KR20020043078A - Oligosaccharide and Probiotics Complex - Google Patents

Abstract

PURPOSE: Provided is a probiotic complex containing oligosaccharide. It improves the growth of animals by increasing a rate of nitrogen accumulation and a rate of weight gain, and immunity by increasing leukocyte count and globulin content. Also, it is used for the prevention or treatment of diabetes and heart disease, and as a health supplementary food. CONSTITUTION: The probiotic complex contains oligosaccharide, Lactobacillus group, Bifidobacterium group, Streptococcus salivarius subsp. Thermophilus as a main ingredient and several organic materials as additives, wherein the oligosaccharide is fructooligosaccharide; Lactobacillus group includes L. rhamnosus A, L. rhamnosus B, L. acidophilus, L. delbrueckii, L. bulgaricus or a combination thereof; Bifidobacterium group includes B. longum, B. breve and a combination thereof; and the additives include alfalfa grass juice powder, wheat grass juice powder, barley grass juice powder, soy lecithin powder, wheat sprout powder(gluten-free), beet juice powder, and bee pollen.

Description

Oligosaccharide-containing complex probiotics {Oligosaccharide and Probiotics Complex}

The present invention relates to a complex probiotic, more specifically, oligosaccharides and Lactobacillus microorganisms ( Lactobacillus group), Bifidobacterium group ( Bifidobacterium group), Streptococcus salivarius subsp. Thermophilus ) and oligosaccharides, characterized in that the addition of several additives in addition to the main component.

Recently, due to digestive disorders caused by excessive intake of instant foods due to the development of industrial civilization, and stressful digestive dysfunction due to diseases and heavy workload, it is difficult to maintain the intestinal flora total and the digestibility of nutrients rapidly decreases.

Therefore, the present invention seeks to promote the health of modern people by developing complex probiotics containing oligosaccharides.

By the way, conventional probiotics (generally probiotics) were some specific single agents or probiotics or prebiotics that have not yet been proved to be used. Therefore, research reports on conventional probiotics and prebiotics will be described in detail below.

Probiotics have been defined as living microorganisms that have a health-promoting effect when consumed in an adequate amount (Guarner and Schaafsma, 1988), suggesting that the longevity of Bulgarian rural people is due to the consumption of fermented milk. It was developed from the theory of the Nobel Prize-winning Russian scientist Mechnikoff (1908).

The mechanism of action of probiotics for improving human health has been proposed by Sanders (2000) as follows.

The mechanism of action on probiotic function is ① antimicrobial activity, ② inhibits colonization of harmful microorganisms, ③ immune effects (adjuvant effect, cytokine secretion, IgA secretion), promotes phagocytosis by peripheral vascular leukocytes, ④ It has been reported that there are antimutagenic effects, ⑤ effect on enzyme activity, ⑥ effect on enzyme transport, and so on.

In addition, the use of probiotics has been demonstrated in many cases in animals. In the following Table 1, the results of the study on the use of probiotics were sorted by breeder.

For young pigs, the reason for this is very stressful, and the addition of lactobacillus to the feed produces a growth-promoting effect, and the intestinal flora is balanced. However, this effect is hardly seen in the growing and finishing pigs in which the intestinal bacteria are settled. Smith and Jones (1963) reported that the addition of probiotics caused changes in the total flora in the intestine or minutes. As part of the experiment, feeding pigs with lactobacillus-containing milk with basic feed reduced the number of E. coli bacteria in the intestine. The increase in the number of lactic acid bacteria greatly improved the growth rate and feed efficiency. It has also been reported that probiotics enhance the utilization of nutrients in pigs. Collington et al. (1988) reported that probiotics not only improved growth and feed efficiency in weaning pigs, but also increased lactose activity, and Newman et al. (1988) reported Lactobacillus faecium. ) Secreted lysine to improve growth rate in growing pigs that lack lysine, while feeding lactobacillus to healthy pigs increased calcium absorption. Han et al. (1982) showed that S. faecium was tested in growing hog pigs for cultivation pigs, which showed 7.3% more probiotics and 13.7% for hog pigs. Improved 8.65% and 9.1%, respectively, and crude protein digestibility and nitrogen accumulation increased. Han et al. (1984a) found that the daily weight gain was 17% and feed efficiency was improved by 3.8% in the experiments with Clostridium butyricum added to weaning piglets. Experiments with the addition of probiotics in pigs showed a 3-15% daily gain and 4-49% feed efficiency, especially in young pigs.

Most of the effects of adding probiotics to poultry have been published with the results on the composition and changes of the intestinal flora, because the gain rate and feed efficiency improvement are related to the change of intestinal flora. The gut of healthy animals is dominated by acid-producing bacteria, which can be broken by stress, disease, and antibiotic treatment. In general, the strains of probiotics added to broiler feed should be able to adhere to the epithelial cells of the vesicles so that they can colonize well, because the poultry's digestive system begins in the vesicles. This is because the transition to the lower organs can prevent the growth of pathogenic bacteria in the entire intestine. To the teotu (Turtuero, 1973) is Lactobacillus ash was made also benefit the young chick's filler (Lactobacillus acidophilus) was added probiotics growth rate and feed efficiency is improved similarly phrase sphere and antibiotics, where the intestinal Escherichia coli (E. coli) The number is said to be significantly reduced. Han et al. (1984c) found that the addition of Lactobacillus sporogens to broiler feeds significantly improved the weight gain and feed efficiency, and led to staphylococci and coliforms. The number of pathogenic microorganisms and the concentration of ammonia are said to decrease.

Han et al. (1984b, c) found that the addition of 0.05% Clostridium butyricum to broiler feed improved 6.2% in weight gain and 6.4% in feed efficiency, and improved L. sporogenes . When added, the weight gain increased by 6.2% and the feed efficiency increased by 6.4%. Liu et al. (1994) reported that when 0.05% of lactic acid bacteria were added to laying hens' feed (62 weeks), egg production increased by 1.28% and feed efficiency improved by 1.4%.

(1994) reported that adding 0.5% lactic acid bacteria in feed for Israeli carp significantly improved daily gain and feed efficiency by 23.7% and 19.4%, respectively, and decreased nitrogen excretion by 11.3%.

Effect of Probiotics on the Growth Rate of Animals in Animals Breeder Age or weight Level of addition Daily gain (%) Feed efficiency (%) source pig 8.0 kg 7.0 kg 7.0 kg 14.7 kg 24.2 kg 34.1 kg 0.36ml / kg^One750.0ppm² 220.0ppm³0.03%⁴0.5%³2.0%⁶ + 5.4 + 3.5 + 9.0 + 7.0 + 2.7 + 18.6 + 10.6 + 41.9 + 49.2 + 3.8 + 8.8 + 16.4 Hale and Newton (1979) Pollmann (1980b) Pollmann (1980b) Han et al. (1984a) Noh et al. (1995) Jeon et al. (1996) Broiler 62.8g62.8g50.7g48.0g42.0g 0.05% ⁴ 0.04% ⁷ 0.1 g / kg ⁸ 0.1% ⁹ 1.0, 0.25 g / kg ¹⁰ + 6.0 + 6.2 + 5.4 + 10.1 + 7.4 + 6.4 + 6.4 + 1.8 + 6.1 + 4.8 Han et al. (1984b) Han et al. (1984c) Mohan et al. (1996) Yeo and Kim (1997) Cabavazzoni et al. (1998) Laying hen 62wks 0.05% ³ +1.28 ^a +1.4 ^b Ryu et al. (1994) carp 4.3 kg 0.5% ³ +23.7 +19.4 Noh et al. (1994) (Han et al. 2000b)^OneLactobacillus fermentation product,²Lactobacilli,³Lactic acid bacteria,⁴Clostridium BoutiqueCl. butyricum),⁵Lactobacillus ashdophilus (L. acidophilus) KC8,⁶Micrococcus species (Micrococcus species)Guasacaromyces spp.Saccharomyces speciesFermentation product).⁷Lactobacillus sporogen (L. sporogenes),⁸Lactobacillus ashdophilus (Lactobacillus acidiphilus),Lactobacillus Cathay (Lactobacillus casei), Bifidobacterium Bifidem (Bifidobacterium bifidum), Aspergillus DuckAspergillus oryzae) And tolulopsis (Torulopsis),⁹Lactobacillus Cathay (Lactobacillus casei), ¹⁰ Bacillus coagulans (Bacillus coagulans) 1000 mg / kg 1 to 7 days; 250 mg / kg 8 days to 49 days,^aEgg weight (g / hen / d),^bEgg weight / feed intake

And Lactobacillus ash FIG filler's (Latobacillus acidophilus), Lactobacillus Cathay (L. casei), Lactobacillus del Brewer station (L. delbruekii) Probiotics are commonly used are microorganisms belonging to the genus Lactobacillus (lactobacilli) in humans, Only one species may be used or in combination with another species. Other microorganisms used as probiotics include Bifidobacterium adolescentis , B. bifidum , B. longum , B. longum , which belongs to the Bifidobacteria microorganism. Streptococcus salivarius subsp.thermophilus belonging to the B. infantis and Streptococci microorganisms, and Streptococcus lactis ( S. lactis ) Gibson and Roberfroid, 1995). It has been hypothesized that these probiotics will have a positive effect on numerous biomedical conditions.Goldin and Gobach (Gordin and Gorbach, 1992) summarize the effects of probiotics, which prevent them from diarrhea and constipation. It plays a positive physiological role in preventing, preventing the dominance of harmful microorganisms and lowering cholesterol levels.

According to the above, the report on the probiotics has progressed considerably, and there are many experimental data. However, the research on prebiotics is still poor. Hereinafter, the prebiotics, which have recently emerged, will be described in detail.

According to Gibson and Roberfroid (Gibson and 1995), prebiotics are "undigested food ingredients, which selectively promote the growth or activity of microorganisms of limited species in the large intestine, thereby enhancing the health of the host." to be. To be classified as a prebiotic in food ingredients, it must: 1) not degrade or absorb at the top of the digestive system, 2) promote the growth or metabolism of one or a limited number of beneficial microorganisms living in the large intestine, and 3) consequently, intestinal The microorganisms should be able to achieve a healthier colony, and 4) promote the health of the host by having a beneficial effect on the intestinal tract and the entire body.

Among the food ingredients, prebiotics are indigestible carbohydrates (oligosaccharides, polysaccharides), several peptides and proteins, and fats (ethers, esters). These constituents are not degraded or absorbed by digestive enzymes in the upper digestive tract because of these structural properties. These ingredients may be referred to as "colon foods". In other words, it enters the large intestine and acts as a substrate for endogenous intestinal microorganisms, indirectly supplying the host with energy, metabolic substrates, and essential microbial nutrients (Gibson and Roberfroid, 1995).

Indigestible carbohydrates in colorectal foods naturally meet the prebiotic criteria defined above. In the case of proteins or peptides in milk or plants, they are not partially digested but mainly have a beneficial effect by helping the intestinal absorption of cations such as calcium and iron and promoting immune function. Rather, anaerobic proteolysis produces mainly the harmful compounds ammonia and amines, and indigestible lipids are not well known for the metabolism of intestinal microflora (Macfarlane and Cummings, 1991).

According to Gibson and Wang (1993), most of the resistant starch, non-starch polysaccharides, and nondigestible oligosaccharides in the case of indigestible carbohydrates were found in Participate in the process but have an unspecified tendency to promote the growth of both beneficial and harmful microbes to humans. As a result, these carbohydrates lack the metabolic selectivity that limits them to beneficial microorganisms such as lactobacilli and bifidobacteria.

As mentioned earlier, not many of the colonic foods meet the requirements of prebiotics. In the case of lactulose, lactobacilli in the children's intestine was used for the manufacture of infant formula forty years ago. Attempts have been made to increase the number of microorganisms (MacGillivary et al., 1959). Recent studies have shown that fructo oligosaccharides increase the number of bifidobacteria microorganisms in adults (Gibson et al., 1995; Buddington et al., 1996; Gibson, 1999; Bouhnik et al., 1999), and galacto oligosaccharides showed similar results when fed to rats (Rowland and Tanaka, 1993).

The oligosaccharides related to the above-described prebiotics will be described in more detail below. Among the oligosaccharides, which are the major constituents of the complex probiotic, to be developed, especially fructooligosaccharides, met oligosaccharides, and the like are attracting much attention in relation to the intestinal microorganisms.

Fructo-oligosaccharide has 2, 3 or 4 fructose bound to glucose, and the binding between fructose is composed of β (1 → 2) (Formula 1). Hidaka et al. (1986) reported that fructooligosaccharides increased the intestinal bacterial flora, and Ota et al. (Ohta et al., 1996) reported that they also influence the absorption of minerals. In addition, there are many reports that fructooligosaccharides exhibit a number of beneficial microorganisms, such as Bifidobacteria (Buddington et al., 1996; Gibson, 1999; Bouhnik et al., 1999).

Mannanoligosaccharides are carbohydrates isolated from the cell walls of yeast. Many reports have found that oligosaccharides are effective in the growth and immunity of animals. The structurally met oligosaccharide primary carbohydrate, D-mannose, is present in most cell walls and acts as a cell surface binding receptor for certain microorganisms such as Escherichia coli and Salmonella (Ofek and Sharon, 1990; Baba et al., 1993). Savage and Zakrezewska (1996) reported that met oligosaccharides isolated from the cell walls of yeast enhance turkey immunity, and MacDornald (1996) reported that met oligosaccharides lowered mortality in broilers. It was. Although the exact mechanism of action has not yet been established, met oligosaccharides enhance protective immune responses and have a beneficial effect on animal growth (Cotter, 1997). Met oligosaccharides promote humoral and cellular immune responses. The addition of met oligosaccharides increased IgG levels in the small intestine and serum, and increased lymphocyte count and white blood cell phagocytosis in the small intestine (Spring and Privulescu, 1998).

Accordingly, the present invention provides a combination of oligosaccharides belonging to a prebiotic and a probiotic that has already been proven to be effective, unlike the conventional single probiotics or single prebiotics described above. The purpose is to develop and provide food supplements, that is, new functional probiotics.

The present invention for achieving the above technical problem,

Oligosaccharides and Lactobacillus microorganisms ( Lactobacillus ), Bifidobacterium microorganisms ( Bifidobacterium ), Streptococcus salivarius subsp. Thermophilus and oligosaccharides characterized in that the additive is added in addition to the main component To a combined probiotic.

In more detail, the configuration of the present invention,

The oligosaccharide is characterized in that the fructooligosaccharide, the Lactobacillus genus microorganisms are Lactobacillus rhamnosus A ( L.rhamnosus A ), Lactobacillus rhamnosus B ( L.rhamnosus B ), Lactobacillus ashdophyllus ( L acidophilus ), Lactobacillus delbrueckii ( L.delbrueckii ), Lactobacillus Bulgaricus ( L.bulgaricus ), characterized in that consisting of one or two or more combinations, the Bifidobacterium microorganisms ( Bifidobacterium group) is Bifidobacteria long gum ( B. longum ), Bifidobacteria is characterized in that consisting of one or two combinations of Bre ( B.breve ).

The present invention will be described in more detail using examples and experimental study charts as follows.

First, in the embodiment of the present invention, in order to investigate the effect of the addition of the oligosaccharide-containing complex probiotics, 28 rats having an average weight of 91 g were disclosed for three weeks. Test fixtures were placed by a full random placement method. Test feeds were formulated using normal feed as shown in Table 2, and crude protein and metabolic energy contents were 19% and 3,176 ㎉ / ㎏, respectively, during the first 1 week (phase I) after the start of the test. During phase II), crude protein and metabolic energy contents were 16% and 3,170 mA / kg, respectively. The feeding rate of the complex probiotics to be developed was 1.76% during the first week of the experiment and 3.0% during the second to three weeks.

Table 3 shows the formulas of the complex probiotics to be developed. Each 5.0 g of the combined probiotic contains 2.5 billion and 2 billion Lactobacillus and Bifidobacterium groups, and Streptococcus salivarius subsp. Thermophilus ) Contains 400 million, oligosaccharides (500 mg).

At the end of the specification test, blood was collected from each of the three rats from the control and test zones, and three rats were treated per treatment to determine intestinal digestive enzymes, intestinal morphology, organ weight, and colonic microbial count. , Pancreas, small intestine and colon were collected.

The environmental temperature of the test animal breeding room was maintained at an average of 22 ± 1 ℃, relative humidity of 65 ± 5%, and the contrast was adjusted by a 12 hour cycle (08:00 ~ 20:00). Water and feed were freely provided for the entire test period. Seven days before the end of the test, three repetitions of each treatment were placed in a metabolic cage, and then, after four days of preliminary periods required for metabolic cage adaptation, foreign matter was collected for three days to remove foreign substances and the total weight was measured. Collected minutes were lyophilized and crushed and stored frozen until analysis, and the general components were analyzed according to the AOAC (1990) method. Urine was collected at the time of sample collection, washed with 0.1% HCl to prevent deterioration of the sample, and stored at -20 ℃ until analysis. Nitrogen accumulation rate was subtracted from intake of nitrogen and discharged from urine. Calculated by value.

The stomach, pancreas and small intestine were opened and removed, and the fat attached to the organs was removed cleanly, washed with physiological saline (0.9% NaCl) to remove blood, and drained with filter paper, and then weighed and measured.

The activity of lactase, maltase and sucrase in the small intestine was measured by Dahlqvist method, and the activity of disaccharide degrading enzyme was expressed as specific activity (unit of activity / protein g) and the protein content of enzyme solution was Bovine serum albumin was used as a standard and quantified by the Lowry method.

The small intestine's villi and choroidal depth are applied to the Pluske method, and the small intestine is excised from three parts (duodenum, jejunum, ileum) and 10% phosphate-buffer formalin (pH 7.4). ) Was fixed for 10 days and observed with an optical microscope.

Intestinal microbial analysis was performed by diluting the contents of the colon in 0.1% peptone water and diluting them at a constant rate. Products 3M Health Care, USA; AOAC, 1995) was inoculated with intestinal bacteria for 24 hours at 35 ℃ and E. Coli for 48 hours at 35 ℃ to count colonies.

In order to examine the blood characteristics of rats, anesthetized with ethyl ether, blood was collected from the heart, placed in a sodium heparin-treated tube, and the blood analyzer was examined for leukocytes and red blood cells. The remaining hematological properties were analyzed by an automated blood analyzer.

Feed Formulation Table and Nutrient Content Raw feed and nutrient content Phase Ⅰ Phase II Control Complex Probiotics Control Complex Probiotics Raw material feed (%): Wheat soybean meal alfalfa corn fish flour soybean milk salt lysine choline calcium phosphate limestone vitamin complex mineral total 47.8024.703.2017.031.501.500.60-0.301.531.340.50-100.00 48.0025.102.2014.671.702.300.60-0.301.531.340.501.76100.00 52.4513.733.7422.172.001.280.600.200.300.882.150.50-100.00 54.1015.151.7816.502.002.800.600.180.300.882.210.503.00100.00 Nutrients content: Metabolic energy (M㎈ / ㎏) crude protein lysine methionine + cystine calcium phosphorus 3176.0519.251.020.601.200.62 3172.0119.211.020.601.200.62 3170.3916.010.930.581.200.62 3170.9016.000.930.581.200.62 (Han et al. 2000b)

Composition of Complex Probiotics (Content per 5.0g) Lactobacillus group 2.5 billion Lactobacillus rhamnosus A (L.rhamnosus A), Lactobacillus rhamnosus B (L.rhamnosus B), Lactobacillus ashdophilus (L.acidophilus), Lactobacillus del Bruieki (L.delbrueckii), Lactobacillus Bulgaricus (L.bulgaricusBifidobacterium group 2.0 billion Bifidobacterium long gum (B.longum), Bifidobacterial Breb (B.breveStreptococcus salivarius subspecies dermaphilusStreptococcus salivarius subsp. Thermophilus)400 million Alfalfa grass juice powder 400 mg Wheat grass juice powder 400 mg Barley grass juice powder 400 mg Soy lecithin powder (99% oil-free) 1000 mg Wheat sprout powder (gluten-free) 450 mg Beet juice powder 300 mg Bee pollen 200 mg Fructooligosaccharides (FOS) 1000 mg (Han et al. 2000b)

Table 4 summarizes the effects of combined probiotic supplementation on the growth of rats. Compared with the control group, the daily weight gain and feed efficiency of the combined probiotics were significantly improved (p <0.05). This result is a result of the combined effect of probiotics and fructooligosaccharides, and showed a significant difference in daily weight gain and feed efficiency even though the stress was minimized by ideally adjusting the specification conditions. In older mice, greater effects can be expected.

Effect of Combined Probiotics on Growth in Rats Treatment Starting weight (g) End weight (g) Daily weight gain (g / day) Feed Intake (g / day) Feed efficiency Control Test Standard Error 90.991.22.57 250.8 ^b 268.0 ^a 5.33 7.62 ^b 8.42 ^a 0.19 22.6324.020.44 2.98 ^a 2.85 ^b 0.04 ^{a, b} Different subscripts are statistically significant between mean values (p <0.05). (Han et al. 2000b)

Increasing nutrient digestibility in rats was observed by adding complex additives to feed (Table 5). The energy and fat digestibility of the test group was significantly higher than that of the control (p <0.05), and the digestibility of dry matter, crude protein, crude ash, and phosphorus also showed higher levels of digestibility of the test group than the control group. The improved digestibility of nutrients seems to be due to the enzymatic activity promotion and enzyme transport function activation of complex probiotics. In this experiment, the digestibility of phosphorus was increased by 7% in the addition group, but there was no significant difference.

Effect of Combined Probiotics on Digestibility of Rats (%) Treatment building energy Crude protein Crude fat View minutes calcium sign Control Test Standard Error 81.8482.770.29 81.48 ^b 83.68 ^a 1.15 78.8881.150.56 77.25 ^b 80.80 ^a 0.81 47.2348.110.83 62.5659.391.02 51.3955.070.56 ^{a, b} Different subscripts are statistically significant between mean values (p <0.05). (Han et al., 2000)

Feeding the combined probiotics was found to enhance the activity of carbohydrate degrading enzymes in the small intestine of rats (Table 6). The enzyme activities of lactase, maltase and sucrase were significantly higher in the test group than in the control group (p <0.05). As shown in Table 5, the increase in energy digestibility in the complex probiotics will be described as the increase in enzyme activity.

Effect of Complex Probiotics on the Activity of Carbohydrate Degrading Enzymes in the Small Intestine of Rats Treatment Lactase (unit / g protein) * Maltase (unit / g protein) Sucrase (unit / g protein) Control Test Standard Error 10.89 ^b 25.69 ^a 3.34 60.68 ^b 106.04 ^a 10.3 10.02 ^b 22.76 ^a 2.91 ^ab Different subscripts are statistically significant between the mean values (p <0.05). * unit: μmol of substrate degraded per minute. (Han et al. 2000b)

On the other hand, the effect of the combination probiotics on the nitrogen accumulation rate in rats is summarized in Table 7. When considering only the differentiation rate, crude protein digestibility tended to be higher in the test group (Table 5), and the nitrogen accumulation rate of the test group was higher than that of the control group (Table 7).

Effect of Combined Probiotics on Nitrogen Accumulation in Rats Treatment N intake (g / day) N in urine (g / day) N in g / day Daily accumulation (g / day) Accumulation rate (%) Control Test Standard Error 0.610.640.01 0.170.170.01 0.130.120.01 0.310.340.01 50.8253.720.92 (Han et al. 2000b)

Table 8 summarizes the effects of the combination probiotics on organ weight and intestinal microflora in rats. Liver and pancreas weights of rats fed the probiotics were heavier than controls, but there was no statistical difference in the length or weight of the small intestine (p> 0.05). Therefore, it was shown that the supplementation of the combined probiotics helped the development of the liver or pancreas in the digestive organs and further the digestion, metabolism and detoxification of nutrients. Enteric microbial count and E. coliE. Coli) The number was significantly lower in the test group (p <0.05). The small number of E. coli in the test plots suggests that the combined probiotic acts to inhibit harmful microbes.

Effects of Combined Probiotics on Organ Weight and Intestinal Microflora in Rats Treatment Stomach (g) Liver (g) Pancreas (g) Intestine microbe Length (cm) Weight (g) Enteric microorganisms (log ₁₀ CFU / mL) E. Coli (log ₁₀ CFU / mL) Control Test Standard Error 1.461.300.09 12.0314.170.95 0.730.830.07 115.3108.04.38 12.1312.230.20 2.351.250.08 2.141.130.16 (Han et al. 2000b)

As shown in Table 9, the addition of the combined probiotics to the feed of young rats showed that the villus height of the duodenum and ileum was much higher than that of the control, so that the absorption of nutrients in this area was much better than that of the control. Melting depth tended to be deeper in the control than in the control, which also showed that the probiotic had a good effect in the gut. Overall, the height of the villi in the test zone was higher, and the depth of the vortex became shallower, and the supply of the complex probiotic was found to provide a favorable condition for the absorption of nutrients. These results indicate that rats fed with multiprobiotics increase the enzyme activity and consequently improve the digestibility of nutrients. In addition to maintaining the total bacterial flora in vivo, this proliferation of nutrients has been shown to contribute to improving the digestibility of nutrients.

Effect of Combined Probiotics on the Height, Melt, and Depth of Villi in Rat Small Intestine Treatment Villi Height (μm) Melting depth (μm) Duodenum factory Chairman Duodenum factory Chairman Control Test Standard Error 456.04541.1241.91 636.56588.4119.80 554.58594.1520.14 261.08179.5927.83 194.19166.7510.13 280.08224.6014.28 (Han et al. 2000b)

It is summarized in Table 10 that the fed probiotics in rat feed resulted in significantly better changes in the blood properties of rats.

Effect of Combination Probiotics on Blood Characteristics in Rats Treatment Leukocytes (10 ³ / μl) Erythrocytes (10 ⁶ / μl) Protein (g / μl) Albumin (g / μl) Globulin (g / μl) Calcium (mg / ㎗) Phosphorus (mg / dl) Magnesium (mg / ㎗) Control furniture standard error 10.21b13.66a0.89 7.08b7.60a0.19 5.735.700.08 3.032.800.06 2.702.900.07 11.7312.070.14 7.037.000.36 1.901.770.04 (Han et al. 2000b)

When probiotics and oligosaccharides were combined, probiotics and leukocyte counts and globulin contents were 34% and 7% higher than controls, respectively (p <0.05). Leukocytes are blood cells that confer immune function in the animal's body, and five subpopulations in leukocytes are involved in phagocytosis, inflammatory response, unspecific and specific cellular defense. Globulin, one of the major plasma proteins, is also important in boosting immune responses in animals and is also involved in the transport of fat and fat-soluble vitamins. In this experiment, we did not inject specific pathogens or antigens, and ideally provided environmental conditions such as temperature, humidity, light, feed, etc. The increased trend and the increased globulin content suggest that the new probiotics supplemented with oligosaccharides will improve disease resistance in animals.

The present invention is not limited to the technical spirit of the above-described specific embodiments or diagrams, and any person of ordinary skill in the art without departing from the gist of the present invention claimed in the claims. Various modifications are possible, of course, and such changes are within the scope of the claims.

When explaining the effect of the configuration of the present invention,

First, the rats of the experimental group to which the combined probiotic was added showed a significant improvement in the growth capacity, especially the feed efficiency. This improvement in feed efficiency appears to be due to increased enzyme activity, increased intestinal villi, and decreased melting and depth.

Second, the leukocyte count of the test group was much higher than that of the control group, and the globulin content tended to be somewhat higher. These phenomena demonstrate that the product's excellent physiological efficacy in combination with oligosaccharide-based probiotics enhances immunity to disease in rats.

Third, the combined probiotics significantly reduced the number of Escherichia coli. This shows the harmful microbial inhibitory effect of the combined probiotic.

Fourth, the liver or pancreas developed more in the digestive system when combined probiotics were added.

Fifth, rats fed the test complex probiotics improved digestibility of various nutrients and improved nitrogen accumulation.

Sixth, the feeding of complex probiotics greatly increases the activity of disaccharide degrading enzymes in the small intestine. The above-mentioned improvement in digestibility is due to this mechanism.

Taken together, the present invention, the combined probiotic added oligosaccharide of the present invention seems to have a synergistic effect of the function of the oligosaccharide to increase the enzyme activity and maintain the intestinal bacterial worm and the function of the probiotic. In the present embodiment, young mice in a healthy state were targeted, but it is estimated that the effect is more effective when the digestive function is poor, such as the elderly, the disabled and dementia patients.

Claims (5)

In probiotics, The main component is oligosaccharide and Lactobacillus microorganism ( Lactobacillus group), Bifidobacterium group ( Bifidobacterium group), Streptococcus salivarius subsp. Thermophilus and additives in addition to the main component is characterized in that the addition Complex probiotics containing oligosaccharides. The method of claim 1, The oligosaccharide is a probiotic complex containing oligosaccharides, characterized in that the fructooligosaccharides. The method of claim 1, The Lactobacillus genus microorganisms are Lactobacillus rhamnosus A ( L.rhamnosus A ), Lactobacillus rhamnosus B ( L.rhamnosus B ), Lactobacillus ashdophyllus ( L.acidophilus ), Lactobacillus del Bruieki ( L. delbrueckii ), L.bulgaricus complex probiotics containing oligosaccharides, characterized in that consisting of one or two or more combinations. The method of claim 1, The bifidobacteria spp bacteria (Bifidobacterium group) are bifidobacteria bacteria ronggeom (B.longum), the combined probiotic bifidobacteria containing the oligosaccharide which is characterized by being a one or two combinations of bacteria breather bracket (B.breve). The method according to any one of claims 1 to 4, The complex probiotic containing the oligosaccharide complex probiotic containing an oligosaccharide, characterized in that the configuration of the following table. Ingredients of Complex Probiotics (Content per 5.0g) Lactobacillus group 2.5 billion Lactobacillus rhamnosus A (L.rhamnosus A), Lactobacillus rhamnosus B (L.rhamnosus B), Lactobacillus ashdophilus (L.acidophilus), Lactobacillus del Bruieki (L.delbrueckii), Lactobacillus Bulgaricus (L.bulgaricusBifidobacterium group 2.0 billion Bifidobacterium long gum (B.longum), Bifidobacterial Breb (B.breveStreptococcus salivarius subspecies dermaphilusStreptococcus salivarius subsp. Thermophilus)400 million Alfalfa grass juice powder 400 mg Wheat grass juice powder 400 mg Barley grass juice powder 400 mg Soy lecithin powder (99% oil-free) 1000 mg Wheat sprout powder (gluten-free) 450 mg Beet juice powder 300 mg Bee pollen 200 mg Fructooligosaccharides (FOS) 1000 mg