JP3173859B2 - Liquid or liquid food - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a liquid or fluid food which is effective for improving intestinal flora. More particularly, the present invention relates to human and animal diseases caused by an imbalance of the intestinal flora, especially humans who are forced to consume liquid or fluid foods as staple foods, for example, patients receiving enteral nutrition The present invention relates to a food which is effective for treating and preventing diarrhea and infectious diseases frequently occurring in premature babies or infants fed with artificial milk.

[0002]

BACKGROUND OF THE INVENTION The composition of the intestinal flora (intestinal flora) of animals is almost constant as long as the environment and diet are stable and the host is healthy. However, the ecosystem of this intestinal flora fluctuates due to various factors such as the age of the host, disease, microbial infection, stress, nutritional components, drug administration, and sometimes an abnormal intestinal flora is formed, and the health of the host May cause adverse effects.

[0003] Indigenous bacteria constituting the intestinal flora are classified into three groups in relation to the host. That is, Group I is a group of bacteria that are the most predominant bacteria in normal intestinal flora and are symbiotic with the host. Group II is a group of bacteria that first appears after the birth of the animal and constitutes the dominant flora once but diminishes shortly thereafter and remains moderate. Group III belongs to a minority in the intestinal flora and is a group of bacteria that can break opponent's defense mechanisms and cause opportunistic infections to exert pathogenicity (Toshimitsu Mitsuoka, Enterobacteriaceae) Classification and ecology of the flora, pp. 51-52, published by The Dietary Study Group, 1986).

[0004] The species belonging to Group I include microorganisms belonging to the genus Bifidobacterium (hereinafter referred to as Bifidobacterium), microorganisms belonging to the genus Bacteroides (hereinafter referred to as Bacteroides), and belonging to the genus Eubacterium. It is an anaerobic bacterium such as a microorganism (hereinafter referred to as Eubacterium) and includes a microorganism belonging to the genus Lactobacillus (hereinafter referred to as Lactobacillus) depending on the animal species. Bifidobacterium and Lactobacillus are typical useful bacteria and are said to be useful bacteria for host health.

[0005] The species belonging to Group II include microorganisms belonging to the genus Enterobacteriaceae (hereinafter referred to as Enterobacteriaceae), microorganisms belonging to the genus Streptococcus (hereinafter referred to as Streptococcus), and the like. It tends to increase when the condition worsens.

[0006] The bacterial species belonging to Group III include microorganisms belonging to the pathogenic strain Enterobacteriaceae (hereinafter referred to as pathogenic enterobacteriaceae) and microorganisms belonging to the genus Staphylococcus (hereinafter referred to as Staphylococcus). ), A microorganism belonging to the genus Clostridium (hereinafter referred to as Clostridium), and a harmful bacterium for the host.

[0007] In order to maintain the health of animals, it is necessary to maintain bifidobacterium and lactobacilli in the intestine at a high bacterial count level and to prevent harmful pathogens such as pathogenic Enterobacteriaceae, Streptococcus, Staphylococcus, and Clostridium. It is important to control the growth of bacteria and keep them low. In order to maintain or improve the composition of intestinal flora normally, usually, antibiotics, lactulose, bifidobacterium growth factors such as fructooligosaccharides, acid bacteria, bifidobacterium, enterococcus, clostridium, etc. Are administered.

On the other hand, lactoferrin in milk is known to have a protective effect against infection, and has already been added to milk powder. Further, this lactoferrin has an effect of inhibiting the adhesion of pathogenic bacteria to teeth, gastrointestinal tract cells and the like, and discloses an agent for inhibiting the adhesion of pathogenic bacteria and its use in foods and drinks containing the same (Japanese Patent Application Laid-Open No. 3-220130). .

[0009]

However, when an antibiotic is used, it is affected by useful and harmful bacteria in the intestine, and certain strains become resistant to drugs. The problem of transmission of drug resistance by the R factor has led to severe restrictions on the use of antibiotics. On the other hand, for Bifidobacterium growth factor and viable bacterial preparations, a certain effect has been observed at the laboratory level, but it is not always possible to obtain a certain effect when applied to human or animal breeding. . This is because in actual breeding, the intestinal and external microbiological conditions are different, the environmental conditions are complicated, and in the case of live bacteria preparations, the bacterial species used, the amount of live bacteria, and the stability in feed This is because there remains a problem in sex, survival in the gastrointestinal tract or fixation. In particular, the strain used varies depending on the species and age of the animal, and selection of an effective strain is an extremely difficult task. Bifidobacterium growth factor is ideally a nutrient substrate for only Bifidobacterium in the intestine, and by oral administration, promotes the growth of Bifidobacterium and thus suppresses harmful bacteria. I do. However, administration of Bifidobacterium growth factor and a viable bacterial preparation does not directly control harmful bacteria, but is expected to have an indirect effect, and its efficacy is weak and has no immediate effect. In addition, regarding the effect of Bifidobacterium growth factor, there is no effect unless Bifidobacterium is present in the intestine of the host, and there is no guarantee that Bifidobacterium cannot serve as a substrate for other bacteria. If the number of bacteria is low, it takes a long time to grow dominantly. Further, some Bifidobacterium growth factors are low molecules, and have an increased water content to increase the osmotic pressure of intestinal contents, and have side effects such as diarrhea.

On the other hand, with respect to the antibacterial activity of lactoferrin, in vitro experiments have been carried out mainly on Escherichia coli, microorganisms belonging to the genus Enterobacteriaceae, Streptococcus, Vibrio (hereinafter referred to as Vibrio), Pseudomonas sp. It is known that microorganisms belonging to the genus C. (hereinafter referred to as Pseudomonas) and microorganisms belonging to the genus Candida (hereinafter referred to as Candida) exhibit antibacterial activity. This antibacterial activity is mainly observed in non-iron-bound apolactoferrin, but in iron-saturated lactoferrin, its action is greatly reduced. It is also known that among strains belonging to bacteria present in the intestine of animals such as enterobacteriaceae, Streptococcus, Staphylococcus, Vibrio, Pseudomonas, etc., there are many strains which are insensitive to lactoferrin. [Arnold, RR et al; Infection and Immunit
y), Vol. 28, pp. 893-898, 1980; Koji Ushijima et al., Journal of Infectious Diseases, Vol. 65, No. 1, pp. 54-60,
1990; and JP-A-3-181421).

[0011] Regarding the physiological activity of lactoferrin in vivo, in a test for the prevention of adherence to E. coli using germ-free mice, pathogenic E. coli was orally administered to germ-free mice and then bovine lactoferrin (1 mg / 0.2 ml / day). ) Was orally administered, and then the mice were dissected and the intestinal tract isolated was homogenized, and the number of bacteria was counted. As a result, it was disclosed that the number of pathogenic Escherichia coli detected was lower than that of the group without bovine lactoferrin. (The aforementioned JP-A-3-220130). Regarding the effect of bovine lactoferrin on the adhesion of pathogenic Escherichia coli, see, eg, Gourd in Rice (25th Annual Meeting of the Japan Society of Aseptic Biological Notology, Abstract, p. 22, p. 1992).
Year). According to the results, when the drinking water containing iron-unsaturated bovine lactoferrin (10 mg / ml) was orally administered, the effect of inhibiting the intestinal mucosa adhesion of toxin Escherichia coli established in the germ-free mouse was observed.

[0012] As described above, bovine lactoferrin is known to have an action of preventing pathogenic Escherichia coli from adhering to the intestinal tract of a germ-free mouse, but is not known to improve the intestinal flora of animals. In the results of the same test conducted by the inventors of the present invention, bovine lactoferrin added to drinking water has no effect of reducing the number of Escherichia coli in the contents of the digestive tract, as long as a normal diet is provided. Was.

Regarding the effect of lactoferrin-added milk,
When low-birth-weight infants are fed lactoferrin-added milk at a concentration of 6.5 mg / 100 ml, the proportion of bifidobacterium in the total bacterial count in feces tends to increase,
It is known that the detection rate of Staphylococcus tended to be low (the 32nd Annual Meeting of the Japanese Society of Premature and Infant Neonates, page 86, 1987). Similarly, one for low birth weight infants
When lactoferrin-added milk is fed at a concentration of 00 mg / 100 ml, the proportion of bifidobacterium in the total bacterial count in feces tends to increase, and the constituent ratio of enterobacteriaceae and Clostridium in the total bacterial count is increased. It is known that there was a tendency to decrease (Perinatal medicine, Vol. 19, No. 4, pp. 125-129, 1989)
Year). However, these reports also include Enterobacteriaceae, Clostridium, Staphylococcus,
It has also been shown that the bacterial count in Streptococcus faeces hardly changes during lactation of lactoferrin-supplemented milk. Lactoferrin in breast milk is 100-500mg
/ 100 ml, especially in colostrum. The literature mentioned above, even if lactoferrin is added at the same concentration as breast milk,
Dramatically reports that the intestinal flora is not improved. It has also been pointed out that the addition of bovine lactoferrin to childcare milk and nursing newborns has little effect on intestinal flora [Balmer et al., Archives of Diseases in Child. Food (Balmer, SE et al; Archives of Disease in Child
hood) 64, 1685-1690, 1989]. That is,
Even if lactoferrin is added to milk at the same concentration as that of breast milk, the absolute value of the number of harmful bacteria cannot be reduced.
On the other hand, the intestinal flora of young animals, premature babies, or newborns in the early stage of development is predominantly harmful bacteria such as Enterobacteriaceae, Clostridium, Staphylococcus, Streptococcus, etc. Inner flow
It is known to be a constituent of La and it is thought to cause diarrhea and infections. In addition, patients who can only consume liquid or fluid foods due to various diseases,
In addition, even in infants fed only with artificial milk, intestinal flora is thought to cause high levels of harmful bacteria and cause diarrhea and infection.

[0014] Diseases of the sick and young animals, which can consume only liquid or fluid foods, are serious problems economically and socially, and are safe for improving intestinal flora abnormalities. Despite the desire to develop effective methods and means for preventing intestinal flora abnormalities, the previously known means have many limitations.

The present invention has been made in view of the above circumstances, and is intended to improve or prevent intestinal flora abnormalities in sick and young animals who can only take liquid or fluid foods. It aims to provide safe and effective means.

[0016]

Means for Solving the Problems The inventors of the present invention have:
In order to solve the above-mentioned problems, as a result of intensive studies to develop a liquid or fluid food having an action to improve the abnormality of the intestinal flora, surprisingly, bovine lactoferrin and various substances were mixed. The combination was found to exhibit completely different effects in vivo (intestinal tract) than in test tubes. That is, even when the concentration of lactoferrin has no effect in a test tube, when ingested as a liquid or fluid food in combination with other substances, the intestinal flora of normal animals (non-sterile animals) and humans Has been found to have a remarkably improving effect, and the present invention has been completed.

That is, the present invention relates to (A) bovine lactoferrin, (B) at least 0.01% (by weight) of a water-soluble indigestible saccharide, and at least 0.5% (by weight) of bovine serum albumin. Or any mixture of these,
The present invention provides a liquid or fluid food effective for improving intestinal flora, comprising:

Hereinafter, the structure of the present invention and preferred embodiments thereof will be described in detail.

In the following description, percentages are values by weight unless otherwise specified, except for the detection rate.

The water-soluble indigestible saccharide used in the present invention is the invention (JP-A-2-222) filed by the present applicant.
No. 659) (Reference Example 1). Lactulose, which is one of the indigestible saccharides, can be commercially available or can be produced by the method described in the invention already filed by the applicant of the present invention (Japanese Patent Publication No. 52-2984). 2).

The food of the present invention is a liquid or fluid food which is effective for improving the intestinal flora, and is constituted so that these foods can be taken as a staple food. Any food that takes a liquid or fluid form at the time of ingestion can be used, including ordinary liquid foods such as milk, processed milk, and whey beverages; It may be a powder used for enteral nutrition and the like. Enteral nutrients include liquid foods and blenders.
Food, full liquid food such as concentrated liquid food, or low protein liquid food,
It may be a special liquid diet such as a low fat liquid diet, a low cholesterol liquid diet, a low sodium liquid diet, and a therapeutic milk, and is administered orally or by tube. A complete liquid diet contains a good balance of protein, fat, sugar, minerals, and vitamins, while a special liquid diet adjusts the composition of nutrients according to the purpose. The bovine lactoferrin, indigestible saccharide, bovine serum albumin, soybean protein, and iron agent used in the present invention are all water-soluble, and can be used by dissolving at the time of use.
In addition, when bovine lactoferrin is added after sterilization, in order to prevent inactivation by heating, for example, pH after dissolution
Can be adjusted to be acidic and sterilized, can be adjusted as it is or the pH can be readjusted, or can be freeze-dried and spray-dried to a powder. In the case of producing a powder containing fat with infant milk, feed milk, enteral nutrition, etc., for example, separately prepare an aqueous phase and an oil phase, add an emulsifier, a stabilizer, and mix uniformly. Then, homogenize with a homogenizer, concentrate and dry to produce an intermediate product powder, and add bovine lactoferrin, indigestible saccharide, bovine serum albumin, soy protein, iron agent and other product raw materials to this intermediate product powder, and homogenize. To produce the final product. Since the indigestible saccharide and the iron agent hardly denature by heating, they can be added at any stage of the production process. The effective addition amount for improving the intestinal flora is 0.1% or more of bovine lactoferrin, 0.01-2% of indigestible saccharide, and 0.5-5 of bovine serum albumin in the concentration in the final product.
%, Soy protein 0.5-5%, iron agent is 10-7 as iron
0 mg / l.

Next, the effects of the present invention will be described in detail with reference to test examples. (Test Example 1) First, in order to examine the action of bovine lactoferrin in vivo (intestinal tract), the initial infection period of neonates and the intestinal flora of premature infants (Goro Tanaka, Therapeutics, Vol. 14, Vol. 5)
No. 594-597, 1985).

5 week old SPF mouse (SLC) Balb
/ C females were divided into 2 groups of 5 females each and placed in a cage containing a feces-prevention net, and one group was bred for 1 week on a normal solid diet, and then allowed to freely take only milk for 1 week Bred.
The other group was kept as a control while always receiving only a solid diet. In addition, both groups were allowed to freely ingest water when bred on a solid diet.

Fresh feces of each mouse were collected with time, and the method of Mitsuoka ("World of Intestinal Bacteria", Sobunbunsha, 1980).
The number of harmful bacteria in feces was measured according to the following method.

As a result, the bacterial counts of enterobacteriaceae, Streptococcus, Staphylococcus and Clostridium in feces were 6.2 ± 0.2 and 6.1 ± 0 in logarithmic values for mice fed a solid diet. .2,5.
2 ± 0.3 and 7.0 (detection limit) or less, whereas in the case of milk-fed mice, 9.0 ± 0.2,
It was 8.2 ± 0.2, 8.1 ± 0.2, 9.8 ± 0.9 (detection rate 60%). The numbers of these bacteria in the stool remained almost stable for several weeks unless the conditions of rearing were changed. From these results, it was found that breeding in milk resulted in one more harmful bacterium than mice bred on a solid diet.
It has become possible to create a mouse (hereinafter, referred to as a milk-breeding mouse) that stably retains an abnormal intestinal flora of 100 to 1000 times or more.

Next, the effect of improving the intestinal flora by bovine lactoferrin-added milk was tested. (Test Example 2) 0%, 0.1%, 0.5%, 1% bovine lactoferrin
%, 2%, 5%, or 7.5%, added to milk, dispensed into a sterilized drinking bottle, and allowed to freely ingest the milk-breeding mouse prepared in Test Example 1 orally from the drinking bottle. Was. Fresh feces were collected over time, and the number of harmful bacteria in the feces was measured by an ordinary method. The administration of 0.1% or more of bovine lactoferrin drastically reduced the number of harmful bacteria in feces, and maintained a low level stably 2 days after administration. The enterobacterial assay in the feces of the 1-5% bovine lactoferrin-administered group dropped to near the level of the bacterial count in the feces of the mice fed a solid diet, and the bacterial count of Clostridium fell below the detection limit. No difference was observed between the group administered with 7.5% bovine lactoferrin and the group administered with 5% bovine lactoferrin.

From these results, it is found that the value is 0.1% or more, preferably 1%.
Oral administration of milk containing ~ 5% bovine lactoferrin has the effect of reducing the number of harmful bacteria in feces of animals having abnormal intestinal flora in a short period of time and improving abnormal intestinal flora. There was found. The same test was performed for iron-saturated lactoferrin and iron-unsaturated lactoferrin, and it was found that the same effect was found to improve intestinal flora.

Next, it is known that dietary fiber or indigestible sugars affect the intestinal flora (Satoshi Innan,
And Shuhei Kiriyama: “Dietary Fibers”, pp. 242-261, Daiichi Shuppan Co., Ltd., 1989), and describe the synergistic action when these substances are used in combination with bovine lactoferrin. (Test Example 3) 0.01 to 2% and 2% bovine lactoferrin hydrolyzate of konjac derived from konjac produced by the same method as Reference Example 1 was added to milk and dispensed into a sterilized drinking bottle.
The milk-bred mice prepared in Test Example 1 were orally ingested orally through a drinking bottle for 2 weeks. Fresh feces from the second week were collected, the intestinal flora in the feces was analyzed by a conventional method, and the bacterial count was measured.

The results are shown in Table 1. 0.01-2%
When the glucomannan hydrolyzate and 2% bovine lactoferrin were administered in combination, the number of harmful bacteria in feces was reduced and maintained at a level even lower than when 2% bovine lactoferrin alone was administered. In particular, when glucomannan hydrolyzate was used in combination with 2% bovine lactoferrin in a concentration range of 0.1 to 1%, harmful bacteria in feces were further reduced to a lower level, and an excellent synergistic effect was observed.

[0030]

[Table 1]

Test Example 4 Lactulose 0.0 produced by the same method as in Reference Example 2
01-2.0% and 2% bovine lactoferrin were added to milk, dispensed into sterilized drinking bottles, and the five milk-breeding mice created in Test Example 1 were orally freely given from the drinking bottle for 2 weeks. Ingested. Fresh feces from the second week were collected, and the intestinal flora in feces was analyzed according to a conventional method.

The results are shown in Table 2. Lactulose, in combination with 2% bovine lactoferrin, in the range of 0.01-2%, synergistically reduced harmful bacteria in mouse feces. However, since lactulose causes diarrhea at concentrations higher than 2%, it is desirable to use lactulose in combination with bovine lactoferrin in the range of 0.01 to 2%, which has been found to have an effect of further improving intestinal flora. .

[0033]

[Table 2]

Next, test examples for screening other substances having an intestinal flora improving effect will be described. (Test Example 5) Various proteins (bovine serum albumin, soy protein, casein, egg albumin, or lysozyme) at a final concentration of 5
%, And a final concentration of 70 mg /
l of the mixture was added to milk, and the milk-breeding mouse prepared in Test Example 1 was allowed to freely ingest. Fresh feces were collected over time, and the number of coliform bacteria was immediately measured on a DHL agar medium.

As a result, even when bovine serum albumin, soybean protein and sodium ferrous citrate were each administered alone, the number of bacteria of Escherichia coli in feces was different, although the degree of reduction was different. It decreased with almost the same tendency as in the case. From these results, it was found that bovine serum albumin, soy protein and iron preparations have an effect of improving the intestinal flora of mice. On the other hand, casein and egg albumin, which are typical proteins, and lysozyme, which is an antibacterial protein, had no effect in vivo.

Therefore, the following test examples describe the synergistic effect when bovine serum albumin, soy protein, or an iron compound is used in combination with bovine lactoferrin. (Test Example 6) 2% of bovine lactoferrin and 0.1 to 7.5% of bovine serum albumin (manufactured by Sigma) were added to milk, dispensed into a sterilized drinking bottle, and the milk-fed mouse 5 prepared in Test Example 1 was added. The animals were allowed free access to a drinking bottle orally for 2 weeks. Fresh feces from the second week were collected, and the intestinal flora in feces was analyzed according to a conventional method.

Table 3 shows the results. When used together with 2% bovine lactoferrin in the range of 0.5 to 5.0% of bovine serum albumin, the harmful bacteria Enterobacteriaceae and Staphylococcus are synergistically reduced as compared with the case of only 2% bovine lactoferrin. did. Therefore, when bovine serum albumin was used in combination with bovine lactoferrin, it was found that 0.5 to 5.0% of bovine serum albumin was effective.

[0038]

[Table 3]

(Reference Test Example 1) 2% of bovine lactoferrin and 0.1 to 7.5% of soybean protein (trademark: "Fujipro CL", manufactured by Fuji Oil Co., Ltd.) were added to milk and dispensed into a sterilized drinking bottle. Then, the five milk-breeding mice prepared in Test Example 1 were orally ingested orally from a drinking bottle for 2 weeks. Fresh feces from the second week were collected, and the intestinal flora in feces was analyzed according to a conventional method.

The results are shown in Table 4. Soy protein 0.
When used in combination with bovine lactoferrin 2% in the range of 5 to 5.0%, compared with bovine lactoferrin 2% alone,
The harmful bacteria enterobacteriaceae and staphylococci have been reduced. In the case of 7.5% soybean protein, a small amount of insolubles precipitated and the drinking bottle was filled up, so that no effect was observed. Therefore, when the soybean protein was used in combination with bovine lactoferrin, it was found that 0.5-5.0% of the soybean protein was effective.

[0041]

[Table 4]

(Reference Test Example 2) Bovine lactoferrin 2% and sodium ferrous citrate (manufactured by Eisai) 50 to 1500 mg / l (5% as iron)
-150 mg / l) was added to milk, dispensed into a sterilized drinking bottle, and the five milk-breeding mice prepared in Test Example 1 were orally ingested orally from the drinking bottle for 2 weeks. Fresh feces from the second week were collected, and the intestinal flora in feces was analyzed according to a conventional method.

Table 5 shows the results. Sodium ferrous citrate 100-700 mg / l (10-7 as iron)
0 mg / l) in combination with bovine lactoferrin 2%, compared to bovine lactoferrin 2% alone.
The harmful bacteria enterobacteriaceae and staphylococci have been reduced. Sodium ferrous citrate 50m
At a concentration of g / l or 1500 mg / l, the harmful bacteria enterobacteriaceae and staphylococcus increased, and no effect was observed. Therefore, when the iron compound is used in combination with bovine lactoferrin, 10 to 70%
A concentration range of mg / l has been found to be effective.

[0044]

[Table 5]

Next, lactoferrin is used in vitro.
Is known to promote the growth of Bifidobacterium, which is a useful bacterium (Akihiko Kodama: Journal of the Japan Pediatric Society, Vol. 87, No. 6, pp. 1000-1013, 1983).
Year. Petschow, B .; Wand Talbot
t, R. D. : Pediatric Research
h, vol. 29, No. 2,208-213,19
91) Therefore, regarding the synergistic effect of bovine lactoferrin and indigestible saccharide, bovine serum albumin, soy protein, or iron compound on intestinal flora improvement, Bifidobacterium, a useful bacterium of intestinal flora, And the effects on Lactobacillus were investigated. (Test Example 7) Bovine lactoferrin 2%, bovine lactoferrin 2 in milk
%, Indigestible saccharide, bovine serum albumin, soy protein, and sodium ferrous citrate at the concentrations that were most effective in Test Examples 3, 4, and 6 to 8, respectively, and dispensed into sterilized drinking bottles. The five milk-breeding mice prepared in Test Example 1 were orally ingested orally from a drinking bottle for 2 weeks. Fresh stool from the second week was collected, and bifidobacterium and lactobacilli in feces were measured according to a conventional method.

The results are shown in Table 6. Compared to the case where only bovine lactoferrin 2% was added, bovine lactoferrin 2% was added with lactulose 0.1%, konjac-derived glucomannan hydrolyzate 0.1%, bovine serum albumin 5%, and soy protein 2. When 5% or 700 mg / l of sodium ferrous citrate (70 mg / l as iron) was added, respectively, Bifidobacterium and Lactobacillus increased. Therefore, by using indigestible saccharides, bovine serum albumin, soy protein, or iron compounds in combination with bovine lactoferrin, the useful bacteria Bifidobacterium and Lactobacillus increase, harmful bacteria decrease, and the intestinal tract decreases. It has been found that the floor is improved.

In addition to milk, commercially available whey drinks, baby milk, and enteral nutrition were tested in the same manner as in each test example, and almost the same results were obtained.

[0048]

[Table 6]

Reference Example 1 Production Example of Water-Soluble Indigestible Sugars Cellulase derived from a microorganism belonging to the genus Aspergillus (trade name: “Cellulase A Amano”, manufactured by Amano Pharmaceutical Co., Ltd.) in 80 l of phosphate buffer (pH 6.5) .Fiber saccharification power 15,000U / g) 67g
And heated to 40 ° C.
4.0 kg was added and dissolved by stirring. Bring the mixture to 40 ° C
After performing the enzyme reaction by holding for 16 hours,
The mixture was heated at 95 ° C. for 10 minutes to stop the enzyme reaction. The reaction solution was centrifuged at 6000 G, and the supernatant was passed through 1,000 ml of cation exchange resin Dowex 50W × 8 (H +) and 4,000 ml of anion exchange resin Dowex 1 × 8 (OH−) in order. After desalting and purification, concentration and spray drying, about 2,800 g of a partial hydrolyzate was obtained. The average molecular weight of this partial hydrolyzate was 2,000. (Reference Example 2) Production example of lactulose Warm water 20 obtained by heating 100 kg of commercially available edible lactose to 90 ° C
The solution was dissolved in 0 kg, and a solution prepared by dissolving 540 g of sodium hydroxide in 15 kg of water was added thereto, heated at 90 ° C. for 5 minutes, and then cooled to 30 ° C. Then, the reaction solution was passed through a cation exchange resin and an anion exchange resin layer to desalinate and decolorize, concentrated to a solid content of 60 to 70%, and lactose crystals precipitated were separated by filtration three times. About 38 kg of lactulose syrup having a content of 69% was obtained.

The resulting lactulose syrup contained about 80% lactulose in solids.

Next, the present invention will be described in more detail and concretely with reference to examples, but the present invention is not limited to the following examples.

[0052]

EXAMPLE 1 100 l of milk was homogenized at 70 ° C. under a pressure of 150 kg / cm ² and sterilized at 90 ° C. for 10 minutes.
1 kg of bovine lactoferrin (manufactured by Oleofina, Belgium) and 0.5 kg of 50% lacturo-syrup (manufactured by Morinaga Milk Industries) were added to 8.5 l, and uniformly dissolved.
Approximately 50 kg of a milk drink containing 2% bovine lactoferrin and 0.5% lactulose at a final concentration was prepared.

This milk drink was dispensed into a sterilized drinking bottle, and the five milk-breeding mice prepared in Test Example 1 were orally freely ingested from the drinking bottle. Fresh feces were collected every two days for two weeks, and the number of bacteria in feces was measured according to a conventional method. The number of bacteria in feces is compared with the number of bacteria before administration as 100%,
On day 2 after administration, the average number of enterobacteriaceae decreased to 1.5%, that of Staphylococcus decreased to 16%, and the number of bacteria of Clostridium fell below the detection limit. Example 2 Whey powder (Morinaga Milk Industry Co., Ltd.) 5 kg,
8.5 kg of sucrose and 0.5 g of konjac-derived glucomannan hydrolyzate prepared in the same manner as in Reference Example 1.
Kg is dissolved in 52.85 Kg of water, cooled to 10 ° C,
3 kg of 10% citric acid solution, 30 kg of 1% pectin solution
And 0.15 Kg of fragrance are added and uniformly dissolved,
The mixture was homogenized at 150 ° C. and a pressure of 150 kg / cm 2 and sterilized at 90 ° C. for 10 minutes to prepare a whey beverage. Separately, bovine lactoferrin (Oleofina, Belgium) 2
1 mol of hydrochloric acid was added to 10 liters of a 0% aqueous solution to adjust the pH to 4, and sterilized at 80 ° C. for 15 minutes. Add 45 l of bovine lactoferrin solution to 45 l of whey beverage and dissolve uniformly,
About 50 l of a whey beverage containing 0.45% of the above glucomannan hydrolyzate and 2% of bovine lactoferrin was obtained.

The whey beverage containing the hydrolyzate of glucomannan and bovine lactoferrin was dispensed into a sterilized drinking bottle, and the five milk-breeding mice prepared in Test Example 1 were orally ingested orally freely from the drinking bottle. I let it. 2 fresh stools every 2 days
Samples were collected weekly, and the number of bacteria in feces was measured according to a conventional method.
Compared with the number of bacteria in the stool assuming that the number of bacteria before administration was 100%, the average number of enterobacteriaceae decreased to 2.0% and staphylococcus decreased to 15% after 2 days after administration.
And the bacterial count of Clostridium fell below the detection limit. Example 3 7.5 kg of lactose (manufactured by Mirai, Germany) and 2.5 kg of skim milk powder (manufactured by Morinaga Milk Industry Co., Ltd.) were dissolved in 50 kg of water to prepare an aqueous phase in a tank. On the other hand, 1.96 kg of vegetable oil (manufactured by Taiyo Yushi Co., Ltd.) and 0.04 kg of lecithin (manufactured by Ajinomoto Co.) were mixed and dissolved to prepare an oil phase. The oil phase was added to the aqueous phase in the tank, mixed by stirring, heated to 70 ° C, homogenized with a homogenizer at a pressure of 150 kg / cm ² ,
For 10 minutes, concentrated, and spray-dried to obtain about 10 kg of an intermediate product powder. To 7.28 kg of this intermediate product powder, 0.5 kg of desalted whey (manufactured by Domo, the Netherlands), 2.0 kg of bovine serum albumin (manufactured by Sigma), 0.02 kg of calcium carbonate, and bovine lactoferrin (manufactured by Oleofina of Belgium) ) 0.2 kg was added and mixed uniformly with a mixer to obtain about 9.5 kg of powdered milk as a final product.

0.2 kg of this milk powder was dissolved in 1.8 kg of sterilized water to prepare 2 kg of a milk preparation containing 0.2% of bovine lactoferrin and 2% of bovine serum albumin at final concentrations. This milk preparation was orally freely ingested from the drinking bottle to the five milk-breeding mice prepared in Test Example 1. Fresh feces were collected two weeks later, and the number of bacteria in feces was measured according to a conventional method. The number of bacteria in the stool was reduced to 1.2% by comparing the number of bacteria on the day before administration to 100%,
Staphylococcus was reduced to 16%, and the bacterial count of Clostridium fell below the detection limit. Reference Example 3 10.8 kg of whey-protein enzyme digest (manufactured by Morinaga Milk Industry Co., Ltd.)
36 kg of dextrin (manufactured by Showa Sangyo KK) and a small amount of water-soluble vitamins and minerals were dissolved in 200 kg of water, and an aqueous phase was prepared in the tank. On the other hand, 3 kg of soybean salad oil (manufactured by Taiyo Yushi Co., Ltd.), 8.5 kg of palm oil (manufactured by Taiyo Yushi Co., Ltd.) and 2.5 kg of safflower oil (manufactured by Taiyo Yushi Co., Ltd.), 0.2 kg of lecithin (manufactured by Ajinomoto Co.), fatty acid An oil phase was prepared by mixing and dissolving 0.2 kg of monoglyceride (manufactured by Kao Corporation) and a small amount of fat-soluble vitamin. The oil phase was added to the water phase in the tank, mixed by stirring, heated to 70 ° C., homogenized with a homogenizer at a pressure of 150 kg / cm ² , and
Sterilized at 10 ° C. for 10 minutes, concentrated and spray-dried to obtain about 59 kg of intermediate product powder. Add sucrose (Hokuren) 8.2
Kg, 7.5 kg of bovine lactoferrin (manufactured by Oreofina of Bergi), amino acid mixed powder (manufactured by Ajinomoto Co.)
2 kg and 0.1 kg of sodium ferrous citrate (manufactured by Eisai Co., Ltd.) were added and mixed uniformly to obtain about 75 kg of an enteral nutritional powder. This enteral nutritional powder was dissolved in water at a concentration of 15%, and 1.5% of bovine lactoferrin and 200 mg / l of sodium ferrous citrate (20 mg as iron)
/ L) was prepared, and fresh stool that had been orally freely ingested orally from a drinking bottle was collected from the milk-breeding mice of 5 mice prepared in Test Example 1 two weeks later, and feces were collected according to a conventional method. The number of bacteria in the was measured. When the number of bacteria in feces was compared with the number of bacteria before administration as 100%, enterobacteriaceae decreased to 1.1% and staphylococcus to 8.5.
%, And the bacterial count of Clostridium has dropped below the detection limit. Reference Example 4 21 kg of sodium caseinate (manufactured by New Zealand Daily Board), soy protein (trademark: Fujipro CL:
4.5 kg of amino acid mixed powder (manufactured by Ajinomoto) 0.4 kg, lecithin (manufactured by Ajinomoto) 0.1 K
g and a small amount of water-soluble vitamins and minerals in 150
It was dissolved in Kg to prepare an aqueous phase. On the other hand, 3.8 kg of corn oil (manufactured by Taiyo Yushi Co., Ltd.) and 15 K of olive oil (manufactured by Taiyo Yushi Corporation)
g and fatty acid monoglyceride (Kao Corporation) 0.2 kg
A small amount of a fat-soluble vitamin was mixed and dissolved to prepare an oil phase. The oil phase is added to the aqueous phase, mixed with stirring, heated to 70 ° C., homogenized with a homogenizer at a pressure of 150 kg / cm ² , sterilized at 90 ° C. for 10 minutes, concentrated, spray-dried, and powdered to about 40 kg. I got To this, 53 kg of sucrose (manufactured by Hokuren) and 7 kg of bovine lactoferrin (manufactured by Oleofina, Belgium) were added and uniformly mixed to obtain about 100 kg of powder for feed.

This powder was dissolved in water at a concentration of 20% to prepare a liquid feed containing 1.4% of bovine lactoferrin and 0.8% of soybean protein. They were orally ingested freely from a drinking bottle. Fresh feces were collected two weeks later, and the number of bacteria in feces was measured according to a conventional method. The number of bacteria in each stool is equal to the number of bacteria before administration by 10
When compared with 0%, enterobacteriaceae was 1.%.
Down to 5%, Staphylococcus down to 10%,
Clostridium bacterial counts fell below the detection limit.

[0057]

As described above, according to the present invention, a liquid or fluid food having the following effects can be obtained. (1) Improve the bacterial composition of abnormal human or animal intestinal flora. (2) It is used as a nutritional diet for humans with reduced digestive function, such as premature babies, newborns, infants, elderly people, or post-operative patients, and enhances its protective function against pathogenic bacteria by continuous use. (3) Effective in preventing and treating diarrhea and infectious diseases caused by pathogenic bacteria in the digestive tract from the stomach to the large intestine.

Continued on the front page (51) Int.Cl. ⁷ Identification code FI A61K 38/00 A23L 2/00 JA61P 1/14 A61K 37/02 (72) Inventor Susumu Teraguchi 2656-4 Naruse, Machida-shi, Tokyo (72) Inventor Kazuhiro Ozawa 2-6-4 Fukamidai, Yamato City, Kanagawa Prefecture (72) Inventor Satoko Yasuda 1-18-3-205, Midori-ku, Midori-ku, Yokohama-shi, Kanagawa Prefecture (56) References JP-A-1-93534 (JP, A) JP-A-4-8269 (JP, A) JP-A-63-56273 (JP, A) Journal of the Japan Pediatric Society, (1983) Vol. 87, No. 6, p. 1000-1003 Food Chemical, (1986) Vol. 10, p. 79-84 (58) Field surveyed (Int. Cl. ⁷ , DB name) A23L 1/305 A23C 9/00-9/16 A61K 38/00 JICST file (JOIS) JAFIC file (JOIS)

Claims (2)

(57) [Claims] 1. (A) bovine lactoferrin, (B) at least 0.01% (by weight) of a water-soluble indigestible saccharide,
A liquid or fluid food effective for improving intestinal flora, comprising at least 0.5% (by weight) of bovine serum albumin, or any mixture thereof. 2. The liquid or fluid food effective for improving the intestinal flora according to claim 1, wherein the water-soluble indigestible saccharide is a glucomannan hydrolyzate.