CN1294253C - 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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- CN1294253C CN1294253C CNB018214851A CN01821485A CN1294253C CN 1294253 C CN1294253 C CN 1294253C CN B018214851 A CNB018214851 A CN B018214851A CN 01821485 A CN01821485 A CN 01821485A CN 1294253 C CN1294253 C CN 1294253C
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Classifications
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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-virulent benefit natural disposition microorganism and benefit life/therepic use 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 therepic use; (iii) a kind of method by the described preparation of described plant origin material produce; The beneficial natural disposition composition that (iv) contains non-virulent benefit natural disposition microorganism of the present invention and/or other beneficial natural disposition microorganisms and preparation of the present invention, and benefit life/therepic use; (v) a kind of composition that is used for industrial application contains preparation of the present invention and the microorganism with industrial applicability; (vi) use the commercial run and the device of a kind of composition in above-mentioned back.
Under normal operation, stomach and intestine (GI) road micro-flora plays an important role to the health and the situation of individuality.As everyone knows, micro-flora is complexity and the multifarious colony of microorganism, and this colony may have useful and deleterious effect 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 gi tract 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 micro-flora 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 pathogenic agent.A kind of phenomenon in back was used ' bacterium antagonism ' already, and ' bacterium interference ', ' barrier effect ', ' building group resistance ', nouns such as ' competition exclusions ' carried out statement.
The pipe intestinal protection micro-flora 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 impact, and probiotic bacterium is recovering gastrointestinal micro-flora and making individual host recover to have potential value aspect normal.
Probiotic bacterium 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 probiotic bacterium can be by the direct antagonistic action mediation to the particular type biology, by causing described biomass to reduce to its metabolic influence or by immune stimulatory power.Probiotic bacterium can suppress the survival quantity of unwanted biology by producing antimicrobial compounds by competition nutrition or competition attachment site.In addition, probiotic bacterium can or reduce the enzyme activity change microbial metabolism by raising, or by improving anti-body contg or improving the macrophage activity stimulating immune system.
WO95/16461 has disclosed the beneficial natural disposition composition of a kind of anaerobic bacterium, it can effectively control or suppress Salmonellas be in the poultry the intravital group of building.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 restraining effect of pathogenic agent is needed the combined action of a large amount of bacterial isolateses.
WO97/35596 has disclosed to use by mixing and has contained the powder of lactobacillus reuteri, Lactobacterium acidophilum and bifidobacteria 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 circumstances.In addition, the biology of surviving is not just to have a metabolic activity at once, and can not survive under extreme, the acidic conditions 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 pre-biotic microorganism associating, strengthen lactic acid-producing 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 substratum.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 commercial run.Described method can be divided into continuation method and batch methods.In continuation method, as continuous aerobic or anaerobically fermenting production line, be used to produce multiple leavened prod, for example, comprise ethanol and methyl alcohol, fermentable starting material are transported on the described production line continuously, and from described production line, collect leavened prod continuously, so that on described production line, be maintained at microbial population described in the whole process of production in check mode.In batch methods, for example, but be not limited in batches aerobic or anaerobically fermenting, in leakage of oil, to the degraded of oil etc., fermentable starting material are mixed with microbial population, 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 pseudomonas or Alcaligenes microorganism begins described process, for example, finishes the biodegradable time of described oil so that reduce.At the used enrichment strainer of microorganism when degrading organic smog or volatile matter, consequently a kind of successive processes wherein, becomes harmless compound with the contaminative degradation.But, the without interruption of organic smog or volatile matter depended in the survival that is present in the microorganism on the described strainer.But, under some occasion, the supply of organic smog or volatile matter is discontinuous, has consequently lost described microorganism.In this case, must recover the survival microorganism of significant quantity in the described strainer at short notice, so that can horse back restore functionality strainer.
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 industrial 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-virulent benefit natural disposition microorganism just can recover the normal gastrointestinal tract flora of people and multiple Mammals 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 milk extracts 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 gastrointestinal tract disease.Be surprised to find that also that in addition described preparation itself can be effectively as the weight increase toughener of Mammals 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 milk extract, it is by under reduced pressure, plant (for example, beet, dill, parsley or natsudaidai) extract is carried out vapor distillation 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 gastro-intestinal infection or imbalance; keep or recover normal gastrointestinal micro-flora; prevention or treatment diarrhoea; prevention or treatment are by enteropathogen; the gastro-intestinal infection that causes as gram negative bacterium or gram positive bacterium; prevention or the Salmonella infection of treatment stomach and intestine; prevention or treatment are by for example; clostridium difficile; salmonella; will Hayes Salmonellas particularly, campylobacter, intestinal bacteria; proteus; the infectious diarrhea that Rhodopseudomonas or fusobacterium cause is by antibiotic therapy; radiotherapy or chronic diarrhoea or diarrhoea that chemotherapy caused, and/or be used to make GI physiologically active normalizing.
According to a further aspect in the invention, the preparation of at least a volatility level part (VF) of containing plant milk extract is provided, and described volatility level part is to prepare by described plant milk extract is carried out vapor distillation by being no more than under reduced pressure under 38 ℃ the bath temperature.Described plant milk extract preferably can obtain from the plant organ that is selected from plant leaf, 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 foodstuff additive, 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 vapor distillation; (d) collect volatility level part by described vapor distillation obtained.Described decompression is preferably the 5-10 millibar.Can be with said preparation as foodstuff additive, fodder additives, 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 milk extract.Described volatility level part prepares by described plant milk extract is carried out vapor distillation 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) significant quantity, and this microorganism has useful biology or therapeutic activity in gi tract; (b) at least a volatility level part (VF) of plant milk extract, this volatility level part are preferably pressed the disclosed method preparation of this paper.Described beneficial natural disposition microorganism can be intestinal bacteria, 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 immunne response, for example, outside the disease that is caused by the immunne response suppression therapy or as an alternative scheme also can be used it for treatment or prevent above-mentioned any imbalance/pathology/disease/syndromes.
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, Arthrobacter, bifidus bacillus, Bacterium lacticum, galactococcus, Nitrosomonas and pseudomonas; (b) at least a volatility level part (VF) of plant milk extract, 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 milk extract, described volatility level part is preferably by under reduced pressure, described plant milk extract is carried out the vapor distillation preparation being no more than under 38 ℃ the bath temperature, for example, described plant milk extract 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 significant quantity, that in gi tract, have useful biology or therapeutic activity in the preparation of at least a volatility level part (VF) of containing plant milk extract.
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 milk extract, 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, Arthrobacter, bifidus bacillus, Bacterium lacticum, galactococcus, Nitrosomonas or pseudomonas.
According to a further aspect in the invention, a kind of microorganism divider is provided, this divider comprises a holder and a gauger 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 milk extract, 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 milk extract, 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 removing oil pollution 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 milk extract, 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 microbial population 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 strainer, described at least a survival microorganism is present in the preparation of at least a volatility level part (VF) of containing plant milk extract, 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 starter of fermenting process, this method may further comprise the steps: the starter microorganism of growth sufficient amount, and with described starter microbial suspension in the preparation of at least a volatility level part (VF) of containing plant milk extract, 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 synoptic diagram of divider of the present invention.
Detailed Description Of The Invention
The present invention is (i) a kind of non-virulent benefit natural disposition microorganism and benefit life/therepic use 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 therepic use; (iii) a kind of method by the described preparation of described plant origin material produce; The beneficial natural disposition composition that (iv) contains non-virulent benefit natural disposition microorganism of the present invention and/or other beneficial natural disposition microorganisms and preparation of the present invention, and benefit life/therepic use; (v) a kind of composition that is used for industrial use contains preparation of the present invention and the microorganism with industrial applicability; (vi) use the commercial run and the device of a kind of composition in above-mentioned back.
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 sheets or the explanation of embodiment in the details that provided.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 used 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 gi tract, 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 to represent to have the isolated bacterial of the characteristic that suppresses at least a pathogenic growth.Employed in this article inhibition test is the in vitro tests of carrying out on solid medium, 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 pathogenic agent 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 liquor has caused the transparent agar of a circle or weakened shows near the pathogenic agent 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 fodder additives, 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) so that 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 milk extract, described volatility level part is by preparing being no more than usually under 38 ℃ the bath temperature described plant milk extract distilled under reduced pressure.
The vegetable material that is used to obtain described volatility level part can obtain from any suitable plant part, 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 as parsley, peppermint and dill.Preparation of the present invention can also optionally contain the agricultural-food of suitable quantity, as volatility level part of honey, propolis or other beehive products, described level part can be with obtaining the identical method preparation of volatility level part with disclosed being used for of this paper from plant milk extract.
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 weight increase, and it has constituted 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 fodder additives 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 fodder additives comprise volatility level part of each 1 part Radix Dauci Sativae, beet, dill and the natsudaidai that are contained in distilled water and the per 100 parts of water.As another kind of alternative, preparation of the present invention or foodstuff additive comprise volatility level part of each 1 part of beet, dill and parsley of being 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 in gi tract, have such as people and/or animal the treatment or beneficial organism learn active beneficial natural disposition biology, so that a kind of beneficial natural disposition composition is provided, or have the biology of industrial application value, so that 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 promptly they are liquid preparations.Under the biologic activity condition, described preparation also can be used as the supportive substratum 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 effect of most of enteropathogen all here takes place, and causes the appearance of bad gastro-intestinal syndrome.Beneficial natural disposition composition of the present invention also can be used as weight increase preparation or foods/feeds additive.Similarly, can be used to have efficient recovery to have the microbial population of industrial applicability industry group compound of the present invention.
Therefore, should be noted that, for beneficial natural disposition purposes, except other beneficial natural disposition 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 phylogeny 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 gastro-intestinal infection.Term ' gastro-intestinal 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 gastro-intestinal infection that caused by yeast, virus and protozoon.
Concerning this paper, term ' treatment significant quantity ' or ' significant quantity ' 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 normalizing of gastrointestinal function.
In a kind of specific embodiments, be used to prevent the beneficial natural disposition composition that gastro-intestinal infection takes place to comprise 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 of treatment significant quantity, be 202226 intestinal bacteria (identical) or any other beneficial natural disposition bacterium preparation as the ATCC preserving number with DSM 12799.
An object lesson of gastro-intestinal infection is the infection that is caused by salmonella, and the present invention is particularly conducive to prevention or treats the gastro-intestinal infection that is caused by salmonella.
In addition, foodstuff additive of the present invention or preparation can be separately or with significant quantity be that 202226 intestinal bacteria (identical with DSM 12799) are united and are used for the treatment of or preventing infection diarrhoea, chronic diarrhoea or the diarrhoea that caused by microbiotic 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 significant quantity.
Described infectious diarrhea can cause by multiple factor, for example, and by being selected from clostridium difficile, salmonella, particularly will Hayes Salmonellas, campylobacter, intestinal bacteria, proteus, Rhodopseudomonas, 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 intravital gastrointestinal micro-flora of object of this treatment of efficient recovery needs, this treatment can cause the normalizing 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 significant quantity 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 immunity system of the object of suffering from the immunity system obstacle, described obstacle may be the dysimmunity that is caused by immunosuppressant 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 damping fluid by volatility level part that vapor distillation 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-food 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 bacteria of the present invention, and it is 202226 intestinal bacteria (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 milk extract is used to prepare the purposes of foods/feeds additive, wherein, described VF is by being no more than the described plant milk extract of distillation under 38 ℃ the bath temperature or preparing such as the agricultural-food 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 milk extract.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, Arthrobacter, bifidus bacillus, Bacterium lacticum, galactococcus, Nitrosomonas and pseudomonas.
As shown in Figure 1, another aspect of the present invention provides a kind of microorganism divider 10.Divider 10 comprises a holder 12 and a gauger 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 milk extract.Described preparation is through selecting, so that at room temperature the viability of described microorganism was kept 2 months at least.Container 12 can be a refrigerating, for example freezes to about 2-20 ℃, preferably freezes to about 4 ℃.Divider 10 can be airborne divider, so that can distribute insect control biotechnological formulation.Described divider can be a hand-held.Under any circumstance, described dispenser designs can be become the solution that distributes pre-determined volume automatically, for example regularly distributed.For this reason, divider 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 milk extract, 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 of removing oil pollution, this method is by will being sprayed at contaminated zone by the oily survival microorganism of degraded such as at least a of pseudomonas.Described survival microorganism is present in the preparation of at least a volatility level part (VF) of containing plant milk extract, 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 microbial population in the biofilter is provided, has comprised at least a survival microorganism such as the organic oil of degrading of pseudomonas, Nitrosomonas or Arthrobacter is assigned on the described strainer.Described survival microorganism is present in the preparation of at least a volatility level part (VF) of containing plant milk extract, 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 starter of fermenting process is provided, the starter microorganism that comprises the sufficient amount of growing, as galactococcus, Escherichia, bifidus bacillus, Bacterium lacticum, galactococcus or pseudomonas, and with described starter microbial suspension in the preparation of at least a volatility level part (VF) of containing plant milk extract, described volatility level part is through selecting, so that the zone of at room temperature survival of described at least a microorganism being tried hard to keep and polluting.Described survival microorganism is present in the preparation of at least a volatility level part (VF) of containing plant milk extract, 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 microbial population in the biofilter is provided, has comprised at least a survival microorganism such as the organic oil of degrading of pseudomonas, Nitrosomonas or Arthrobacter is assigned on the described strainer.Described survival microorganism is present in the preparation of at least a volatility level part (VF) of containing plant milk extract, 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 starter of fermenting process is provided, the starter microorganism that comprises the sufficient amount of growing, as galactococcus, Escherichia, bifidus bacillus, Bacterium lacticum or pseudomonas, and with described starter microbial suspension in the preparation of at least a volatility level part (VF) of containing plant milk extract, 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 sheets.
Embodiment 1
Prepared plant extracts or from " volatility level part " of the extract of agricultural-food
Fully clean with the fresh vegetables that tap 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 rotatory evaporator, and under the following conditions in decompression (5-10 millibar) evaporation down: bath temperature is no more than 38 ℃, and condenser inlet is described in more detail.The biological intestinal bacteria 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 substratum: (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 glucose concn 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 fermentor 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 Mammals 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 foodstuff additive of human gi-tract function normalizing
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-10ml/L.Carry out balance with distilled water.With 10
7The cell concn of cells/ml is suspended in this mixture with intestinal bacteria (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 Mammals and bird gastrointestinal tract 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 concn of cells/ml is suspended in this mixture with intestinal bacteria (ATCC 202226) cells (embodiment 2).Can optionally add NaCl (for example 4-10g/L) in order to improve mouthfeel.
Embodiment 6
Intestinal bacteria (ATCC 202226) are right
The antagonistic activity of Salmonella typhimurium (ATCC 14028)
The culture dish of the Brilliant Green agar (the selective growth substratum of salmonella) of modification is housed with the Salmonella typhimurium inoculation.On agar, form the hole of 9 mm dias.The described foodstuff additive (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 foodstuff additive, but will filter the liquid deposition that obtains in described hole by microorganism filter membrane (0.45 micron in aperture).
Observed the inhibitory area (10-17 millimeter) that lacks the Salmonella typhimurium bacterium colony around in each hole of containing described foodstuff additive.The inhibitory area does not appear around in the hole of containing the filtrate that does not contain described beneficial natural disposition biology.
Embodiment 7
Intestinal bacteria ATCC202226 and
Parental generation intestinal bacteria M-17 is to the antagonistic activity of Shigellae
Shigella flexneri, Shigella sonnei, intestinal bacteria (ATCC 202226) and M-17 were grown 18-20 hour down at 37 ℃ on nutrient agar medium respectively.All cultures are all gathered in the crops in the salt solution, and are diluted to the optical density(OD) of 10Klett unit.Shigellae is diluted 1 milliliter of the sample aliquot of culture itself or be inoculated in the ventilation cup-shaped test tube that nutritive medium (5 milliliters) is housed with the dilution cultures of intestinal bacteria (ATCC 202226) combination (1 milliliter).Under 37 ℃, hatched described test tube 24 hours.By with described culture bed board to nutrient agar medium, determine colony forming unit (CFU) quantity of described pathogenic agent and intestinal bacteria (ATCC 202226).Two kinds of Shigellae 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 Shigellae | |
| Shigella flexneri | Shigella sonnei | |
| The pure Shigellae culture of intestinal bacteria ATCC 202226 intestinal bacteria M-17 | ＜5×10 4 1×10 6 6×10 6 | ＜5×10 4 2×10 6 2×10 6 |
Embodiment 8
With foodstuff additive (embodiment 4) and Colicine
(drying agent of intestinal bacteria M-17) is used for gastroenteritis patient in hospital
One group of 60 patient that gastroenteritis takes place after being in hospital are divided into three groups at random, make each group have similar distribution aspect sex, age and the gastroenteritis severity of symptom.All patients accept normal supportive treatment, comprise rehydrated, VITAMIN etc.The severity of symptom does not need to use antibiotic therapy.20 patient every days of first group 3 times were accepted 10 milliliters foodstuff additive (embodiment 5) 30 minutes ante cibum and treat 7 day time altogether.Scheme (2 30 minutes ante cibum of the every day use) Colicine (Colibacterinum 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 gastroenteritis symptom normalizing of all groups, and shown in the table 2 below.
Table 2
| Symptom | The beginning of symptom normalizing, fate | ||
| Foodstuff additive | 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 intestinal bacteria 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
Table 1
| Benefit natural disposition biology | The growth of Shigellae | |
| Shigella flexneri | Shigella sonnei | |
| The pure Shigellae culture of intestinal bacteria ATCC 202226 intestinal bacteria M-17 | ＜5×10 4 1×10 6 6×10 6 | ＜5×10 4 2×10 6 2×10 6 |
Embodiment 8
With foodstuff additive (embodiment 4) and Colicine
(drying agent of intestinal bacteria M-17) is used for gastroenteritis patient in hospital
One group of 60 patient that gastroenteritis takes place after being in hospital are divided into three groups at random, make each group have similar distribution aspect sex, age and the gastroenteritis severity of symptom.All patients accept normal supportive treatment, comprise rehydrated, VITAMIN etc.The severity of symptom does not need to use antibiotic therapy.20 patient every days of first group 3 times were accepted 10 milliliters foodstuff additive (embodiment 4) 30 minutes ante cibum and treat 7 day time altogether.Scheme (2 30 minutes ante cibum of the every day use) Colicine (Colibacterinum 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 gastroenteritis symptom normalizing of all groups, and shown in the table 2 below.
Table 2
| Symptom | The beginning of symptom normalizing, fate | ||
| Foodstuff additive | 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 intestinal bacteria M-17 of the work of producing, and recommended degree of being used for the treatment of rushes down, and [in these 2 groups of the Vidal Handbook:Pharmaceutical preparations in, all patients' symptom of diarrhea all disappeared in 2-3 days.After using described additive, in 84.5% patient, observed the complete normalizing of gastrointestinal microorganisms fauna.This is the remarkable increase by lactobacillus and genus bifidobacterium, the minimizing of intestinal bacteria sum, and haemolysis intestinal bacteria and prove such as the completely dissolve of Staphylococcus, proteus vulgaris or even oidiomycetic other pathogenic agent.In remaining patient of 15.5%, observed remarkable improvement.
Embodiment 11
The patient who foodstuff additive (embodiment 5) is used to suffer from radiotherapy later stage gastroenteritis
Use described foodstuff additive (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 gastroenteritis occurred after the combination therapy of accepting radiotherapy or radiation and chemotherapy.
Before using described foodstuff additive, 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.Hemanalysis shows that physiochemical indice is significantly improved.
In the control group of similar scale, continue to exist described symptom.
Embodiment 12
Foodstuff additive (embodiment 4) are used for AIDS patient
The patient who suffers from AIDS chronic diarrhoea occurs through regular meeting.Allow one group of such patient use 10 milliliters of foodstuff additive (embodiment 5), every day 3 times, use 30 minutes ante cibum, used altogether 20 days, and control group is not accepted additive.Treatment result is shown in the following table 4.
Every day 3 times is used described foodstuff additive (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 normalizing of gastrointestinal microorganisms fauna.This is the remarkable increase by lactobacillus and genus bifidobacterium, the minimizing of intestinal bacteria sum, and haemolysis intestinal bacteria and prove such as the completely dissolve of Staphylococcus, proteus vulgaris or even oidiomycetic other pathogenic agent.In remaining patient of 15.5%, observed remarkable improvement.
Embodiment 11
The patient who foodstuff additive (embodiment 4) is used to suffer from radiotherapy later stage gastroenteritis
Use described foodstuff additive (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 gastroenteritis occurred after the combination therapy of accepting radiotherapy or radiation and chemotherapy.
Before using described foodstuff additive, 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.Hemanalysis shows that physiochemical indice is significantly improved.
In the control group of similar scale, continue to exist described symptom.
Embodiment 12
Foodstuff additive (embodiment 4) are used for AIDS patient
The patient who suffers from AIDS chronic diarrhoea occurs through regular meeting.Allow one group of such patient use 10 milliliters of foodstuff additive (embodiment 4), every day 3 times, use 30 minutes ante cibum, used altogether 20 days, and control group is not accepted additive.Treatment result is shown in the following table 4.
Table 4
| Parameter | Foodstuff additive | 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 normalizing the average time opening | 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 foodstuff additive shows the microbiotic normalizing of intestines: the intestinal bacteria sum reduces, and the haemolysis intestinal bacteria disappear, and the quantity of lactobacillus and genus bifidobacterium increases, and candidiasis 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 fodder additives, the weight average of every piggy has increased by 1.0 kilograms than control group when wean.
Embodiment 14
Described fodder additives (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 fodder additivess.Compare mortality ratio 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 fodder additives has increased by 0.39 kilogram than control group.When more described fodder additives and antibiotic therapy in brood son, the weight increase of the little piglet of the described fodder additives of discovery acquisition is higher 2.4 kilograms than control group.
Embodiment 15
Fodder additives (embodiment 5) is applied to show the little piglet of diarrhoea
Use 5 milliliters fodder additives for performance hundreds of piggy son every piggys every day that out-degree is rushed down.The control group antibiotic treatment of identical scale: advocin, gentamicin, amoxycilline Trihydrate bp.The symptom of diarrhea of accepting the group of described fodder additives disappeared in 1-2 days.Death occurs, and the piggy son grows normal.Microbiotic 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 fodder additives 3 days.All survivals except that 2 in these little piglet.
Embodiment 16
Fodder additives (embodiment 5) is used for healthy calf
Use 5 milliliters of fodder additivess for the healthy calf of birth just by emulsion, use 7 day time.Calf 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 sickness rate of suffering from diarrhoea in this farm is 20-30%.
Embodiment 17
Fodder additives (embodiment 5) is used to the calf that occurs suffering from diarrhoea
Use 10 milliliters of fodder additivess 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.
Embodiment 18
Fodder additives (embodiment 5) is used for healthy sheep lamb and goat kids
Use 3 milliliters fodder additives for the sheep lamb and the goat kids 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 fodder additivess (5 milliliters).For sheep lamb and goat kids, the sickness rate of suffering from diarrhoea in described farm is generally 50%.
Embodiment 19
Sheep lamb and goat kids that fodder additives (embodiment 5) is used to show diarrhoea
The described fodder additives of test in the herd that suffers from the pathogenic colon bacillus infection.In the previous year of test, die from diarrhoea for nearly 90 in 120 sheep lambs and the goat kids.Breed by PHA inductive splenocyte with anti-:
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
Foodstuff additive (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 foodstuff additive 2 times every day during treating.In accepting all patients of foodstuff additive, significant immunostimulation has appearred, and it shows as:
| Gynecological tumor | Abdominal tumor | |||
| Contrast | Foodstuff additive | Contrast | Foodstuff additive | |
| T-lymphocyte (CD3+, HLA-, DR+) lymphoblastic transformation | 0.03±0.01 | 0.10±0.03 | 1.0±0.1 | 1.6±0.2 |
| A. spontaneous b.PHA-induces phagocytic cell quantity (%) | 530±110 1600±360 60.7±2.9 | 880±80 5720±2100 76.5±2.2 | 700±120 4400±1340 57.7±1.5 | 1070±100 13370±3720 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 nutritive medium 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 reference of this specification sheets, just as each part document, patent, patent application by special and specialize to receive and do this paper reference.In addition, any reference 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 (32)
1. beneficial natural disposition composition, contain ATCC preserving number as activeconstituents and be the survival coli strain BU-230-98 of 202226 (DSM 12799) and be used to keep the preparation of the viability of described coli strain, wherein said preparation comprises at least a volatility level part (VF) of plant milk extract, wherein said volatility level part is by under reduced pressure, under 38 ℃ the bath temperature described plant milk extract is carried out vapor distillation and prepare being no more than, and wherein said at least a volatility level part is through selecting, so that at room temperature will reach 10
7The viability of the described at least a microorganism of cells/ml kept 2 months at least.
2. beneficial natural disposition composition as claimed in claim 1 also comprises at least a flavouring agent.
3. beneficial natural disposition composition as claimed in claim 1, wherein said plant milk extract is to obtain from the plant organ that is selected from blade, stem, root and fruit.
4. beneficial natural disposition composition as claimed in claim 1, wherein said plant milk extract obtains from being selected from vegetables and herbal plant.
5. beneficial natural disposition composition as claimed in claim 4, wherein said vegetables are selected from soybean, clover, garlic, beet and wild cabbage.
6. beneficial natural disposition composition as claimed in claim 4, wherein said herbaceous plant is selected from parsley, peppermint and dill.
7. beneficial natural disposition composition as claimed in claim 1 also contains the beehive product.
8. beneficial natural disposition composition as claimed in claim 7, wherein said beehive product is a propolis.
9. beneficial natural disposition composition as claimed in claim 1 is identified to be used for prevention or treatment gastro-intestinal infection or imbalance.
10. beneficial natural disposition composition as claimed in claim 1 is identified to be used for keeping or recovering normal gastrointestinal micro-flora.
11. beneficial natural disposition composition as claimed in claim 1 is identified to be used for prevention or treatment diarrhoea.
12. beneficial natural disposition composition as claimed in claim 9 is identified to be used to prevent or treat the gastro-intestinal infection that is caused by enteropathogen.
13. beneficial natural disposition composition as claimed in claim 12, wherein said pathogenic agent is gram negative bacterium or gram positive bacterium.
14. beneficial natural disposition composition as claimed in claim 1 comprises volatility level part of distilled water and beet, dill and parsley.
15. beneficial natural disposition composition as claimed in claim 1 comprises volatility level part of distilled water and beet, dill and natsudaidai.
16. beneficial natural disposition composition as claimed in claim 1 is identified to be used for prevention or the Salmonella infection of treatment stomach and intestine.
17. beneficial natural disposition composition as claimed in claim 1, by evaluation be used to prevent or treat infectious diarrhea, by antibiotic therapy, radiotherapy or chronic diarrhoea or diarrhoea that chemotherapy caused.
18. beneficial natural disposition composition as claimed in claim 1 is identified to be used for the treatment of infectious diarrhea, comprises volatility level part of distilled water and beet, dill and parsley.
19. beneficial natural disposition composition as claimed in claim 18, wherein said infectious diarrhea is by clostridium difficile, salmonella, particularly will Hayes Salmonellas, and campylobacter, intestinal bacteria, proteus, Rhodopseudomonas or fusobacterium cause.
20. composition as claimed in claim 1 is identified to be used to make GI physiologically active normalizing.
21. 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.
22. 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 at room temperature will reach 10
7The viability of the described coli strain of cells/ml kept 2 months at least.
23. method as claim 22, wherein said preparation comprises at least a volatility level part (VF) of plant milk extract, and described plant milk extract obtains by water extraction, and described volatility level part is by under reduced pressure, under 38 ℃ the bath temperature described plant milk extract is carried out vapor distillation and prepares being no more than.
24. as the method for claim 23, wherein said plant milk extract is available from the plant organ that is selected from blade, stem, root and fruit.
25. as the method for claim 23, wherein said plant milk extract obtains from being selected from vegetables and herbal plant.
26. as the method for claim 25, wherein said vegetables or herbaceous plant are selected from soybean, parsley, peppermint, dill, clover, garlic, beet and wild cabbage.
27., also comprise and add the beehive product as the method for claim 23.
28. as the method for claim 27, wherein said beehive product is a propolis.
29. as the method for claim 23, wherein said at least a volatility level part is from clover, soybean, beet, dill and peppermint.
30. one kind has been added the ATCC preserving number is the food of the survival coli strain BU-230-98 of 202226 (DSM 12799).
31., contain at least a milk-derived material as the food of claim 30.
32., be selected from cheese and sour milk as the food of claim 31.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| 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 |
| US09/725,846 | 2000-11-30 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1494374A CN1494374A (en) | 2004-05-05 |
| CN1294253C true CN1294253C (en) | 2007-01-10 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB018214851A Expired - Fee Related CN1294253C (en) | 2000-11-30 | 2001-11-27 | Preparation of compositions comprising bacterial strains and volatile plant extracts and therapeutic and industrial applications thereof |
Country Status (14)
| Country | Link |
|---|---|
| EP (1) | EP1343370A4 (en) |
| JP (1) | JP4410992B2 (en) |
| KR (1) | KR20030082549A (en) |
| CN (1) | CN1294253C (en) |
| AU (2) | AU2100402A (en) |
| BR (1) | BR0116178A (en) |
| CA (1) | CA2430292A1 (en) |
| IL (1) | IL156213A0 (en) |
| MX (1) | MXPA03004851A (en) |
| NZ (1) | NZ526748A (en) |
| PL (1) | PL366363A1 (en) |
| RU (1) | RU2281326C2 (en) |
| WO (1) | WO2002043649A2 (en) |
| ZA (1) | ZA200304747B (en) |
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| RU2639544C1 (en) * | 2016-12-16 | 2017-12-21 | Общество с ограниченной ответственностью "БИОРЕАКТОР" | Means for treatment and prevention of bird gastrointestinal tract diseases |
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| 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 |
| CN111212655A (en) | 2017-08-14 | 2020-05-29 | 赛里斯治疗公司 | Compositions and methods for treating cholestatic diseases |
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- 2001-11-27 KR KR10-2003-7007337A patent/KR20030082549A/en not_active Application Discontinuation
- 2001-11-27 BR BR0116178-4A patent/BR0116178A/en not_active IP Right Cessation
- 2001-11-27 RU RU2003119151/13A patent/RU2281326C2/en not_active IP Right Cessation
- 2001-11-27 EP EP01998302A patent/EP1343370A4/en not_active Ceased
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- 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 MX MXPA03004851A patent/MXPA03004851A/en not_active Application Discontinuation
- 2001-11-27 CN CNB018214851A patent/CN1294253C/en not_active Expired - Fee Related
- 2001-11-27 JP JP2002545628A patent/JP4410992B2/en not_active Expired - Fee Related
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| Publication number | Publication date |
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| ZA200304747B (en) | 2004-09-03 |
| MXPA03004851A (en) | 2004-05-04 |
| PL366363A1 (en) | 2005-01-24 |
| KR20030082549A (en) | 2003-10-22 |
| CN1494374A (en) | 2004-05-05 |
| BR0116178A (en) | 2003-10-14 |
| WO2002043649A8 (en) | 2004-04-29 |
| WO2002043649A3 (en) | 2002-08-29 |
| CA2430292A1 (en) | 2002-06-06 |
| RU2281326C2 (en) | 2006-08-10 |
| EP1343370A2 (en) | 2003-09-17 |
| WO2002043649A2 (en) | 2002-06-06 |
| NZ526748A (en) | 2006-03-31 |
| AU2002221004B2 (en) | 2007-01-25 |
| IL156213A0 (en) | 2003-12-23 |
| JP4410992B2 (en) | 2010-02-10 |
| JP2004534512A (en) | 2004-11-18 |
| AU2100402A (en) | 2002-06-11 |
| EP1343370A4 (en) | 2004-02-11 |
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