polyester bag, samples were taken at 20 cm height from the bottom at 10 o'clock to 11 o'clock on the left and right sides of the center, entrance, and inside of the pig house, and analyzed using GC (gas chromatography) equipped with FPD Respectively. All the test results were expressed as mean ± SD. Comparisons between the groups were analyzed by one-way ANOVA followed by Duncan's multiple range test.

The results are shown in Tables 4 to 6 below.

Elapsed time (days) Ammonia concentration (ppm) Deodorization rate
(%) Control Treatment Early 24 24 - 7 27 10 62.9 14 35 11 68.6 21 30 8 73.3 28 31 5 83.8

Elapsed time (days) Hydrogen sulfide concentration (ppm) Deodorization rate
(%) Control Treatment Early 0.39 0.39 - 7 0.45 0.12 73.3 14 0.53 0.08 84.9 21 0.50 0.05 90.0 28 0.48 0.05 89.6

Elapsed time (days) Methyl mercaptan (ppm) Deodorization rate
(%) Control Treatment Early 5.1 5.1 - 7 10.0 2.1 79.0 14 11.0 0.3 97.3 21 10.5 0.2 98.1 28 10.4 0.3 97.1

As can be seen from the above results, after the 7th day of the specification test, the odor of the treated group was reduced by about 60% or more compared to that of the control group, and the effect of reducing the odor was reduced by 90% at the end of the test (4 weeks).

2. Daily gain , Daily feed intake and feed efficiency

Body weight and feed intake were measured for the treatment and control until the end of the experiment, and daily gain, daily feed intake and feed efficiency were calculated. The results are shown in Table 7 below.

Item Control Treatment Initial weight (kg) 5.61 + - 0.80 ^* 5.60 ± 0.77 Final weight (kg) 24.58 ± 6.04 26.28 + - 4.80 Daily weight gain (kg) 0.386 ± 0.12 0.402 + 0.09 Total feed intake (kg) 498.19 532.64 Daily feed intake (kg) 0.73 0.75 Total weight gain (kg) 264.82 295.63 Feeding rate (FC) 1.88 1.78

{ ^* Mean (log cfu / g feces) + SD}

As can be seen from the results of the specification test in Table 7, the daily treatments of the microbial agent of the present invention showed a tendency to improve the daily gain of body weight and the feed conversion ratio as compared with the control.

Agricultural Biotechnology Research Institute KACC92014P 20141218

Claims (5)

Separated from the soil,
Ammonia, hydrogen sulfide, methyl mercaptan, and trimethylamine,
Which inhibit pathogenic bacteria including Bacillus cereus, Clostridium perfringens, E. coli, Listeria monocytogenis, Listeria innocia, Salmonella typhimurium, Salmonella entericidis and Staphylococcus aureus,
( Rhodobactersphaeroides CB8521) (accession number: KACC92014P) having immunostimulatory activity. A microorganism composition for animal husbandry comprising Rhodobacter sphaeroides CB 8521 (accession number: KACC92014P) as an active ingredient,
Ammonia, hydrogen sulfide, methyl mercaptan, and trimethylamine,
Which inhibit pathogenic bacteria including Bacillus cereus, Clostridium perfringens, E. coli, Listeria monocytogenis, Listeria innocia, Salmonella typhimurium, Salmonella entericidis and Staphylococcus aureus,
Wherein the microbial composition has an immunological activity. 3. The method of claim 2,
Wherein said composition comprises Rhodobacter sphaeroides CB 8521 in the range of 1.0 × 10 ⁶ cfu / g to 1.0 × 10 ¹¹ cfu / g. Liquid culture of Rhodobacter sphaeroides CB 8521 (accession number: KACC92014P) for 40 to 48 hours;
The cultured Rhodobacter sphaeroides CB 8521 (Accession No .: KACC92014P) was added to a culture medium containing 0.1 to 2.0% casein peptone or vegetable protein, 0.5 to 3.0% yeast extract, 1.0 to 3.0% sodium acetate, 0.1 to 2.0% ammonium sulfate _{{(NH 4) 2 SO 4} }, 0.1 ~ 2.0% glutamic acid, 0.1% to 2.0% sodium succinate, 0.1% to 1.0% J. potassium phosphate _{(K 2 HPO 4), 0.001} ~ 0.05% of minerals (iron, magnesium, manganese, etc. ) And the rest of distilled water at 25 to 37 ° C for 40 to 48 hours under a humid condition, and then centrifuging the cells to recover the cells; And
Mixing the recovered cells with a cryoprotectant at a weight ratio of 1: 1 to 1: 2, followed by lyophilization to obtain a final volume of 5.0 × 10 ¹⁰ cfu / g to 3.0 × 10 ¹² cfu / g of the microorganism Gt; 5. The method of claim 4,
Wherein the cryoprotectant is at least one selected from skim milk powder, maltodextrin, lactose, wheat flour, trehalose, and mannitol.

Images