CN1543355A - Immunogen adherence and method of making and using same - Google Patents
Immunogen adherence and method of making and using same Download PDFInfo
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- CN1543355A CN1543355A CNA018235352A CN01823535A CN1543355A CN 1543355 A CN1543355 A CN 1543355A CN A018235352 A CNA018235352 A CN A018235352A CN 01823535 A CN01823535 A CN 01823535A CN 1543355 A CN1543355 A CN 1543355A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/12—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria
- C07K16/1203—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-negative bacteria
- C07K16/1228—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-negative bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
- C07K16/1232—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-negative bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia from Escherichia (G)
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
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- A61K39/00—Medicinal preparations containing antigens or antibodies
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- A61K39/08—Clostridium, e.g. Clostridium tetani
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- A—HUMAN NECESSITIES
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
- C07K2317/11—Immunoglobulins specific features characterized by their source of isolation or production isolated from eggs
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Abstract
A microbial adherence inhibitor in the form of fowl egg antibodies is disclosed, along with the method of making it and methods of using it. The inhibitor functions by substantially preventing the attachment or adherence of colony-forming immunogens in the rumen and intestinal tracts of host food animals. The inhibitor is made by inoculating female birds with the immunogen, harvesting the eggs which contain antibodies to the immunogen, drying the egg contents and adding to the feed or water for the host animals. Dependent upon the particular immunogen with which the female bird is inoculated, the egg antibody is used to promote the growth of food animals by improving feed conversion rates by decreasing the waste of dietary protein caused by the presence of certain colony-forming organisms in the animals, and to substantially reduce or eliminate the incidence of illnesses caused by the presence of certain illness-causing colony-forming immunogens, such as E. coli 0157:H7), in meat from food animals, and in other food stuffs.
Description
Invention field
The present invention relates to the microorganism adhering inhibitor of a stud bird ovum antibody formation, it is used for preventing in fact that the clone from forming property immunogen or hapten and adhering to or adhere in host's consumption animal (foodanimal) cud and the intestinal, relate to a kind of method for preparing such adhesion inhibitors, and the method for using such inhibitor with: (1) promotes the growth of consumption animal by improving food conversion ratio, described improving food conversion ratio is by reducing by there is some clone's formation property in the consumption animal body, consuming the proteic loss of consumption that proteic biology causes realizes; (2) reduce or eliminate the generation of disease in fact, described disease is that origin comes from and has in the meat of consumption animal and other food that some is pathogenic, the clone forms the property immunogen or hapten causes that these consumption animals itself do not suffer from described target disease.
Background of invention
Can significantly reduce animal includes, but are not limited to the common bacterial immunogen of the proteic ability of utilization of consumption
Peptostreptococcus anaerobius (Peptostreplococcus anaerobius),
Have a liking for the amine shuttle Bacterium (Clostridium aminophilum)And
Si Shi clostridium (Clostridium sticklandii)According to the Russell report in (USDA-ARS, in May, 1993), the other biological that these are biological and wherein be disclosed has caused nearly the albumen in 25% the feed stripped to be wasted jointly.This every year for Livestock Production person, similar loss was 25,000,000,000 dollars, and this is especially obvious for " even the absorption of their albumen is seemingly substantial, but still often lacking proteic grazing animal ".When the host digested and assimilated albumen in the food, these deleterious biologies were ammonia in a kind of mode of spending freely property with protein degradation, and it is converted into carbamide by liver and kidney, and the host loses when therefore discharging as urine.These deleterious biologies are also competed with beneficial organism, and it is essential that described beneficial organism effectively utilizes ammonia for the host.In addition, they need other beneficial organisms in the cud with good utilisation ammonia more.
A main target of the present invention be prevent harmful organism such as peptostreptococcus anaerobius, Si Shi clostridium in fact and have a liking for the clostridial gathering of amine and this class biology in cud of consumption animal and the growth in the intestinal, it causes eliminating them in fact in animal body by the fowl ovum antibody of administration at particular organisms.
The common bacterial immunogen that causes the food-borne disease of human body (food borne illeness) comprise escherichia coli (
E.coli), the Listera genus (
Listeria), Salmonella (
Salmonella) and campylobacter (
Campylobacter), they all produce the symptom of similar influenza, as nausea,vomiting,diarrhea and/or fever, cause kidney injury or death in some cases.In recent years, the food that pollutes these antibacterials has caused having caused gastrointestinal damage and having recalled and destroyed millions of pound food in 100,000 people at ten hundreds of or number.Consequent economic loss is startling.What especially threaten public health is escherichia coli
0157:H7, this is the strain pathogenic bacterium of common intestinal bacteria, is identified first in nineteen eighty-two.This antibacterial is present in the intestinal of consumption animal, and excretes with feces.Thereby this antibacterial enters into food supply, not only in the meat of those animals, and in food such as milk, fruit juice, Caulis et Folium Lactucae sativae, Herba Medicaginis bud and Radix Raphani etc.
Hapten is part or incomplete immunogen, such as some toxin, itself can not cause antibody to form, but can with specific antibodies.This class hapten can comprise bacteriotoxin, yeast mycete (yeast mold) toxin, virus, parasitic toxins and algae toxin etc.
Other clone's formation property biologies comprise actinomycetes, streptococcus, bacterioid class, as
B. Ruminicola, CrytococcusWith the yeast mycete.
Another main purpose of the present invention be by to animals administer at specific immunogen or haptenic fowl ovum antibody, to prevent immunogen such as escherichia coli in the cud of consumption animal or intestinal in fact
0157:H7, or haptenic adhesion and former or haptenic gathering of this para-immunity and growth and from animal wastes, remove immunogen or hapten in fact.
Prior art
The preparation that is used to diagnose or treats the bird ovum antibody of specified disease is widely known by the people.Hint is not used for suppressing biological, especially nonpathogenic, consumes the gathering of proteic biology and the preparation of pathogenic immunogenic adhesion and accumulative bird ovum antibody.
Representational prior art patent comprises:
Polson, U.S. Patent number 4,550,019
Stolle etc., U.S. Patent number 4,748,018
Tokoro, U.S. Patent number 5,080,895
Carroll, U.S. Patent number 5,196,193
Lee, U.S. Patent number 5,367,054
Coleman, U.S. Patent number 5,585,098
Stolle etc., U.S. Patent number 5,753,268
Raun, U.S. Patent number 3,794,732 is just used the polyester antibiotic and is discussed with the utilization that improves feedstuff in the ruminant aspect the rate of ruminating.It has specifically described uses antibiotic as growth promoter in ruminant.
Raun, U.S. Patent number 3,947,836, just when to the animal oral administration, the application that specific Antibiotique composition is ruminated feed conversion for raising is discussed.Particularly, animal has been developed the function of rumen that produces more propionates with respect to acetate, has therefore improved feed conversion.
Ivy etc., U.S. Patent number 4,933,364 promotes the method for mammiferous growth of food production and feed conversion ratio to discuss with regard to another.They propose to use zinc antibiotic, and it can insoluble form be added the zinc antibiotic complex that strengthens the mammiferous feed conversion ratio of food production to produce.They are with reference to two patents of monensin having been carried out detailed protection: U.S. Patent number 3,501,568 and 3,794,732.
Other have mentioned the reasonable needs that use these materials about the list of references that uses additive such as monensin, because they may be toxic to some animal such as saddlery.These antibiotic, they are not licensed for milch cow, must careful administration.In addition, because it is in the syrup based additive of typical additive that monensin can not be added to for feed stripped (fee), originally reduced feedstuff absorption (Pate, F., " Ionophores Do Not Appear To Work In MolassesSupplements ", ONA Reports, in November, 1966,2 pages, Florida Cattleman andLivestock Journal; Lona, R.P. etc., J.Anim.Sci.75 (1): 2571-2579,1979.).
Polson, U.S. Patent number 4,550,019 relates to being used to prepare and is used for the preparation and the application of fowl yolk antibody of immune formulation that animal comprises people's passive immunity, described antibody is as being used for immunoadsorption process (immunosorbitive processes), and especially quantitative analysis test is especially for diagnosis, pathology, the immunoreagent of the micro-scale testing of legal medical expert and pharmacokinetics (pharmacokinectic) research.
Stolle etc., U.S. Patent number 4,748,018 relate to a kind of in mammal first immunologic tolerance appears and after, use anti-various antigenic any fowl yolk antibodies, use various medications under various conditions and use in conjunction with the various compositionss of this antibody method to the mammal passive immunity.
Tokoro, U.S. Patent number 5,080,895 relate to a kind of specific antibody and its preparation method and application that contains the material that derives from ovum, and described application is to be used for the treatment of infectious disease or other diseases, with additive, be used for the serodiagnosis field as livestock and poultry feedstuff, cosmetics and medicine.Although not clearly statement clearly, is used ovum antibody and is treated the infectious intestinal disease of livestock or poultry so that the method that is easy to oral administration antibody to be provided in feedstuff.
Carroll, U.S. Patent number 5,196,193 and the U.S. Patent number 5,443,976 of division relate to the antitoxin compositions that contains horse antibody or fowl yolk antibody come in and Serpentis, Aranea, Scorpio or Jellyfish poisonous substance.
Lee, U.S. Patent number 5,367,054 relates to the method for the ovum immunoglobulin of large scale purification treatment infectious disease.
Coleman, U.S. Patent number 5,585,098 relates to the little chicken yolk immunoglobulin of a kind of oral administration to reduce somatic number purpose method in the excretory milk of ruminant.
U.S. Patent numbers such as Stolle 5,753,268 relate to a kind of Anticholesterolemic (anti-cholesterolemic) ovum vaccine and preparation method thereof and treat the purposes of humans and animals angiopathy as food additive.
Summary of the invention
Extensively statement the present invention relates to a kind of method that specific target immunogen is produced the microorganism adhering inhibitor of inoculating first by in the egg-laying season of hen or near their egg-laying season the time, this inhibitor is used for host's consumption animals administer to prevent to clone formation property immunogen or hapten in fact in cud of consumption animal and/or the adhesion in the intestinal.Then, be enough to allow in the bird body, after the time of generation at the immunogenic antibody of target, gather in the crops the ovum that these birds produce at one section.All bird ovum contents that contain antibody are separated with chorion, and carry out drying.Bird ovum content also can carry out drying on feed additive or carrier material.In order to use when the needs, these can be stored or load onto ship transportation through super-dry, isolating ovum antibody adhesion inhibition thing.
The desired use of this inhibitor is depended in the immunogenic selection of target that is used for bird inoculation, and selecting non-pathogenic, consuming proteic biology is in order to improve feed conversion ratio, is in order to reduce in fact or to eliminate a disease and select to cause the biology of the disease of targeting.
Be mixed with dry ovum content at the immunogenic specific antibody of target and be by with antibody materials and animal feed mixing in fact, the animal feed supplement that contains antibody that will obtain then comes the consumption animals administer for the consumption animal.If to improve efficiency of feed utilization is purpose, the animal feed that contains antibody should be supplied to the consumption animal during fattening (finishing) timetable normally before butchering.Stoping the target microorganism to assemble these microorganisms the most at last in fact in the cud of animal or intestinal clears out of in the animal body.The degraded that is wasted of consumption albumen that objective microbe will effectively reduce the feeding produce natural disposition animal of feeding is assembled and is removed in this inhibition.In addition, the competition effect that weakens Yu do not produce the ammonia biology that obtains will further be strengthened the host and utilize feedstuff (May 1993. for Russell, USDA-ARS) most effectively.If so that pathogenic biology is eliminated from the meat of consumption animal is purpose, sufficient supplies contains the feedstuff of antibody before butchering, with stop in fact target immunogen or hapten in this animal intestinal adhesion and remove immunogen or hapten from animal.
The invention particularly relates to produce and be specific to escherichia coli
0157:H7Adhesion inhibitors and reduce or eliminate the bacterial disease of stomach of this kind in fact.Although with reference to eliminating by escherichia coli
0157:H7The disease description that causes the present invention, but should be appreciated that the present invention is not limited thereto, but it is equally applicable to eliminate the disease that is caused by other clone's formation property immunogens and hapten.
The description of preferred embodiment
The present invention is based on such notion: specificity suppresses the clone and forms property, consumes proteic biology, and as peptostreptococcus anaerobius, Si Shi clostridium and have a liking for the amine clostridium and the clone forms the property pathogenic organisms is as escherichia coli
0157:H7, Listera belongs to, and Salmonella and campylobacter in cud of consumption animal or the adhesive capacity in the intestinal, thereby reduce its breeding, growth and accumulative ability.Can design diet in the life of animal modifies so that cud and intestinal more are difficult for holding these biologies.Although Producer is administration microbial inhibitor of the present invention arbitrarily,, should on the scale of feed lot,, formulate and carry out timetable through the careful administration process of determining and designing at fattening period preferably for effective animal feed utilization rate.Utilized the low dosage of this inhibitor, more secular build-up effect, but also this predetermined cycle that is easy to combine with existing production practices will provide most economical attracting return rate by the animal performance of improving.
In order to eliminate pathogenic biology, can be before butchering administration inhibitor immediately, or in some important cycle (substantial period) administration of the life cycle of animal.Preferably in the scale of feed lot, employing is careful definite and design, the administration process in moderate cycle.As described, the cycle is utilized low dosage before butchering, and more secular build-up effect is easy to combine with existing production practices, and is most economical.It also makes mud and the fecaluria of microorganism Lock-out in the animal outside, and it is potential important polluter aspect butchering.In existing feeding system, use ionophore such as monensin (monesin) usually, the feed additive so that trade name Rumensin sells improves the feed conversion ratio of consumption animal.Have a kind polyester antibiotic to be approved for the feedstuff of supplying with beef cattle and newborn replacement heifer (dairy heifer), but approval is not used for lactescent milk cattle (diarycow).Most Gram-positive biologies are to ionophore, the special sensitivity of antibiotic right and wrong, described ionophore is if the inappropriate use of antibiotic also is quite virose to host animal.Because these antibiotic right and wrong are special, the required cud biology of cellulose of the vegetable material that a lot of digestion are taken in also can be affected.Follow the appearance of the drug resistance bacterial strain of storage (carry over) problem and biology also to pay close attention to for commercial production institute is main.Use broad ectrum antibiotic to have further defective, comprise vulnerability, extra cost and consumer boycott etc. human error.In addition, monensin class additive can not with general together administration of syrup based additive.
Any its host cud or digestive tract in accumulative biology necessarily have the ability that adheres to or be attached to the surface so that breeding and growth.The specific biology that the present invention mentions is no exception for this rule.About other factors for example beneficial organism also must consider the needs of certain enzyme, need special reagent to reduce the quantity of target organism in cud or the intestinal, do not disturb other normal floras simultaneously.Biostatic agent of the present invention is in the mode of high special, and on the basis that accumulates, the strong interference adheres to, thereby prevents the target biological self reproducing, growth and gathering.Media by simple daily feed additive, product offers a kind of antibody preparation of host in fact, described antibody preparation is that design is used for not being in order to treat any disease in the animal, but for any resident antibacterial in specificity removing cud or the digestive tract, with the adhesion of the many identical antibacterials that prevent any new invasion.This microbial inhibitor can not have a direct impact final food fully, and can never leave over unfavorable residual substance in animal or ultimate food.In addition, owing to prevented the breeding of harmful organism, as time goes by, for example at 120 days fattening periods of feed lot, they will disappear by natural degradation in the environment of feed lot, help to eliminate the important potential pollution source that pollutes once more.Inhibitor product itself can be classified in the natural materials of animal origin, can be used to the almost raising program of any kind of equally.Because active component is fully natural, they can together use with most of feedstuffs and feed additive, and described feed additive comprises syrup based additive.
All mammals and birds are all adopted similar protection mechanism at juvenile stage, and it is used for immunoreation at once, obtain the ability of preparation antibody until itself.More specifically be called the passive antibody protection, this defense mechanism is by Placenta Hominis, and breast milk perhaps passes to the mammal cub by both.But birdling then is the passive antibody protection that obtains them in ovum by the antibody of storing that occurs the period of embryo.Particularly, birds have the ability of in ovum " loading " a large amount of antibody when laying eggs, and this antibody is than the concentrated manyfold of the antibody that exists in the maternal serum.In addition, compare with mammal antibody, bird antibody is more stable, and the anti-deactivation that causes by digestion, especially under unfavorable conditions.In case immunity, hen produce unique IgY type immunoglobulin in yolk, in albumin, deposit general chicken IgM and IgA immunoglobulin simultaneously.Albumin helps the resistance to whole ovum preparation, and helps protected birds antibody.In addition, the lot of antibodies in the ovum is more special those, and they are specific to the parent antigen of the immunity of contact in the recent period.All these cause that the bird ovum provides in a large number, the ideal source of cheap, high special and stable antibody.Although illustrate the present invention by utilizing chicken to prepare bird antibody, other comprise that poultry such as turkey, duck and goose also can use.
Particularly, in the amount of the finished product of producing flow by ideal generation stable continuous and time and the predetermined moment, obtained young female chicken, Rhode Island Reds typically, White Leghorns, the cohort of the kind that Sex linkage heterozygote (hybrid crosses) or other can be produced huge ovum, egg laying amount is high and be easy to raise, they approximately arrive the egg-laying season, are 19 weeks approximately for chicken.After the isolation of the appropriate time in about 2-4 week and complying with, each cohort all will enter the rehydrated vaccine program that owns (proprietary) preparations of specific antigen that uses antibody to wish.These antigens can obtain from commercial channels, such as American type culture collection (theAmerican Type Culture Collection:ATCC).Antigen can intramuscular injection, but preferred subcutaneous injection.In about 4-5 week, the average ovum of collecting will contain many required specific antibodies, and described antibody is to exist with available and stable form.In the whole egg-laying season, can inoculate chicken once more to keep high antibody horizontal with target antigen.
From after ovum is broken by the gross of predetermined chicken group, content is separated and mixed with shell, preferred pasteurization (removing the pathogenic microorganism that potential chicken is originated, thereby reducing potential food pollution).Adopt the business method of standards to carry out drying whole ovum contents, such as utilizing environment or hot-air spray drying, and detect to determine whole titre or antibody horizontal up to 50 ℃.The ovum content can carry out drying separately, also can carry out drying on Testa oryzae or the analog harmless feed additive such as dried Semen sojae atricolor.The trace routine of use standard, as ELISA, perhaps coagulation experiment, perhaps similar method.Then produce on other average level of production of many standard activity components with representational batch with batch mixing from the chicken cohort.Exsiccant ovum antibody microbial inhibitor can store on carrier mass such as bean shell, pill and/or tablet and transport.Needs and appointment according to feed formula teacher and final consumer, final antibody product can comprise the additive that some are harmless, as exsiccant milk surum or exsiccant soybean protein powder according to ration (feed ration) preparation, exsiccant Semen sojae atricolor or Testa oryzae, or analog.To produce enough active and stable product according to an ovum of said procedure production and processing, so that the protection with the accumulative control of the as many anti-specified microorganisms of 350-700 daily dose to be provided at least.This method provides a kind of economy, safety and effective and efficient manner first, is used for controlling beef cattle and dairy herd (dairy herds) feed conversion ratio biology, and a kind of economy, safety and effective and efficient manner, is used for controlling cows
Greatly Enterobacteria 0157:H7With other pathogenic biologies.
The present invention stresses feed conversion ratio in particular, is because itself and beef cattle, with to expand to the galactopoiesis cattle relevant with dairy herd, also relevant with the problem that pathogenic biology is eliminated from cattle.Yet the notion that stops microorganism adhering has very large for multiple different food safety and production application, potential significance economically.The application in one of them this field is special in the specific unfavorable microorganism in feedstuff and the water.An example of this application comprises that effective inhibition prepares to pathogenic in the animal feed of chicken and other poultry or even the product of corruptibility microorganism.The application in this field of another one is as the rinsing auxiliary element at special unfavorable microorganism.The example of this application comprise effective removing pathogenic or even the product of corruptibility microorganism, it is used in the solution that cleans stain and rinsing cattle trunk (beef carcasses) or be used for poultry the cold preservation after being cleaned.
The accumulative microorganism of most of successes, antibacterial, virus and parasite etc., a large amount of dissimilar molecule of having evolved on their surface, it is called " adhesin (adherins) ", and it can very closely adhere to the specific molecular of one or more types of a part on host's various surfaces.Adhesion inhibitors is a kind of bird antibody with very high specific activity, its can be very closely in conjunction with, bag by, cover and eliminate these adhesin, described adhesin itself is suitable for very with the lock ﹠ key type that the mode of unique chemical structure adheres on their host.Except this direct aggression, be included in most biological fluids, as blood, lymph, saliva, tear and in a way the component identification antibody of the complement system of intestinal secretion thing adhere to as the active initiator of the defence of they a lot of types.Therefore, the activatory specific antibody that combines very possible a lot of other cytophylaxis systems adheres to and wraps by very fast and reaching optimum effect aspect chemical ablation and the microorganism of final destruction target.
By following embodiment, will further illustrate the present invention.
Embodiment 1: the selection of the hen that lays eggs
The strain of hen of laying eggs is different with purposes as required.Any female poultry that lays eggs comprises chicken, turkey, duck, emus or any other poultry, can carry out immunoprophylaxis.The common strain of preferred laying hen is normally selected according to the egg laying amount in every year, the size of ovum and the difficulty or ease of raising.Rhode Island Red, White Leghorn and red Sex linkage heterozygote are based on the ovum size, and (big-super large, the 50-65gm) animal of Xuan Zeing is used for immune schedule.Observe the raising difficulty or ease of animal and the size and the homogeneity of ovum, and every hen quantity of laying eggs every year.Although can adopt any hen that lays eggs,, show that these chickens are best for cost uses with being easy to.The homogeneous of the ovum of red Sex linkage heterozygote, the ovum amount of every animal is higher.These animal outputs are big-other ovum of super large level (50-65gm), and every hen every year can output 300 ovum nearly.
Embodiment 2: store the preparation of culture
The American type culture collection colon bacillus 0157: H7 preserving number #43895 is used as the mode trickle biology.This microorganism is separated from give birth to beef (raw hamburger), and breeds in cattle.Adopt the method for ATCC that this storage thing is carried out aquation once again.With antibacterial rehydration (Tryptase Soy Broth, Becton Dickinson) in 1.0ml TSB meat soup, and switching is in 5ml TSB sterilization meat soup, in 37 ℃ of incubated overnight (about 18 hours).Observe satisfied muddiness growth.Stand-by used as the storage thing in storing.This culture is rule on Sorbitol-MacConkey agar (Difco), produces to confirm colony clone.
Embodiment 3: preparation H antigen is as immunogen
H antigen is selected to the preparation immunogen, is used for the immunity hen that lays eggs.In incubation, use certain condition keeping the antigenic optimum growh of H, with in order to prepare the concentration that increase is provided.(VIS) production is preferred to calf perfusion agar (Veal Infusion Agar) for H antigen with calf perfusion meat soup (Veal InfusionBroth) (VIB, Becton Dickinson).The storage thing TSB that has inoculated VIB was 22 ℃-24 ℃ or incubated at room temperature 18 hours.This stimulates the flagellum of antibacterial to form.In being covered with the arrow-necked bottle of VIA, inoculation VIB culture.After 22 hours, can observe good growth.After 4 days, gather in the crops product.By using Dulbecco ' s PBS solution (pH7.3-7.4) flushing agar surface to merge arrow-necked bottle.With product collection in pipe.Use spectrophotometer counting and McFarland scopometer quality testing to survey density.Concentration is approximately 3 * 10/12/ml in the storage thing.Use motion agar slant (Northeast Laboratory Services) to detect the mobility.To store thing and in PBS, be diluted to 1 * 10
9/ ml, and stirred at ambient temperature 1 hour.Flagellum is just removed by the appearance from antibacterial like this.Use centrifugal collection supernatant.And full bacterium precipitation separated with supernatant.Determine the about 14.7mg/ml of dry weight, and material is used as the antigenic storage immunogen of H.In PBS, be diluted to 1mg/ml, again 60-70 ℃ of heating 30 minutes.Help pollution is reduced to minimum like this.Inoculation Thiogylcollate meat soup is grown to detect, and with the animal inoculation immunogen.
Embodiment 4: preparation O antigen is as immunogen
(BHI acumedia) is used to stimulate O antigen to the brain heart perfusion on antibacterial.Form and store thing TSB inoculation BHI meat soup, and cultivated 18 hours at 37 ℃.This stimulates organizator antigen on antibacterial.The arrow-necked bottle inoculation that will contain BHI meat soup is with the BHI broth culture.Follow slow stirring, at 37 ℃ of inoculation arrow-necked bottles.Observe good growth after 22 hours.Merge arrow-necked bottle and utilize centrifugal 30 minutes and sterile saline (0.9%) results material under about 3000rpm.Cutting is collected in the pipe.Use spectrophotometer counting and McFarland scopometer reference material to detect density.Material is diluted to is about 1 * 10
9/ ml.With with 0.9% Sterile Saline (Herzberg, 1972) in 1: 1 ratio of culture add 4% NaTDC (Difco) solution, and (22-24 ℃) stir about 18 hours at room temperature.Centrifugal material is to remove whole cells.Supernatant is taken as O antigen and stores thing.Dry weight is confirmed as about 14.9mg/ml.The sterilization PBS, pH 7.4, middle cut back to 1mg/ml as the O immunogen.
Embodiment 5: preparation WC antigen is as immunogen
Tryptone bean peptone meat soup (Tryptic Soy Broth) (TSB, Northeast Laboratory Services) and yeast extract (BBL) are used to prepare full cell (WC) antigen.TSB adds that the inoculation of yeast extract 0.6% meat soup stores thing with TSB, cultivates 18 hours for 37 ℃.Isoantigen and other surface antigen appear in this stimulation on antibacterial.Add yeast extract meat soup inoculation arrow-necked bottle with TSB.Follow slow stirring, 37 ℃ of cultivations.Observe good growth after 22 hours.Merge arrow-necked bottle, and utilize, and be collected in the pipe at centrifugal 30 minutes results of about 3000rpm product.Product is resuspended in sterilization PBS, among the pH7.4.Use spectrophotometer counting and McFarland scopometer reference material to detect density.Dry weight is about 19.7mg/ml.Product is diluted to about 2 * 10
9/ ml or 2mg/ml dry weight, and adding the PBS solution of 0.6% formaldehyde with 1: 1 ratio of culture, and room temperature (22-24 ℃) stir about 18 hours with fixed cell.Inoculation Thiogylcollate meat soup is grown to detect, and detects the pH value (pH7-7.4) of preparation.Supernatant is used as the antigenic storage thing of WC.At PBS, should store thing among the pH 7.4 and be diluted to 1mg/ml, as the WC immunogen.
Embodiment 6: preparation A antigen is used as immunogen
The Minca culture medium is used to the antigenic preparation of A.This is a kind of stimulation pili (pilii) and the antigenic standard medium of associated adhesion.(676-678) inoculation TSB stores thing to the Minca broth bouillon, cultivates 18 hours at 37 ℃ for Inf.Immun., Feb.1977.This stimulates Adhesion Antigen to occur on antibacterial.With Minca broth bouillon inoculation arrow-necked bottle, follow and slowly be stirred in 37 ℃ of cultivations.Observe good growth after 18 hours.Merge arrow-necked bottle, adopt at centrifugal 30 minutes results of about 2500rpm product, and be collected in the pipe.Precipitation is resuspended among the PBS, and uses stirring rod to stir 1 hour 22-24 ℃ (room temperature).This removes flagellum.Product collection in pipe, and is resuspended in PBS and 0.01%Tween20 with precipitation
TMIn, transfer to Waring blender (4 ℃) stirring at low speed 30 minutes in cold.Use spectrophotometer counting and McFarland scopometer reference material to detect density.This product is centrifugal to remove full cell.Supernatant is as the antigenic storage thing of A.If desired, can carry out deactivation in 40 minutes 60 ℃ of heating.Add 50 μ g/ml gentamycins (Gentamycin) as antiseptic.Inoculation Thiogylcollate meat soup detects growth.Measuring dry weight is about 10.6mg/ml.Use PBS, pH 7.4 these products of dilution are to 1mg/ml, as the A immunogen.
Embodiment 7: preparation P antigen is as immunogen
Reinforced clostridial medium is used to prepare P antigen.This is the standard medium of a kind of stimulation peptococcus anaerobius (Peptostreptococcus anaerobius) Adhesion Antigen.These cultures must be grown under the condition of strict anaerobic.Cultivate the storage culture according to ATCC#49031.For other is biological, to cultivate subculture in a small amount.Thiogylcollate culture medium (Difco) has been inoculated the storage thing and had been cultivated 48 hours.Inoculate arrow-necked bottle with reinforced clostridial medium.And cover culture medium with the mixture of oxygenless gas.Merge arrow-necked bottle, and utilize, be collected in the pipe at centrifugal 30 minutes results of 25000rpm product, and low-speed running 30 minutes.Measure density.Centrifugal product is to remove full cell.Supernatant is used as the antigenic storage thing of P.If desired, 60 ℃ are heated 40 minutes with deactivation.Measure dry weight.Be about 20.5mg/ml.Use PBS, pH 7.4 these products of dilution are used as the P immunogen to 1mg/ml.
Embodiment 8: preparation CS antigen is as immunogen
Reinforced clostridial medium is used to prepare CS antigen.This be a kind of stimulation Si Shi clostridium (
Clostridium sticklandii) standard medium of Adhesion Antigen.These cultures must be grown under the condition of strict anaerobic.Cultivate the storage culture according to ATCC#12662.For other is biological, to cultivate subculture in a small amount.Thiogylcollate culture medium (Difco) has been inoculated the storage thing and had been cultivated 48 hours.Inoculate arrow-necked bottle with reinforced clostridial medium.And cover culture medium with the mixture of oxygenless gas.Merge arrow-necked bottle, and utilize at centrifugal 30 minutes results of 25000rpm product.With product collection in pipe, and low speed rotation 30 minutes.Use spectrophotometer counting and McFarland scopometer inspection reference material to detect density.Product is centrifugal to remove full cell.Supernatant is used as the antigenic storage thing of CS.If desired, 60 ℃ are heated 40 minutes with deactivation.Measure dry weight and be about 22mg/ml.Use PBS, pH 7.4 these products of dilution are to 1mg/ml, as the CS immunogen.
Embodiment 9: preparation CA antigen is as immunogen
Reinforced clostridial medium is used to prepare CA antigen.This is a kind of stimulation
Clostridium AminophiliusThe standard medium of Adhesion Antigen.These cultures must be grown under the condition of strict anaerobic.Cultivate the storage culture according to ATCC#49906.For other is biological, to cultivate subculture in a small amount.Thiogylcollate culture medium (Difco) has been inoculated the storage thing and had been cultivated 48 hours.Inoculate arrow-necked bottle with reinforced clostridial medium.And cover culture medium with the mixture of oxygenless gas.Merge arrow-necked bottle, and utilize at centrifugal 30 minutes results of 25000rpm product.With product collection in pipe, and low speed rotation 30 minutes.Use spectrophotometer counting and McFarland scopometer inspection reference material to detect density.Product is centrifugal to remove full cell.Supernatant is used as the antigenic storage thing of CA.If desired, 60 ℃ are heated 40 minutes with deactivation.Measure dry weight and be about 20.5mg/ml.Use PBS, pH 7.4 these products of dilution are to 1mg/ml, as the CA immunogen.
Embodiment 10: use H, and O, WC and A antigen preparation elisa plate are used for monitoring the antibody of ovum, chicken and feedstuff
H, O, WC and A ELISA: (O, or WC or unite use: carbonate buffer solution 10 μ g-200 μ g/ml), pH9.6 wraps by 96 hole bread boards (flat, Costar ) for H, A to use the antigen of 100 μ l/ml variable concentrations.Bread board was hatched 18 hours at 22-37 ℃.Aspirate each hole to prevent cross-contamination.With 390 μ l/ every hole 0.5%BSA blocking test plate, and hatched 1 hour at 37 ℃.Select the optional row of bread board to contrast as positive or negative.Use contains Tween
TM20 lavation buffer solution decantation test plate once.Xiang Kongzhong adds the every hole of 100 μ l/ dilute sample, adds two holes, hatches 1 hour for 37 ℃.Horseradish peroxidase (the Kirkegardand Perry Laboratories that adds goat-anti chicken IgG; 1: 1000-1: 3000) conjugate.After hatching 1 hour, (TMB KPL), and used 0.1M phosphoric acid cessation reaction after 10 minutes to add substrate according to the description of manufacturer.Use Dynatech ELISA reader to measure the optical density in each hole, and recorded information is used for further data analysis at 450nm.
Embodiment 11: discrete ovum and serum are along with the analysis of time
Different phase selection ovum between duration of immunity is monitored the antibody response to specific antigen.Chicken in that monitoring in the 0th day is selected carried out once after four month in every month.Complete ovum is separated with shell, draw the 1ml sample.Then this sample is used the buffer extracting to analyze antibody content.H, O, the immunogenic standard ELISA of WC and A is used for analyzing.From the OD reading, deduct the negative control reading.Every animal serum two weeks after the 4th immunogen injection collect.
The listed data of following table are the examples through the result of acquisition in first four months.
| The ovum sample date | H, chicken | O, chicken | WC, chicken | A, chicken |
| First day: for the first time after the injection | 0.03OD | ?Neg | ?0.05OD | ?Neg |
| 1 month | 0.60OD | ?Neg | ?0.05OD | ?Neg |
| 5 weeks | 0.74 | ?ND | ?ND | ?ND |
| 2 months | 1.22OD | ?1.11OD | ?0.88OD | ?0.79OD |
| 3 months | 1.00OD | ?1.4OD | ?0.99OD | ?1.4OD |
| 4 months | 1.16OD | ?1.4OD | ?0.94OD | ?1.22OD |
| Serum: 1 month | 1.4OD | ?0.91OD | ?1.17OD | ?0.97OD |
Embodiment 12: use P, CS and CA antigen preparation elisa plate are used for monitoring the antibody of ovum, chicken and feedstuff.
P, CS and CA ELISA: (CA or unite use: carbonate buffer solution 10 μ l-200 μ g/ml), pH9.6 wraps by 96 hole bread boards (flat, Costar ) for P, CS to use the antigen of 100 μ l/ml variable concentrations.Sample board was hatched 18 hours at 22-37 ℃.Aspirate each hole to prevent cross-contamination.With 390 μ l/ every hole 0.5%BSA blocking test plate, and hatched 1 hour at 37 ℃.Select the optional row of bread board to contrast as positive or negative.Use contains Tween
TM20 lavation buffer solution decantation test plate once.Xiang Kongzhong adds the every hole of 100 μ l/ dilute sample, adds two holes, hatches 1 hour for 37 ℃.Horseradish peroxidase (the Kirkegard andPerry Laboratories that adds goat-anti chicken IgG; 1: 1000-1: 3000) conjugate.After hatching 1 hour, (TMB KPL), and used 0.1M phosphoric acid cessation reaction after 10 minutes to add substrate according to the description of manufacturer.Use Dynatech ELISA reader to measure the optical density in each hole, and recorded information is used for further data analysis at 450nm.
Embodiment 13: with H immunogen immune chicken
The hen that lays eggs of six selections, three white Leghorns and three Rhode Island Reds were about for 19 ages in week, with storing the injection of thing H immunogen.Carry out four injections (500 μ g, 100 μ g, 200 μ g and 250 μ g), carry out week about.In initial the last time injection two weeks of back, collect blood serum sample.Booster immunization if desired, each booster immunization 100 μ g (per six months).Around in, produce excellent antibody in four the ovum in six hens.ELISA H antigen reading average out to: 1: 10,000 times of dilution, 1.00OD; 1: 50,000 times of dilution, 0.265OD.The antibody production of Leghorn hen is bad, but the antibody production of all three Rhode Island Reds is good.After six weeks, follow all chickens to reply, for 1: 20,000 times of dilution, average H antigen ELISA reading is 1.40OD.
Embodiment 14: with O immunogen immune chicken
The hen that lays eggs of six selections, six white Leghorns were about for 19 ages in week, and injection is to store thing O immunogen.Carry out four injections (500 μ g, 100 μ g, 200 μ g and 250 μ g), carry out week about.In initial the last time injection two weeks of back, collect blood serum sample.Booster immunization if desired, each booster immunization 100 μ g (per six months).Around in, produce excellent antibody in five the ovum in six hens.O antigen ELISA reading average out to: 1: 10,000 times of dilution, 1.42OD; 1: 50,000 times of dilution, 0.68OD.After six weeks, be accompanied by and still have five chickens to reply, average O antigen ELISA reading is: 1: 20, and 000 times of dilution, 1.15OD.
Embodiment 15: with WC immunogen immune chicken
The hen that lays eggs of six selections, six Rhode Island Reds were about for 19 ages in week, and injection is to store thing WC immunogen.Carry out four injections (500 μ g, 100 μ g, 200 μ g and 250 μ g), carry out week about.In initial the last time injection two weeks of back, collect blood serum sample.Booster immunization if desired, each booster immunization 100 μ g (per six months).Around in, produce excellent antibody in four the ovum in six hens.WC antigen ELISA reading average out to: 1: 10,000 times of dilution, 0.95OD; 1: 50,000 times of dilution, 0.250OD.After six weeks, be accompanied by and still have five chickens to reply, average WC antigen ELISA reading is: 1: 20, and 000 times of dilution, 0.95OD.
Embodiment 16: with A immunogen immune chicken
The hen that lays eggs of six selections, six white Leghorns were about for 19 ages in week, and injection is to store thing A immunogen.Carry out four injections (500 μ g, 100 μ g, 200 μ g and 250 μ g), carry out week about.In initial the last time injection two weeks of back, collect blood serum sample.Booster immunization if desired, each booster immunization 100 μ g (per six months).Around in, produce excellent antibody in five the ovum in six hens.A antigen ELISA reading average out to: 1: 10,000 times of dilution, 1.40OD; 1: 50,000 times of dilution, 0.576OD.After six weeks, be accompanied by still all chickens and reply, average A antigen ELISA reading is: 1: 20, and 000 times of dilution, 1.15OD.
Embodiment 17: with P immunogen immune chicken
The hen that lays eggs of six selections, white Leghorns was about for 19 ages in week, and injection is to store thing P immunogen.Carry out four injections (500 μ g, 100 μ g, 200 μ g and 250 μ g), carry out week about.In initial the last time injection two weeks of back, collect blood serum sample.Booster immunization if desired, each booster immunization 100 μ g (per six months).Around in, produce excellent antibody in five the ovum in six hens.
Embodiment 18: with CS immunogen immune chicken
The hen that lays eggs of six selections, white Leghorns was about for 19 ages in week, and injection is to store thing CS immunogen.Carry out four injections (500 μ g, 100 μ g, 200 μ g and 250 μ g), carry out week about.In initial the last time injection two weeks of back, collect blood serum sample.Booster immunization if desired, each booster immunization 100 μ g (per six months).Around in, produce excellent antibody in five the ovum in six hens.
Embodiment 19: with CA immunogen immune chicken
The hen that lays eggs of six selections, red Sex linkage heterozygote was about for 19 ages in week, with storing the injection of thing CA immunogen.Carry out four injections (500 μ g, 100 μ g, 200 μ g and 250 μ g) week about.In initial the last time injection two weeks of back, collect blood serum sample.Booster immunization if desired, each booster immunization 100 μ g (per six months).In all around, all produce excellent antibody in the ovum of all six hens.
Embodiment 20: store the preparation of produce product whole egg reagent
Will from for
Colon bacillus 0157: H7All four kinds of immunogen groups, perhaps for the hen combination of the selection of three kinds of immunogen groups of Anaerobe, be used to prepare the product batches of whole egg reagent.Sterling (U.S. Patent number 5,753,228) has carried out fabulous review to the selection and the storage of ovum.With the ovum randomization, and remove shell.With whole egg mixing in fact, adopt the condition (60 ℃ (140) carried out 3.5 minutes) of standard to carry out Pasteur sterilization, Charley, H. and C.Weaver, the 3rd edition, Foods:a scientific approach, Merrill-Prentice Hall, 350 pages, 1998).In case sterilization, the activity of specimen, and 4 ℃ of storages are until drying, or spray on the carrier.Getting the sample of 250 μ l analyzes.
ELISA result's example is provided:
Pasteur disinfectant whole egg:
Colon bacillus 0157: H7
| Immunogen | Dilution factor | ??O.D. |
| ?WC | ?500 | ??0.532 |
| ?WC | ?2500 | ??0.113 |
| ?H | ?500 | ??0.466 |
| ?H | ?2500 | ??0.115 |
| ?O | ?500 | ??0.338 |
| ?O | ?2500 | ??0.128 |
| ?A | ?500 | ??0.588 |
| ?A | ?2500 | ??0.155 |
Pasteur disinfectant whole egg: Anaerobe
| Immunogen | Dilution factor | Batch #1 | Batch #2 | Batch #3 |
| ?CA | ?100 | ?0.339 | ?0.275 | ?0.627 |
| ?CA | ?500 | ?0.104 | ?0.296 | ?0.201 |
| ?P | ?100 | ?0.724 | ?0.882 | ?0.576 |
| ?P | ?500 | ?0.248 | ?0.594 | ?0.651 |
| ?CS | ?100 | ?0.457 | ?0.268 | ?0.650 |
| ?CS | ?500 | ?0.304 | ?0.143 | ?0.476 |
Embodiment 21: the bag quilt of feed additive carrier
Although whole egg can be dispersed in the water stockpile, or, use carrier to help to distribute with a kind of unified method with the ovum of exsiccant form as full powder.This makes it be easier to mix with standard feed.If desired, can use a large amount of carriers so that the media as feed additive to be provided.The bean shell of rough, purified and particle form (pelted format), rice husk, corn, cotton seed hulls, the corn of evaporate to dryness, beet pulp (beet pulp) or any other material.The pretreated product of Pasteur disinfectant whole egg is applied on the carrier and or directly feed give animal, perhaps be dried to the moisture content of residue 10-15%.About 1000ml Pasteur disinfectant whole egg is sprayed on the spherical bean shell of 50lbs.The carrier that preferably is used for domestic animal is globular bean shell, but for the son pig, preferred meticulous graininess bean shell.Feed additive mixes with the standard animal feed.Preferred level is the 10-15lbs feed additive-2000lbs of animal feed.
Embodiment 22: use pack by the analysis of back feed additive sample
After being applied to feed additive on the carrier, collect sample from a plurality of batches feed additives.Analytic sample, the result is as follows:
ProductName humidity % albumen % fat % fiber, thick %
| Rough bean shell does not wrap quilt | 11.59 | ?26.76 | ?9.10 | ?18.63 |
| CAMAS EYE 0157, rough bean shell | 12.35 | ?25.67 | ?8.26 | ?19.46 |
| CAMAS?EYE *Contrast rough bean shell | 12.06 | ?24.89 | ?9.92 | ?20.38 |
| The bean granule does not wrap quilt | 11.65 | ?9.89 | ?2.43 | ?33.47 |
| CAMAS EYE efficient, granule | 12.37 | ?10.19 | ?2.57 | ?33.12 |
*CAMAS EYE identifies the inhibitor according to the present invention's preparation
Embodiment 23: the ovum product is along with analysis-colon bacillus 0157 of time: H7
Employing is at H, O, and the sample of the immunogenic ELISA system anlysis of WC and A whole egg preparation finishes the activity of back along with the time with monitoring in the initial immunity schedule.The animal of choosing from every cohort places and produces cohort.The average ELISA OD reading (feminine gender deducts) in 4-June is listed in the following table.The ovum of sampling 250 μ l is also 1: 500 and 1: 2 from whole egg, and 500 are diluted in the PBS buffer, carry out suitable ELISA then to measure each dilution average OD reading.Deduct the negative control reading from each reading.These immunogens show replying of different good specificity in animal body.In these detected, the A immunogen provided best replying.Below the time dependent data of these immunogens are listed in:
| Immunogen | April | May | June |
| ?H：1∶500 ????1∶2500 | ?0.388 ?0.085 | ?0.848 ?0.237 | ?0.718 ?0.195 |
| ?O：1∶500 ????1∶2500 | ?0.593 ?0.147 | ?0.792 ?0.294 | ?0.704 ?0.184 |
| ?WC：1∶500 ?????1∶2500 | ?0.398 ?0.062 | ?0.730 ?0.273 | ?0.578 ?0.130 |
| ?A：1∶500 ????1∶2500 | ?0.700 ?0.102 | ?1.014 ?0.305 | ?0.909 ?0.224 |
Embodiment 24: the ovum product is along with analysis-feed efficiency of time
Employing is at P, and the sample of the immunogenic ELISA system anlysis of CS and CA whole egg preparation finishes the activity of back along with the time with monitoring in the initial immunity schedule.The animal of choosing from every cohort places and produces cohort.The average ELISA OD reading in 4-June is listed in the following table.Utilize 250 μ l whole eggs as the sample of ovum and 1: 500 and 1: 2,500 are diluted in the PBS buffer, carry out suitable ELISA then to measure each dilution average OD reading.Deduct the negative control reading from each reading.These immunogens show replying of different good specificity in animal body.
| Immunogen | April | May | June |
| ?P：1∶500 ????1∶2500 | ?1.182OD ?0.785OD | ?1.128OD ?0.489OD | ?0.942OD ?0.343OD |
| ?CS：1∶500 ?????1∶2500 | ?0.843OD ?0.318OD | ?0.989OD ?0.356OD | ?0.582OD ?0.187OD |
| ?CA：1∶500 ?????1∶2500 | ?1.156OD ?0.409OD | ?1.087OD ?0.282OD | ?0.998OD ?0.507OD |
Embodiment 25: the antibody of feed additive analytical-colon bacillus 0157: the H7 that lives
Employing is at H, O, the sample of the bean shell of the immunogenic ELISA system anlysis of WC and A bag quilt, detecting in the Pasteur sterilization, spraying, dry and store after activity.The whole egg product batches is being handled and after drying on the rough bean shell, write down good antibody response.Two batches data are as follows:
| Batch: the bean shell of bag quilt | The WC immunogen | The H immunogen | The O immunogen | The A immunogen |
| Batch #1:1: 10 1: 100 | ?0.673OD ?0.106OD | ?1.103OD ?0.236OD | ?1.105OD ?0.229OD | ?1.229OD ?0.302OD |
| Batch #2:1: 10 1: 100 | ?1.174OD ?0.177OD | ?1.291OD ?0.396OD | ?1.180OD ?0.327OD | ?1.224OD ?0.458OD |
Embodiment 26: the antibody activity analysis-feed conversion ratio of feed additive
Employing is at P, and the immunogenic ELISA system anlysis of CS and CA bag in the Pasteur sterilization, is sprayed with detection by the sample of the bean shell of crossing, the activity after drying and the storage.The whole egg product batches is being handled and after drying on the rough bean shell, write down good antibody response.Extract 1 gram 15lbs bag by the sample of bean shell and analysis.Three batches data record is as follows:
| Batch: the bean shell of bag quilt | The P immunogen | The CS immunogen | The CA immunogen |
| Batch #1:1: 100 1: 500 | ?0.067OD ?0.057OD | ?0.289OD ?0.131OD | ?0.051OD ?0.037OD |
| Batch #2:1: 100 1: 500 | ?0.028OD ?0.049OD | ?0.039OD ?0.015OD | ?0.095OD ?0.021OD |
| Batch #3:1: 100 1: 500 | ?0.046OD ?0.012OD | ?0.115OD ?0.055OD | ?0.136OD ?0.012OD |
Embodiment 27: after feedstuff mixes, and the recovery of active antibodies and egg protein
Anti-with containing
Colon bacillus 0157: H7The whole egg reagent product bag of adhesion inhibitors is become the refining bean shell of the bag quilt of bag.One bag of feed additive (15lbs) is added in the standard feed stripped of 2000lbs.Use feed additive from the whole egg preparation contrast of raising chicken scattered.Also mixed by the bean shell with the said preparation bag, as the test feed additive that contains specific antibody.Collect the sample of mixed fodder, and analyze the immunogenic active antibodies of ELISA WC and carry out commercial ELISA detect egg protein in the food (Vertatox Quantitative Egg Allergen Test, Neogen).The data of two batches ration are listed in the following table.
| Mixed fodder | The 1st batch | The 2nd batch |
| Detect feed additive: 1: 6,000 1: 12000 | ?0.172OD ?0.009OD | ?0.112OD ?0.036 |
| Control feed-additive-free 1: 6,000 1: 12000 | ?0.049 ?0.005 | ?Neg ?Neg |
| Detection feedstuff-additive-free: egg protein | 0.958OD?17ppm | ?1.268OD＞20ppm |
| Control feed-additive-free: egg protein | 0.800OD?15ppm | ?1.050OD＞20ppm |
Embodiment 28: the raising of domestic animal
Domestic animal to two cohorts is used
Colon bacillus 0157: H7Feed additive (bag by to purified bean shell) and control feed additive (bag by to contrast ovum and non-special adhesion inhibitors) are raised.Use 15lbs feed additive/2000lbs feedstuff letting animals feed.The heavily about 1000lbs of initial animal heavily surpasses 1400lbs when putting on market.All animals look all in the pink of condition, and test animal is edible more feedstuff in 87 days.Right
Colon bacillus 0157: H75 of initial experimental session test animal are positive, and control animal is 1 positive only.In 30 days of use feed additive, all test animals are
Colon bacillus 0157: H7Feminine gender, and three continuous samples keep surpassing 30 days feminine gender.The scheme of employing standard is taken a sample.All animals have all been cooked labelling on ear, and place different fences.In the sampling weekly of first moon, the sampling of two weeks is until being transported to market then.The grab sample is taken from rectum, and is placed in the markd sterilization sack.All samples are all placed on ice until handling at laboratory.All samples is handled in four hours of collection every day.Use contains the TSB dilution faecal samples of 0.6% yeast extract.The mixture line of dilution is being contained or do not containing on the Sorbitol-MacConkey ' s agar of cefixime-tellurite fill-in (Dynal ).Choosing colourless clone further detects.The latex agglutination test is used to identification of escherichia coli serotype 0157 (Oxoid dry Spot
TM Escherichia coli 0157).If the positive of detection then selects single being cloned on the SMC agar streak plate further to separate.Isolating clone's commodity in use EIA detects EH
Colon bacillus 0157(Binax, NOW EH
Colon bacillus 0157).API-20E method (AnalytabProducts) (Appl.Environ.Microbiol., 62 (7) 2567-2570,1966 are adopted in the affirmation of biochemical property; J.Clin.Micro.36 (10): 3112,1998.).
Colon bacillus 0157: H7One of amazing and the most troubling character be: need to produce very a spot of microorganism and just can cause human diseases.For example, need at least 10,000 more pathogenic
SalmonellaSerotype, but few to 10
Colon bacillus 0157: H7Cause the people symptom to occur.Therefore, a kind of parasitism has or the outside animal that has infected this microorganism, can influence 16 tons of beef ends of as many as (ground beef) to a kind of degree when butchering, if so that inappropriate preparation, independent a product just can cause disease.Have the probability of this microorganism very low although a kind of animal is parasitic at any one time, it is still very high at the probability that any feed lot occurs.
The protection consumer that has obtained at present the official recognition is not subjected to escherichia coli
0157:H7 threatens three kinds of diverse ways.These three kinds of methods are that cook completely (1), the sterilization of (2) steam Pasteur, and (3) irradiation, these three kinds of methods all have shortcoming separately, comprise mistake artificial or machinery, cost, the resistance of consumer etc.
Microorganism in any host's who accumulates in it the esophagus is in order to breed, and must have in its surface mount or the ability of adhering to.
Colon bacillus 0157: H7No exception.Adhesion inhibitors of the present invention is strong on the accumulation basis to disturb adhesive attraction, thereby prevents the gathering and the breeding of specific target microorganism.By the media of simple day feed additive, this product essence provides a kind of specific antibody preparation for the host, and its design is not to be used for treating any disease of animal (domestic animal essence is not subjected to
Colon bacillus 0157: H7Influence only be its temporary host), and only be any resident antibacterial of removing and prevent that any new invading bacteria from adhering to intraesophageal.This adhesion inhibitors can never stay unfavorable residue in animal body for not directly influence of host itself, therefore in no case can final food product be had any impact.In addition, owing to prevent this microbial reproduction, so As time goes on (at 120 days fattening periods of feed lot), it will disappear by natural degradation from the fecaluria of mud and coating animal, eliminate this important potential pollution source when butchering.If handle suitably, by the slaughterhouse effluent or use other food materials that the fecaluria of as fertilizer sources causes cross-contamination chance also essence eliminate.
Clearly, do not breaking away under the condition of the spirit and scope of the present invention and can much revise and change above the present invention who illustrates.Only described specific embodiment by embodiment, the present invention only limits by accompanying Claim.
Claims (92)
1. the ability by suppressing immunogen adherence animal cud or intestinal is to reduce the ability of described immunogen breeding, thereby reduce by the clone's formation property that in consumption animal cud or intestinal, exists, consume proteic immunogen and the proteic consumption of consumption that causes, promote the method for consumption growth of animal, described method comprises:
A. with described particular target consume proteic immunogen or approximately reach their egg-laying season inoculation hen;
B. be enough to allow in described bird, after the time of generation at the immunogenic antibody of described target, gather in the crops the ovum of giving birth to by described bird at one section;
C. the content that contains antibody with described ovum separates with shell;
D. with described isolating ovum antibody materials drying;
E. by animal feed or the water described exsiccant ovum antibody product that obtains that distributes in fact equably; With
F. described animal feed that contains antibody that obtains or water are supplied with the consumption animal, to prevent the adhesion of described target immunogen in described animal intestinal in fact.
2. according to the method for claim 1 herein, the proteic immunogen of described consumption is selected from peptostreptococcus anaerobius (P.anaerobius), Si Shi clostridium (C.sticklandii) and have a liking for amine clostridium (C.aminophilum).
3. the method for claim 1, it comprises: a kind of exsiccant feed vector material is provided, by being come the dry described content that contains antibody by described carrier mass with the described content bag that contains antibody, thereby in animal feed or water, distribute to wrap and promoted the growth of described animal to prevent the adhesion of described immunogen in described animal intestinal in fact with the carrier mass of the described content that contains antibody and animal feed or water with the described carrier mass of the described content bag quilt that contains antibody and supply.
4. the method for claim 3, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
5. the ability by suppressing immunogen adherence consumption animal cud or intestinal is to reduce the ability of described immunogen breeding, thereby reduce owing in consumption animal cud or intestinal, existing and consume the proteic consumption of consumption that proteic immunogen causes, promote the method for consumption growth of animal, the proteic immunogen of described consumption is the P antigen from Streptococcus anaerobius, and described method comprises:
A. use from the P antigen of Streptococcus anaerobius or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird and the ovum given birth to by described bird, produce at from the antigenic antibody of the P of Streptococcus anaerobius;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described results ovum separates with chorion;
E. with the described content drying that contains antibody;
F. the described exsiccant content that contains antibody in fact equably distributes in animal feed or water; With
G. the described content that contains antibody that obtains and animal feed or water are supplied with the consumption animal, preventing the adhesion of described immunogen in described animal intestinal in fact, thereby promote the growth of described animal.
6. the method for claim 5, comprise: a kind of exsiccant feed vector material is provided, by being come the dry described content that contains antibody by described carrier mass with the described content bag that contains antibody, in animal feed or water, distribute to wrap and prevented that with essence thereby the adhesion of described immunogen in described animal intestinal from promoting the growth of described animal with the carrier mass of the described content that contains antibody and animal feed or water with the described carrier mass of the described content bag quilt that contains antibody and supply.
7. the method for claim 6, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
8. the ability by suppressing immunogen adherence consumption animal cud or intestinal is to reduce the ability of described immunogen breeding, thereby reduce owing in consumption animal cud or intestinal, existing and consume the proteic consumption of consumption that proteic immunogen causes, promote the method for consumption growth of animal, the proteic immunogen of described consumption is that described method comprises from the clostridial CS antigen of Si Shi:
A. use from the clostridial CS antigen of Si Shi or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird and the ovum given birth to by described bird, produce at from the antigenic antibody of the clostridial CS of Si Shi;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described results ovum separates with chorion;
E. with the described content drying that contains antibody;
F. the described exsiccant content that contains antibody in fact equably distributes in animal feed or water; With
G. the described content that contains antibody that obtains and animal feed or water are supplied with the consumption animal, preventing the adhesion of described immunogen in described animal intestinal in fact, thereby promote the growth of described animal.
9. the method for claim 8, comprise: a kind of exsiccant feed vector material is provided, by being come the dry described content that contains antibody by described carrier mass with the described content bag that contains antibody, in animal feed or water, distribute to wrap and prevented that with essence thereby the adhesion of described immunogen in described animal intestinal from promoting the growth of described animal with the carrier mass of the described content that contains antibody and animal feed or water with the described carrier mass of the described content bag quilt that contains antibody and supply.
10. the method for claim 9, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
11. the ability by suppressing immunogen adherence consumption animal cud or intestinal is to reduce the ability of described immunogen breeding, thereby reduce owing in consumption animal cud or intestinal, existing and consume the proteic consumption of consumption that proteic immunogen causes, promote the method for consumption growth of animal, the proteic immunogen of described consumption is from having a liking for the clostridial CA antigen of amine, and described method comprises:
A. use from have a liking for the clostridial CA antigen of amine or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird and the ovum given birth to by described bird, produce at from having a liking for the antigenic antibody of the clostridial CA of amine;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described results ovum separates with chorion;
E. with the described content drying that contains antibody;
F. the described exsiccant content that contains antibody in fact equably distributes in animal feed or water; With
G. the described content that contains antibody that obtains and animal feed or water are supplied with the consumption animal, preventing the adhesion of described immunogen in described animal intestinal in fact, thereby promote the growth of described animal.
12. the method for claim 11, comprise: a kind of exsiccant feed vector material is provided, by being come the dry described content that contains antibody by described carrier mass with the described content bag that contains antibody, in animal feed or water, distribute to wrap and prevented that with essence thereby the adhesion of described immunogen in described animal intestinal from promoting the growth of described animal with the carrier mass of the described content that contains antibody and animal feed or water with the described carrier mass of the described content bag quilt that contains antibody and supply.
13. the method for claim 12, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
14. the ability by suppressing immunogen adherence animal cud or intestinal is to reduce the ability of described immunogen breeding, thereby reduce owing in consumption animal cud or intestinal, there being the proteic consumption of consumption clone's formation property, that the proteic immunogen of consumption causes, promote the method for consumption growth of animal, described method comprises:
A. with described particular target consume proteic immunogen or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird, produce at the immunogenic antibody of described target;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described ovum separates with shell;
E., a kind of exsiccant feed vector material is provided;
F. with the content bag that contains antibody of described results ovum by described exsiccant feed vector material;
G. distribute bag in fact equably by the described carrier mass that obtains by animal feed with the content that contains antibody of described ovum; With
H. the carrier mass and the animal feed of the content bag quilt that contains antibody of the described results ovum of the described usefulness that obtains are supplied with the consumption animal, preventing the adhesion of described target immunogen in described animal intestinal in fact, thereby promote the growth of described animal.
15. the method for claim 14, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
16. one kind by reducing the ability of described immunogen breeding, thereby reduce owing in consumption animal cud or intestinal, existing and consume the proteic consumption of consumption that proteic immunogen causes, promote the method for consumption growth of animal, the proteic immunogen of described consumption is the P antigen from Streptococcus anaerobius, and described method comprises:
A. use from the P antigen of Streptococcus anaerobius or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird and the ovum that produces by described bird, produce at from the antigenic antibody of the P of Streptococcus anaerobius;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described results ovum separates with shell;
E., a kind of exsiccant feed vector material is provided;
F. with the content bag that contains antibody of described results ovum by described exsiccant feed vector material;
G. in animal feed, distribute bag in fact equably by described carrier mass with the content that contains antibody of described results ovum; With
H. the dry support material and the animal feed of the content bag quilt that contains antibody of the described results ovum of the described usefulness that obtains are supplied with the consumption animal, preventing the adhesion of described immunogen in described animal intestinal in fact, thereby promote the growth of described animal.
17. the method for claim 16, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
18. the ability by suppressing immunogen adherence consumption animal cud or intestinal is to reduce the ability of described immunogen breeding, thereby reduce owing in consumption animal cud or intestinal, existing and consume the proteic consumption of consumption that proteic immunogen causes, promote the method for consumption growth of animal, the proteic immunogen of described consumption is that described method comprises from the clostridial CS antigen of Si Shi:
A. use from the clostridial CS antigen of Si Shi or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird and the ovum that produces by described bird, produce at from the antigenic antibody of the clostridial CS of Si Shi;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described results ovum separates with shell;
E., a kind of exsiccant feed vector material is provided;
F. with the content bag that contains antibody of described results ovum by described exsiccant feed vector material;
G. in animal feed, distribute bag in fact equably by described carrier mass with the content that contains antibody of described ovum; With
H. the described content that contains antibody that obtains and animal feed are supplied with the consumption animal, preventing the adhesion of described immunogen in described animal intestinal in fact, thereby promote the growth of described animal.
19. the method for claim 18, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
20. the ability by suppressing immunogen adherence consumption animal cud or intestinal is to reduce the ability of described immunogen breeding, thereby reduce owing in consumption animal cud or intestinal, existing and consume the proteic consumption of consumption that proteic immunogen causes, promote the method for consumption growth of animal, the proteic immunogen of described consumption is from having a liking for the clostridial CA antigen of amine, and described method comprises:
A. use from have a liking for the clostridial CA antigen of amine or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird and ovum, produce at having a liking for the antigenic antibody of the clostridial CA of amine by described bird product;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described results ovum separates with chorion;
E., a kind of exsiccant feed vector material is provided;
F. with the content bag that contains antibody of described results ovum by described exsiccant feed vector material;
G. in animal feed, distribute bag in fact equably by described carrier mass with the content that contains antibody of described ovum; With
H. the described content that contains antibody that obtains and animal feed are supplied with the consumption animal, preventing the adhesion of described immunogen in described animal intestinal in fact, thereby promote the growth of described animal.
21. the method for claim 20, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector.
22. one kind by suppress immunogen adherence to the ability of the cud of consumption animal or intestinal to reduce described immunogen fertility reduces or eliminates the disease incidence that is caused by the target clone formation property that exists, pathogenic immunogen in fact in meat method, described method comprises:
A. with the described specific pathogenic immunogen of target or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird, produce at the immunogenic antibody of described target;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described ovum separates with shell;
E. with the described isolating content drying that contains antibody of described ovum;
F. by animal feed or the water described exsiccant ovum antibody product that obtains that distributes in fact equably; With
G. described animal feed that contains antibody that obtains or water are supplied with the consumption animal, to prevent the adhesion of described target immunogen in described animal intestinal in fact.
23. the method for claim 22, wherein: described pathogenic immunogen is selected from escherichia coli, and Listera belongs to (Listeria), Salmonella (Salmonella) and campylobacter (Campylobacter).
24. the method for claim 22, it comprises: a kind of exsiccant feed vector is provided, and the described drying of the content that contains antibody of described ovum is to be realized by described exsiccant feed vector by the described isolating content bag that contains antibody with described ovum.
25. the method for claim 24, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
26. one kind by suppress the escherichia coli immunogen adherence to the ability of the cud of consumption animal or intestinal to reduce described escherichia coli immunogen fertility reduces or eliminates the disease incidence that is caused by the clone's formation property that exists, Escherichia coli immunogen in fact in meat method, described method comprises:
A. with described Escherichia coli immunogen or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird, produce at the immunogenic antibody of described escherichia coli;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described ovum separates with shell;
E. with described isolating ovum antibody materials drying;
F. by animal feed or the water described exsiccant ovum antibody product that obtains that distributes in fact equably; With
G. described animal feed that contains antibody that obtains or water are supplied with the consumption animal, to prevent the adhesion of described escherichia coli immunogen in described animal intestinal in fact.
27. the method for claim 5, it comprises: a kind of exsiccant feed vector is provided, and the described drying of the content that contains antibody of described ovum is to be realized by described exsiccant feed vector by the described isolating content bag that contains antibody with described ovum.
28. the method for claim 27, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
29. one kind belongs to immunogen adherence and belongs to the immunogen fertility and reduce or eliminate the method that is belonged to the disease incidence that immunogen causes in meat by the clone's formation property that exists, pathogenic Listera in fact to reduce described Listera to the ability of the cud of consumption animal or intestinal by suppressing Listera, described method comprises:
A. with described pathogenic Listera belong to immunogen or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird, produce and belong to immunogenic antibody at described Listera;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described ovum separates with shell;
E. with described isolating ovum antibody materials drying;
F. by animal feed or the water described exsiccant ovum antibody product that obtains that distributes in fact equably; With
G. described animal feed that contains antibody that obtains or water are supplied with the consumption animal, belong to the adhesion of immunogen in described animal intestinal to prevent described Listera in fact.
30. the method for claim 29, it comprises: a kind of exsiccant feed vector is provided, and the described drying of the content that contains antibody of described ovum is to be realized by described exsiccant feed vector by the described isolating content bag that contains antibody with described ovum.
31. the method for claim 30, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
32. one kind is come essence to reduce or eliminate the method that by clone the forming property, the pathogenic salmonella that in meat exist belonged to disease incidence that immunogen cause to the ability of the cud of consumption animal or intestinal to reduce described Salmonella immunogen fertility by suppressing the Salmonella immunogen adherence, described method comprises:
A. with described pathogenic salmonella belong to immunogen or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird, produce at the immunogenic antibody of described Salmonella;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described ovum separates with shell;
E. with described isolating ovum antibody materials drying;
F. by animal feed or the water described exsiccant ovum antibody product that obtains that distributes in fact equably; With
G. described animal feed that contains antibody that obtains or water are supplied with the consumption animal, to prevent the adhesion of described Salmonella immunogen in described animal intestinal in fact.
33. the method for claim 32, it comprises: a kind of exsiccant feed vector is provided, and the described drying of the content that contains antibody of described ovum is to be realized by described exsiccant feed vector by the described isolating content bag that contains antibody with described ovum.
34. the method for claim 33, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
35. one kind is come essence to reduce or eliminate the method for the disease incidence that by clone the forming property that in meat exist, pathogenic campylobacter immunogen caused to the cud of consumption animal or the ability of intestinal to reduce described campylobacter immunogen fertility by suppressing the campylobacter immunogen adherence, described method comprises:
A. with described pathogenic campylobacter immunogen or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird, produce at the immunogenic antibody of described campylobacter;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described ovum separates with shell;
E. with described isolating ovum antibody materials drying;
F. by animal feed or the water described exsiccant ovum antibody product that obtains that distributes in fact equably; With
G. described animal feed that contains antibody that obtains or water are supplied with the consumption animal, to prevent the adhesion of described campylobacter immunogen in described animal intestinal in fact.
36. the method for claim 35, it comprises: a kind of exsiccant feed vector is provided, and the described drying of the content that contains antibody of described ovum is to be realized by described exsiccant feed vector by the described isolating content bag that contains antibody with described ovum.
37. the method for claim 36, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
38. one kind by suppressing immunogen adherence consumption animal cud or the ability of intestinal come essence to reduce or eliminate the method for the disease incidence that causes by the target clone formation property that in meat, exists, pathogenic immunogen with the ability that reduces described immunogen breeding, described method comprises:
A. with the pathogenic immunogen of described particular target or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird, produce at the immunogenic antibody of described target;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described ovum separates with shell;
E., a kind of exsiccant feed vector material is provided;
F. with the content bag that contains antibody of described ovum by described exsiccant feed vector material;
G. in animal feed, distribute bag in fact equably by described carrier mass with the content that contains antibody of described ovum; With
H. the exsiccant carrier mass and the animal feed of the content bag quilt that contains antibody of the described described ovum of usefulness that obtains are supplied with the consumption animal, to prevent that in fact described immunogen is in cud of described animal or the adhesion in the intestinal.
39. the method for claim 38, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
40. a method of producing the microorganism adhering inhibitor, described microorganism adhering inhibitor are used for the consumption animals administer is prevented the adhesion of target clone formation property immunogen at cud or the intestinal of described consumption animal with essence that this method comprises:
A. with described particular target clone form the property immunogen or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird, produce at the immunogenic antibody of described target;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described ovum separates with shell; With
E. the described isolating content that contains antibody of dry described ovum.
41. the method for claim 40, wherein: the ability that described clone's formation property immunogen is known reduction animal use edible protein a kind of.
42. the method for claim 41, wherein: described clone's formation property immunogen is from peptostreptococcus anaerobius, Si Shi clostridium and have a liking for the amine clostridium.
43. according to the method for claim 40, wherein: described clone's formation property immunogen is known a kind of of food-borne disease that cause in the people.
44. the method for claim 43, wherein: described clone's formation property immunogen is from escherichia coli, and Listera belongs to, Salmonella and campylobacter.
45. the method for claim 40, it comprises: a kind of exsiccant carrier mass is provided, and the isolating described drying that contains the content of antibody of described ovum is to be realized by described exsiccant carrier mass by the isolating content bag that contains antibody with described ovum.
46. the method for claim 45, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
47. method of producing the microorganism adhering inhibitor, described microorganism adhering inhibitor is used for the consumption animals administer is prevented to clone the adhesion of formation property immunogen at cud or the intestinal of described consumption animal with essence, described immunogen is the P antigen from peptostreptococcus anaerobius, and this method comprises:
A. use from the P antigen of peptostreptococcus anaerobius or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird and the ovum that produces by described bird, produce at from the antigenic antibody of the P of peptostreptococcus anaerobius;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with the ovum of described results separates with chorion; With
E. the dry described content that contains antibody.
48. the method for claim 47, it comprises: a kind of exsiccant carrier mass is provided, and the isolating described drying that contains the content of antibody of described ovum is to be realized by described exsiccant carrier mass by the isolating content bag that contains antibody with described ovum.
49. the method for claim 48, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
50. method of producing the microorganism adhering inhibitor, described microorganism adhering inhibitor is used for the consumption animals administer is prevented to clone the adhesion of formation property immunogen at cud or the intestinal of described consumption animal with essence, described immunogen is that described method comprises from the clostridial CS antigen of Si Shi:
A. use from the clostridial CS antigen of Si Shi or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird and the ovum that produces by described bird, produce at from the antigenic antibody of the clostridial CS of Si Shi;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with the ovum of described results separates with chorion; With
E. the dry described content that contains antibody.
51. the method for claim 50, it comprises: a kind of exsiccant carrier mass is provided, and the isolating described drying that contains the content of antibody of described ovum is to be realized by described exsiccant carrier mass by the isolating content bag that contains antibody with described ovum.
52. the method for claim 51, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
53. method of producing the microorganism adhering inhibitor, described microorganism adhering inhibitor is used for the consumption animals administer is prevented to clone the adhesion of formation property immunogen at cud or the intestinal of described consumption animal with essence, described immunogen is from having a liking for the clostridial CA antigen of amine, and described method comprises:
A. use from have a liking for the clostridial CA antigen of amine or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird and the ovum that produces by described bird, produce at from having a liking for the antigenic antibody of the clostridial CA of amine;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with the ovum of described results separates with chorion; With
E. the dry described content that contains antibody.
54. the method for claim 53, it comprises: a kind of exsiccant carrier mass is provided, and the isolating described drying that contains the content of antibody of described ovum is to be realized by described exsiccant carrier mass by the isolating content bag that contains antibody with described ovum.
55. the method for claim 54, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
56. method of producing the microorganism adhering inhibitor, described microorganism adhering inhibitor is used for the consumption animals administer is prevented to clone the adhesion of formation property immunogen at cud or the intestinal of described consumption animal with essence, described immunogen is from colibacillary bacillus coli antigen, and described method comprises:
A. with described escherichia coli cloning form the property immunogen or approximately reach their egg-laying season inoculation hen;
B. one section permission produces time at the immunogenic antibody of described escherichia coli in described bird after, the ovum that results are produced by described bird;
C. the content that contains antibody with the ovum of described results separates with shell; With
D. dry described isolating ovum antibody adhesion inhibition material.
57. the method for claim 56, it comprises: a kind of exsiccant carrier mass is provided, and the isolating described drying that contains the content of antibody of described ovum is to be realized by described exsiccant carrier mass by the isolating content bag that contains antibody with described ovum.
58. the method for claim 57, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
59. method of producing the microorganism adhering inhibitor, described microorganism adhering inhibitor is used for the consumption animals administer is prevented to clone the adhesion of formation property immunogen at cud or the intestinal of described consumption animal with essence, described immunogen is to belong to antigen from the Listera that Listera belongs to, and described method comprises:
A. with described Listera belong to the clone form the property immunogen or approximately reach their egg-laying season inoculation hen;
B. one section permission in described bird, produce belong to the time of immunogenic antibody at described Listera after, the ovum that results are produced by described bird;
C. the content that contains antibody with the ovum of described results separates with shell; With
D. dry described isolating ovum antibody adhesion inhibition material.
60. the method for claim 59, it comprises: a kind of exsiccant carrier mass is provided, and the isolating described drying that contains the content of antibody of described ovum is to be realized by described exsiccant carrier mass by the isolating content bag that contains antibody with described ovum.
61. the method for claim 60, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
62. method of producing the microorganism adhering inhibitor, described microorganism adhering inhibitor is used for the consumption animals administer is prevented to clone the adhesion of formation property immunogen at cud or the intestinal of described consumption animal with essence, described immunogen is the Salmonella antigen from Salmonella, and described method comprises:
A. with described Salmonella clone form the property immunogen or approximately reach their egg-laying season inoculation hen;
B. one section permission produces time at the immunogenic antibody of described Salmonella in described bird after, the ovum that results are produced by described bird;
C. the content that contains antibody with the ovum of described results separates with shell; With
D. dry described isolating ovum antibody adhesion inhibition material.
63. the method for claim 62, it comprises: a kind of exsiccant carrier mass is provided, and the isolating described drying that contains the content of antibody of described ovum is to be realized by described exsiccant carrier mass by the isolating content bag that contains antibody with described ovum.
64. the method for claim 63, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
65. method of producing the microorganism adhering inhibitor, described microorganism adhering inhibitor is used for the consumption animals administer is prevented to clone the adhesion of formation property immunogen at cud or the intestinal of described consumption animal with essence, described immunogen is the campylobacter antigen from campylobacter, and described method comprises:
A. with described campylobacter clone form the property immunogen or approximately reach their egg-laying season inoculation hen;
B. one section permission produces time at the immunogenic antibody of described campylobacter in described bird after, the ovum that results are produced by described bird;
C. the content that contains antibody with the ovum of described results separates with shell; With
D. dry described isolating ovum antibody adhesion inhibition material.
66. the method for claim 65, it comprises: a kind of exsiccant carrier mass is provided, and the isolating described drying that contains the content of antibody of described ovum is to be realized by described exsiccant carrier mass by the isolating content bag that contains antibody with described ovum.
67. the method for claim 66, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
68. a method of producing the microorganism adhering inhibitor, described microorganism adhering inhibitor are used for the consumption animals administer is prevented the adhesion of target clone formation property immunogen at cud or the intestinal of described consumption animal with essence that this method comprises:
A. with described specific target clone form the property immunogen or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough in described bird, produce at the immunogenic antibody of described target;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described ovum separates with shell;
E., a kind of exsiccant carrier mass is provided; With
F. with the content bag that contains antibody of described ovum by described exsiccant carrier mass.
69. the method for claim 68, wherein: from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp provides a kind of exsiccant feed vector material.
70. one kind is used for the consumption animals administer is prevented that with essence target clone from forming the property immunogen at the cud of described consumption animal or the adherent microorganism adhering inhibitor of intestinal, it is by the method production of following step:
A. with described specific target clone form the property immunogen or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough in described bird, produce at the immunogenic antibody of described target;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described ovum separates with shell; With
E. the described isolating content that contains antibody of dry described ovum.
71. the microorganism adhering inhibitor of claim 76, wherein: the ability that described clone's formation property immunogen is known reduction animal use edible protein a kind of.
72. the microorganism adhering inhibitor of claim 71, wherein: described clone's formation property immunogen is from peptostreptococcus anaerobius, Si Shi clostridium and have a liking for the amine clostridium.
73. the microorganism adhering inhibitor of claim 70, wherein: described clone's formation property immunogen is known a kind of of food-borne disease that cause in the people.
74. the microorganism adhering inhibitor of claim 73, wherein: described clone's formation property immunogen is from escherichia coli, and Listera belongs to, Salmonella and campylobacter.
75. the microorganism adhering inhibitor of claim 70, wherein: the isolating described drying that contains the content of antibody of described ovum is to realize by the feed vector material that the content bag that contains antibody with described ovum is dried.
76. the microorganism adhering inhibitor of claim 75, wherein: described exsiccant feed vector material is from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp.
77. microorganism adhering inhibitor, it is used for consumption animals administer essence is prevented the adhesion of target clone formation property immunogen at cud or the intestinal of described consumption animal that described inhibitor comprises the exsiccant ovum content that is specific to the immunogenic antibody of described target that combines.
78. the microorganism adhering inhibitor of claim 77, wherein: the ability that described clone's formation property immunogen is known reduction animal use edible protein a kind of.
79. the microorganism adhering inhibitor of claim 78, wherein: described clone's formation property immunogen is from peptostreptococcus anaerobius, Si Shi clostridium and have a liking for the amine clostridium.
80. the microorganism adhering inhibitor of claim 78, wherein: described clone's formation property immunogen is known a kind of of food-borne disease that cause in the people.
81. the microorganism adhering inhibitor of claim 80, wherein: described clone's formation property immunogen is from escherichia coli, and Listera belongs to, Salmonella and campylobacter.
82. one kind by suppress immunogen adherence to the ability of the cud of consumption animal or intestinal to reduce the ability that described immunogen is bred, thereby reduce because the proteic consumption of consumption that in the cud of described consumption animal or intestinal, exists the proteic immunogen of consumption to cause, promote the microorganism adhering inhibitor of consumption growth of animal, the proteic immunogen of described consumption is the P antigen from Streptococcus anaerobius, and described microorganism adhering inhibitor is by comprising the method production of following step:
A. use from the P antigen of Streptococcus anaerobius or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird and the ovum that produces by described bird, produce at from the antigenic antibody of the P of Streptococcus anaerobius;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described ovum separates with shell; With
E. the described content that contains antibody of dry described ovum.
83. the microorganism adhering inhibitor of claim 82, wherein: the isolating described drying that contains the content of antibody of described ovum is to realize by the feed vector material that the content bag that contains antibody with described ovum is dried.
84. the microorganism adhering inhibitor of claim 83, wherein: described exsiccant feed vector material is from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp.
85. one kind by suppress immunogen adherence to the ability of the cud of consumption animal or intestinal to reduce the ability that described immunogen is bred, thereby reduce because the proteic consumption of consumption that in the cud of described consumption animal or intestinal, exists the proteic immunogen of consumption to cause, promote the microorganism adhering inhibitor of consumption growth of animal, the proteic immunogen of described consumption is from the clostridial CS antigen of Si Shi, and described microorganism adhering inhibitor is by comprising the method production of following step:
A. use from the clostridial CS antigen of Si Shi or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird and the ovum that produces by described bird, produce at from the antigenic antibody of the clostridial CS of Si Shi;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described ovum separates with shell; With
E. the described content that contains antibody of dry described ovum.
86. the microorganism adhering inhibitor of claim 85, wherein: the isolating described drying that contains the content of antibody of described ovum is to realize by the feed vector material that the content bag that contains antibody with described ovum is dried.
87. the microorganism adhering inhibitor of claim 86, wherein: described exsiccant feed vector material is from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp.
88. one kind by suppress immunogen adherence to the ability of the cud of consumption animal or intestinal to reduce the ability that described immunogen is bred, thereby reduce because the proteic consumption of consumption that in the cud of described consumption animal or intestinal, exists the proteic immunogen of consumption to cause, promote the microorganism adhering inhibitor of consumption growth of animal, the proteic immunogen of described consumption is from having a liking for the clostridial CA antigen of amine, and described microorganism adhering inhibitor is by comprising the method production of following step:
A. use from have a liking for the clostridial CA antigen of amine or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird and the ovum that produces by described bird, produce at from having a liking for the antigenic antibody of the clostridial CA of amine;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described ovum separates with shell; With
E. the described content that contains antibody of dry described ovum.
89. the microorganism adhering inhibitor of claim 88, wherein: the isolating described drying that contains the content of antibody of described ovum is to realize by the feed vector material that the content bag that contains antibody with described ovum is dried.
90. the microorganism adhering inhibitor of claim 89, wherein: described exsiccant feed vector material is from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp.
91. one kind is used for the consumption animals administer is prevented cud or the intestinal adherent microorganism adhering inhibitor of target clone formation property immunogen described consumption animal with essence, described microorganism adhering inhibitor is by comprising the method production of following step:
A. with described specific target clone form the property immunogen or approximately reach their egg-laying season inoculation hen;
B. allow a period of time to be enough to allow in described bird, produce at the immunogenic antibody of described target;
C. gather in the crops the ovum that produces by described bird;
D. the content that contains antibody with described ovum separates with shell;
E., a kind of exsiccant feed vector material is provided; With
F. with the content bag that contains antibody of described ovum by described exsiccant feed vector material.
92. the microorganism adhering inhibitor of claim 91, wherein: described exsiccant feed vector material is from comprising the bean shell, rice husk, corn, cotton seed hulls, one group of material of the corn of evaporate to dryness and beet pulp.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/US2001/049588 WO2003061693A1 (en) | 2001-12-28 | 2001-12-28 | Immunogen adherence and method of making and using same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1543355A true CN1543355A (en) | 2004-11-03 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA018235352A Pending CN1543355A (en) | 2001-12-28 | 2001-12-28 | Immunogen adherence and method of making and using same |
Country Status (8)
| Country | Link |
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| EP (1) | EP1476184A4 (en) |
| JP (1) | JP2006504393A (en) |
| KR (1) | KR20040096491A (en) |
| CN (1) | CN1543355A (en) |
| AU (1) | AU2002246753B8 (en) |
| BR (1) | BR0117062A (en) |
| CA (1) | CA2438698C (en) |
| WO (1) | WO2003061693A1 (en) |
Family Cites Families (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4748018A (en) * | 1984-02-07 | 1988-05-31 | Stolle Research & Development Corp. | Method of passive immunization of mammals using avian antibody |
| EP0225254B1 (en) * | 1985-11-25 | 1994-03-16 | Ghen Corporation | Specific antibody-containing substance from eggs and method of production and use thereof |
| US5753228A (en) * | 1992-08-25 | 1998-05-19 | Arizona Board Of Regents On Behalf Of The University Of Arizona | Method of treating parasitosis by the enteral administration of hyperimmune hen egg yolk antibodies |
| JPH11506424A (en) * | 1995-03-24 | 1999-06-08 | オフィディアン ファーマシューティカルズ インコーポレーテッド | Treatment for Verotoxin-producing Escherichia coli |
| US5741489A (en) * | 1996-05-24 | 1998-04-21 | Anitox Corporation | Passively administered antibody that enhances feed conversion efficiency |
| US5725873A (en) * | 1996-07-22 | 1998-03-10 | Wisconsin Alumni Research Foundation | Method of improving the growth or the efficiency of feed conversion of an animal and compositions for use therein |
| WO1998014209A1 (en) * | 1996-10-02 | 1998-04-09 | Ovimmune, Inc. | Oral administration of chicken yolk antibodies to treat disease |
| US6083500A (en) * | 1997-10-06 | 2000-07-04 | University Of Georgia Research Foundation, Inc. | Biological control of food pathogens in livestock |
| CZ287989B6 (en) * | 1998-03-20 | 2001-03-14 | Medipharm Cz, S. R. O. | Peroral preparation intended for prevention and treatment of infectious gastroenteritis in pigs |
| DE19910159A1 (en) * | 1999-02-26 | 2000-09-14 | Rosemarie Heis | Specific IgY egg yolk antibodies, their extraction and their use |
| US6162441A (en) * | 1999-12-15 | 2000-12-19 | Republic Of Korea (Management: Rural Development Administration) | Method for the production of anti-escherichia. coli O157 : H7 antibody |
| CA2395840A1 (en) * | 1999-12-27 | 2001-07-05 | The University Of Manitoba | Genetic vaccines for the production of chicken egg-yolk antibodies against enterotoxigenic escherichia coli and other pathogens |
-
2001
- 2001-12-28 WO PCT/US2001/049588 patent/WO2003061693A1/en active Application Filing
- 2001-12-28 JP JP2003561636A patent/JP2006504393A/en active Pending
- 2001-12-28 CA CA2438698A patent/CA2438698C/en not_active Expired - Lifetime
- 2001-12-28 EP EP01994346A patent/EP1476184A4/en not_active Withdrawn
- 2001-12-28 KR KR10-2004-7006486A patent/KR20040096491A/en not_active Application Discontinuation
- 2001-12-28 AU AU2002246753A patent/AU2002246753B8/en not_active Expired
- 2001-12-28 BR BR0117062-7A patent/BR0117062A/en not_active Application Discontinuation
- 2001-12-28 CN CNA018235352A patent/CN1543355A/en active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| CA2438698A1 (en) | 2003-07-31 |
| JP2006504393A (en) | 2006-02-09 |
| CA2438698C (en) | 2013-12-17 |
| EP1476184A4 (en) | 2006-01-04 |
| BR0117062A (en) | 2005-12-13 |
| WO2003061693A1 (en) | 2003-07-31 |
| AU2002246753A1 (en) | 2003-09-02 |
| AU2002246753B2 (en) | 2009-05-28 |
| KR20040096491A (en) | 2004-11-16 |
| AU2002246753B8 (en) | 2009-09-24 |
| EP1476184A1 (en) | 2004-11-17 |
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