Background Art Malt is that a budded barley is dried, which is called as"Yeotgireum"or "Yeotgilgeum". The malt is used for manufacturing a thick soypaste mixed with red pepper or a paste for wrapped rice, and a row material of beer. When manufacturing the malt, it is preferable that a length of a bud thereof is about one and half times of the barley. It is known that malt has an abundance of digestive enzymes, which is therefore used as a digestive or a peptic for symptoms of indigestion in Chinese Medicine.

And a shiitake mushroom is called as Lentinus edodes in a scientific name.

It is known that Eritadenine existing in the shiitake mushroom has a decreasing effect of the contents of cholesterol and neutral fatty acid in blood of a mouse, and it is reported that an extract of the shiitake mushroom has a multiplication operation

of lactic ferments (Journal of Korean Society of Food Science and Technology, 28 (5), 947,1996).

Rotavirus belongs to a family of Reoviridae, and it is known that the rotavirus is an important cause of an acute child enteritis in developing countries and undeveloped countries. The rotavirus is composed of 6 groups (A-F), different each other antigenically, wherein A-C groups of rotavirus are shown up in both of human and animal, and D-F groups thereof are shown up in animal only.

And a group classified according to a serum has a common or crossing-over antigen, and among these, the A group has been properly investigated and researched since the existence of rotavirus has been reported, and the A group is most portion of strains separated from human.

It is known that rotavirus is a main cause of an acute diarrhea and enteritis in a child animal of cow, a horse, a pig, a monkey, etc. According to a statistical data of World Health Organization on a diarrhea disease occurring sporadically to an baby and a child, the number of under 5 years old child's death caused by diarrhea is reached to five million a year, and one million, 20% of them, is dead due to diarrhea caused by rotavirus.

In the United States America, 35-50% of baby diarrhea is caused by rotavirus, and in case of developing countries, the problem is much more serious because a diarrhea caused by rotavirus is occurred about 10 times in 6-11 months old babies, and it is known that 90% of babies have an antibody of rotavirus before 24 months. According to a research report examining more than 12 species of a cause of diarrhea during 12 months for baby's diarrhea group of Seoul area in Korea, rotavirus has been detected as 47% of diarrhea group, which was the highest appearance-frequency, and was over twice of 20%, a statistical date of World Health Organization.

Rotavirus is infected through the way of oral from the feces, which is a cause of a contagiousness acute diarrhea to babies less than 2 years old. This kind of infection state is called as infantile gastroenteritis or acute gastroenteritis. Acute gastroenteritis caused by rotavirus is accompanied with clinical symptoms such as pyrexia, vomiting, diarrhea, <BR> <BR> respiratory symptom etc. , and is generally occurred between late autumn and early winter in the temperature regions including Korea, and baby diarrhea patients caused by rotavirus is 70-80% of diarrhea patients during this term.

Rotavirus is excreted as the feces on about 8 days after showing symptoms, and 1011 of virus per gram of the feces is found. Babies between 6 months old and 2 years old are easily infected, and when infection, dehydration and a metabolic acidosis are occurred. It is known that a cause of diarrhea is an intestinal trouble of an absorption function of a mucous cell due to rotavirus infection. Rotavirus is infected in a villus cell of the small intestine and multiplied in a protoplasm of the cell, which causes a trouble in a transportation system. A damaged cell is dropped into the intestine, and a lot of viruses are excreted, and then rotavirus is found in the feces. A diarrhea caused by rotavirus is caused by a damage of an absorption function of sodium and glucose by substituting a damaged cell with an immature crypt cell, not having absorption ability.

From the foregoing reasons, a vaccine for rotavirus has developed and employed, but the use thereof has stopped, therefore there is no appropriate medicine. Accordingly, the development of a new material being able to restrain rotavirus infection has been needed.

Lactic ferments is classified into Lactobacillus genus, Bifidobacterium genus, and Streptococcus genus. A variation of lactic ferments in the intestine according to age shows that bifidobacterium is shown up on 3rd-4th day after birth, and on 5th day, it becomes the most superior virus. Also, after finishing the weaning period, anaerobes and

bifidobacterium is superior, and other lactic ferments such as lactobacillus, etc is considerably diminished. In case of becoming nearly an old person, a balance of intestinal virus is aged, so that the strength of bifidobacterium is weakening, therefore the number of lactobacillus virus becomes larger than that of bifidobacterium. Accordingly, when getting old, if taking the bifidobacterium virus, the number thereof becomes similar to intestinal flora of babyhood, adolescence or manhood.

Bifidobacterium breve K-110, Korean lactic ferments, has a main characteristic in a curative effect of large intestine cancer, bladder cancer and ascites cancer (Korean Patent No. 0232639 and J. of Microbiol. Biotecbnol. 9 (3), 368,1999), and inhibits a growth of helicobacter pylori that is a virus causing a stomach ulcer, so that the bifidobacterium brave K-110 can be used for the treatment of stomach disease due to having an preventive effect of a stomach ulcer (J. Microbiol. Biotechnol. 10 (4), 532, 2000). Generally, a physical activity of lactic ferments is, for example, a multiplication restraint of harmful bacteria, an alleviation of lactose intolerance, a lowering-effect of serum cholesterol, an anti-cancer effect, an immune response, an improvement of diarrhea and constipation, a vitamin synthesis, etc.

Bifidobacteria is shown up on around 3rd-4th day after the birth and is grown as a superior virus on around 5th day, so that the number of E. coli or other lactic ferments such as lactobacillus, streptococcus, etc. is diminished to one-hundredth. It is inferred that this reason is due to a factor which accelerates a multiplication of bifidobacteria existing in mother's milk only. Therefore, it can be known that bifidovacteria is more important virus than other lactic ferments such as lactobacillus.

A conventional technology is focused on the manufacture of lactic-acid fermented product such as yoghurt by using lactic ferments. This technology is a common method comprising the steps of inoculating lactic ferments in milk, adding skim

milk, multiplying lactic ferments, and preparing a lactic ferments beverage. A technology for producing a lactic ferments beverage or a lactic acid using cereals except milk is, for example, a lactic-acid fermented beverage made of vegetables and Chinese herb using lactobacillus plantarum (Korean Pat. Publication No. 2000-0055832), a rice's embryo bud beverage (Korean Pat. Application No. 1996-042486), a method for preparing a lactic-acid fermented product using a cereal-type rice (Korean Pat. Publication No.

1992-16030), a method for preparing a liquid phase of a lactic-acid fermented product using cereals powder (Korean Pat. Publication No. 1997-14592), a method for producing a lactic-acid using a component made of bean (Korean Pat. Application No. 1999-0017375), a production of lactic ferments crude powder using lactobacillus acidophilus strain (Korean Pat. No. 148340), lactic ferments capable of restraining a multiplication of hericobacter pylori infective bacteria in the stomach (Korean Pat. Publication No. 2002- 0072486,2002-0061040 & 2002-0045496), lactic ferments strain having a high absorbability of a heavy metal and a product using the same (Korean Pat. Publication No.

2000-0065010), a raw material for a metabolic bone disease prevention and improvement food (Korean Pat. Publication No. 2002-0048959). As shown above, there are no researches or patents as to lactic ferments having an effect for rotavirus.

And, as a prior art for a lactic-acid fermented material using a mushroom, there is a lactic ferments culture medium including mushroom powder or a mushroom extract and a lactic-acid fermented composition using the said culture medium (Korean Pat.

Publication No. 2002-0085159). But, the present invention using malt or shiitake mushroom is big different from a prior art which ferments a lactic ferments culture medium with a culture medium consisting of a skim milk powder and a mushroom powder (Cortinellus shiitake, Ganoderma lucidum, Agaricuses subfunereus, Oligosaccharide, Dextrin).

Disclosure of Invention An object of the present invention is to prepare an extract by adding water to malt or mushroom, and carrying out hydrothermal extraction.

Another object of the present invention is to prepare a lactic-acid fermented material having a restraint effect of rotavirus infection by transplanting lactic ferments to hydrothermal extraction of malt or mushroom, or malt or mushroom itself, which is then fermented.

A further object of the present invention is to provide a food having a restraint effect of rotavirus infection as an additive material that is able to be added to a baby food, a powdered milk, a sunsik, a uncooked food and a functional food, etc, wherein the additive material is prepared by transplanting lactic ferments to hydrothermal extract of malt or mushroom, or malt or mushroom itself, which is then fermented. Hereinafter, the above word'sunsik'means Korean traditional health food which is a mixture of brown rice, glutinous rice, barley, black soybean, black sesame, Perilla japonica, and Coix mayuen Roman.

In the present invention, a fermented material having a restraint effect for rotavirus infection, a cause of diarrhea, is prepared by fermenting malt or shiitake mushroom by using bifidobacterium breve K-110 (Korean lactic ferments), bifidobacterium longum (B. longum) KCTC3215 (available in the market), bifidobacterium breve (B. breve) JCM1192, bifidobacterium infantis (B. infantis) JCM7007. This material can be used as a raw material for a baby food, a powdered milk, or a functional food.

Best mode for Carrying Out the Invention In order to accomplish the above objects, the present invention is characterized in

that an extract having a restraint effect for rotavirus infection is prepared by adding water to malt or a mushroom in which the amount of water is about decuple of malt or mushroom, and then carrying out hydrothennal extraction.

Also, the present invention is characterized in that a lactic-acid fermented material having a restraint effect for rotavirus infection is prepared by fermenting the extract, or malt or mushroom itself with lactic ferments in the temperature range of 30~38°C for about 24 hours.

Also, the present invention is characterized in that these extracts or fermented materials can be used as a raw material capable of inhibiting rotavirus infection by adding those to a baby food, a powdered milk, a infant food, a sunsik, a uncooked food, a <BR> functional food, etc. , wherein the raw material is an extract having a restraint effect for rotavirus infection, or a fermented material that is prepared by fermenting these extracts with lactic ferments.

Example 1: Preparation of an extract of malt or mushroom Water was added to 500g of malt or shiitake mushroom, wherein the amount of water was decuple of malt or shiitake mushroom, and then carried out hydrothermal extraction with heating. The above extraction process was carried out twice, and an extracted supernatant was concentrated and used for a sample having a restraint effect for rotavirus.

Example 2: Preparation of malt fermented material and mushroom fermented material using lactic ferments Lactic ferments was transplanted to a malt or mushroom extract to become 108 of lactobacilli per m, which was then fermented at 35~38°C for about 24 hours, and a lactic ferments solution was obtained, which was used for a sample.

Example 3: Culture of MA-104 cell MA-104 cell was cultured using 10% FBS, 1% antibiotics-antimycotics mixture and DMEM (Dulbecco's Modified Eagle's Medium) culture medium in which 3.5g/L sodium bicarbonate was added. When multiplying to about 80% length of a flask, the cultured MA-104 was treated with 0.25% trypsin, and thereafter a cell was separated, and then passage-cultured. The culture of cell was carried out in C02 incubator saturated with 5% carbon dioxide.

Example 4: Preparation of rotavirus stock solution 2X1O6 of MA-104 cells were aliquoted in a tissue culture flask, which was then cohered to CO2 incubator at about 37°C for about 1 hour, and thereafter a culture medium was removed, and the surface of a cell was washed twice with DMEM culture medium, not including FBS. And 200. of infective DMEM culture medium containing 400, nlQ of Wa virus solution having 100ßg/mQ of trypsin was added therein and a virus solution was activated at about 37°C for about 30 minutes. The activated virus solution was spread on the surface of a cell, which was then infected at about 37°C for about 1 hour and removed, and 5mQ of infective DMEM culture medium containing 5, ug/m. of trypsin, not including FBS was added therein and cultured in C02 incubator for 5 days. After the culture, a cytophatic effect was confirmed. And a cell membrane of the cell was destroyed by repeating three times a process of refrigerating and rapid melting, which was then centrifuged with 5000rpm at 4°C for 20 minutes, and obtained a supernatant, virus solution, and the virus solution was frozen and kept at-70 C.

Example 5: Measurement of an restraint effect for rotavirus infection of lactic ferments and a fermented material MA-104 cell was treated with 0.25% trypsin and removed from the flask, and was washed twice with a washed DMEM culture medium, and the number of cells was controlled to be 3X105. The stock solution of Wa virus was diluted with an infected DMEM culture medium to be five hundred times thereof. Malt or shiitake mushroom extract having a concentration of 0. 4mg/mQ (Example 1), 50, ze of malt or shiitake mushroom fermented material (Example 2), 50uQ of Wa virus diluted solution and 100go of MA-104 cell having 3x10 cell/m-C were spread and cultured in C02 incubator at 37°C for 5 days, and thereafter a cytophatic effect was examined, and then a restraint activity for rotavirus infection was measured. As shown in Table 1, the results of a restraint effect for rotavirus infection of lactic ferments showed that bifidobacterium breve K-110 has most excellent effect in bifidobacterium longum KCT3215, bifidobacterium breve JCM1192 and bifidobacterium infantis JCM7007.

Table 1: Restraint effect for rotavirus infection of lactic ferments Lactic ferments Human Infection Wa rotavirus Animal Infection SA11 rotavirus 50% infective restraint 50% infective restraint effect concentration (mg/ml) Bifidobacterium >0.1 >0.1 Longum KCTC3215 Bifidobacterium >0. 1 >0. 1 Breve JCM1192 Bifidobacterium >0.1 >0. 1 Infantis JCM7007 Bifidobacterium 0.074 0.072 Breve K-110 108/mu of Lactic ferments was transplanted to a hydrothermal extract of malt or shiitake mushroom prepared by Example 1 and a malt or mushroom extract of Example 2, which was then fermented at 35~38 C for about 24 hours. And an anti-rotavirus effect of

lactic-acid fermented solution was measured. The results thereof were shown as the following Table 2. It was shown that a restraint effect for rotavirus infection of a fermented solution prepared by fermenting malt or shiitake mushroom with bifidobacterium breve K-110 lactic-acid strain showing most excellent effect for restraining rotavirus infection was more excellent than that of sole malt or sole lactic ferments.

Although it was not shown in Tables, a restraint effect of lactobacillus acidophilus, lactobacillus bulgarius, lactobacillus casei, bifidobacterium infantis, bifidobacterium longum, etc were lower than that of the fermented solution of the present invention.

Table 2: Restraint effect for rotavirus infection of lactic-acid fermented material Sample Human infection Wa rotavirus Animal infection SA11 50% infective restraint rotavirus concentration (mg/ml) 50% infective restraint effect Malt 0. 08 0. 09 Malt fermented 0.05 0.04 material Shiitake mushroom 0. 12 0. 11 Fennented material of 0. 06 0.05 shiitake mushroom An infective restraint effect of these lactic ferments was measured in the state that the final concentration of a cell wall and a cytoplasm produced by lactic ferments is O. 1mg/mQ when measuring a restraint effect for infection. The results are shown in the following Table 3. As shown in Table 3, the restraint effect of a cytoplasm is more excellent than that of a cell wall. These results show that lactic ferments can use a cytoplasm being able to extract with water or organic solvent, even though the lactic ferments are alive-bacteria or dead-bacteria.

Table 3: Restraint effect for Wa rotavirus infection of lactic ferments Lactic ferments Inhibiting rate (%) Cytoplasm fractionation Cell wall fractionation Bifidobacterium 14.3 28.6 Lon um KCTC3215 Bifidobacterium 28.6 14.3 Breve JCM1192 Bifidobacterium 14.4 14.3 Infantis JCM7007 Bifidobacterium 85.7 28.6 Breve K-110

It can be inferred that these lactic ferments or lactic-acid fermented solution can help a prevention of rotavirus infection, or a treatment of rotavirus type diarrhea. In view that rotavirus type diarrhea is generally occurred in the weaning period, the rotavirus type diarrhea, which is 50% of the weaning period diarrhea, can be prevented by adding a lactic-acid fermented material of the present invention to a baby food.

Industrial Applicability As described above, the present invention can provide an extract of malt or shiitake mushroom and a lactic-acid fermented material thereof, which has a restraint effect of rotavirus infection causing of diarrhea. And also, these fermented materials according to the present invention can be used as a raw material for a baby food, a powdered milk, a sunsik, an uncooked food, a functional food, etc.

Although the preferred embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that various modifications, additions and substitutions are possible, without departing from the scope and spirit of the invention as disclosed in the accompanying claims.