KR101164512B1 - Probiotic composition for animal comprising bifidobacterium pseudocatenulatum spm1204 or its culture - Google Patents

Abstract

The present invention relates to Bifidobacterium pseudocatenulatum ( Bifidobacterium pseudocatenulatum ) SPM1204 lactic acid bacteria (Accession No .: KCTC 10616BP) or a culture thereof as a probiotic composition for livestock.

The present invention is a Bifidobacterium pseudocatenulatum ( Bifidobacterium pseudocatenulatum ) SPM1204 lactic acid bacteria (Accession Number: KCTC 10616BP) or a livestock probiotic composition using the culture thereof as an active ingredient for various intestinal pathogenic bacteria present in the intestine of the animal As well as exhibiting a good dressing effect, it can be usefully used in the manufacture of feed, food by promoting the growth of livestock.

Bifidobacterium pseudocatenuratum, formal preparation, probiotic composition

Description

Bifidobacterium Pseudocatechinulatium SPM12204 Probiotic composition for livestock comprising Lactobacillus or its culture as an active ingredient {PROBIOTIC COMPOSITION FOR ANIMAL COMPRISING BIFIDOBACTERIUM PSEUDOCATENULATUM SPM11020 OR ITS CULTURE}

The present invention relates to Bifidobacterium pseudocatenulatum ( Bifidobacterium pseudocatenulatum ) SPM1204 lactic acid bacteria (Accession No .: KCTC 10616BP) or a culture thereof as a probiotic composition for livestock.

In the intestines of livestock, many kinds of microorganisms coexist and form an ecosystem. Intestinal bacteria are fermented anaerobicly or microaerobically using feed that is continuously supplied, and the body's body contains temperature, pH, osmotic pressure and oxidation. Physical as reduction? Since the chemical environment is almost constant, the reproductive density and metabolism activity of the microorganisms are active. However, extreme stress, environmental stress such as emissary, and hyperacidity due to over-feeding of livestock of the livestock, destroys homeostasis in the intestine of the livestock, and consequently causes a lot of deterioration of productivity of livestock such as disease and mortality due to lower immunity. Affect These arguments common infectious bacteria include Salmonella spp typically cause food poisoning (Salmonella spp . ), Staphylococcus aureus , enterotoxigenic E. coli , Listeria monocytogenes ( Listeria) monocytogenes ), Campylobacter jejuni) and Clostridium perfringens (Clostridium perfringens ).

Drugs such as antibiotics and chemotherapy have been used successfully for prevention and treatment. Antibiotic growth-promoting mechanisms, as reported by Visek (1978) and Walton (1980), showed good growth rates and improved productivity in harsh environments. The addition of antibiotics to livestock feed improves nutrient utilization and productivity, and in addition to suppressing and treating various disease outbreaks, it has recently been used for the purpose of growing livestock, promoting fattening and improving feed efficiency. have.

However, the use of antibiotics reduces the number of effective microorganisms in the intestine, and increases the microorganisms that are resistant to antibiotics due to misuse, which decreases their efficacy or remains in the body of livestock, adversely affecting the human body when ingested by humans. After the report, further suppression of the use of antibiotics for the purpose of promoting growth is being strengthened. For example, according to the veterinarian's diagnosis of the disease symptoms, antibiotics that need to be administered in an appropriate amount are produced by mutant pathogenic microorganisms that have become antibiotic-resistant due to the inadequate use of formulated feed or livestock farms. In addition, there is a medical report that one of the causes of antibiotic resistance is more than four times higher than that of developed countries due to the absence of antibiotics.

In Europe, the use of zinc bacitracin, Spiramycin, tylosin phosphate and virginiamycin among antibiotics that have been allowed to be used as feed additives have been banned. This ban prevents the treatment of human diseases by making bacteria resistant to antibiotics used to promote livestock growth to the parasitic bacteria in the human body, making them resistant to drugs used in the human body. It was carried out on the premise that there is a risk of harm.

Against this backdrop, we began to develop probiotics that do not have antibiotic residue in meat, are safe for human health while consuming, and promote livestock growth, and prevent and adjuvant bacterial diseases such as antibiotics.

Commonly used definitions for probiotics are described in R. Fuller, Journal of Applied Bacteriology 1989, 66, 365-378], according to which probiotics are administered as feed additives and, due to the "equilibrium" of the intestinal flora, have a beneficial effect on host animals. Include. That is, probiotics are 'substances that help intestinal bacterial balance by microorganisms or microorganisms that promote the growth of other microorganisms', or' live or microorganisms alone or mixed which improve the balance of human or animal intestinal bacteria and effect the host. Cultures', 'microbial agents or components of microorganisms having a beneficial action on the host' and the like.

Since probiotics have been reported to be the cause of longevity, many types of probiotics have been isolated and identified since it has been reported that the administration of probiotics in the intestines of livestock acts as a competitive relationship between microorganisms. These bacteria act to maintain the number of beneficial microorganisms in the intestine, can maximize the efficiency of the beneficial microorganisms in competition with harmful bacteria, and has been identified as a factor that can improve productivity in large scale farming test.

The antimicrobial effects of probiotic agents on pathogenic microorganisms can be explained by several mechanisms, for example the production of inhibitors that can kill pathogens (Vanderberg, PA, FEMS Microbiol. Rev., 12, 221-238). , 1993), nutrient competition with pathogens at settlement sites (Bernet, MF et al., Gut., 35, 483-489, 1994; Majamaa, H. et al., J. Pediatr. Gastroenterol. Nutr., 20, 333-338, 1995; and Sato, K. et al., Microbiol. Immunol., 32, 1189-1200, 1988), host immunity enhancement (Perdigon, ME et al., Immunology, 63, 17-23, 1988; Hatcher, GE et al., J. Dairy Sci., 76, 2485-2492, 1993; and Paubert, B. et al., Int. J. Immunother., 11, 153-161, 1995), and inhibition of bacterial toxin production and activity (Czerucka, D. et al., Gastroenterol, 106, 65-72, 1994; Brandqed, RL et al., Appl. Environ. Microbiol. 64: 564-568, 1998).

Examples of microorganisms that can be used as livestock probiotics include lactic acid bacteria, Bacillus bacteria, fungi and yeasts. Among these, lactic acid bacteria can be divided into five genera, which are divided into Streptococcus, Lactobacillus, Leukonostoc, Bifidobacteria, and Pediococcus.

Among the lactic acid bacteria, Lactobacillus sp. Microorganisms produce acidophillin to inhibit the growth of harmful bacteria such as dysentery, Salmonella and staphylococcus, and inhibit the proliferation of diarrhea causative bacteria to normalize the intestinal flora to stop diarrhea. It is known to do. In addition, Pediococcus pentosaceus secretes a strain-specific antimicrobial peptide, Helicobacter pylori ) and Listeria sp. have been shown to have excellent ability to inhibit the growth of harmful bacteria in the body.

Bifidobacterium was first discovered by Tissier as a microorganism that lives in the intestines of humans, animals and insects. Bifidobacterium has the characteristics of gram positive, absolute anaerobic, GC-rich, etc. and is found in the intestinal dominant flora of the mother's infant. Bifidobacteria is an actinomycetes family that differs from the well-known lactobacillus lactobacilli, lactococci, and others. (Mycobacteria) and taxonomically close. It also produces lactic acid: acetic acid in a 3: 2 ratio as a metabolite as a lactic acid-producing strain, and plays a role in preventing foreign invading pathogens by adjusting pH. These properties are known to play a key role in maintaining the balance of normal intestinal flora. Ingesting milk fermented with Bifidobacterium has the effect of alleviating lactose intolerance. Recent studies have revealed useful functions such as lowering cholesterol, enhancing immune function, and anticancer activity. In addition, the adsorption power to the intestinal mucosa is excellent, and there is increasing interest in the world.

Until now, the research direction of Bifidobacterium has been to identify and count strains, develop screening methods, screen for strains that secrete acid, bile, and bacteriocins, analyze their own biochemical properties, and plasmids derived from Bifidobacterium. Analysis, cloning and biochemical analysis of characteristic enzymes are the mainstream. In addition, studies on the expression of genes, the development of vector systems, and their genetic characteristics in the intestinal lactic acid strains of humans and animals have been published.

The present inventors identified lactic acid bacteria by collecting feces from healthy adults aged 20 to 30 years in the intestine of Koreans, and the Bifidobacterium pseudocatenulatum was cross-linked of dendritic cells to exogenous antigens. Bifidobacterium Bifidobacterium ( Bifidobacterium) was found to have an immune-increasing activity that not only promotes cross-presentation but also directly activates macrophages to induce the production of NO and TNF-α. pseudocatenulatum ) was designated as SPM1204 dated March 31, 2004 and deposited in the Korea Institute of Biotechnology and Biotechnology Center (Accession No .: KCTC 10616BP), filed on August 26, 2004, and filed on August 1, 2006. Registration (registration number 10-0610379) was received.

The present inventors inhibit the growth of Bifidobacterium pseudocatenulatum SPM1204 by inhibiting the growth of harmful bacteria such as heterologous bacteria, Salmonella, Staphylococcus aureus, and inhibiting the growth of diarrhea causative bacteria to stop the diarrhea by normalizing the intestinal flora. ash are known to act also to secrete pilrin (acidophillin) and strain-specific antimicrobial peptide pylori (Helicobacter pylori ), Listeria sp . When used in combination with Pediococcus pentosaceus, which is known for its ability to inhibit the growth of harmful bacteria in the body, it shows excellent suitability against various intestinal pathogenic bacteria present in the intestines of animals. It was found that it can be usefully used for the manufacture of feed, food, etc., and developed a mixed lactic acid bacterium composition.

The present invention exhibits an excellent suiting effect on various intestinal pathogenic bacteria present in the intestine of the livestock, and also has an excellent inhibitory effect against harmful bacteria that can be usefully used in the manufacture of feed and food by promoting the growth of livestock. An object of the present invention is to provide a livestock probiotic composition comprising the lactic acid bacteria or a culture thereof as an active ingredient.

The present invention Bifidobacterium Pseudocatechinulatum ( Bifidobacterium) in order to achieve the above technical problem pseudocatenulatum ) SPM1204 lactic acid bacteria (Accession Number: KCTC 10616BP) or a culture thereof provides a probiotic composition for livestock as an active ingredient.

The livestock probiotic composition of the present invention is Lactobacillus Lactobacillus acidophilus ) or Pediococcus pentosaceus ( Fediococcus pentosaceus) is characterized in that the composition further comprises one or more lactic acid bacteria or a culture thereof selected from the group consisting of.

The livestock probiotic composition of the present invention is Bifidobacterium pseudocatenuratum ( Bifidobacterium) as an active ingredient based on the total weight of the composition. pseudocatenulatum ) SPM1204 lactic acid bacteria (Accession Number: KCTC 10616BP) or its culture is characterized in that it comprises a 10 ⁷ to 10 ¹⁵ cfu / g preferably 10 ¹⁰ to 10 ¹³ cfu / g.

The livestock probiotic composition of the present invention is Lactobacillus Lactobacillus acidophilus) or Phedi O Lactococcus pen content of at least one lactic acid bacteria or a culture selected from the group consisting of gravel-aged mouse (Pediococcus pentosaceus) 10 ⁷ to 10 ¹⁵ cfu / g, preferably from 10 ¹⁰ to 10 ¹³ cfu / g It characterized by including as.

Probiotic composition for livestock of the present invention is Bifidobacterium pseudocatenulatum ( Bifidobacterium pseudocatenulatum ) SPM1204 lactic acid bacteria (Accession No .: KCTC 10616BP) or its culture is 10 ⁷ to 10 ¹⁵ cfu / g preferably 10 ¹⁰ to 10 ¹³ cfu / g, Lactobacillus acidophyllus ( Lactobacillus acidophilus) or a culture thereof, and Phedi O Lactococcus pen soil three mouse (Pediococcus pentosaceus ) or a culture thereof is characterized in that it comprises a content of 10 ⁷ to 10 ¹⁵ cfu / g preferably 10 ¹⁰ to 10 ¹³ cfu / g.

Livestock probiotic composition of the present invention is Salmonella gallinarum ( Salmonella gallinarum ), Salmonella enteritidis ( Salmonella enteritidis ), Salmonella typhimurium ( Salmonella) typhimurium ), Staphylococcus aureus ), E. coli , Listeria monocytogenes ( Listeria) monocytogenes ), Campylobacter jejuni ), Clostridium perfringens ( Clostridium perfringens ) is characterized by having an intestinal effect on the intestinal pathogenic bacteria.

Livestock probiotic composition of the present invention is E. coli ATCC 1056, E. coli ATCC 25922, Salmonella typhimurium ( Salmonella typhimurium ) ATCC 13311, Salmonella typhimurium ) KCTC 2053, Staphylococcus aureus ) ATCC 6538p characterized in that it has a formal effect.

Probiotic composition for livestock of the present invention is characterized in that it is used for livestock feed.

In the present invention, the livestock is characterized in that the calf.

Hereinafter, the probiotic formal preparation composition for animals using the SPM1204 lactic acid bacteria or a culture thereof of the present invention as an active ingredient will be described in more detail.

Bifidobacterium pseudocatenulatum SPM1204 used in the present invention is a lactic acid bacterium identified by the present inventors by collecting feces from healthy adults aged 20 to 30 years in the intestine of Koreans. As of March 31, it was deposited with the Korea Institute of Biotechnology and Biotechnology Center (Accession No .: KCTC 10616BP).

Animal probiotic dressing composition of the present invention is Bifidobacterium Pseudocatechinulatum ( Bifidobacterium Ruth is also required (Lactobacillus Lactobacillus know the pseudocatenulatum) SPM1204 acidophilus ) or Pediococcus pentosaceus ( Pediococcus pentosaceus) may further comprise one or more lactic acid bacteria or cultures thereof, and provides a livestock probiotic composition comprising these mixed lactic acid bacteria or their cultures as an active ingredient do.

In this case, the probiotic composition composed of the three microorganisms may include various antimicrobial organic acids and nonprotein antimicrobial substances produced by the microorganism, and when included as an active ingredient in the formal formulation, may have the same effect as the composition including the strain. .

The livestock probiotic composition of the present invention has the following new characteristics. First, the lactic acid bacteria according to the present invention exhibits the effect of suppressing harmful microorganisms in the intestine, showing a formal effect. Second, the lactic acid bacteria according to the present invention can be mixed culture with other lactic acid bacteria, the mixed culture liquid has the effect of inhibiting the growth of harmful microorganisms.

The probiotic composition according to the present invention is Salmonella gallinarum ( Salmonella gallinarum ), Salmonella enteritidis (Salmonella enteritidis ), Salmonella typhimurium ( Salmonella) typhimurium ), Staphylococcus aureus ), E. coli , Listeria monocytogenes ( Listeria) monocytogenes ), Campylobacter jejuni), Clostridium perfringens (Clostridium Intestinal pathogenic bacteria such as perfringens ) can be suppressed, and in particular, it is useful as a probiotic lactic acid bacterium because it exhibits excellent anti-lethal effect on E. coli in the calf in vivo, and shows superior acid resistance and bile resistance.

In the present invention, the probiotic composition is Bifidobacterium pseudocatenulatum ( Bifidobacterium) as an active ingredient. pseudocatenulatum ) SPM1204 (Accession No .: KCTC 10616BP) or its culture is characterized in that it comprises a content of 10 ⁷ to 10 ¹⁵ cfu / g, preferably 10 ¹⁰ to 10 ¹³ cfu / g.

The livestock probiotic composition of the present invention is Lactobacillus Lactobacillus acidophilus) or Phedi O Lactococcus pen content of at least one lactic acid bacteria or a culture selected from the group consisting of gravel-aged mouse (Pediococcus pentosaceus) 10 ⁷ to 10 ¹⁵ cfu / g, preferably from 10 ¹⁰ to 10 ¹³ cfu / g It characterized by including as.

The probiotic composition in the present invention, Bifidobacterium pseudocatenurlatum ( Bifidobacterium pseudocatenulatum ) SPM1204 (Accession No .: KCTC 10616BP) or its culture, Lactobacillus acidophilus) or its culture and Phedi O Lactococcus pen soil three mouse (Pediococcus pentosaceus ) or a culture thereof may be included in an amount of 10 ⁷ to 10 ¹⁵ cfu / g, preferably 10 ¹⁰ to 10 ¹³ cfu / g.

The 'suit' refers to the use for treating and ameliorating the symptoms caused by abnormal fermentation of the intestinal bacterial flora of livestock. The symptoms include, but are not limited to, for example, infectious diarrhea caused by harmful microorganisms, gastroenteritis, inflammatory bowel disease, neuro enteritis syndrome, small intestine microbial hypergrowth, intestinal acute diarrhea and the like.

The above-mentioned compositions may contain the crushed cell wall fraction of lactic acid bacteria, live bacteria, dead bacteria, dried bacteria or cultures thereof according to the present invention as an active ingredient, and may further include a suitable excipient or carrier. The culture is culture medium itself cultured lactic acid bacteria according to the present invention in a suitable liquid medium, the filtrate (filtered or centrifuged supernatant) from which the strain was removed by filtration or centrifugation, the culture solution by sonication or to the culture medium Cell lysates obtained by treating lysozyme, but are not limited thereto.

In addition, the individual or mixed culture of the SPM1204 according to the invention can also be provided in the form of a feed composition. The feed composition can be used as a substitute for the existing antibiotics to inhibit the growth of harmful intestinal bacteria and maintain the intestinal flora in a stable condition to improve the health of the livestock, improve the weight gain and meat quality of the livestock and increase the milk yield and immunity . Feed composition of the present invention may be prepared in the form of fermented feed, compound feed, pellet form, silage (silage) and the like, but is not limited thereto. The fermented feed may be prepared by fermenting an organic material by adding the lactic acid bacteria and various microorganisms or enzymes of the present invention, the blended feed may be prepared by mixing various kinds of lactic acid bacteria of the general feed and the present invention. In addition, the pellet-type feed may be prepared by applying heat and pressure to the fermented feed or blended feed in a pellet machine.

On the other hand, since the probiotic composition of the present invention additionally exhibits branch action, digestion promotion and anticancer activity, as a probiotic composition such as fungicides, digestive agents and antibacterial agents, antimicrobial compositions, fermentation products and food additives for promoting the growth of livestock May also be provided.

In addition, the probiotic composition according to the present invention is suitable for ingestion by livestock together with the lactic acid bacteria of the present invention, it is possible to suppress the growth of harmful microorganisms when ingesting other types of known microorganisms having the activity of improving the balance of the intestinal flora It may further comprise.

In addition, the microorganism according to the present invention may be improved or improved to have equivalent activity or better activity by conventional physicochemical mutation methods and the like known in the art.

Bifidobacterium pseudocatenulatum ( Bifidobacterium pseudocatenulatum ) SPM1204 lactic acid bacteria (Accession No .: KCTC 10616BP) according to the present invention or a livestock probiotic composition using the culture thereof as an active ingredient is a variety of intestinal pathogenic bacteria present in the intestines of livestock In addition to showing an excellent dressing effect, it has an excellent inhibitory effect against harmful bacteria that can be useful for the production of feed, food, etc. by promoting the growth of livestock, can be useful for feed, food, cosmetics, medicines, etc. .

Example 1 Preparation of Mixed Lactic Acid Bacteria

<1-1> Bifidobacterium pseudo Carthage Press ratum (Bifidobacterium pseudocatenulatum ) SPM1204 lactic acid bacteria (Accession Number: KCTC 10616BP).

The present inventors have identified Bifidobacterium pseudocatenulatum SPM1204 lactobacillus identified from feces of healthy adults aged 20 to 30 years in the intestines of Koreans. Deposited in the BRC (Accession No .: KCTC 10616BP), and used a newly identified lactic acid bacteria in Korea Patent Registration 0610379.

<1-2> Preparation of mixed lactic acid bacteria.

The lactic acid bacteria used to prepare the probiotic composition of the present invention are the SPM1204 lactic acid bacteria, Lactobacillus acidophilus CBT and Pediococcus pentosaceus ( Pediococcus ), as shown in Table 1. pentosaceus ) CBT. The Lactobacillus asidophilus and Pediococcus pentosaceus were used to purchase strains deposited in the Korea Collection for Type Culture (KCTC). In addition, although not shown in the present embodiment, it may further include other known microorganisms having an intestinal effect on intestinal harmful bacteria.

The lactic acid bacteria were respectively inoculated in sterilized GAM broth, followed by incubation at 37 ° C. for 16 hours, and then inoculated with 1% of each of the spawn cultures and sterilized by GAM (General anaerobic medium) (Nissui Pharm. Co. Ltd.). , Japan) under an anaerobic system (Bactron Anaerobic Chamber, Sheldon Manufacturing Inc., USA) filled with a mixed gas consisting of N ₂ (90%), H ₂ (5%) and CO ₂ (5%) in broth. Incubated for 18 hours at ℃. Thereafter, the culture solution was centrifuged at 4000 rpm for 5 minutes to remove the supernatant and only the cells were recovered. The cells obtained by inoculating and culturing the three strains were orally administered to the calf for the experiment in the following Examples at a concentration of 3.0 × 10 ¹¹ cfu / g.

Example 2 Intestinal Activity of Mixed Lactic Acid Bacteria

In order to determine the activity of the mixture of lactic acid bacteria suits the total E. coli (E. coli), which before and after oral administration of the mixture of lactic acid bacteria in the calf, present in the feces of the calf, Salmonella (Salmonella spp .) and Staphylococcus spp .) number change was measured.

Two-month-old female calves were orally administered 4 g of mixed lactic acid bacteria (3.0 × 10 ¹¹ cfu / g) once a day for a week each morning. Fecal samples were taken before and after administration of the mixed lactic acid bacteria, and total coliform ( E. coli ) and Salmonella ( Salmonella ) in the feces of the calf before and after administration. spp .) and Staphylococcus The change of the number of spp .) was measured, and the result is shown in FIG.

As shown in Figure 1, the total coliform ( E. coli ) , Salmonella ( Salmonella ) present in the feces of the calf before the mixed lactobacillus administration spp.) The number of 6.4 log ₁₀ cfu / g and 3.0 log ₁₀ after the administration of the mixture of lactic acid bacteria in cfu / g log ₁₀ cfu / g value is the total E. coli 0 (E. coli), Salmonella (Salmonella of spp .) decreased significantly.

Next, according to whether or not the growth of the calf shows the above-described effect in the calf, and the concentration of the mixed lactic acid bacteria, total coliform ( E. coli ) , Salmonella ( Salmonella ) present in the calf spp .) and Staphylococcus To determine if the number of spp .) was different , seven-month-old calves were divided into three groups, and the group orally administered 4 g of mixed lactic acid bacteria (3.0 x 10 ¹¹ cfu / g) was group 2-1; Groups orally administered 8 g of mixed lactic acid bacteria (3.0 x 10 ¹¹ cfu / g) were classified into group 2-2. In addition, the group which did not administer mixed lactic acid bacteria was made into the comparative example 2-1. Total E. coli (E. coli) to separate the feces from these were then administered orally to the mixture of lactic acid bacteria in a concentration as described above to three groups once a day for five months every morning for the calf of the, Salmonella (Salmonella spp .) and Staphylococcus The change in the number of spp .) was measured and shown in FIG. 2.

The total number of E. coli, as indicated in Figure 2a is that of Comparative Example 2-1 6.1 In the case of log ₁₀ cfu / g, when the group 2-1 3.2 log ₁₀ cfu / g, the group 2-2 3.3 log ₁₀ cfu / g, in the case of group 2-1, 2-2 to which the mixed lactic acid bacteria were administered, it was confirmed that the total number of E. coli was reduced by half compared to Comparative Example 2-1 regardless of the concentration of the mixed lactic acid bacteria administered above. In addition, as shown in Figure 2b Total Salmonella ( Salmonella) For spp.) In the case of Comparative Example 2-1 by a mixture of lactic acid bacteria administration of 5.4 log ₁₀ cfu / g, when the group 2-1 2.0 log ₁₀ cfu / g, when the group 2-2 2.0 log ₁₀ cfu / g In the case of the group 2-1, 2-2 to which the mixed lactobacillus was administered, the total Salmonella bacteria ( Salmonella) compared to the comparative example 2-1 regardless of the concentration of the mixed lactic acid bacteria administered above The number of spp .) was reduced by half. In addition, as shown in Figure 2c Total Staphylococcus ( Staphylococcus) spp .) by the administration of mixed lactic acid bacteria 4.4 log ₁₀ cfu / g for Comparative Example 2-1, 2.6 log ₁₀ cfu / g for Group 2-1, 2.0 log ₁₀ cfu / for Group 2-2 as g, one group 2-1 administering a mixture of lactic acid bacteria, in the case of Staphylococcus 2-2 compared to Comparative example 2-1 by the administration of the administration of a mixture of lactic acid bacteria (Staphylococcus The number of spp .) was reduced by half.

Example 3 Antibacterial Activity of Mixed Lactic Acid Bacteria Culture

Example 1-2 above SPM1204 lactic acid bacteria, Lactobacillus know even field produced in loose (Lactobacillus consisting of acidophilus) and Pedy five years Tosa Caucus pen mouse (Pediococcus pentosaceus) In order to measure the antimicrobial activity of the mixed lactic acid bacteria probiotic composition was tested by the following method.

E. coli ATCC 1056 and E. coli ATCC 25922 were used as strains for testing antimicrobial activity. First, the mixed lactic acid bacteria probiotic composition was prepared in the same manner as in Example 1-2 in GAM (General anaerobic medium) (Nissui Pharm. Co. Ltd., Japan) broth N ₂ (90%), H ₂ (5%) and CO ₂ ( Pre-incubated for 18 hours at 37 ° C. under an anaerobic system filled with mixed gas (Bactron Anaerobic Chamber, Sheldon Manufacturing Inc., USA) consisting of 5%). E. coli ( E. coli ATCC 1056 and E. coli ATCC 25922) were preincubated for 18 hours at 37 ° C. under anaerobic systems in NA (Nutrient Agar) broth.

Table 2 shows the mixing ratio of the E. coli cultured in NA and GAM broth and the mixed lactic acid bacteria culture medium. For Groups 3-1 and 3-1 ', ⁹ ml of E. coli ATCC 1056 and E. coli ATCC 25922 were mixed with 1 ml of mixed lactic acid bacteria preculture with a concentration of 2.0 x 10 ¹¹ cfu / ml. 1: 9, and in groups 3-2 and 3-2 'of 5 ml of mixed lactic acid bacteria preculture with a concentration of 2.0 x 10 ¹¹ cfu / ml, E. coli ATCC 1056 and E. coli ATCC 25922 5 ml of each of the precultures were mixed well at a ratio of 1: 1 and then cultured under anaerobic conditions. In Comparative Examples 3-1 and 3-2, only 10 ml of E. coli ATCC 1056 and E. coli ATCC 25922 were used without mixing the mixed lactic acid bacteria.

After incubation under anaerobic conditions, bacteria were taken at 3, 6, 9 and 24 hour intervals, respectively, and diluted with saline and plated on EMB agar plates. After counting live colonies after 24 hours of culture, it was examined whether the mixed culture solution had a direct antimicrobial effect against E. coli and the results are shown in Table 3. The number of counted colonies was expressed in log ₁₀ cfu / ml.

As a result, as shown in Table 3, Comparative Examples 3-1 and 3-2 in which only E. coli ATCC 1056 and E. coli ATCC 25922 were cultured alone without mixed culture with the mixed lactic acid bacteria culture medium were E. coli numbers (log ₁₀ cfu / ml). In addition, the concentration of the mixed lactic acid bacteria culture medium used in the groups 3-1 and 3-1 'is for E. coli ATCC 1056 and E. coli ATCC 25922 It did not show antimicrobial activity.

However, for groups 3-2 and 3-2 ', these two E. coli strain In particular, the E. coli ATCC 1056 showed a log ₁₀ cfu / ml value of 0 only 6 hours after the mixed culture, and the E. coli ATCC 25922 showed a log ₁₀ after 9 hours. The cfu / ml value represented zero. In the case of groups 3-1 and 3-1 ', the concentration of lactic acid bacteria used was low, and thus did not show antimicrobial effect. Using 5 times the concentration of the mixed lactic acid bacteria showed excellent antibacterial effect.

On the other hand, Figure 3 is a view showing the antimicrobial effect of the mixed lactic acid bacteria against S. typhimurium ATCC 13311, S. typhimurium KCTC 2053 and S. aureus ATCC 6538p. Mixing lactic acid bacteria of the present invention exhibited the Tie Salmonella blood head Titanium (S. typhimurium) ATCC 13311, Salmonella tie blood head Titanium (S. typhimurium) KCTC 2053 and Staphylococcus aureus (S. aureus) antimicrobial activity even in the ATCC 6538p. For E. coli ATCC 1056 and E. coli ATCC 25922, the log ₁₀ cfu / ml value was 0 in 6 and 9 hours for groups 3-2 and 3-2 ', but the Salmonella typhimurium (S. typhimurium) ATCC 13311, Salmonella tie blood head Titanium (S. typhimurium) KCTC 2053 and Staphylococcus aureus (S. aureus) ATCC 6538p is E. coli It showed low antimicrobial activity compared to the strain.

The E. coli strain used in this embodiment would be to have to cause a variety of diseases such as colitis, including diarrhea with bacteria widely present in the gut of the animal, including the calf induces death of the calf, the bacterial, including the E. coli Mixed culture experiments with strains, it was confirmed that the mixed lactic acid bacteria culture medium exhibits a direct antimicrobial effect against the bacterial strains, including the E. coli .

Example 4 Growth Promotion Experiment of Calf

E. coli in Example 2 and Example 3 by the mixed lactic acid bacteria treatment The number was markedly reduced. To determine whether this effect directly affected the growth of calves, twelve calves aged 7 months were categorized and weighed.

Group 4-1 orally administered 4 g mixed lactic acid bacteria culture medium (3.0 x 10 ¹¹ cfu / g) was group 4-1; Groups orally administered 8 g of mixed lactic acid bacteria culture medium (3.0 × 10 ¹¹ cfu / g) were classified into group 4-2. In addition, the group which did not administer the said mixed lactic acid bacteria culture liquid was made into the comparative example 4. Mixed lactobacillus cultures administered in groups 4-1 and 4-2 were orally administered to seven-month-old calves once daily every morning for five months, and the calf weights were measured at regular intervals, as shown in Table 4. 4 and FIG. 4.

As shown in FIG. 4 and Table 4, in the case of Comparative Example 4, the total weight was increased by 65.6 kg, but by 80.6 kg in the group 4-1 and 94.3 kg in the group 4-2. In particular, in the case of Group 4-2, it can be seen that the concentration increased by about 1.5 times or more.

Thus, in a comprehensive analysis of the present embodiments, the composition according to the present invention reduced the number of intestinal E. coli in the calf, and also suppressed the growth of harmful microorganisms, including the intestinal E. coli , and thus required the calf in the intestine. It will serve to improve the balance of total bacteria. In addition, this will protect the animal from diseases caused by harmful bacteria in the intestine of the animal, including calves, thereby maintaining animal health and ultimately promoting animal growth.

Figure 1 shows the total E. coli , Salmonella spp . And staphyloccus spp . The change in number is shown.

Figure 2 shows the total E. coli , Salmonella present in the feces of calves after oral administration of mixed lactobacillus for 5 months for calf 7 months old. spp . And staphyloccus spp . The change in number is shown.

3 is E. coli ATCC 1056, E. coli ATCC 25922, Salmonella tie blood head Titanium (Salmonella typhimurium ) ATCC 13311, Salmonella typhimurium ( Salmonella typhimurium ) KCTC 2053 and S. aureus shows the antimicrobial activity of mixed lactic acid bacteria against ATCC 6538p.

Figure 4 measures the weight change of the calf after oral administration of the mixed lactic acid bacteria culture.

Claims (9)

Bifidobacterium pseudocatenulatum SPM1204 Lactobacillus (Accession Number: KCTC 10616BP) or its culture, Lactobacillus acidophilus or its culture, and Pediococcus pentosaceus ( Pediococcus pentosaceus) or its cultures in amounts of 10 ¹⁰ to 10 ¹³ cfu / g, respectively, Has a formal effect on E. coli ATCC 1056, E. coli ATCC 25922, Salmonella typhimurium ATCC 13311, Salmonella typhimurium KCTC 2053, Staphylococcus aureus ATCC 6538p , Probiotic composition for livestock, characterized in that it is used for calf feed. delete delete delete delete delete delete delete delete

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