CN101172118A - Probiotics as alternative medicines against infectious diseases - Google Patents
Probiotics as alternative medicines against infectious diseases Download PDFInfo
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Abstract
A novel representational system that is a method for improving feeding probiotics is provided. The probiotics, especially the Pediococcus acidilactici and the Saccharomyces boulardii are mixed in different colloid net including xanthan, chitosan or gelatin. By using said mixed colloid the activity of the probiotics can be protected, and multiple execrable surroundings and disadvantageous temperature for growth can be prevented. The novel representational method also provides many kinds of method for treating infectious disease for birds and mammals by using activated probiotics. Furthermore, the probiotics is usually acted as adjuvants for improving the generation of antibody and the cellularity immunity effect. In particular, probiotics were shown to reduce the number of viable oocytes from fecal samples, stimulate antibody production, and stimulate of proliferation of splenocytes in chickens infected with Elimeria. In addition, probiotics were shown to relieve symptoms of parvovirus infection in dogs.
Description
Technical field
The present invention relates to a kind of probiotic microorganisms compositions, and this probiotic microorganisms compositions of a kind of usefulness is regulated immunoreactive method.
Background technology
The description of pertinent literature comprises that information is published in 37 CER1.97 and 37 CER1.98.
Probiotic bacteria is a kind of activated microorganism, should give an amount of probiotic bacteria, and it can authorize " host " wholesome effect (select from the United Nations's food and agricultural organization and world health organization, replace medicine 2001).Probiotic bacteria is popularized and is used and the mankind, and the Animal nutrition additive, for example says, yeast Chang Zuowei gives the supplementary of poultry.Excellent happy breast and lactobacillus, lactobacillus (Lactobacillus) and bacillus bifidus (Bifidobacterium) are commonly used to prevention and treatment and diarrhoea, with disease (Alvaez, etal, 2001 of relevant gastrointestinal infection; Fuller1989; Majamaa, et al 1995).
The characteristics of many probiotic bacteria uniquenesses, for example anti-infective character is regulated immunoreation, increases epidemic prevention function, metabolic function and the wriggling and the absorption function that change intestinal.These functions all make probiotic bacteria become alternative medicine (Walker and Buckley, 2006) to the mankind and animal.
Though the product of probiotic bacteria, as the short fatty acid (SCFA) that connects, intoxicated sugar of the albumen of cell wall (peptidoglycan) and the short deoxyribonucleic (DNA) that connects contain the CPG arrangement, function that can profitable probliotics.Using activated microorganism, eat to the human and animal, is the main at present policy (Walker andBuckley, 2006) of using and studying probiotic bacteria.In order to make probiotic bacteria to the maximum effect of the mankind and animal performance, we must be sent to the antibacterial that lives enteral and in enteral growth, breeding (Kailasapatha and Chin 2000).
Lactobacillus (Lactobacillus) and bacillus bifidus (Bifidobacterium) are two kinds of modal lactic acid probiotic bacterias, discuss in the scientific literature of being everlasting, and see in the general merchandise of also being everlasting.
This two genus antibacterial is to belong to the antibacterial that is fit to anaerobic environment, and most kind and microspecies are to oxygen and high temperature all very sensitive (Gomes et al, 1995 in this two genus antibacterial; Talwakar and Kailasapathy, 2004), so, always under the environment of not only opening but also closing, be that very difficulty is kept the activity of this two bacterioid in room temperature.Therefore, for present commodity, the method for long preservation, and, often have different results to deliver (Tuomola et al, 2001) in the shipping conditions of different temperatures.
United States Patent (USP) number 5,968,569, statement colloid in pet food, the starch net includes probiotic microorganisms, and special declaration is Saccharomycodes (Saccharomyces) and pediococcus acidilactici (Pediococcus acidilactici).
United States Patent (USP) number 6,551,663, statement are given in the milk powder of house pet and are contained lactic acid ferment and lactic acid, and comprise patent 5,968,569 probiotic bacterias of being stated.
The microorganism that United States Patent (USP) number 6,780,447 statement animal foods comprise sorbic acid (sorbic acid) and activity and non-activity are arranged, these microorganisms are meant pediococcus acidilactici (P.acidilactici).
Special combination in United States Patent (USP) number 6,827, the 957 statement animal foods, the shell and the probiotic bacteria that comprise soft internal layer of one deck and hard mix, and are meant Saccharomycodes (Saccharomyces) especially.
United States Patent (USP) number 6,827,397 comprises yeast with fat, comprises various different probiotic bacteria, comprises saccharomyces boulardii (Saccharomyces boulardii) and pediococcus acidilactici (Pediococcus acidilactici).
In the U.S. Patent application 2003/0049240, disclosing a kind of method, is with lactobacillus (Lactobacillus) and bacillus bifidus (Bifidobacterium), the infection of treatment Helicobacter pylori (helicobacter).
In the U.S. Patent application 2003/0109025, claim that a kind of method is used for making a kind of microgranule, the outside is coated with activated microorganism.
In the U.S. Patent application 2004/0197352, claim predecessor's composition wherein of probiotic bacteria and reduce muscle element (creatine) and BUN comprises various microorganisms not of the same race.
In the U.S. Patent application 2003/0165472, claim a kind ofly to be used for storing and the transportation microorganism.
In the U.S. Patent application 2006/0008511, claim that probiotic microorganisms is included in xanthan gum (xanthangum), in the mixture of chitosan (chitosan) and gelatin (gelatin).
Example phase WICCON whether in cus toms clearance or not in the past all is to be used for explanation.
Summary of the invention
Following system cooperates various system tools and method to explain with idea, illustrate representatively, is not limited thereto space a little.In different systems, there are some above-mentioned problems just to reduce relatively or solution, when we solve problem with the diverse ways system.
The specific embodiment that is disclosed comprises that a kind of house pet uses goods, and these goods comprise the probiotic bacteria in the mixture that is encapsulated in xanthan gum and chitosan.The specific embodiment that is disclosed comprises that preparation to the edible food of house pet, comprises probiotic microorganisms and is packaged in a kind of gelatine capsule again, and this probiotic microorganisms is yeast (Saccharomyces) and lactic acid bacteria.In this disclosed, yeast was meant Saccharomycodes (Saccharomyces), and lactic acid bacteria is meant pediococcus acidilactici (Pediococcus acidilactici).Yeast is to refer in particular to Bradley saccharomyces cerevisiae (Saccharomyces cereviase boulardii), is Saccharomyces boulardii again.So-called lactobacillus is meant pediococcus acidilactici (Pediococcus acidilactici).In this announcement, the concentration of xanthan gum (xanthangum) is at 0.2-2.0% (weight percent/volume), and the concentration of chitosan is in 0.1-1.0% (weight/volume), and acid-base value (PH) is between 2-7.More more specifically, the concentration of xanthan gum (xanthan gum) is about 1.25% (weight/volume), and the concentration of chitosan is (weight/volume) about 0.4%, and soda acid (PH) degree is about 4.15.The specific embodiment that is disclosed comprises treatment bird and the relevant infectious disease of mammal the intestines and stomach.In the process of this treatment, comprise probiotic bacteria is packaged in the inside, or probiotic bacteria is added in the feedstuff and snacks of animal, when probiotic bacteria is fed in bird and mammals, can strengthen birds and mammiferous immunologic function in mode so, resist cause of disease.
Description of drawings
Fig. 1 represents, the relation of acid-base value (PH) and xanthan gum and chitosan capsules hardness.
Fig. 2 represents, the activity of probiotic bacteria and the relation that has or not xanthan gum and chitosan capsules at desuperheat.
Fig. 3 represents, the activity of probiotic bacteria and the relation that has or not xanthan gum and chitosan capsules heating.
Fig. 4 represents, the activity of probiotic bacteria and the relation that has or not xanthan gum and chitosan capsules at sour environment.
Fig. 5 represents, the effect that the probiotic bacteria antagonist produces.
Fig. 6 represents, the effect of probiotic bacteria pair cell immunity.
Fig. 7 represents, by the cyst worm in (E.acervullina) infected chicken feces.
Fig. 8 represents, by (E.acervullina) or the chicken that (E.tenella) infects, the cyst worm in the feces.
The specific embodiment
Activated lactobacillus and yeast are used for feeding and give the probiotic bacteria of cat and dog house pet, are packed by a kind of biological heteropolymer xanthan gum (xanthan gum) of microorganism and chitosan (Chiosan) hybrid protection to form.Xanthan gum (xanthan gum) is a kind of polysaccharide glue, and the amount with seldom promptly may be dissolved in the water; add a little stirring; just become very dense condensed solution, it can form the tough protecting film of one deck on solution, can help probiotic bacteria to improve pyrolysated patience.Chitin (chitin) also is a kind of Polysaccharides, and major part is Fructus Vitis viniferae amine (N-acetyl-glucosamine) unit bodies that is connected by branch not.Chitosan (Chitosan) is the chitin of a kind of removal amido (acetyl), and this polymer is commonly used to treating water, Emulsion that flushing is taken a picture and improvement artificial synthetic fiber, the dyeing capacity of weaving cotton cloth and the preparation of wound healing.
Probiotic microorganisms is wrapped in the aqueous solution, contain 0.5-2.5% (percentage by weight, weight/volume) chitosan (chitosan) capsule of xanthan gum (Xanthan gum) and 0.2-0.8% (weight/volume), the acid-base value of this solution (PH) is between 2 to 7, solution more preferably is to contain 1.25% xanthan gum (xanthan) and 0.4% chitosan (chitosan) capsule (weight/volume), acid-base value (PH) was at 4: 15, and this kind is wrapped and still activated microbial cell still can form 10 for every milliliter
10Bacterial strain unit bodies (cfu/ml).
Activated probiotic microorganisms is wrapped in the mixed liquor of xanthan gum (xanthan) and chitosan (chitosan), it can preserve activation of microorganism, and with the dosage of correct active probiotic, deliver in the intestines and stomach of house pet, so make the feeding probiotic bacteria can reach maximum benefit for cat and dog.Usually cat and dog can not refused edible probiotic bacteria, are wrapped in the mixed liquor of xanthan gum (xanthan) and chitosan (chitosan).
The inventor wishes not to be limited in this explanation, their suggestion is given birth to little living bacterium when benefit and is included in xanthan gum (xanthan gum) and chitosan (chitosan) solution, and special effect is because this capsule mixed liquor forms a place special xanthan gum chitosan (xanthan chitosan) in conjunction with complex.This complex has two kinds of crossing electrolyte of electrodes in solution.Owing to different electrodes is attracted each other and is formed polyeletrolyte.According to surveying in neutral solution, xanthan gum (xanthan gum) major part is the electronegative utmost point, and chitosan (chitosan) major part is to have anelectrode, and these two kinds of polymer have different electrodes in aqueous solution, will be combined into a stable polymerization complex and tough capsule.Solution becomes will make the amido of chitosan partly remove (deionization) in alkalescence when pH-value (PH) raises just, makes to produce more unsettled combination between two kinds of polymer, causes becoming more weak protection capsule.
The relation of pH-value (PH) between 2 to 8 of the capsular hardness of Fig. 1 xanthan gum (xanthan) and chitosan (chitosan).Capsule forms under above-mentioned suitable normal solution environmental.Utilize TA.XT25 can measure capsular hardness and toughness, with the relation of different pH-values, with 5 kilograms of measured value devices, measure under apart from 1 millimeter during measurement, Fig. 1 represents that the most tough spike of capsule is to drop between the pH-value 3 to 4.When pH-value between 6 to 8, capsular toughness also weakens relatively, the data of Fig. 1 also meet theoretic discussion the last period.To inhale the stability that forms the polymerization complex mutually relevant with pH-value because of positive and negative polarities for the xanthan gum of xanthan gum (xanthan) and chitosan (chitosan) and chitosan Xanthan chitosan.
Fig. 2 demonstration, under the cryogenic effect, the activity of probiotic bacteria and the capsular relation that has or not xanthan gum (xanthan) and chitosan (chitosan).Capsule will be arranged and do not have capsular probiotic bacteria to put 1 hour at 0 ℃, the active probiotic number that does not have the capsule protection is from 10
9.3Bacterial strain body/milliliter (cfu/ml) reduces to 10
8.3, strain body/milliliter.Capsular active microorganism number is arranged from 10
9.3Bacterial strain body/milliliter reduces to 10
9Bacterial strain body/milliliter, Fig. 2 show that at low temperatures capsule is to the protection effect of probiotic microorganisms.
Fig. 3 demonstration, under the high-temperature effect, the activity of probiotic bacteria and the capsular relation that has or not xanthan gum (xanthan) and chitosan (chitosan).Capsule will be arranged and do not have capsular probiotic bacteria to put 150 seconds at 60 ℃, the active little aerobic plate count order that does not have the capsule protection is from 10
9Bacterial strain body/milliliter reduces to 10
7Bacterial strain body/milliliter.The little aerobic plate count order of capsular activity is arranged from 10
9Bacterial strain body/milliliter reduces to 10
8.9Bacterial strain body/milliliter.Fig. 3 shows that at high temperature, capsule is to the protection effect of probiotic microorganisms.
Fig. 4 shows, when soda acid matter low (acid solution), and the capsular relation of mixed liquor that the activity of probiotic bacteria and xanthan gum colloid (xanthan) that has or not xanthan gum (xanthan) and chitosan (chitosan) and chitosan gather chitose (chitosan).Capsule will be arranged and do not have the probiotic bacteria of cutose, be placed under the environment of pH-value 20 to 60 fen not clock etc., after 30 minutes, not have the little living bacterium of capsular activity from original 10
9Reduce to 10
5.7Bacterial strain body/milliliter after 60 minutes, reduces to 10
5.5Bacterial strain body/milliliter.The little living bacterium of capsular activity is arranged from original 10
9Reduce to 10
7.8Bacterial strain body/milliliter, after 30 minutes and 60 minutes, Fig. 4 shows that at the high solution of acidity, capsule is to the protection effect of probiotic microorganisms!
Probiotic bacteria can be used as and substitutes medicine, the infectious disease that can cause for the chicken defence parasite of suitable barbecue.
The coccidiosis of birds is the most common at poultry farming, and topmost parasitic disease seriously can cause death, and the lighter causes dyspepsia, absorbs not goodly, causes poor growth, and reduces the chance (Lillehoj et al., 2004) of laying eggs.The most tangible condition of illness of coccidiosis suppresses growth exactly, slows down the increase of body weight, in more serious situation, can lose weight, and these all can have influence on the economic benefit (Dalloul and Lillehoj, 2006) of domestic fowl farming.At present, medicine and live bacterial vaccines are the methods that coccidiosis treated in two kinds of main Chinese, but owing to use the medication management formality of prevention eqpidemic disease to increase, and the raising of vaccine cost, the demand of replacement therapy method also just improves relatively, for the chicken that is infected by eimeria tenella, though observing the intravital antibody of chicken increases, but the increase of cellular immunity is only the basic foundation (Dalloul and Lillehoj, 2004) for the treatment of disease.Nearest probiotic bacteria progress of research, the antibacterial that proof is lived can influence host's humoral immunity to be increased, especially to cause disease (Isolauri E., 2003 of relevant intestinal by rotavirus (rotavirus), coliform (E.coli) and salmonella (Salmonella); Majamaa, etal 1995; Perdigon et al 2001).For more effective use probiotic bacteria becomes a kind of alternative medicine, the infectious disease that antagonism is caused by eimeria tenella, we must prove the chicken of using the probiotic bacteria feeding and being infected by eimeria tenella, and intravital humoral immunization of its chicken and cellular immunization are improved, and good effect is arranged.
Check to use the chicken of probiotic bacteria, produce any possible toxicity in that its body is any being infected by eimeria tenella.
The chicken that the 15-20 just one day is big is classified one group as, has 6 groups.Fed from first day and to give different feedstuffs, one group is the general feedstuff on the market of a matched group feeding, and feeding adds probiotic bacteria in addition.The feedstuff of pediococcus acidilactici (P.acidilactici) is divided into five kinds of different feedstuffs again in the feedstuff of feeding profitable probliotics, be distinguished into 0,0.01%, 0.05%, 0.1% and 0.4% according to the probiotic bacteria of variable concentrations.
At the 10th day, except the variation of matched group (does not have feeding pediococcus acidilactici Pediococcus acidilactici, do not infect), other all chickens all give the egg capsule worm of the germination of 5000 eimeria tenellas of oral infection (Eimeriaacervulina).After metainfective the 0th, 6,9 days, measure the weight of all chickens, and write down the increment number of their body weight.In infection back 6 days, begin to collect the feces of chicken, collect 4 days (infecting the back the 6th day to the 9th day) continuously, collect feces and be put in the keg, and the egg capsule worm in the record feces.
With different experiments group and the collected data difference of matched group, (ANOVA) to analyze with " variation analysis " with the formula GRAPHPAD INSTAT of statistics, this is the registered trade mark of statistics software formula, and is designed by GraphPad software company.Company is positioned at the Santiago, markon Fo Niya state.Data are after this formula is analyzed, and finding has evident difference with matched group between the experimental group, and its probability P＜0.05 is used the Dunnett comparison test in addition, comes more every group intermediate value.
Chart 1 (TABLE-US-00001) shows that pediococcus acidilactici (Podiococcus acidilactici) but has tangible influence and effect to the number of egg capsule worm in the growth of the chicken that infected by the coccidiosis bacterium and the chicken manure.At first inspect chicken by the egg capsule insect infection of 5000 eimeria tenellas after the comparison that increases of 1-9 days body weight, one has 6 groups, is 1,2,3,4,5,6 in regular turn.Give 0,5000,5000,5000,5000 egg capsule worms for every group.And the pediococcus acidilactici (P.acidilactici) that feeds different weight percentage, be respectively 0%, 0%, 0.01%, 0.05%, 0.1% and 0.4%.
Average every group body weight increment is followed successively by 396 grams, 343 grams, and 362 grams, 366 grams, 395 grams and 382 grams, its standard deviation is 45,51,58,36,44.Secondly the every cell mean of egg capsule worm of collected eimeria tenella is about 0 * 10 successively in feces
8, 2.65 * 10
8, 1.97 * 10
8, 2.39 * 10
8, 1.39 * 10
8, 2.10 * 10
8, its standard deviation is 0.67,0.50,0.27,0.23,0.60.
A. by chicken 1-9 days body weight recruitments behind 5000 germination egg capsule insect infections, be unit with the gram.
| Group | Germination egg capsule worm dosage | Probiotic bacteria (%) | Average weight recruitment (gram) | |
| 1 | 0 | 0 | 396 | 45 |
| 2 | 5000 | 0 | 343 | 45 |
| 3 | 5000 | 0.01 | 362 | 51 |
| 4 | 5000 | 0.05 | 366 | 58 |
| 5 | 5000 | 0.1 | 395 | 36 |
| 6 | 5000 | 0.4 | 382 | 44 |
B. the similar Table A of egg capsule insect infection chicken germinates
| Group | Germination egg capsule worm dosage | Probiotic bacteria (%) | Egg capsule worm meansigma methods (X10 8) | |
| 1 | 0 | 0 | 0 | 0 |
| 2 | 5000 | 0 | 2.65 | 0.67 |
| 3 | 5000 | 0.01 | 1.97 | 0.50 |
| 4 | 5000 | 0.05 | 2.39 | 0.27 |
| 5 | 5000 | 0.1 | 1.39 | 0.23 |
| 6 | 5000 | 0.4 | 2.10 | 0.60 |
When want reagent used become a new medicine before, must remove this reagent and can produce any probability that the side effect of toxin is arranged, could move then and use this reagent, do various numbers can researchs.Pediococcus acidilactici (P.acidilactici) is the microorganism of a kind of self-sow at animal and human's class enteral, never puts down in writing on any document, and this pediococcus acidilactici has tangible toxic action.In order to guarantee that investigating pediococcus acidilactici might cause the side effect that is fit to tender chicken generation murder by poisoning to this, we feed the pediococcus acidilactici of 0.1-0.4% and give tender chicken or the chicken that is infected by eimeria tenella germination egg capsule worm (E.acervulina).
1A shows as table, and the tender chicken that is infected by eimeria tenella no matter whether is not as long as the feeding pediococcus acidilactici just loses weight or narrow dead phenomenon.Further the result shows, the tender chicken that is infected by eimeria tenella feeds the probiotic bacteria that can mix kind, that is to say that pediococcus acidilactici (P.acidilactic) and saccharomyces boulardii (S.boulardii) concentration are respectively 0.01% and 1.0%.These infected chickens are between infection period, and body weight increases often on the contrary, and in the middle of the process of whole experiment, we are after hello probiotic bacteria is eaten to chicken, and not observing any major organs has any variation.These organs comprise, liver, heart, kidney, spleen.This experiment proves the tender chicken that is infected by eimeria tenella with similar experimental result, can feed with the young ball lactobacillus higher 40 times and (that is to say than normal concentration, concentration is 0.01% to 0.4%) or feed with mixed lactic sheet coccus and saccharomyces boulardii concentration and can not spy out any modal variation from 0.01 up to 1.0%.
In order to prove that pediococcus acidilactici (P.acidilactic) can be used for defending chicken to be infected the disease that is produced by eimeria tenella.Whether our observation uses pediococcus acidilactici to feed chicken, and then the germination egg capsule worm (Eimeria tellena) of oral a kind of eimeria tenella.What is interesting is the chicken of infected eimeria tenella, feed again with the number minimizing of pediococcus acidilactici demonstration egg capsule worm and approximately lack 20-40% percentage ratio (referring to Fig. 7) than the egg capsule borer population that the chicken that does not have the feeding pediococcus acidilactici produces.Under similar situation, we observe when chicken is infected by E.acervulina, if feed to mix two kinds of probiotic lactic acid sheet coccuses and saccharomyces boulardii, the number of egg capsule worm can reduce to 30-50% percentage ratio.When chicken is infected by E.tellena, to feed again with same mixing probiotic bacteria, the number of egg capsule worm is than the chicken minimizing 10-20% (referring to Fig. 8) that does not have the feeding probiotic bacteria, so these results show that probiotic bacteria can lower cause of disease or parasite to the intravital infringement of chicken.
The feeding probiotic bacteria can make the tender chicken that is infected by eimeria tenella, stimulates the intravital humoral immune reaction of chicken.
Fig. 5 is presented at the different probiotic bacteria of feeding concentration and mixes kind of (pediococcus acidilactici and a saccharomyces boulardii).There not to be probiotic bacteria to mix kind of (with a NOR representative), 0.1% or 0.2% ratio blending probiotic bacteria mixes plants feeding 21 days (with MG0.1 and MG0.2 representative) in general feedstuff, and the antibody (Anti-ETMICZ) that produces anti-eimeria tenella in the check chicken body reacts.Relatively hatching does not have infected chicken and by the egg capsule insect infection chicken of 5000 eimeria tenellas (E.tenella) after 12 days, and after infecting the egg capsule worm 10 days, get the serum of chicken, check the antibody of anti-eimeria tenella.Antibody (Anti-ETMICZ) reaction that produces anti-eimeria tenella in the chicken body is different, each ribbon is represented intermediate value plus-minus standard deviation (N=3), the chicken of infected eimeria tenella and do not have infected chicken, their intermediate value has tangible difference, and probability is less than 0.05 (P＜0.05).
In order to assess antibody to eimeria tenella antigen, the reaction of ETMIC2.ETMIC2 is a kind of Eimeria albumen in the eimeria tenella, and its gene is separated, and analysis and evaluation (Dalloul et al, 2006) and with the intravital serum of ELISA calibrating chicken in addition.By the chicken that eimeria tenella (E.tentlla) infects, significantly all cause that generation is to the proteic antibody of eimeria tenella at all in their serum.Furtherly, using feeding probiotic lactic acid sheet coccus (P.acidilactici) again by the chicken of insect infection, his intravital serum is to the antibody of eimeria tenella, and significantly exceeding does not have the feeding probiotic bacteria again by the chicken of insect infection, antibody in the body in the serum, its probability P＜0.05 (as Fig. 5).This result proves that significantly probiotic lactic acid sheet coccus (P.acidilactici) is to stimulate the antibody that produces humoral immunity, resists the tender chicken of specific parasitic infection.
The cellular immunity reaction of general, lymphocytic breeding.
Fig. 6 shows that ConcanavalineA (conA) can guide the breeding of splenocyte.After tender chicken infects eimeria tenella, feeding normal diet or contain variable concentrations 0.01% again, 0.1% or 1.0% mixing probiotic lactic acid sheet coccus and saccharomyces boulardii are hatched 14 posteriori chickens with 5000 eimeria tenella E.acervulina (EA) or the egg capsule insect infection of E.tenella (ET), taking out splenocyte is cultivated, and add conA and continue to cultivate 24 hours, then active with containing [
3H]-thymidine, measure cell proliferation, each ribbon is represented intermediate value plus-minus standard deviation (N=3).Infect the chicken of eimeria tenella (EA or ET) not of the same race, the intermediate value difference of their cell proliferation.
Probability (P) is less than 0.05 (P＜0.05).
We utilize the tender chicken that is infected by eimeria tenella, one group of feeding normal diet, another group feeding contains the feedstuff of profitable probliotics, utilize conA to impel the detection of splenocyte breeding reaction, determine whether to be produced the cell immune response of general, the infection that resists eimeria tenella by the chicken that feeding contains profitable probliotics such as pediococcus acidilactici.In the tender chicken that is infected by the EA eimeria tenella, to feed containing the tender chicken of 0.1% probiotic bacteria, their splenocyte reproduction speed is significantly fed with the splenocyte of the tender chicken of DIFFERENT FEED all fast (P＜0.05) than other any one group.These different feedstuffs are meant the common probiotic feed that does not contain probiotic bacteria or contain variable concentrations, in the tender chicken that is infected by the ET eimeria tenella, feeding is to contain the tender chicken group of 0.01% and 0.1% probiotic bacteria, the splenocyte that significantly shows them is under the stimulation that is subjected to conA, reproduction speed is faster than other experimental grouies, and feed with the splenocyte reproduction speed of the chicken of 0.1% probiotic bacteria than 0.01% fast.
Fig. 7 shows the tender chicken that comparison is infected by eimeria tenella E.tenella (ET), by eimeria tenella ET, and the number of egg capsule worm in the chicken manure of infection.The feedstuff (REG) that feeding is common or contain 0.1% probiotic bacteria (MG0.1) or contain 0.2% probiotic bacteria (MG0.2).Metainfective 6 to 9 days, collect the egg capsule worm in the chicken manure every day.The MG representative contains the feedstuff of chicken ball lactobacillus, and each ribbon is represented intermediate value plus-minus standard deviation (N=5 cage).
Fig. 8, egg capsule worm (EA or ET) with 5000 eimeria tenellas, infect the number of egg capsule worm in the chicken chicken manure after hatching in 12 days, infecting, these infected chicken feeding normal diets (REG), contain 0.01% mixing probiotic bacteria (M0.01), 0.1% (M0.1) or 1.0% (M1.0).M represents mixed lactic sheet coccus (P.acidilactici) and saccharomyces boulardii (S.boulardii), and each ribbon is to represent intermediate value plus-minus standard deviation (N=5 cage).
The alternative medicine that probiotic bacteria can be infected by the virus as Canis familiaris L. dyspepsia or Canis familiaris L..
The application of probiotic bacteria---for the dyspeptic eqpidemic disease of Canis familiaris L..
Use the example of probiotic bacteria success: Mai Teling MITOMAX
TM, be a kind of mixed lactic sheet coccus (P.acidilactici) and saccharomyces boulardii S.boulardii) and probiotic bacteria.The infection that it can help chicken to resist eimeria tenella makes us think further to do field test and whether assesses Mai Teling MITOMAX
TMCan help to solve the dyspeptic problem of Canis familiaris L..Step Trane MITOMAX
TMBe the registered trade mark of probiotic bacteria, belong to ImagilinTechnology, LLC is positioned at Potomac, the Maryland State.It is that mixed lactic sheet coccus (P.acidilactici) and saccharomyces boulardii (S.boulardii) are packaged in the capsule of gelatin.The assessment of this field is the experiments (table 2) of cooperating of 4 veterinarys with St. Paul,Brazil 3 tame pets hospitals, and the body weight of Canis familiaris L. is distributed in 2 kilograms to 26 kilograms, and Canis familiaris L. was not waited in about 1 years old to 15 years old of age.These Canis familiaris L.s all always have the dyspeptic disease of the intestines and stomach in various degree, feed the probiotic bacteria with 1 or 2 every day, decide 1 or two by the body weight of Canis familiaris L..Through using the treatment of probiotic bacteria in 14 days, most Canis familiaris L. all has tangible improvement, no longer is subjected to dyspeptic phenomenon.The demonstration that this result is very clear and definite, probiotic bacteria has good effect for the dyspeptic Canis familiaris L. of treatment.Table 2 (TABLE-US-0002) is done probiotic bacteria to treatment Canis familiaris L. dyspeptic field test summary, this field test are that the pets hospital at St. Paul,Brazil carries out.
The field assessment that table 2 probiotic bacteria is suffered from the Canis familiaris L. that the dyspepsia disease is arranged, assessment is that the pets hospital in the Sao Paulo, Brazil carries out.
| Name | Age (year) | Body weight (kilogram) | Sex (man) or (woman) | Disease before probiotic bacteria is handled (D, V, C, OC, F, LA) * | Benefit is given birth to. the bacterium processing time (my god) | The effect that probiotic bacteria is handled ** | |
| |
9 | 16 | The | D+ | 4 | Recover | |
| Pai Ruoji | |||||||
| 4 | 10 | The man | D+，V+， |
4 | Recover good | ||
| Breathe out |
9 | 13 | The | OC | 10 | Alleviate | |
| Benny | |||||||
| 5 | 12 | The | D+.F++ | 9 | Recover | ||
| Ba Erni | |||||||
| 1 | 4 | The | LA+ | 4 | Alleviate disease | ||
| Group is sharp | 6 | 12 | The | OC+ | 9 | Alleviate disease | |
| Send out |
3 | 9 | The | LA+ | 4 | Alleviate disease | |
| Anti-card | 2 | 6 | The woman | D+， |
4 | Alleviate disease | |
| Tom Si | 11 | 24 | The | LA+ | 9 | Alleviate | |
| Te Te | |||||||
| 10 | 5 | The woman | D++， |
9 | Recover | ||
| A Deni | |||||||
| 10 | 25 | The | D+ | 15 | Recover | ||
| Kate | |||||||
| 2 | 6 | The | D++++ | 7 | Recover | ||
| Tang Li | |||||||
| 15 | 26 | The man | D++， |
4 | Recover good | ||
| Agree sieve | 13 | 4 | The | D++ | 6 | Recover good | |
| Mallier | 13 | 4 | The man | D++ | 14 | Recover | |
| Haake | |||||||
| 10 | 4 | The woman | D++，V++，F+， |
9 | Recover good | ||
| Pai Like | 2 | 3 | The man | D+，F+ | 12 | Recover | |
| Qiao Fei | |||||||
| 1 | 7 | The man | V+， |
4 | Recover | ||
| Macha | |||||||
| 1 | 2 | The man | D++++， |
9 | Recover good |
*: dyspepsia has different diseases, is divided into D=diarrhoea, the V=vomiting, and the C=constipation, the OC=body odor, the F=flatulence, LA=loses special appetite, severity extent: ++ ++=very serious, ++ +=serious, ++=slight between serious, +=slight.
*: the probiotic bacteria treatment is meant oral Trane MITOMAX advanced in years
TMCapsule comprised mixed lactic sheet coccus and saccharomyces boulardii.If the body weight of Canis familiaris L. is less than 20 kilograms, every day an oral Trane MITOMAX advanced in years
TMIf the body weight of Canis familiaris L. surpasses more than 20 kilograms, every day 2 Trane MITOMAX advanced in years
TM
Probiotic bacteria is a kind of alternative medicine, can be used to stop Canis familiaris L. and infects the bloody diarrhea that causes because of small virus.
By the Canis familiaris L. that small virus infects, have serious intestinal, stomach symptom, as vomiting and bloody diarrhea.Infected doggie, if there is not suitable medical treatment, will die of illness in several days, because this disease that to be virus cause, antibiotic can not effectively be pounced on and subtract virus, the rehabilitation of Canis familiaris L. is to rely on the infection that the immune system of Canis familiaris L. is resisted small virus, diffusion, so this can be that good example is described probiotic bacteria and is used for doing and substitutes the doggie that pharmaceutical treatment is infected by small virus.Inspected the doggie that confirms to be subjected to the small virus infection for four, though through after the comprehensive antibiotic processing, bloody diarrhea still can't stop, these four doggie feeding probiotic bacteria mixed lactic sheet coccuses (P.acidilactici) and saccharomyces boulardii (S.boulardii), bloody diarrhea stops fully after 2-3 days, it is normal block that stool also recovers, and process is taken probiotic bacteria in continuous 2 week, also without any recurrence case (table 3).
Table 3 probiotic bacteria infects the curative effect of Canis familiaris L. to small virus
*
| Name | Age (moon) | Body weight (kilogram) | Small virus detects | Infect processing through the standard small virus ＊＊After the card shape | Use probiotic microorganisms | |
2 weeks back recurrence |
| Kai Li | |||||||
| 4 | 5 | Positive | Bloody diarrhea is more than a day in 6 |
2 days | Antidiarrheal, block stool | No | |
| Dawn lives | 5 | 6.4 | Positive | Bloody diarrhea, one day more than 6 |
3 days | Antidiarrheal, block stool | No |
| Fo Laxi | 12 | 20.5 | Positive | Bloody diarrhea, one day more than 6 |
2 days | Antidiarrheal, block | No |
| Morley | |||||||
| 9 | 20.5 | Positive | Bloody diarrhea, one day more than 6 |
3 days | Antidiarrheal, block stool | No |
*: by the Dr.Volkenbourgh of the animal first aid clinic of California Lancaster, DVM is performed.
*: the processing that the standard small virus infects Canis familiaris L. is with comprehensive antibiotic (Normosol R, Reglan, Cefazolin, Metronidazole or Ampiliccin, Famotidine) in the doggie that infects, some Canis familiaris L. can be accepted polysaccharide starch (hetastarch) or blood plasma transports in+/-.The processing of probiotic bacteria is to feed with 1 day 1 Trane MITOMAX advanced in years
TMThe comprehensive agent of-pediococcus acidilactici P.acidilactici and saccharomyces boulardii S.boulardii.
Probiotic bacteria depends on the activity of probiotic bacteria to the effectiveness of animal and human's body, the probiotic bacteria of colloid capsule packing, through being housed in 25 ℃ of 4 ℃ of low temperature or room temperature, after 2 years, the active number that the number of the active probiotic after the storage is inspected with original probiotic bacteria is similar.The tender chicken that eimeria tenella infects, the feeding feedstuff, comprise 0.01% to 0.4% probiotic lactic acid sheet coccus P.acidilactici after, on the form or on the body weight and do not have infected tender chicken, do not have tangible difference.Similarly the result obtains the tender chicken that different kind eimeria tenella (EA or ET) infects too.
Probiotic bacteria (the comprehensive agent of pediococcus acidilactici P.acidilactici or pediococcus acidilactici P.acidilactici and saccharomyces boulardii S.boulardii) is for the effectiveness of the tender chicken of eimeria tenella infection, depend on: 1. promote chicken to make the breeding that antibody 3. promotes the chicken splenocyte, probiotic bacteria (the comprehensive agent of pediococcus acidilactici P.acidilactici or pediococcus acidilactici P.acidilactici and saccharomyces boulardii S.boulardii) from stool sample extraction egg sac number purpose reduction 2..From the field test assessment of animal medical clinic, prove the intestines and stomach digestion disease, as have loose bowels, vomiting, forfeiture appetite, halitosis is resultful.The most important thing is, the comprehensive agent of probiotic oral (pediococcus acidilactici P.acidilactici and saccharomyces boulardii S.boulardii) is after 2-3 days, and by the dog class that small virus infects, health can be recovered immediately, these results describe probiotic bacteria and can be used to do replacement medicine, tackle infectious disease.
Optimal probiotic bacteria is yeast and lactobacillus, and lactobacillus is Mycosphaerella (Pediococcus), Lactobacillus (Lactobacillus), Bacillus (Bacillus), Bifidobacterium (Bifidobacterium), Streptococcus (Streptococcus), Enterococcus (Enterococcus) or Saccharomycodes.Saccharomycodes is Saccharomyces, and capsule bag probiotic bacteria has medical effect to morbific antibacterial, virus, fungus, parasite and unit cell body.Capsule bag probiotic bacteria is to ancylostome, ascarid, whipworm, and cestode has medical effect, and capsule bag probiotic bacteria is resultful to coccidiosis as Giardia.
After the people infects the infectiousness gastroenteropathy, take the capsule bag probiotic bacteria of suitable dosage, capsule bag probiotic bacteria antagonism human body infectiousness gastroenteropathy is effectively, and it is that effectively the powdery probiotic bacteria is that similar medical effect is arranged with capsule bag probiotic bacteria that capsule bag probiotic bacteria resists fishes infectious gastroenteropathy.
Claims (17)
1. probiotic microorganisms compositions, it comprises: at least a be encapsulated in the gelatin or the mixture of xanthan gum and chitosan in probiotic microorganisms.
2. compositions as claimed in claim 1, wherein said probiotic microorganisms are yeast or lactobacillus.
3. compositions as claimed in claim 2, wherein said probiotic microorganisms are Mycosphaerella (Pediococcus), Lactobacillus (Lactobacillus), Bacillus (Bacillus), Bifidobacterium (Bifidobacterium), Streptococcus (Streptococcus) or Enterococcus (Enterococcus) antibacterial or Saccharomycodes (Saccharomyces) yeast.
4. compositions as claimed in claim 3, wherein said Mycosphaerella antibacterial are pediococcus acidilactici (Pediococcus acidilactici), and perhaps described Saccharomycodes yeast is Bradley saccharomyces cerevisiae (Saccharomyces cerevisiae boulardii).
5. method of infectious disease of regulating the host to antigenic immunoreation and treatment birds, Fish and mammal, described method comprises: strengthen described immunoreactive host or use probiotic microorganisms compositions as claimed in claim 1 to birds, Fish or the mammal of the described treatment of needs to needs.
6. method as claimed in claim 5, wherein said probiotic microorganisms are included in the food of animal, the snacks of house pet, the local flavor material of animal, the milk product that fermented or the yoghourt.
7. method as claimed in claim 5, wherein said birds are chicken, Psittacula alexandri fasciata, parakeet, pigeon, turkey, duck or goose.
8. method as claimed in claim 5, wherein said mammals are Canis familiaris L., cat, guinea pig, Asia rat, mouse, rabbit, monkey, orangutan, pig or people.
9. method as claimed in claim 5, wherein said infectious disease is caused by pathogenic bacteria, virus, fungus, parasite or protozoacide.
10. method as claimed in claim 9, wherein said virus are parvovirus (parvovirus), rotavirus (rotavirus) or influenza virus (influenza virus).
11. method as claimed in claim 9, wherein said parasite are ancylostome, ascarid, whipworm or cestode.
12. method as claimed in claim 9, wherein said protozoacide are eimeria tenella (Eimeria), giardia lamblia (Giardia), Cryptosporidium (Cryptosporidum), toxoplasma (Toxoplasma), plasmodium (Plasmodium), Lai Shiman protozoon (Leishmania) or ring sporozoon (Cyclospora).
13. method as claimed in claim 5, wherein said adjusting immunoreation are the breedings that increases production of antibodies or increase T cell and helper T cell.
14. one kind strengthens birds, Fish and mammal to antigenic immunoreactive method, described method comprises to described birds, Fish or mammal uses probiotic microorganisms compositions as claimed in claim 1.
15. method as claimed in claim 14, wherein said enhance immunity reaction comprises the breeding that increases production of antibodies or increase the T cell.
16. method as claimed in claim 14, wherein said probiotic microorganisms are included in the food of animal, the snacks of house pet, the local flavor material of animal, the milk product that fermented or the yoghourt.
17. a probiotic microorganisms compositions, it comprises at least a powdery probiotic microorganisms.
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| US11/493,859 US7935334B2 (en) | 2005-07-07 | 2006-07-26 | Probiotics as alternative medicines against infectious diseases |
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