CN100577035C - Lactic-acid fermented materials able to restrain rotavirus infection and preparation method thereof - Google Patents

Lactic-acid fermented materials able to restrain rotavirus infection and preparation method thereof Download PDF

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CN100577035C
CN100577035C CN200380108052A CN200380108052A CN100577035C CN 100577035 C CN100577035 C CN 100577035C CN 200380108052 A CN200380108052 A CN 200380108052A CN 200380108052 A CN200380108052 A CN 200380108052A CN 100577035 C CN100577035 C CN 100577035C
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rotavirus
lactic acid
fructus hordei
hordei germinatus
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CN1731937A (en
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李庆守
金东玹
金尚郁
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BARUMIN Co Ltd
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L31/00Edible extracts or preparations of fungi; Preparation or treatment thereof
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135Bacteria or derivatives thereof, e.g. probiotics
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L7/00Cereal-derived products; Malt products; Preparation or treatment thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66Microorganisms or materials therefrom
    • A61K35/74Bacteria
    • A61K35/741Probiotics
    • A61K35/744Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/745Bifidobacteria
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06Fungi, e.g. yeasts
    • A61K36/07Basidiomycota, e.g. Cryptococcus
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/88Liliopsida (monocotyledons)
    • A61K36/899Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
    • A61K36/8998Hordeum (barley)

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Abstract

The present invention relates to a lactic-acid fermented materials of the malt or shiitake fungus able to restrain rotavirus infection and thereof producing method. The characteristic of the invention is utilizing the extraction of the malt or shiitake fungus and the lactic-acid fermented material from the extraction fermented by lactic-acid to produce effective material for rotavirus, the rotavirus is one reason for causing diarrhea. And these extracts or fermented materials can be used as a material capable of inhibiting rotavirus infection by adding those to a baby food, a powdered milk, a infant food, a sunsik, a functional food, and the like.

Description

Suppress lactobacillus fermentation of rotavirus infection and preparation method thereof
Technical field
The present invention relates to suppressing the preparation method of effective extract of rotavirus infection and fermentate.This extract is to make by Fructus Hordei Germinatus or mushroom are carried out extraction heat, and this fermentate is to make by these extracts of lactobacillus-fermented.Specifically, add the water of about 10 times of amounts in Fructus Hordei Germinatus or mushroom, extraction heat is carried out in heating, transplants lactic acid bacteria in this Fructus Hordei Germinatus or mushroom extract, in 35 ℃~38 ℃ fermentations 24 hours, obtains lactic acid bacteria Fructus Hordei Germinatus fermented product or mushroom fermentate.These fermentates are effective to suppressing rotavirus infection.
Background technology
So-called Fructus Hordei Germinatus, the seed that the is meant wheat dried product that sprouts, it can be called as " Fructus Hordei Germinatus " or " wheat bean sprouts ".Fructus Hordei Germinatus is used to make the thick chilli sauce or the tartar sauce of Korea S, and the Fructus Hordei Germinatus of barley also is used as the raw material of beer.As everybody knows, when producing Fructus Hordei Germinatus, the length of bud is the most suitable when being self 1.5 times of left and right sides.Fructus Hordei Germinatus is well-known to be rich in digestive ferment, and the past uses as the digestion appetizer when running into indigestion in the traditional Chinese medical science.
The formal name used at school of mushroom is called as Lentinus edodes.Contained eritadenine (eritadenine) has the cholesterol that reduces in the mouse blood and the effect of neutral fat acid in the mushroom.And, report that the mushroom extract has the effect (for example, with reference to following non-patent literature 1) of propagation lactic acid bacteria.
Rotavirus (Rotavirus) belongs to Reoviridae (Reoviridae), and is known as bring out most important factor in the acute infantile enteritis reason in developing country or undeveloped country.Rotavirus is made up of different 6 types (A-F) of antigen.The rotavirus of A, B, C type is found and is present in the human and animal, and D, E, F type only are found in animal.Press the different virus types of distinguishing of serum, common each other, or have cross-reacting antigen.Since the existence of reporting rotavirus the earliest, the most active about the research of A type wherein, the investigation of its character is also obtained bigger progress, almost be distributed in all and separate and bacterial strains of coming from the people.Rotavirus is to cause the acute diarrhea of young animal such as children's and ox, horse, pig, monkey and the main cause of enteritis.In conjunction with scattered complain of diarrhea illness among child and the children, statistics according to world health institution shows, because of the dead children's number of suffering from diarrhoea reaches 5,000,000 every year, wherein have 20% among the children of the earth below last 5 year old, promptly die from the diarrhoea that causes by rotavirus for more than 100 ten thousand.In the U.S., the 35%-50% in the infantile diarrhea results from rotavirus.And in developing country, this problem is then more serious, the diarrhoea that the about generation of 6-11 month child is caused by rotavirus for 10 times.And 90% child just had rotavirus antibody before 24 months.With the baby diarrhea crowd in Seoul, South Korea area is that object lasts 12 months the bringing out research report that diarrhoea reason body investigates more than 12 kinds is shown, from the diarrhoea crowd 47% detect rotavirus, the frequency of occurrences is the highest.This value reaches more than 2 times well beyond the statistical value 20% of world health institution.
Rotavirus is by the ight soil peroral infection, is the reason that acute infection diarrhoea takes place for the child that caused discontented 2 years old, and this kind Infection Status is commonly referred to child's gastroenteritis or acute gastroenteritis.The acute gastroenteritis that is caused by rotavirus is attended by clinical symptoms such as heating, vomiting, diarrhoea, respiratory apparatus be unusual, with the area, temperate zone headed by the Korea S, generation is arranged nearly all in late fall to early winter, accounts for the 70%-80% of baby diarrhea patient during this.From finding that symptom virus about the 8th day the time drains in a large number as ight soil, find 10 in every 1g ight soil 11Individual virus.And it is the most frequent that 6 months to 2 years old child infects this virus, causes dehydration and metabolic acidosis during infection.Diarrhoea normally occurs when causing the disturbance of absorptive function of intestinal tube mucomembranous cell after infecting rotavirus.The villus cell of virus infections small intestine is bred in the cytoplasm of these cells, causes the obstacle of the system of transporting.Because damaged cell comes off from intestinal tube, discharges a large amount of virus, so just can find virus in the ight soil.By the diarrhoea that rotavirus causes, be that the absorption function of the sodium that do not had the prematurity gland nest cell of absorption function to replace to produce owing to damaged cell and glucose damages and causes.
Therefore, although exploitation and used vaccine corresponding to rotavirus, because side effect interrupts still not having gratifying alternative medicine up to now after it uses.Therefore, people look forward to developing as early as possible the material that can suppress rotavirus infection.
Lactic acid bacteria is broadly divided into lactobacillus, Bifidobacterium, streptococcus.If the variations of lactic acid bacteria can be found in the age-based different investigation intestines, Bifidobacterium appearred on the 3rd~4 in the birth back, became the bacterium that has superiority most on the 5th.And after weaning period finished, anaerobic bacteria and Bifidobacterium became advantage, and be then considerably less as the lactobacillus of other lactic acid bacterias etc.In addition, with advancing age, the disorder of intestinal bacterium balance, the Bifidobacterium force of once having the advantage weakens, and the force of lactobacillus then surpasses Bifidobacterium.Therefore, along with the development of aging, more and more be necessary by taking Bifidobacterium so that keep par in intestinal flora and child or between twenty and fifty period.Main feature as the bifidobacterium breve K-110 of Korea S lactic acid bacteria is (for example effectively to treat the carrying out of colorectal cancer, carcinoma of urinary bladder, ascites carcinoma, with reference to following patent documentation 1 and non-patent literature 2), can the growth of the helicobacter pylori that causes gastric ulcer be suppressed, thereby effect (for example, with reference to following non-patent literature 3) with treatment disease of stomach.Usually, as the physiological activity of lactic acid bacteria, can exemplify, relax lactose intolerance, reduce serum cholesterol to the propagation that suppresses harmful bacterium, anticancer, immune, improve diarrhoea and constipation, synthetic vitamin etc.Bifidobacterium began to occur after birth on the 3rd~4, just grew into the bacterium of tool advantage about the 5th day, original Escherichia coli (E.coli) or reduce to one of percentage as the quantity of the lactobacillus of other lactic acid bacterias, streptococcus etc.This is because have only the factor that has promotion Bifidobacterium propagation in the breast milk, thereby as can be known, compares with the lactobacillus as other lactic acid bacterias, and Bifidobacterium is a most important bacterium in the intestinal flora.
In technology in the past, just efforts be made so that and prepare lactic acid fermentation product such as sour milk with lactic acid bacteria, a lot of technology have been developed, but these technology nearly all are inoculating lactic acid bacteriums in milk, add the common method that skimmed milk power is made the sour milk beverage of having bred lactic acid bacteria again.Use the sour milk beverage of the cereal beyond the milk or the technology of producing lactic acid can also be exemplified below: utilize lactic acid fermentation beverage that lactobacillus germ gets (for example from plant and Chinese medicine, with reference to following patent documentation 2), rice germ beverage (for example, with reference to following patent documentation 3), use method that the graniform metric system is equipped with the lactic fermentation product (for example, with reference to following patent documentation 4), use method that flour prepares fluid milk acid fermentation product (for example, with reference to following patent documentation 5), use produce lactic acid from the composition of beans method (for example, with reference to following patent documentation 6), the method of producing the lactic acid bacteria material powder with strain of lactobacillus acidophilus (for example, with reference to following patent documentation 7), the lactic acid bacteria of the helicobacter pylori propagation in the inhibition stomach and intestine (for example, with reference to following patent documentation 8 to 10), lactic acid bacteria strains that the heavy metal energy of adsorption is higher and the goods that use this lactic acid bacteria strains are (for example, with reference to following patent documentation 11), prevention and improve the metabolic bone disorder use the food material of lactobacillus-fermented (for example, with reference to following patent documentation 12).Yet, but do not carry out yet relevant patent about the effective lactic acid bacteria of rotavirus is studied.As the patent of the lactobacillus fermentation that has used mushroom, can exemplify the lactic acid bacteria culture solution that contains mushroom powder or mushroom extract, the mushroom lactic fermentation composition (for example, with reference to following patent documentation 13) that uses this nutrient solution.In this communique, disclose and used the lactic acid bacteria culture solution that comprises skimmed milk power, mushroom powder (mushroom powder, purple sesame powder, champignon powder), oligosaccharides, dextrin to ferment.Technology when but, using Fructus Hordei Germinatus or mushroom extract and powder self thereof among this technology and the present invention differs widely.
Maximum characteristics of the present invention are, by using lactobacillus-fermented Fructus Hordei Germinatus and mushrooms such as Korea S lactic acid bacteria bifidobacterium breve K-110, the bifidobacterium longum KCT3215, the bifidobacterium breve JCM1192 that sell on the market, bifidobacterium infantis JCM7007, preparation has the fermentate of beneficial effect fruit to the virus-rotavirus infection that causes suffering from diarrhoea.And these fermentates also can be used for the raw material of ablactational food or milk powder, the raw material of baby food, can also be as the raw material of functional food.
[patent documentation 1] Republic of Korea's patent is logined communique No. 0232639
[patent documentation 2] Republic of Korea patent disclosure 2000-0055832 communique
[patent documentation 3] Republic of Korea patent application 1996-042486 communique
[patent documentation 4] Republic of Korea patent disclosure 92-16030 communique
[patent documentation 5] Republic of Korea patent disclosure 97-14592 communique
[patent documentation 6] Republic of Korea patent application 10-1999-0017375 communique
No. 148340 communiques of [patent documentation 7] Republic of Korea patent login
[patent documentation 8] Republic of Korea patent disclosure 2002-0072486 communique
[patent documentation 9] Republic of Korea patent disclosure 2002-0061040 communique
[patent documentation 10] Republic of Korea patent disclosure 2002-0045496 communique
[patent documentation 11] Republic of Korea patent disclosure 2000-0065010 communique
[patent documentation 12] Republic of Korea patent disclosure 2002-0048959 communique
[patent documentation 13] Republic of Korea patent disclosure 2002-0085159 communique
Association of [non-patent literature 1] Korean foods section will, 28 (5), 947, (1996)
[non-patent literature 2] J.Microbiol.Biotechnol.9 (3), 368 (1999)
[non-patent literature 3] J.Microbiol.Biotechnol.10 (4), 532 (2000)
Summary of the invention
The object of the present invention is to provide by in Fructus Hordei Germinatus or mushroom, adding water and carried out extraction heat and prepare suppressing the method for the effective extract of rotavirus infection.
Other purposes of the present invention are to provide by fermenting to water hot extraction's thing of Fructus Hordei Germinatus or mushroom or to Fructus Hordei Germinatus mushroom transplanting lactic acid bacteria itself and prepare suppressing the method for the effective lactobacillus fermentation of rotavirus infection.
Other purposes of the present invention also be to provide contain to suppress effective Fructus Hordei Germinatus of rotavirus infection and mushroom extract and lactobacillus fermentation thereof as ablactational food, milk powder, baby food, the vegetarian food of material, eat something rare food, functional food.
For achieving the above object, what the present invention had is characterized as, and carries out extraction heat by the water and the heating that add 10 times of amounts in Fructus Hordei Germinatus or mushroom, and preparation is to suppressing the effective extract of rotavirus infection.
In addition, feature of the present invention also is, by to Fructus Hordei Germinatus or mushroom extract or to Fructus Hordei Germinatus or mushroom itself is transplanted lactic acid bacteria and in 35 ℃~38 ℃ fermentations 24 hours, preparation was to the effective lactobacillus fermentation of inhibition rotavirus infection.
In addition, the invention provides with above-mentioned to rotavirus infection have the Fructus Hordei Germinatus that suppresses effect and mushroom extract and lactobacillus fermentation thereof as ablactational food, milk powder, baby food, the vegetarian food of raw material, eat something rare food, functional food.
The present invention can utilize Fructus Hordei Germinatus or mushroom extract and the lactobacillus fermentation that obtains through the above-mentioned extract of lactobacillus-fermented prepares the effective goods of rotavirus, and wherein said rotavirus is the virus that causes the diarrhoea reason.And these fermentates also can and be eaten something rare the raw material of food as raw material, the vegetarian food of the raw material of ablactational food or milk powder, baby food, can also be as the raw material of functional food.
The specific embodiment
About specific embodiments of the invention, be exemplified below.And, simple deformation of the present invention or change also are contained within the scope of the present invention, so protection scope of the present invention is carried out clearly according to claim.
The preparation of embodiment 1 Fructus Hordei Germinatus or mushroom extract
Add 10 times water in 500g Fructus Hordei Germinatus or mushroom, extraction heat is carried out in heating.This operation is carried out twice repeatedly.Then, concentrated supernatant uses as the test portion that suppresses rotavirus with this.
Embodiment 2 adopts the preparation of Fructus Hordei Germinatus fermented product and the mushroom fermentate of lactic acid bacteria
In the Fructus Hordei Germinatus that the method according to embodiment 1 prepares or the extract of pestering, transplant lactic acid bacteria, transplant 10 in the extract of every 1ml 8Individual lactic acid bacteria just obtained streptococcus acidi lactici fermented solution in 24 hours in 35 ℃~38 ℃ fermentations.Use as test portion with this zymotic fluid.
The cultivation of embodiment 3MA-104 cell
DMEM culture medium (Dulbecco with the sodium acid carbonate that has added 10% FBS, antibiotic-antifungal agent mixed liquor of 1% and 3.5g/L TS modified eagle TS medium) cultivates the MA-104 cell.When cell was bred to 80% left and right sides in flask, the trypsin treatment with 0.25% continued behind the isolated cell to cultivate.During cultured cell, use to be full of 5%CO 2CO 2Incubator.
Embodiment 4 rotavirus are preserved the preparation of liquid
To tissue culture flasks (25cm 2) middle five equilibrium adding 2 * 10 6Individual MA-104 cell is positioned over 37 ℃ CO 2Incubator made it adherent in 1 hour, removed nutrient solution then, and is clean with DMEM nutrient solution washed cell surface 2 times with what do not contain FBS.Then, in Wa virus (Wa virus) solution of 400 μ l, add and contain the tryptic infectious DMEM culture medium 20 μ l of 100 μ g/ml, in 37 ℃ the activation 30 minutes after, viral solution is coated in equably the surface of cell, remove after 1 hour in 37 ℃ of infection, interpolation 5ml does not contain FBS but contains the tryptic infectious DMEM culture medium of 5 μ g/ml, in 37 ℃ CO 2Cultivated 5 days in the incubator.Cultivate the back and confirm cell degeneration effect (CPE), cell membrane breaks fully behind 3 freezing and quick-thawings repeatedly, and centrifugation is 20 minutes under 4 ℃, the condition of 5000rpm, get contain virus supernatant in-70 ℃ of freezing preservations back uses.
Embodiment 5 lactic acid bacterias and fermentate are to the mensuration of the inhibition effect of rotavirus infection
From flask, to separate the MA-104 cell after 0.25% the trypsin treatment,, regulate cell number to 3 * 10 to clean with after the DMEM culture medium washing 2 times 5Individual, will store 500 times of liquid dilutions by the Wa virus of the same preparation of embodiment 4 methods with infectious DMEM culture medium.In 96 orifice plates, coating concentration transfers to the Fructus Hordei Germinatus of the Fructus Hordei Germinatus of 0.4mg/ml or mushroom extract (embodiment 1), 50 μ l or Lentinus edodes fermented product (embodiment 2), 50 μ l Wa viral dilution liquid and 3 * 10 5The MA-104 cell 100 μ l of cell/ml are in 37 ℃ CO 2Incubator was cultivated after 5 days, and observation of cell sex change effect (CPE) and mensuration are to the inhibition activity of rotavirus infection.Lactic acid bacteria is to the result of the inhibition effect of rotavirus infection, as shown in table 1, compare with lactic acid bacterias such as the bifidobacterium longum KCT3215, the bifidobacterium breve JCM1192 that sell on the market, bifidobacterium infantis JCM7007, bifidobacterium breve K-110 has demonstrated outstanding effect.
[table 1]
Lactic acid bacteria is to the inhibition effect of rotavirus infection
Lactic acid bacteria The people infects Wa rotavirus 50% and infects inhibition concentration (mg/ml) Zoogenetic infection SA11 rotavirus 50% infects and suppresses effect
Bifidobacterium longum KCT3215 >0.1 >0.1
Bifidobacterium breve JCM1192 >0.1 >0.1
Bifidobacterium infantis JCM7007 >0.1 >0.1
Bifidobacterium breve K-110 0.074 0.072
Transplant lactic acid bacteria in the Fructus Hordei Germinatus of the Fructus Hordei Germinatus that in the preparation of the Fructus Hordei Germinatus of embodiment 1 or the extract of pestering, obtains or the extract of mushroom and embodiment 2 or the mushroom extract, transplant 10 in the extract of every 1ml 8Individual lactic acid bacteria obtained streptococcus acidi lactici fermented solution in 24 hours in 35 ℃~38 ℃ fermentations.Measure the anti-rotavirus effect of this streptococcus acidi lactici fermented solution, in table 2, represented its result.And, with the zymotic fluid that rotavirus the most effective bifidobacterium breve K-110 fermented Fructus Hordei Germinatus and mushroom are made, demonstrate the effect of the inhibition rotavirus infection outstanding than independent Fructus Hordei Germinatus or independent lactic acid bacteria.In addition, though do not show in the table that the effect of lactobacillus acidophilus, lactobacillus bulgaricus (L.bulgaricus), Lactobacillus casei (L.casei), bifidobacterium infantis (B.infantis) and bifidobacterium longum (B.longum) etc. then is lower than zymotic fluid of the present invention.
[table 2]
Lactobacillus fermentation is to the inhibition effect of rotavirus infection
Sample The people infects Wa rotavirus 50% and infects inhibition concentration (mg/ml) Zoogenetic infection SA11 rotavirus 50% infects and suppresses effect
Fructus Hordei Germinatus 0.08 0.09
Fructus Hordei Germinatus fermented product 0.05 0.04
Mushroom 0.12 0.11
Lentinus edodes fermented product 0.06 0.05
The cell wall constituent of lactic acid bacteria production and the ultimate density of cytoplasm composition are transferred to 0.1mg/ml, measure the inhibition infectious effect of these lactic acid bacterias, table 3 has been represented its result.As shown in table 3, to compare with the lactobacillus cell wall, the cytoplasm composition demonstrates outstanding effect.From then on the result as can be seen, no matter lactic acid bacteria is viable bacteria or dead bacterium, can use can water or the cytoplasm composition of organic solvent extraction.
[table 3]
Lactic acid bacteria is to the inhibition effect of Wa rotavirus infection
Figure C20038010805200091
We think that such lactic acid bacteria or streptococcus acidi lactici fermented solution are of value to prevention rotavirus infection and treatment rotavirus diarrhoea.And, consider that rotavirus diarrhoea betides the children of weaning period, absorb by in ablactational food, adding this fermentation dried thing, just can prevent rotavirus diarrhoea, and this rotavirus diarrhoea accounts for 50% of weaning period diarrhoea.

Claims (2)

1. can suppress the preparation method of the lactobacillus fermentation of rotavirus infection, it is characterized by, extraction heat is carried out in the water and the heating that add 10 times of amounts in Fructus Hordei Germinatus, and inoculating lactic acid bacterium in the extract of preparation thus fermented 24 hours in 35 ℃~38 ℃ then.
2. can suppress the lactobacillus fermentation of rotavirus infection, it is following being prepared: extraction heat is carried out in the water and the heating that add 10 times of amounts in Fructus Hordei Germinatus, and inoculating lactic acid bacterium in the extract of preparation thus fermented 24 hours in 35 ℃~38 ℃ then.
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婴幼儿轮状病毒肠炎的抗病毒治疗概述. 刘奋忠等.《海峡药学》,第14卷第6期. 2002 *

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