CN1494374A - Preparation of compositions comprising bacterial strains and volatile plant extracts and therapeutic and industrial applications thereof - Google Patents
Preparation of compositions comprising bacterial strains and volatile plant extracts and therapeutic and industrial applications thereof Download PDFInfo
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- CN1494374A CN1494374A CNA018214851A CN01821485A CN1494374A CN 1494374 A CN1494374 A CN 1494374A CN A018214851 A CNA018214851 A CN A018214851A CN 01821485 A CN01821485 A CN 01821485A CN 1494374 A CN1494374 A CN 1494374A
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Abstract
The present invention discloses: (i) a non-pathogenic probiotic microorganism and its probiotic/therapeutic uses; (ii) a formulation comprising an aqueous solution of a volatile fraction (VF) prepared from the extract of at least one plant derived material and its therapeutic uses; (iii) a process of manufacturing the formulation from the plant derived material; (iv) a probiotic composition comprising the non-pathogenic probiotic microorganism of the invention and/or other probiotic microorganism(s) and the formulation of the invention, and its probiotic/therapeutic uses; (v) a composition for industrial applications comprising the formulation of the invention and microorganism(s) of industrial applicability; and (vi) industrial processes and apparatuses in which the latter composition is used.
Description
Invention field and background
The present invention relates to (i) a kind of non-pathogenic benefit natural disposition microorganism and benefit life/therapeutical uses thereof; The preparation that (ii) contains the aqueous solution of the volatility level part (VF) for preparing by at least a plant origin material extraction thing, and therapeutical uses; (iii) a kind of method by the described preparation of described plant origin manufacture of materials; The beneficial natural disposition composition that (iv) contains non-pathogenic benefit natural disposition microorganism of the present invention and/or other beneficial natural disposition microorganisms and preparation of the present invention, and benefit life/therapeutical uses; (v) a kind of composition that is used for commercial Application contains preparation of the present invention and the microorganism with industrial applicability; (vi) use the industrial method and the device of a kind of composition in above-mentioned back.
Under normal operation, stomach and intestine (GI) road microflora plays an important role to the health and the situation of individuality.As everyone knows, microflora is complexity and the multifarious colony of microorganism, and this colony may have useful and illeffects to individuality.Under the normal pregnancy condition, the fetus in the uterus is aseptic, and but, during being born, fetus can obtain microorganism by vagina the time, causes the formation of gastrointestinal micro-flora.Finally be stabilized in intrinsic gastrointestinal microorganisms fauna in the stomach and intestine and be very complex set body, it has 1000 kinds of dissimilar microorganisms of surpassing, and comprises about 400 kinds of dissimilar bacteriums [Fuller R.J.AppliedBacteriology 66:365-378 (1989)].The composition of gastrointestinal micro-flora although there are a lot of bacteriums to survive in intestines and stomach and to grow, has a variety of bacteriums not survive and to grow by host and microbiological factor decision.In addition, the biology of survival must be avoided creeping effect, and this effect can rinse out described bacterium by food usually.This purpose can be by can realizing by combine the microorganism of fixing it with gastrointestinal wall, and/or by the speed breeding of removing speed realizes to surpass initiatively.
Generally, the existence of gastrointestinal micro-flora is a symbiosis, because described microflora not only helps to decompose the food that some can't digest, and can also protect individuality to avoid the infection that is caused by pathogene.A kind of phenomenon in back was used ' bacterium antagonism ' already, and ' bacterium interference ', ' barrier effect ', ' building group resistance ', nouns such as ' competition exclutions ' carried out statement.
The pipe intestinal protection microflora is very stable.But, it is so ineffective in juvenile, old and patient's body of being endangered.In addition, it may be subjected to some diet and Effect of Environmental, and three kinds of most important factors are excessive cleaning, antibiotic therapy and anxiety.
Balance at gastrointestinal micro-flora is subjected under the condition of negative effect, and probio is recovering gastrointestinal micro-flora and making individual host recover to have potential value aspect normal.
Probio is a quasi-microorganism, is defined as to produce the microorganism of the work of favourable influence to animal and human's class host.Described beneficial effect comprises the microbial balance that improves intestinal microflora or improves the characteristic of autochthonous microorganism fauna.The described beneficial effect of probio can be by the direct antagonism mediation to the particular type biology, by causing described biomass to reduce to the influence of its metabolism or by immune stimulatory power.Probio can suppress the survival quantity of unwanted biology by producing antimicrobial compound by competition nutrition or competition attachment site.In addition, probio can or reduce the enzyme activity change microbial metabolism by raising, or by improving antibody content or improving the macrophage activity stimulating immune system.
WO95/16461 has disclosed the beneficial natural disposition composition of a kind of anaerobic bacteria, and it can effectively control or suppress be in the group that builds in the carcass of salmonella.This benefit natural disposition composition comprises 29 kinds and is essentially pure bacterium, particularly colibacillary colony or culture biologically.But, at the beneficial natural disposition composition disclosed in this PCT open source literature the inhibitory action of pathogene is needed the synergy of a large amount of bacterial isolateses.
WO97/35596 has disclosed to use by mixing and has contained the powder of lactobacillus reuteri, lactobacillus acidophilus and bifidobacterium infantis and the freshly prepd beneficial natural disposition mixture that a kind of liquid obtains.It is said that this mixture can effectively prevent human infectious diarrhea or by diarrhoea that antibiotic therapy caused.But, the bacterium of cryodesiccated work is an anabiotic state.Must make described microorganism moistening before using, so that recover its viability, it is disadvantageous doing like this, because a lot of bacteriums can not survive in rehydrated process under the normal condition.In addition, the biology of surviving is not just to have a metabolic activity at once, and can not survive under extreme, the acid condition of stomach.In addition, when using for the diarrhoea acceptor, the speed that they are discharged from stomach and intestine has surpassed the speed of recovering viability, causes rare or does not have useful effect.
Preserving the viability of beneficial natural disposition biology and keep its activity by its preparation is the theme of a lot of documents.WO98/26787 has disclosed by beta glucan is provided, and optionally with prebiotics and/or probiotic microorganisms associating, strengthen production of lactic acid microorganism in the animal gastrointestinal tract, preferred lactobacillus retain colony.
WO97/34591 also disclosed by the selectivity modification is provided to individuality or unmodified starch or its mixture strengthen retaining colony or suppressing the unwanted colony that retains of microorganism at the GI selected position of individuality, described modification or unmodified starch play one or more beneficial natural disposition microorganism carrier effects, and as described microbial growth or keep medium.Described prebiotics spare is combined on the described carrier in one way, so that protect described microorganism by large intestine or GI other positions the time.
Microorganism is used in the multiple industrial method.Described method can be divided into continuation method and batch methods.In continuation method, as continuous aerobic or anaerobic fermentation production line, be used to produce multiple fermented product, for example, comprise ethanol and methyl alcohol, fermentable raw material are transported on the described production line continuously, and from described production line, collect fermented product continuously, so that on described production line, be maintained at micropopulation described in the whole process of production in check mode.In batch methods, for example, but be not limited in batches aerobic or anaerobic fermentation, in leakage of oil, to the degraded of oil etc., fermentable raw material are mixed with micropopulation, so that ferment.Under some occasion, continuously fermenting and all needing in the fermentation process in batches a large amount of colony with related microorganisms to begin or recover described process with work.For example, when biodegradation oil, preferably the big as far as possible colony with pseudomonad or Alcaligenes microorganism begins described process, for example, finishes the biodegradable time of described oil so that reduce.At the used enrichment filter of microorganism when degrading organic smog or volatile materials, consequently a kind of continuous process wherein, becomes harmless compound with the contaminative degradation.But, the without interruption of organic smog or volatile materials depended in the survival that is present in the microorganism on the described filter.But, under some occasion, the supply of organic smog or volatile materials is discontinuous, has consequently lost described microorganism.In this case, must recover the survival microorganism of effective dose in the described filter at short notice, so that can horse back restore functionality filter.
Therefore, generally believe needs, and highly preferred a kind of preparation, in said preparation, comprise that beneficial natural disposition microorganism and the microorganism that can be used for the microorganism of multiple commercial Application can keep viability for a long time and be the metabolic activity form.
When the present invention is put into practice, be surprised to find that already that the single culture itself that comes from colibacillary non-pathogenic benefit natural disposition microorganism just can recover the normal gastrointestinal tract flora of people and multiple mammal and bird.Be surprised to find already, this microorganism, and other microorganisms can preserve with survival and metabolic activity form in the preparation of the aqueous solution of the volatility level part that comprises various plant extractss for a long time.Have found that comprising the biological beneficial natural disposition composition of benefit that is suspended in the described preparation can effectively treat and prevent various enterogastric diseases.Be surprised to find that also that in addition described preparation itself can be effectively as the weight increase reinforcing agent of mammal and bird with as immunostimulant.
The present invention's general introduction
According to an aspect of the present invention, providing the ATCC preserving number is the coli strain BU-230-98 of 202226 (DSM12799).
According to a further aspect in the invention, providing a kind of ATCC of containing preserving number is the preparation that the beneficial natural disposition composition and being used to of the survival coli strain BU-230-98 of 202226 (DSM 12799) is kept the viability of described coli strain.Described beneficial natural disposition composition preferably also comprises at least a flavouring agent, described preparation preferably comprises at least a volatility level part (VF) of plant extracts, it is by under reduced pressure, plant (for example, beet, dill, parsley or grape) extract is carried out steaming and prepares preferably being no more than under 38 ℃ the bath temperature.
Beneficial natural disposition composition of the present invention can be used for prevention or treatment alimentary infection or imbalance; keep or recover normal gastrointestinal micro-flora; prevention or treatment diarrhoea; prevention or treatment are by enteropathogen; the alimentary infection that causes as Gram-negative bacteria or Gram-positive bacteria; prevention or the salmonella infection of treatment stomach and intestine; prevention or treatment are by for example; clostridium difficile; Salmonella; will Hayes salmonella particularly, campylobacter, Escherichia coli; proteus; the infectious diarrhea that pseudomonas or fusobacterium cause is by antibiotic therapy; radiotherapy or chronic diarrhea or diarrhoea that chemotherapy caused, and/or be used to make GI physiologically active normalization.
According to a further aspect in the invention, the preparation of at least a volatility level part (VF) of containing plant extracts is provided, and described volatility level part is to prepare by described plant extracts is carried out steaming by being no more than under reduced pressure under 38 ℃ the bath temperature.Described plant extracts preferably can obtain from the plant organ that is selected from plant leaf blade, stem, root and fruit.Described plant can be vegetables, as soybean, clover, garlic, beet and wild cabbage or herbaceous plant, as parsley, peppermint and dill.Described preparation also comprises the beehive product, for example, but is not limited to propolis.Described preparation can be used as animal doctor's feed or food additives, is used to strengthen the weight increase of receptor.As what further disclose in this article, also can use it for and preserve the survival bacterium.
Therefore, according to a further aspect in the invention, provide a kind of method for preparing volatility level part of plant, may further comprise the steps (a) and grind vegetable-derived materials, obtained a kind of plant biological material; (b) described plant biological material is mixed with water, and stir at ambient temperature; (c) under reduced pressure, and the mixture that is obtained in to step (b) under being no more than 38 ℃ bath temperature carries out steaming; (d) collect volatility level part by described steaming obtained.Described decompression is preferably the 5-10 millibar.Can be with said preparation as food additives, feed addictive, and have beyond thought therapeutic action, and favourable bacterium preservation characteristics.Also disclosed the treatment preparation of volatility level part of containing plant, said preparation can prepare with physiology or animal doctor's acceptable additive, carrier or mixing diluents by at least a volatility level part (VF) with plant extracts.Described volatility level part prepares by described plant extracts is carried out steaming being no more than under 38 ℃ the bath temperature preferably under reduced pressure.
Therefore, according to a further aspect in the invention, provide a kind of beneficial natural disposition composition, said composition comprises the beneficial natural disposition microorganism of at least a survival of (a) effective dose, and this microorganism has useful biology or therapeutic activity in intestines and stomach; (b) at least a volatility level part (VF) of plant extracts, this volatility level part are preferably pressed the disclosed method preparation of this paper.Described beneficial natural disposition microorganism can be Escherichia coli, preferably the ATCC preserving number is the bacterial strain BU-230-98 of 202226 (DSM12799), this microorganism is except can be used for treatment or prevention indigestion symptom, be used to strengthen the patient's who suffers from dysimmunity immune response, for example, outside the disease that is caused by the immune response suppression therapy or as an alternative scheme also can be used it for treatment or prevent above-mentioned any imbalance/pathology/disease/syndrome.
According to a further aspect in the invention, provide a kind of composition, said composition comprises (a) at least a microorganism, as Escherichia, and Alcaligenes, arthrobacterium, Bifidobacterium, lactobacillus, galactococcus, nitrosomonas and pseudomonad; (b) at least a volatility level part (VF) of plant extracts, described at least a volatility level part is through selecting, so that at room temperature the viability of described at least a microorganism was preserved 2 months at least, preferably at least 3-6 month, more preferably 6-12 month or longer time at least.
According to a further aspect in the invention, it is the method for the coli strain BU-230-98 of 202226 (DSM 12799) that a kind of growth ATCC preserving number is provided, this method may further comprise the steps: with the ATCC preserving number is that the starting culture of the coli strain BU-230-98 of 202226 (DSM 12799) is inoculated in the growth medium, and the optical density (OD) that described coli strain is grown under 650nm is 15-30.
According to a further aspect in the invention, provide a kind of production beneficial natural disposition method for compositions, this method may further comprise the steps: with the ATCC preserving number is that the survival coli strain BU-230-98 of 202226 (DSM 12799) is suspended in a kind of preparation, so that keep the viability of described coli strain.Described preparation preferably comprises at least a volatility level part (VF) of plant extracts, described volatility level part is preferably by under reduced pressure, described plant extracts is carried out the steaming preparation being no more than under 38 ℃ the bath temperature, for example, described plant extracts is to obtain from being selected from such as the vegetables of soybean, parsley, peppermint, dill, clover, garlic, beet or wild cabbage or the plant organ of herbal blade, stem, root and fruit.
According on the other hand, the invention provides the beneficial natural disposition method for compositions of a kind of preparation, this method may further comprise the steps: with the beneficial natural disposition microbial suspension of at least a survival effective dose, that in intestines and stomach, have useful biology or therapeutic activity in the preparation of at least a volatility level part (VF) of containing plant extracts.
According to a further aspect in the invention, provide a kind of preparation to contain to survive the method for compositions of microorganism, this method may further comprise the steps: with at least a microbial suspension in the preparation of at least a volatility level part (VF) of containing plant extracts, described at least a volatility level part is through selecting, so as can be at room temperature will described at least a microorganism viability maintenance at least 2 months.For example, described microorganism can be Escherichia, Alcaligenes, arthrobacterium, Bifidobacterium, lactobacillus, galactococcus, nitrosomonas or pseudomonad.
According to a further aspect in the invention, a kind of microorganism distributor is provided, this distributor comprises a holder and a meter that is connected with this holder, this holder is equipped with at least a microorganism in the preparation that is present at least a volatility level part (VF) of containing plant extracts, described at least a volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
According to a further aspect in the invention, a kind of method of biology being carried out BIOLOGICAL CONTROL is provided, this method may further comprise the steps: can be sprayed at epidemic regions to the survival microorganism that described biology carries out BIOLOGICAL CONTROL with at least a, described at least a survival microorganism is present in the preparation of at least a volatility level part (VF) of containing plant extracts, described at least a volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
According to a further aspect in the invention, a kind of method that greasy dirt dyes of removing is provided, this method may further comprise the steps: can be sprayed at contaminated zone by the oily microorganism of degraded with at least a, described at least a survival microorganism is present in the preparation of at least a volatility level part (VF) of containing plant extracts, described at least a volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
According to a further aspect in the invention, a kind of method of recovering the organic smoke degradation micropopulation in the biofilter is provided, this method may further comprise the steps: at least a survival microorganism that can degrade organic smog is distributed on the described filter, described at least a survival microorganism is present in the preparation of at least a volatility level part (VF) of containing plant extracts, described at least a volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
According to a further aspect in the invention, provide a kind of preparation to be used for the method for the leavening of fermentation process, this method may further comprise the steps: the leavening microorganism of growth sufficient amount, and with described leavening microbial suspension in the preparation of at least a volatility level part (VF) of containing plant extracts, described at least a volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
The summary of accompanying drawing
In this article, only the present invention will be described in conjunction with the accompanying drawings with way of example.Following mask body is in detail referring to accompanying drawing, should be noted that, details shown in the figure is the character of giving an example, and be only used for the discussion of describing property of the preferred embodiments of the invention, such accompanying drawing is provided, and is in order to provide principle of the present invention and conceptional features to be carried out explanation the most useful and that be convenient to understand most.Thus, the demonstration of CONSTRUCTED SPECIFICATION of the present invention is not exceeded functional understanding needs of the present invention,, those skilled in the art can be understood to implement the present invention with some kinds of forms described the description of the drawings.
In the accompanying drawings:
Fig. 1 is the schematic diagram of distributor of the present invention.
Detailed Description Of The Invention
The present invention is (i) a kind of non-pathogenic benefit natural disposition microorganism and benefit life/therapeutical uses thereof; (ii) contain preparation by the aqueous solution of volatility level part (VF) of at least a plant origin material extraction thing preparation, and therapeutical uses; (iii) a kind of method by the described preparation of described plant origin manufacture of materials; (iv) contain the beneficial natural disposition composition of non-pathogenic of the present invention benefit natural disposition microorganism and/or other beneficial natural disposition microorganisms and preparation of the present invention, and benefit life/therapeutical uses; (v) a kind of composition for industrial use contains preparation of the present invention and the microorganism with industrial applicability; (vi) commercial run and the device of the above-mentioned rear a kind of composition of application.
With reference to the accompanying drawings with the description of the drawings principle that the present invention may be better understood and operation.
Before in detail explaining at least a embodiment of the present invention, it must be understood that, application of the present invention not and be confined to below specification or the explanation of embodiment in the details that provides. The present invention can have other embodiments, perhaps implements in every way or execution. In addition, should be understood that the phraseology and terminology used herein are for illustration purpose, and should being considered limiting property.
On the one hand, the present invention relates to a kind of beneficial natural disposition microorganism, it is to come from colibacillary non-pathogenic bacteria, for example, having useful physiology and/or therapeutic activity in intestines and stomach, and be № 202226 at the preserving number of ATCC, is No.12799 at the preserving number of DSMZ.
" beneficial natural disposition " is used as adjective in this article, is used for the bacterium that expression has the separation of the characteristic that suppresses at least a pathogenic growth. Employed inhibition test is the in vitro test of carrying out at solid medium in this article, wherein, the culture supernatant of candidate's separation of bacterial is coated on the surface of described solid medium, observes the characteristic that it suppresses pathogenic growth. Usually, will be placed on the agar plate surface of having inoculated described pathogen with the scraps of paper that the culture supernatant of candidate's benefit natural disposition bacterial strain was soaked. The stand density that benefit natural disposition bacterium supernatant has caused the transparent agar of a circle or weakened shows near the pathogen that has suppressed the described scraps of paper. Having other inhibition tests can use maybe can design, and comprises external or the interior direct growth competition experiments of body, and it can produce a category and be similar to the disclosed beneficial natural disposition bacterium of this paper. Usually the bacterial isolates identified of above-mentioned any test all belongs to the type of beneficial natural disposition bacterium, as employed this term in this article.
Beneficial natural disposition bacterium of the present invention can be used as food or feed addictive, and to have added the ATCC preserving number be food or the feed of the survival coli strain BU-230-98 of 202226 (DSM 12799) in order to provide. Food of the present invention can also comprise at least a milk-derived material, and can be selected from cheese and sour milk.
The invention still further relates to the preparation of at least a volatility level part (VF) of containing plant extracts, described volatility level part is by preparing usually being no more than under 38 ℃ the bath temperature described plant extracts distilled under reduced pressure.
Can obtain from any suitable plant part for the vegetable material that obtains described volatility level part, such as fruit, blade, stem or root. A lot of plants all are suitable as the source of described volatility fraction, and for example apple, oranges and tangerines, soybean, beet, wild cabbage, garlic and clover, and herbaceous plant are such as parsley, peppermint and dill. Preparation of the present invention can also optionally contain the agricultural product of suitable quantity, such as volatility level part of honey, propolis or other beehive products, described level part can be used with disclosed being used for of this paper and obtain the identical method preparation of volatility level part from plant extracts.
As what will be explained in more detail in the following embodiments, can be with described preparation itself as the foods/feeds additive. The inventor has found that said preparation has the activity that strengthens the body weight increase, and it has consisted of one aspect of the present invention. A kind of specific embodiments of this aspect according to the present invention, described the weight of animals increase enhancing preparation or feed addictive comprise volatility level part of distilled water and clover, soybean, beet and dill, and the volume ratio of described volatility level part is preferably 2: 8: 1: 4. In addition, preparation of the present invention or feed addictive comprise volatility level part of each 1 part carrot, beet, dill and grape contained in distilled water and the per 100 parts of water. As another kind of alternative, preparation of the present invention or food additives comprise volatility level part of each 1 part of beet, dill and parsley contained in distilled water and the per 100 parts of water.
On the other hand, the present invention relates to a kind of composition, said composition contains above-mentioned preparation of the present invention and at least a survival microorganism, described microorganism can be to have treatment or the active beneficial natural disposition biology of beneficial organism in the intestines and stomach such as people and/or animal, in order to a kind of beneficial natural disposition composition is provided, or have the biology of industrial application value, in order to the composition with industrial applicability is provided.
The special benefits of beneficial natural disposition of the present invention and industry group compound comes from the following fact, and namely they are liquid preparations. Under the BA condition, described preparation also can be used as the supportive culture medium of bacterium alive, and cryodesiccated preparation is then different, and wherein, described bacterium is to be under the anaerobic condition. As a result, for instance, beneficial natural disposition composition of the present invention just works after oral at once, and this effect starts from GI top, and the Main Function of most of enteropathogen all here occurs, and causes the appearance of bad gastro-intestinal syndrome. Beneficial natural disposition composition of the present invention also can be used as body weight increases preparation or foods/feeds additive. Similarly, industry group compound of the present invention can be used for having efficient recovery to have the micropopulation of industrial applicability.
Therefore, should be noted that, for beneficial natural disposition purposes, except other benefit bacteriums that belong to bacterial strain BC-230-98 (ATCC preserving number 202226, equally with 12799 numbering in the DSM preservation) can be used with the preparation in vegetable material of the present invention source. Described beneficial natural disposition bacterium has the very antagonistic activity of wide spectrum. They also belong to identical systematic growth type with most of enteropathogen, and have identical survival system. Therefore, the inhibition of enteropathogen can comprise multiple different mechanism with discharging, and for example secretion of antagonistic substance is to the competition utilization of nutrient with to the competition of adhesion receptor. Therefore, any non-pathogenic bacteria that meets above-mentioned standard all can be used in the beneficial natural disposition composition of the present invention.
According to a further aspect in the invention, beneficial natural disposition composition of the present invention can be can be used for prevention or treatment alimentary infection. Term ' alimentary infection ' is used to indicate any infection that is caused by enteropathogen, includes but not limited to Gram-negative and gram-positive bacterium. In order to improve GI population equilibrium and health, preparation of the present invention and beneficial natural disposition composition also can be used for preventing the alimentary infection that caused by yeast, virus and protozoan.
Concerning this paper, term ' treatment effective dose ' or ' effective dose ' are the consumptions of being determined by factor well known in the art. Described consumption must be enough to that the efficient recovery gastrointestinal micro-flora is arranged, thereby causes the normalization of gastrointestinal function.
In a kind of specific embodiments, the beneficial natural disposition composition that is used for the generation of prevention alimentary infection comprises preparation of the present invention, said preparation is to be for example 5: 1: 5 with distilled water and volume ratio: volatility level part of 15: 1 clover defined herein, soybean, beet, dill and peppermint, and the beneficial natural disposition bacterium for the treatment of effective dose, be 202226 Escherichia coli (identical with DSM 12799) or any other beneficial natural disposition bacterium preparation such as the ATCC preserving number.
An object lesson of alimentary infection is the infection that is caused by Salmonella, and the present invention is particularly conducive to prevention or treats the alimentary infection that is caused by Salmonella.
In addition, food additives of the present invention or preparation can be separately or with effective dose be that 202226 Escherichia coli (identical with DSM 12799) are united and are used for the treatment of or prevention infection diarrhoea, chronic diarrhea or the diarrhoea that caused by antibiotic or chemotherapy such as the ATCC preserving number.
According to another kind of specific embodiments of the present invention, the described beneficial natural disposition composition that is used for the treatment of diarrhoea can contain distilled water and volume ratio is for example 5: 1: 5: volatility level part of 15: 1 clover, soybean, beet, dill and peppermint, and the ATCC preserving number 202226 (DSMZ 12799) of treatment effective dose.
Described infectious diarrhea can cause by multiple factor, for example, and by being selected from clostridium difficile, Salmonella, particularly will Hayes salmonella, campylobacter, Escherichia coli, proteus, pseudomonas, fusobacterium, intestines staphylococcus.More than be in the multiple infectious agent a few.
In addition, beneficial natural disposition composition of the present invention can be used to have the gastrointestinal micro-flora in the subject of this treatment of efficient recovery needs, this treatment can cause the normalization of gastrointestinal function.For example, described composition can comprise volatility level part of distilled water and clover, soybean, beet, dill, peppermint, parsley and wild cabbage, the ratio of volatility level part is preferably for example 1: 5: 5: 2: 2: 1, and the ATCC preserving number of treatment effective dose is the beneficial natural disposition bacterium of 202226 (DSM 12799).
Other purposes that contain at least a the present invention's benefit natural disposition composition the beneficial natural disposition bacterium that has therapeutic action such as GI position comprise alleviates the lactose intolerance of suffering from the patient who does not tolerate lactose, the treatment enterocolitis, the treatment constipation, be used for reducing the cholesterol content of blood, be used for the treatment of indigestion symptom, and/or being used to stimulate the immune system of the object of suffering from the immune system obstacle, described obstacle may be the dysimmunity that is caused by immunosuppressive therapy.
On the other hand, the present invention relates to a kind of method that is used for volatility level part of prepared plant extracts, this method may further comprise the steps: (a) grind the plant origin material, so that obtain the plant biological material; (b) preferably be blended in the plant biological material and the water of acquisition in the step (a), and stirred this mixture at ambient temperature at least 2 hours with the weight ratio of 3 parts of water and 1 part of plant biological material; (c) under reduced pressure, and at the mixture that preferably is being no more than distillation acquisition in step (b) under 38 ℃ the bath temperature; (d) collect, can further dilute described level part with suitable buffer solution by volatility level part that steaming obtained.
Volatility level part of the present invention can be mixed with water, so that obtain foods/feeds additive of the present invention or preparation.Can also prepare foods/feeds additive of the present invention or preparation by mixing more than one the phytovolatilization level part that obtains with the inventive method.This mixture can further mix with water.
Described volatility level part can be with any suitable fruit, vegetables, the leaf of plant, stem or root preparation.For example, described plant can be apple, citrusfruit, soybean, beet, garlic, wild cabbage or clover, or herbaceous plant, as parsley, peppermint or dill.Should be pointed out that described preparation can also contain from agricultural product if suitable, as volatility level part of honey or propolis or other beehive products.Described volatility level part can prepare with the identical method of prepared plant extracts volatility level part.
Method according to the present invention, described distilation steps preferably carries out under the reduced pressure of 5-10 millibar.
Just as noted above, foods/feeds additive of the present invention or preparation can also make up with at least a beneficial natural disposition or industrial preparation, so that beneficial natural disposition of the present invention or industry group compound are provided.Therefore, preparation method of the present invention can also comprise and the beneficial natural disposition of at least a survival or industrial microorganism is suspended in the volatility level part that obtains or is suspended in step in the mixture of the volatility level part that obtains as stated above in above-mentioned steps (d).
According to a kind of concrete grammar of the present invention, the beneficial natural disposition microorganism of described suspension can be a novel bacterial of the present invention, and it is 202226 Escherichia coli (preserving number at DSM is 12799) that this bacterium comes from preserving number by the ATCC preservation.
At last, the present invention relates to volatility level part (VF) of plant extracts is used to prepare the purposes of foods/feeds additive, wherein, described VF is by being no more than the described plant extracts of distillation under 38 ℃ the bath temperature or preparing such as the agricultural product of honey or propolis.
Another aspect of the present invention comprises that a kind of preparation contains the method for compositions of the microorganism that survives.This method is by at least a microbial suspension is realized in the preparation of at least a volatility level part (VF) of containing plant extracts.Described preparation is through selecting, so that can be at room temperature the viability of described microorganism be kept 2 months at least.Selected microorganism can be beneficial natural disposition or have an industrial use.The biology that can keep in preparation of the present invention comprises Escherichia, Alcaligenes, arthrobacterium, Bifidobacterium, lactobacillus, galactococcus, nitrosomonas and pseudomonad.
As shown in Figure 1, another aspect of the present invention provides a kind of microorganism distributor 10.Distributor 10 comprises a holder 12 and a meter that is connected with it 14.Holder 12 accommodates at least a microorganism in the preparation of at least a volatility level part (VF) of containing plant extracts.Described preparation is through selecting, so that at room temperature the viability of described microorganism was kept 2 months at least.Container 12 can freeze, and for example freezes to about 2-20 ℃, preferably freezes to about 4 ℃.Distributor 10 can be airborne distributor, so that can distribute insect control biologic product.Described distributor can be a hand-held.Under any circumstance, described dispenser designs can be become the solution that distributes predetermined automatically, for example periodic allocation.For this reason, distributor 10 can comprise a programmable or default controlling organization 16, valve 18 etc.
Therefore, according on the other hand, the invention provides a kind of biological control method of biology, this method is by being sprayed at epidemic regions to the survival microorganism that described biology carries out BIOLOGICAL CONTROL and realizing at least a.Described survival microorganism is present in the preparation of at least a volatility level part (VF) of containing plant extracts, and described volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
According to a further aspect in the invention, provide a kind of method that greasy dirt dyes of removing, this method is by will being sprayed at contaminated zone by the oily survival microorganism of degraded such as at least a of pseudomonad.Described survival microorganism is present in the preparation of at least a volatility level part (VF) of containing plant extracts, and described volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
According to a further aspect in the invention, a kind of method of recovering the organic smoke degradation micropopulation in the biofilter is provided, has comprised at least a survival microorganism such as the organic oil of degrading of pseudomonad, nitrosomonas or arthrobacterium is assigned on the described filter.Described survival microorganism is present in the preparation of at least a volatility level part (VF) of containing plant extracts, and described volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
According to a further aspect in the invention, a kind of method for preparing the leavening of fermentation process is provided, the leavening microorganism that comprises the sufficient amount of growing, as galactococcus, Escherichia, Bifidobacterium, lactobacillus, galactococcus or pseudomonad, and with described leavening microbial suspension in the preparation of at least a volatility level part (VF) of containing plant extracts, described volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
By checking following examples, those of ordinary skills can understand other objects, advantages and novel features of the present invention, and described embodiment is not used to limit purpose.In addition, various embodiments of the invention described above and each of aspect, and following claim part can find experimental basis from following examples.
Embodiment
Referring to following embodiment, these embodiment describe the present invention in non-limiting mode with above-mentioned specification.
Embodiment 1
Prepared plant extracts or from " volatility level part " of the extract of agricultural product
Fully clean with the fresh vegetables that running water will be bought from the market, chopping, and in commercial blenders, wear into fine powder.Weight ratio with 3 parts of water and 1 part of vegetables biological substance is added distilled water in the described vegetables biological substance to, and stirs at least 2 hours at ambient temperature.
Described mixture is transferred in the evaporation flask of rotary evaporator, and under the following conditions in decompression (5-10 millibar) evaporation down: bath temperature is no more than 38 ℃, and the condenser inlet temperature is 2-5 ℃.Per 2.5 kilograms of plant biological materials are collected about 1 liter of condensate (volatility level part).As what will further specify hereinafter, this material can be preserved 12 months under refrigerated condition and do not lose its characteristic at least.
Can use agricultural product to replace described vegetable material, handle with identical method such as honey or propolis.
Usually, each " volatility level part " is with the preparation of one type vegetables, and is used to prepare the different mixture that is used for various purposes with different dilution ratios.
Embodiment 2
The ATCC preserving number is 202226 biological colibacillary separation of beneficial natural disposition and growth
The biological Escherichia coli of benefit natural disposition are (on May 3rd, 1999, preserving number with 202226 is preserved in ATCC, on May 4th, 1999, preserving number with 12799 is preserved in DSM) from Escherichia coli M-17, separate, comprise that the survival thing from the preparation that is kept at volatility level part of containing embodiment 1 described plant origin material 37 ℃ down long-term (2 months) begins the series transfer of separator, and screening can with the separator of pathogene competition, as what be described in more detail below.The biological Escherichia coli of beneficial natural disposition (on May 3rd, 1999, the preserving number with 202226 was preserved in ATCC, and on May 4th, 1999, the preserving number with 12799 was preserved in DSM) are grown in following medium: (NH
4)
2SO
4-5g/L, KH
2PO
4-13g/L, Na
2HPO
4-13g/L, MgCl
2-3g/L, CaCl
2-0.3g/L, yeast extract-10g/L, soy peptone-10g/L and glucose-5g/L.Along with extra nutrient (yeast extract-1g/L, soy peptone-2.5g/L and glucose-90g/L), the concentration of glucose in the fermentation culture is remained on the level of 2g/L are added in the growth of described culture continuously.By continuous interpolation 4N NH
4OH remains neutrality with the pH of fermentation culture.Cultivation is under 30 ℃, carries out 16 hours with the ventilation speed of 0.5vvm in the standard fermentation tank, and growth reached and converged this moment.This method can obtain 10
10-10
11Cells/ml.By centrifugal results Bacillus coli cells, be suspended in the salt solution, and precipitate once more.Described microorganism biological material can be placed in the salt solution and to keep 48 hours in refrigerator and do not lose viability.
Embodiment 3
Preparation can increase the foods/feeds additive formulations of mammal and bird body weight fast
This mixture contains the following volatility level part by the foregoing description 1 described method preparation: clover 50ml/L, soybean-200ml/L, beet-25ml/L and dill-100ml/L.Carry out balance with distilled water.
Embodiment 4
Preparation is used to make the food additives of human gi-tract function normalization
This mixture contains the following volatility level part by the foregoing description 1 described method preparation: clover-50ml/L, soybean-10ml/L, beet-50ml/L, dill-50ml/L, peppermint-20ml/L, parsley-20ml/L and wild cabbage-10m1/L.Carry out balance with distilled water.Cell concentration with 107 cells/ml is suspended in this mixture with Escherichia coli (ATCC 202226) cells (embodiment 2).Can optionally add NaCl (for example 4-10g/L) in order to improve mouthfeel.
Embodiment 5
Preparation is used to prevent the feed of mammal and bird alimentary infection to add preparation
This mixture contains the following volatility level part by the foregoing description 1 described method preparation: clover-50ml/L, soybean-10ml/L, beet-50ml/L, dill-150ml/L and peppermint-10ml/L.Carry out balance with distilled water.With 10
7The cell concentration of cells/ml is suspended in this mixture with Escherichia coli (ATCC 202226) cells (embodiment 2).Can optionally add NaCl (for example 4-10g/L) in order to improve mouthfeel.
Embodiment 6
Escherichia coli (ATCC 202226) are right
The antagonistic activity of salmonella typhimurium (ATCC 14028)
The culture dish of the Brilliant Green agar (the selective growth medium of Salmonella) of modification is housed with the salmonella typhimurium inoculation.On agar, form the hole of 9 mm dias.The described food additives (embodiment 4) of certain volume are deposited in each hole, and under 35 ℃, hatched described dull and stereotyped 24 hours.Repeat identical process, but need not described food additives, but will filter the liquid deposition that obtains in described hole by microorganism filter membrane (0.45 micron in aperture).
Observed the inhibition zone (10-17 millimeter) that lacks the salmonella typhimurium bacterium colony around in each hole of containing described food additives.The inhibition zone does not appear around in the hole of containing the filtrate that does not contain described beneficial natural disposition biology.
Embodiment 7
Escherichia coli ATCC202226 and
Parental generation Escherichia coli M-17 is to the antagonistic activity of Shigella
Shigella flexneri, Shigella sonnei, Escherichia coli (ATCC 202226) and M-17 were grown 18-20 hour down at 37 ℃ on nutrient agar respectively.All cultures are all gathered in the crops in the salt solution, and are diluted to the optical density of 10Klett unit.Shigella is diluted 1 milliliter of the sample aliquot of culture itself or be inoculated in the ventilation cup-shaped test tube that nutrient solution (5 milliliters) is housed with the dilution cultures of Escherichia coli (ATCC 202226) combination (1 milliliter).Under 37 ℃, hatched described test tube 24 hours.By with described culture bed board to nutrient agar, determine colony forming unit (CFU) quantity of described pathogene and Escherichia coli (ATCC 202226).Two kinds of Shigella species have been shown at pure culture with have CFU quantity in the colibacillary mixed culture of beneficial natural disposition below in the table 1.
Table 1
| Benefit natural disposition biology | The growth of Shigella | |
| Shigella flexneri | Shigella sonnei | |
| The pure Shigella culture of Escherichia coli ATCC 202226 Escherichia coli M-17 | ＜5×10 41×10 66×10 6 | ＜5×10 42×10 62×10 6 |
Embodiment 8
With food additives (embodiment 4) and colicine
(drying agent of Escherichia coli M-17) is used for enterogastritis patient in hospital
One group of 60 patient that enterogastritis takes place after being in hospital are divided into three groups at random, make each group have similar distribution aspect sex, age and the enterogastritis severity of symptom.All patients accept normal supportive treatment, comprise rehydrated, vitamin etc.The order of severity of symptom does not need to use antibiotic therapy.20 patient every days of first group 3 times were accepted 10 milliliters food additives (embodiment 5) 30 minutes ante cibum and treat 7 day time altogether.Scheme (2 30 minutes ante cibum of the every day use) colicine (Colibacteriilum siccum) that the patient of second group of same quantity recommends according to the manufacturer, and the 3rd group do not accepted additive.
Write down the beginning (in fate) of the enterogastritis symptom normalization of all groups, and shown in the table 2 below.
Table 2
| Symptom | The beginning of symptom normalization, fate | ||
| Food additives | Colicine | Contrast | |
| Body temperature | 2.8±0.2 | ?2.7±0.2 | ?3.7±0.2 |
| Poison | 2.5±0.1 | ?3.6±0.1 | ?4.6±0.1 |
| Stomachache | 3.3±0.2 | ?5.1±0.2 | ?6.1±0.2 |
| Diarrhoea | 1.8±0.2 | ?3.4±0.2 | ?4.4±0.2 |
Colicine (Colibacterinum siccum) is by BIOMED company (Moscow, Russia) freeze-dried preparation of the Escherichia coli M-17 of the work of producing, and recommended diarrhoea [the Vidal Handbook:Pharmaceutical preparations inRussia (N.B.Nikolaeva that is used for the treatment of, B.P.Alperovich and V.N.Sovinov, Eds.) AstrapharmService, 1997, Moscow, p.275].
Embodiment 9
The patient who food additives (embodiment 4) is used to suffer from acute alimentary infection
This research comprises the patient of the severe alimentary infection of suffering from the various causes of disease: the food poisoning of salmonellosis, Escherichia disease, shigellosis, staphy lococcus infection and unknown etiology.In all patients, be in hospital with the indication that appears as of the acute attack of disease and acute gastroenteritis.With described food additives 186 patients are altogether treated, and 102 patients accept standard care like the category.
Write down the beginning (in fate) of the enterogastritis symptom normalization of all groups, and shown in the table 3 below.
Table 3
| Symptom | Normalization begins, fate | |
| Seminar | Contrast | |
| Weak abdominal pain diarrhea bed fate generates heat | ?1.2±0.3 ?1.6±0.2 ?1.5±0.2 ?2.3±0.2 ?4.8±0.3 | ?2.7±0.3 ?2.9±0.3 ?2.4±0.3 ?4.7±0.4 ?6.7±0.4 |
In one is independently studied, with described food additives one group of 30 patient that suffer from ileotyphus to be treated, 80% disease of patient symptom disappeared in 3 days.3 routine described additive treatments are only arranged owing to the development of more serious chronic enteritis stops.
Food additives (embodiment 4) are used for the gastro intestinal disorders patient that causes by antibiotic
The patient that will suffer from severe GI imbalance is divided into 2 groups.First group comprises 48 patients that suffer from gastric ulcer, and these patients are carrying out gastro intestinal disorders having occurred after the antibiotic therapy at helicobacter pylori.Second group comprises 22 patients, and these patients are carrying out gastro intestinal disorders having occurred after the antibiotic therapy at pneumonia.
Every day 3 times is used described food additives (5 milliliters) in ante cibum, uses altogether 7 days.In these 2 groups, all patients' symptom of diarrhea all disappeared in 2-3 days.After using described additive, in 84.5% patient, observed the complete normalization of gastrointestinal microorganisms fauna.This is the remarkable increase by lactobacillus and Bifidobacterium, the minimizing of Escherichia coli sum, and haemolysis Escherichia coli and prove such as the complete obiteration of staphylococcus, proteus vulgaris or even oidiomycetic other pathogene.In remaining patient of 15.5%, observed remarkable improvement.
Embodiment 11
The patient who food additives (embodiment 5) is used to suffer from radiotherapy later stage enterogastritis
Use described food additives (10 milliliters, every day 3 times, use 30 minutes ante cibum, used altogether 14 days) for 24 patients that suffer from uterus, colon and cancer of the stomach, described patient enterogastritis occurred after the therapeutic alliance of accepting radiotherapy or radiation and chemotherapy.
Before using described food additives, all patients have defecation desire regular and misery, liquid-like excrement (every day 4-12 time), the main suit of mucous membrane (9 example) and blood (3 example) occurs in ight soil.
Taking in described additive 2 or after 3 days, all patients have noticed the painful minimizing that alleviates with defecation frequency, and stool has normal outward appearance.After symptom of diarrhea disappears 4-5 days, the appearance of blood and mucous membrane stopped.Blood analysis shows that physiochemical indice is significantly improved.
In the control group of similar scale, continue to exist described symptom.
Food additives (embodiment 4) are used for AIDS patient
The patient who suffers from AIDS chronic diarrhea occurs through regular meeting.Allow one group of such patient use 10 milliliters of food additives (embodiment 5), every day 3 times, use 30 minutes ante cibum, used altogether 20 days, and control group is not accepted additive.Treatment results is shown in the following table 4.
Table 4
| Parameter | Food additives | Contrast |
| Patient's number mean age, when every day in when beginning year experiment, the defecation frequency experiment finished after defecation frequency experiment every day a month every day defecation frequency normalization the average time started | 30 38±1 3.4±0.3 1.1±0.1 1.5±0.2 6.0±0.7 | 20 36 ± 2 3.6 ± 0.3 3.2 ± 0.3 3.7 ± 0.3 is still undesired |
The patient who accepts described food additives shows the microbiotic normalization of intestines: the Escherichia coli sum reduces, and the haemolysis Escherichia coli disappear, and the quantity of lactobacillus and Bifidobacterium increases, and candida albicans reduces.
Embodiment 13
The weight increase that foods/feeds additive (embodiment 3) is used to quicken healthy piglet.
Use 3 milliliters of described foods/feeds additives for every day every little piglet, up to wean.Accept the group of described feed addictive, the weight average of every piggy has increased by 1.0 kilograms than control group when wean.
Described feed addictive (embodiment 5) is used for healthy little piglet
The little piglet of hundreds of health the 1st and the 3rd day of birth and when wean every little piglet use 3 milliliters of described feed addictives.Compare lethality with the control group of accepting the standard precautions antibiotic therapy and reduced 50%.The weight average of accepting group every piggy when wean of feed addictive has increased by 0.39 kilogram than control group.When more described feed addictive and antibiotic therapy in brood son, the weight increase of the little piglet of the described feed addictive of discovery acquisition is higher 2.4 kilograms than control group.
Embodiment 15
Feed addictive (embodiment 4) is applied to show the little piglet of diarrhoea
Use 5 milliliters feed addictive for hundreds of piggy son every piggys every day that show diarrhoea.The control group antibiotic treatment of identical scale: advocin, gentamicin, Amoxicillin.The symptom of diarrhea of accepting the group of described feed addictive disappeared in 1-2 days.Death occurs, and the piggy son grows normal.Antibiotic can make most of little piglet stop diarrhoea, but these little piglet are still grown slowly.
Give and to accept antibiotic therapy about 70 hairs 1 week, that generally be considered to die of suffering from diarrhoea and educate slow little piglet and used feed addictive 3 days.All survivals except that 2 in these little piglet.
Feed addictive (embodiment 4) is used for healthy calf
Use 5 milliliters of feed addictives for the healthy calf of birth just by emulsion, use 7 day time.Veal above 95% diarrhoea all occurs up to 14 ages in days, has diagnosed the rotavirus positive of a small amount of case simultaneously.Usually, general, the incidence of disease of suffering from diarrhoea in this farm is 20-30%.
Embodiment 17
Feed addictive (embodiment 6) is used to the calf that occurs suffering from diarrhoea
Use 10 milliliters of feed addictives for every calf that diarrhoea occurs every day by emulsion.The symptom of diarrhea of the calf of 90-95% disappeared in 1-2 days.The diarrhoea of all the other 5-10% is caused by virus.Obtained relatively poor result with these calves of antibiotic therapy.
Feed addictive (embodiment 6) is used for healthy sheep lamb and kid
Use 3 milliliters feed addictive for the sheep lamb and the kid of birth just by emulsion, use 7 day time altogether.In some cases, can occur slightly to the moderate symptom of diarrhea.These symptoms can disappear usually automatically, or can successfully treat by enough heavy dose of feed addictives (5 milliliters).For sheep lamb and kid, the incidence of disease of suffering from diarrhoea in described farm is generally 50%.
Embodiment 19
Sheep lamb and kid that feed addictive (embodiment 5) is used to show diarrhoea
The described feed addictive of test in the herd that suffers from the enteropathogenic E infection.In the previous year of test, die from diarrhoea for nearly 90 in 120 sheep lambs and the kid.Inoperative with antibiotic therapy, because this disease is to take place suddenly, and very fast death.In 3-5 days time, use 5 milliliters feed addictive every day for every sheep lamb that shows diarrhoea and kid by emulsion.Animals of control group is according to normal recommendation antibiotic therapy.About 90% the symptom of diarrhea of accepting the sheep lamb of feed addictive and kid disappeared in 1-2 days.The further growth of these lambs is acted normally.The control group of accepting antibody (gentamicin) shows relatively poor result.The diarrhoea of control group continues to exist, and need treat repeatedly with antibiotic.The growth of control animals is seriously slow.
Embodiment 20
Feed addictive (embodiment 5) is used for poultry
In the phase of breeding (42-49 days) feed addictive is added in the drinking water, make every chicken take in 0.01 milliliter feed addictive every day.In the in check test of carrying out with young chicken, found that weight increase is 3.2%, also be accompanied by food conversion simultaneously and improve 4%.
Turkey with age in 1 day-6 week has obtained good result equally.Every fowl is accepted 0.01 milliliter additive every day.Observed weight increase, reduced lethality simultaneously above 10%.Still the poultry every day that shows diarrhoea is with 0.1 milliliter food additives treatment, and need not any antibiotic therapy, and compares with the contrast bird with antibiotic therapy only, shows better recovery situation.
Embodiment 21
Feed addictive (embodiment 5) is used for dog and cat
Feed addictive is used for doggie, causes in 24-48 hour, making symptom of diarrhea to stop.
Embodiment 22
Feed addictive (embodiment 5) is used for mouse; Immunostimulating evidence
The C571B1 mouse in 2 ages in week is divided into 4 groups, every group of 10 animals.Do not allow mouse drink water from 8 in the morning at 4 in afternoon to next day, 15 milliliters water (the 1st and 3 group) or feed addictive (the 2nd and 4 group) are provided then.Repeat this and handled for 2 weeks.When this processing finishes, under the etherization condition, carry out laparotomy ventrotomy from the 3rd and 4 group mouse.On stomach wall, form 1.5 centimetres otch.Carrying out intestines by dry gauze stimulates.All mouse were all slaughtered after laparotomy ventrotomy in 24 hours; The excision spleen.Separating Morr. cell, and under the condition that has phytolectin (PHA), on the RPMI medium, cultivated 48-72 hour.In the group of handling with feed addictive, the weakening of splenocyte proliferation activity that is caused by laparotomy ventrotomy is restored.
Breed by the splenocyte that PHA induces:
1. control group/water 3060 ± 290
2. control group/water/laparotomy ventrotomy 2120 ± 300
3. test group/BioCocktail 2740 ± 370
4. test group/BioCocktail/ laparotomy ventrotomy 3040 ± 520
Embodiment 23
Food additives (embodiment 4) are used for the people; Immunostimulating evidence
The multidigit patient is implemented uterus, uterine neck, stomach and intestines malignant tumor operation.Gynecologic patient (20 patients) is accepted radiotherapy subsequently.Belly patient (30 patients) accepts chemotherapy.About 1/2 patient uses food additives 2 times every day during treating.In accepting all patients of food additives, significant immunostimulation has appearred, and it shows as:
The gynecological tumor abdominal tumor
Control food additive control food additive
T-lymphocyte 0.03 ± 0.01 0.10 ± 0.03 1.0 ± 0.1 1.6 ± 0.2
(CD3+，HLA-，DR+)
Lymphoblastic transformation
A. spontaneous 530 ± 110 880 ± 80 700 ± 120 1070 ± 100
B.PHA-induces 1600 ± 360 5720 ± 2,100 4400 ± 1,340 13370 ± 3720
Phagocyte quantity (%) 60.7 ± 2.9 76.5 ± 2.2 57.7 ± 1.5 74.2 ± 1.9
Embodiment 24
The bacterium preservation effect of described preparation
Bacillus coli cells is dispersed in the preparation for preparing by the foregoing description 1 method, and at room temperature preserved 6 months.When beginning in described 6 months and end, will be seeded on the nutrient solution agar plate with suitable serial dilution ratio from the sample of described dispersion liquid.Measure the colony forming unit quantity of these two kinds of samples.The concentration of being measured is 1 * 10 in both cases
7, confirmed the effect of said preparation aspect the preservation microorganism.Above-mentioned experiment with pseudomonas putida carries out has simultaneously obtained identical result.
Although described the present invention in conjunction with specific embodiments of the present invention already, clearly, a variety of changes, improvement and variation are apparent to those skilled in the art.Therefore, all such changes, improvement and variation all are considered to belong to the design of appended claims and scope widely.All documents, patent and the patent application mentioned in this manual, all by with identical degree in full form receive to make the list of references of this specification, just as each part document, patent, patent application by special and specialize to receive and do this paper reference.In addition, any list of references of quoting or mention in this application should not be considered to admit that described document can be used as prior art of the present invention.
Claims (92)
1.ATCC preserving number is the coli strain BU-230-98 of 202226 (DSM 12799).
One kind to contain the ATCC preserving number be the preparation that the beneficial natural disposition composition and being used to of the survival coli strain BU-230-98 of 202226 (DSM 12799) is kept the viability of described coli strain.
3. beneficial natural disposition composition as claimed in claim 2 also comprises at least a flavouring agent.
4. beneficial natural disposition composition as claimed in claim 2, wherein said preparation comprise at least a volatility level part (VF) of plant extracts.
5. beneficial natural disposition composition as claimed in claim 4, wherein said volatility level part is by under reduced pressure, under 38 ℃ the bath temperature described plant extracts is carried out steaming and prepares being no more than.
6. beneficial natural disposition composition as claimed in claim 4, wherein said plant extracts is to obtain from the plant organ that is selected from plant leaf blade, stem, root and fruit.
7. beneficial natural disposition composition as claimed in claim 4, wherein said plant extracts obtains from being selected from vegetables and herbal plant.
8. beneficial natural disposition composition as claimed in claim 7, wherein said vegetables are selected from soybean, clover, garlic, beet and wild cabbage.
9. beneficial natural disposition composition as claimed in claim 7, wherein said herbaceous plant is selected from parsley, peppermint and dill.
10. beneficial natural disposition composition as claimed in claim 4 also contains the beehive product.
11. beneficial natural disposition composition as claimed in claim 10, wherein said beehive product is a propolis.
12. beneficial natural disposition composition as claimed in claim 2 is identified to be used for prevention or treatment alimentary infection or imbalance.
13. beneficial natural disposition composition as claimed in claim 2 is identified to be used for keeping or recovering normal gastrointestinal micro-flora.
14. beneficial natural disposition composition as claimed in claim 2 is identified to be used for prevention or treatment diarrhoea.
15. beneficial natural disposition composition as claimed in claim 14 is identified to be used to prevent or treat the alimentary infection that is caused by enteropathogen.
16. beneficial natural disposition composition as claimed in claim 15, wherein said pathogene is Gram-negative bacteria or Gram-positive bacteria.
17. beneficial natural disposition composition as claimed in claim 2 comprises volatility level part of distilled water and beet, dill and parsley.
18. beneficial natural disposition composition as claimed in claim 2 comprises volatility level part of distilled water and beet, dill and grape.
19. beneficial natural disposition composition as claimed in claim 2 is identified to be used for prevention or the salmonella infection of treatment stomach and intestine.
20. beneficial natural disposition composition as claimed in claim 2, by evaluation be used to prevent or treat infectious diarrhea, by antibiotic therapy, radiotherapy or chronic diarrhea or diarrhoea that chemotherapy caused.
21. beneficial natural disposition composition as claimed in claim 2 is identified to be used for prevention or treatment infectious diarrhea, comprises volatility level part of distilled water and beet, dill and parsley.
22. beneficial natural disposition composition as claimed in claim 21, wherein said infectious diarrhea is by clostridium difficile, Salmonella, particularly will Hayes salmonella, and campylobacter, Escherichia coli, proteus, pseudomonas or fusobacterium cause.
23. composition as claimed in claim 2 is identified to be used to make GI physiologically active normalization.
24. contain the preparation of at least a volatility level part (VF) of plant extracts, described volatility level part is by under reduced pressure, under 38 ℃ the bath temperature described plant extracts is carried out steaming and prepares being no more than.
25. preparation as claimed in claim 24, wherein said plant extracts are to obtain from the plant organ that is selected from plant leaf blade, stem, root and fruit.
26. preparation as claimed in claim 24, wherein said plant extracts obtains from being selected from vegetables and herbal plant.
27. preparation as claimed in claim 26, wherein said vegetables are selected from soybean, clover, garlic, beet and wild cabbage.
28. preparation as claimed in claim 26, wherein said herbaceous plant is selected from parsley, peppermint and dill.
29. preparation as claimed in claim 24 also contains the beehive product.
30. preparation as claimed in claim 29, wherein said beehive product is a propolis.
31. animal doctor's food additives that are used to strengthen the weight increase of animal, at least a volatility level part (VF) of containing plant extracts, described volatility level part is by under reduced pressure, under 38 ℃ the bath temperature described plant extracts is carried out steaming and prepares being no more than.
32., comprise volatility level part of distilled water and beet, dill and parsley as animal doctor's feed addictive of claim 31.
33. food additives that are used to strengthen the acceptor weight increase, at least a volatility level part (VF) of containing plant extracts, described volatility level part is by under reduced pressure, under 38 ℃ the bath temperature described plant extracts is carried out steaming and prepares being no more than.
34., comprise volatility level part of distilled water and beet, dill and parsley as the food additives of claim 33.
35. a beneficial natural disposition composition comprises:
(a) the beneficial natural disposition microorganism of at least a survival of effective dose, this microorganism has useful biology or therapeutic activity in intestines and stomach; With
(b) at least a volatility level part (VF) of plant extracts.
36. the beneficial natural disposition composition as claim 35 also comprises at least a flavouring agent.
37. as the beneficial natural disposition composition of claim 35, wherein said volatility level part is by under reduced pressure, under 38 ℃ the bath temperature described plant extracts is carried out steaming and prepares being no more than.
38. as the beneficial natural disposition composition of claim 35, wherein said beneficial natural disposition microorganism is Escherichia coli.
39. as the beneficial natural disposition composition of claim 37, wherein said beneficial natural disposition microorganism is that the ATCC preserving number is 202226 coli strain BU-230-98.
40. beneficial natural disposition composition as claimed in claim 35 is identified to be used for prevention or treatment alimentary infection or imbalance.
41. beneficial natural disposition composition as claimed in claim 35 is identified to be used for keeping or recovering normal gastrointestinal micro-flora.
42. beneficial natural disposition composition as claimed in claim 35 is identified to be used for prevention or treatment diarrhoea.
43. beneficial natural disposition composition as claimed in claim 35 is identified to be used to prevent or treat the alimentary infection that is caused by enteropathogen.
44. beneficial natural disposition composition as claimed in claim 43, wherein said pathogene is Gram-negative bacteria or Gram-positive bacteria.
45. beneficial natural disposition composition as claimed in claim 43 is used for the treatment of indigestion symptom.
46. beneficial natural disposition composition as claimed in claim 43 is used to strengthen the immune response of suffering from the dysimmunity patient.
47. beneficial natural disposition composition as claimed in claim 46, wherein said dysimmunity is caused by the immune response suppression therapy.
48. beneficial natural disposition composition as claimed in claim 35 comprises volatility level part of distilled water and beet and dill.
49. beneficial natural disposition composition as claimed in claim 35 is identified to be used for prevention or the salmonella infection of treatment stomach and intestine.
50. beneficial natural disposition composition as claimed in claim 35, by evaluation be used to prevent or treat infectious diarrhea, by antibiotic therapy, radiotherapy or chronic diarrhea or diarrhoea that chemotherapy caused.
51。Beneficial natural disposition composition as claimed in claim 35 is identified to be used for prevention or treatment infectious diarrhea, comprises volatility level part of distilled water and beet and dill.
52. beneficial natural disposition composition as claimed in claim 51, wherein said infectious diarrhea is by clostridium difficile, Salmonella, particularly will Hayes salmonella, and campylobacter, Escherichia coli, proteus, pseudomonas or fusobacterium cause.
53. beneficial natural disposition composition as claimed in claim 35 is identified to be used to make GI biologic activity to recover normal.
54. beneficial natural disposition composition as claimed in claim 53, comprise distilled water, and wherein said volatility level part is from clover, soybean, beet, dill, peppermint, parsley and wild cabbage, and described beneficial natural disposition microorganism is that the ATCC preserving number is 202226 coli strain Bu-230-98.
55. a composition comprises:
(a) at least a microorganism; With
(b) at least a volatility level part (VF) of plant extracts, described at least a volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
56. beneficial natural disposition composition as claimed in claim 55, wherein said at least a microorganism is selected from Escherichia, Alcaligenes, arthrobacterium, Bifidobacterium, lactobacillus, galactococcus, nitrosomonas or pseudomonad.
57. a method for preparing volatility level part of plant may further comprise the steps:
(a) grind vegetable-derived materials, obtain a kind of plant biological material;
(b) described plant biological material is mixed with water, and stir at ambient temperature;
(c) under reduced pressure, and the mixture that is obtained in to step (b) under being no more than 38 ℃ bath temperature carries out steaming; With
(d) collect volatility level part by described steaming obtained.
58. as the method for claim 57, wherein said plant extracts is available from plant leaf blade, stem, root or fruit.
59. as the method for claim 58, wherein said plant is vegetables or herbaceous plant.
60. as the method for claim 59, wherein said vegetables or herbaceous plant are selected from soybean, clover, garlic, beet, dill, peppermint, parsley and wild cabbage.
61. as the method for claim 57, wherein said decompression is the 5-10 millibar.
62. one kind prepares the method for the treatment of preparation, comprises the step of acceptable additive, carrier or mixing diluents at least a volatility level part (VF) of plant extracts and the physiology or veterinarily.
63. as the method for claim 62, wherein said volatility level part is by under reduced pressure, under 38 ℃ the bath temperature described plant extracts is carried out steaming and prepares being no more than.
64. a growth ATCC preserving number is the method for the coli strain BU-230-98 of 202226 (DSM 12799), this method may further comprise the steps: with the ATCC preserving number is that the starting culture of the coli strain BU-230-98 of 202226 (DSM 12799) is inoculated in the growth medium, and the optical density (OD) that described coli strain is grown under 650nm is 15-30.
65. produce beneficial natural disposition method for compositions for one kind, this method may further comprise the steps: with the ATCC preserving number is that the survival coli strain BU-230-98 of 202226 (DSM 12799) is suspended in a kind of preparation, so that keep the viability of described coli strain.
66. as the method for claim 65, wherein said preparation comprises at least a volatility level part (VF) of plant extracts.
67. as the method for claim 66, wherein said volatility level part is by under reduced pressure, under 38 ℃ the bath temperature described plant extracts is carried out steaming and prepares being no more than.
68. as the method for claim 66, wherein said plant extracts is available from the plant organ that is selected from plant leaf blade, stem, root and fruit.
69. as the method for claim 66, wherein said plant extracts obtains from being selected from vegetables and herbal plant.
70. as the method for claim 69, wherein said vegetables or herbaceous plant are selected from soybean, clover, garlic, beet, dill, peppermint, parsley and wild cabbage.
71., also comprise and add the beehive product as the method for claim 66.
72. as the method for claim 71, wherein said beehive product is a propolis.
73. as the method for claim 65, wherein said at least a volatility level part is from clover, soybean, beet, dill and peppermint.
74. one kind prepares beneficial natural disposition method for compositions, this method may further comprise the steps: with the beneficial natural disposition microbial suspension of at least a survival effective dose, that in intestines and stomach, have useful biology or therapeutic activity in the preparation of at least a volatility level part (VF) of containing plant extracts.
75. as the method for claim 74, wherein said volatility level part is by under reduced pressure, steaming prepares by described plant extracts is carried out being no more than under 38 ℃ the bath temperature.
76. as the method for claim 74, wherein said beneficial natural disposition microorganism is Escherichia coli.
77. as the method for claim 75, wherein said beneficial natural disposition microorganism is that the ATCC preserving number is 202226 coli strain BU-230-98.
78. as the method for claim 74, wherein said at least a volatility level part comprises volatility level part of clover, soybean, beet, dill, peppermint, parsley and wild cabbage.
79. one kind prepares the method for compositions that contains the microorganism that survives, this method may further comprise the steps: with at least a microbial suspension in the preparation of at least a volatility level part (VF) of containing plant extracts, described at least a volatility level part is through selecting, so as can be at room temperature will described at least a microorganism viability maintenance at least 2 months.
80. as the method for claim 79, wherein said at least a microorganism is selected from Escherichia, Alcaligenes, arthrobacterium, Bifidobacterium, lactobacillus, galactococcus, nitrosomonas or pseudomonad.
81. microorganism distributor, this distributor comprises a holder and a meter that is connected with this holder, this holder is equipped with at least a microorganism in the preparation that is present at least a volatility level part (VF) of containing plant extracts, described at least a volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
82. as the distributor of claim 81, wherein said distributor is airborne.
83. as the distributor of claim 81, wherein said distributor is a hand-held.
84. as the distributor of claim 81, wherein said distributor is designed to distribute automatically predetermined volume.
85. as the distributor of claim 81, wherein said distributor is designed to regularly distribute automatically predetermined volume.
86. method of biology being carried out BIOLOGICAL CONTROL, this method may further comprise the steps: can be sprayed at epidemic regions to the survival microorganism that described biology carries out BIOLOGICAL CONTROL with at least a, described at least a survival microorganism is present in the preparation of at least a volatility level part (VF) of containing plant extracts, described at least a volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
87. remove the method that greasy dirt dyes for one kind, this method may further comprise the steps: can be sprayed at contaminated zone by the oily microorganism of degraded with at least a, described at least a survival microorganism is present in the preparation of at least a volatility level part (VF) of containing plant extracts, described at least a volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
88. method of recovering the organic smoke degradation micropopulation in the biofilter, this method may further comprise the steps: at least a survival microorganism that can degrade organic smog is distributed on the described filter, described at least a survival microorganism is present in the preparation of at least a volatility level part (VF) of containing plant extracts, described at least a volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
89. one kind has been added the ATCC preserving number is the food of the survival coli strain BU-230-98 of 202226 (DSM 12799).
90., contain at least a milk-derived material as the food of claim 89.
91., be selected from cheese and sour milk as the food of claim 90.
92. method for preparing the leavening that is used for fermentation process, this method may further comprise the steps: the leavening microorganism of growth sufficient amount, and with described leavening microbial suspension in the preparation of at least a volatility level part (VF) of containing plant extracts, described at least a volatility level part is through selecting, so that viability maintenance that at room temperature will described at least a microorganism at least 2 months.
Applications Claiming Priority (2)
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|---|---|---|---|
| US09/725,846 | 2000-11-30 | ||
| US09/725,846 US6500423B2 (en) | 1999-06-03 | 2000-11-30 | Bacterial strain, processed plant extracts, compositions containing same, processes for their preparation and their therapeutic and industrial applications |
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| JP (1) | JP4410992B2 (en) |
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| US3908028A (en) * | 1973-04-19 | 1975-09-23 | Procter & Gamble | Soft drink composition containing paramethoxycinnamaldehyde as a flavoring agent and sweetner |
| GB1501098A (en) * | 1974-06-11 | 1978-02-15 | Brewing Patents Ltd | Preparation of hop oil |
| US4986985A (en) * | 1987-11-02 | 1991-01-22 | Bar Ilan University | Method of treating skin virus infections |
| CH675685A5 (en) * | 1988-06-24 | 1990-10-31 | Flachsmann Ag Emil | |
| US5443826A (en) * | 1988-08-02 | 1995-08-22 | Borody; Thomas J. | Treatment of gastro-intestinal disorders with a fecal composition or a composition of bacteroides and E. Coli |
| US5082489A (en) * | 1990-02-06 | 1992-01-21 | The Royal Institution For The Advancement Of Learning (Mcgill University) | Composition for biocontrol of wild buckwheat |
| US5478557A (en) * | 1992-07-29 | 1995-12-26 | The United States Of America, As Represented By The Secretary Of Agriculture | Probiotic for control of salmonella |
| US5486367A (en) * | 1992-10-13 | 1996-01-23 | Kansas State University Research Foundation | Enzymatic method for accelerating fermentation of comestible products |
| US5376374A (en) * | 1993-05-24 | 1994-12-27 | Zelaya; Luz M. | Oral rinse composition |
| US5733774A (en) * | 1995-02-02 | 1998-03-31 | Ecoscience Corporation | Method and composition for producing stable bacteria and bacterial formulations |
| AU714310B2 (en) * | 1995-05-12 | 1999-12-23 | Japan Pharmaceutical Development Co., Ltd | Plant extract |
| BR9608841A (en) * | 1995-05-26 | 1999-12-07 | Ropapharm B V | Pharmaceutical composition, use of composition, and process to recover oil from plants |
| US5679364A (en) * | 1995-06-07 | 1997-10-21 | Lee County Mosquito Control District | Compositions and methods for reducing the amount of contaminants in aquatic and terrestrial environments |
| GB2303799B (en) * | 1995-07-29 | 1999-03-10 | Siemens Plc | Improvements in or relating to aqueous sample testing apparatus |
| US5888395A (en) * | 1996-08-30 | 1999-03-30 | The Regents Of The University Of California | Method for enhanced longevity of in situ microbial filter used for bioremediation |
| US5773075A (en) * | 1996-12-13 | 1998-06-30 | Kalamazoo Holdings, Inc. | High temperature countercurrent solvent extraction of Capsicum solids |
| US5965128A (en) * | 1997-08-13 | 1999-10-12 | University Of Georgia Research Foundation Inc. | Control of enterohemorrhagic E. coli 0157:H7 in cattle by probiotic bacteria and specific strains of E. coli |
| IL130303A0 (en) * | 1999-06-03 | 2000-06-01 | M G Novobiotech Ltd | A bacterial strain processed plant extracts and probiotic compositions for human and veterinary use |
-
2001
- 2001-11-27 WO PCT/IL2001/001088 patent/WO2002043649A2/en active Application Filing
- 2001-11-27 CA CA002430292A patent/CA2430292A1/en not_active Abandoned
- 2001-11-27 BR BR0116178-4A patent/BR0116178A/en not_active IP Right Cessation
- 2001-11-27 NZ NZ526748A patent/NZ526748A/en unknown
- 2001-11-27 RU RU2003119151/13A patent/RU2281326C2/en not_active IP Right Cessation
- 2001-11-27 AU AU2100402A patent/AU2100402A/en active Pending
- 2001-11-27 AU AU2002221004A patent/AU2002221004B2/en not_active Ceased
- 2001-11-27 MX MXPA03004851A patent/MXPA03004851A/en not_active Application Discontinuation
- 2001-11-27 JP JP2002545628A patent/JP4410992B2/en not_active Expired - Fee Related
- 2001-11-27 CN CNB018214851A patent/CN1294253C/en not_active Expired - Fee Related
- 2001-11-27 IL IL15621301A patent/IL156213A0/en unknown
- 2001-11-27 PL PL01366363A patent/PL366363A1/en unknown
- 2001-11-27 KR KR10-2003-7007337A patent/KR20030082549A/en not_active Application Discontinuation
- 2001-11-27 EP EP01998302A patent/EP1343370A4/en not_active Ceased
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2003
- 2003-06-19 ZA ZA200304747A patent/ZA200304747B/en unknown
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109561710A (en) * | 2016-06-14 | 2019-04-02 | 普瑞特克微生物有限责任公司 | Animal feed pellets comprising feed addictive, its preparation and application |
| CN107319103A (en) * | 2017-08-01 | 2017-11-07 | 河南牧业经济学院 | Promote feed addictive of growth of animals or poultry and intestinal growth and its preparation method and application |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2002043649A3 (en) | 2002-08-29 |
| CA2430292A1 (en) | 2002-06-06 |
| MXPA03004851A (en) | 2004-05-04 |
| BR0116178A (en) | 2003-10-14 |
| ZA200304747B (en) | 2004-09-03 |
| WO2002043649A2 (en) | 2002-06-06 |
| IL156213A0 (en) | 2003-12-23 |
| RU2281326C2 (en) | 2006-08-10 |
| AU2002221004B2 (en) | 2007-01-25 |
| WO2002043649A8 (en) | 2004-04-29 |
| AU2100402A (en) | 2002-06-11 |
| JP2004534512A (en) | 2004-11-18 |
| EP1343370A2 (en) | 2003-09-17 |
| NZ526748A (en) | 2006-03-31 |
| EP1343370A4 (en) | 2004-02-11 |
| CN1294253C (en) | 2007-01-10 |
| PL366363A1 (en) | 2005-01-24 |
| JP4410992B2 (en) | 2010-02-10 |
| KR20030082549A (en) | 2003-10-22 |
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