CN102174408B - Method for batch preparation of chicken Eimeria coccidium oocysts - Google Patents
Method for batch preparation of chicken Eimeria coccidium oocysts Download PDFInfo
- Publication number
- CN102174408B CN102174408B CN2011100292716A CN201110029271A CN102174408B CN 102174408 B CN102174408 B CN 102174408B CN 2011100292716 A CN2011100292716 A CN 2011100292716A CN 201110029271 A CN201110029271 A CN 201110029271A CN 102174408 B CN102174408 B CN 102174408B
- Authority
- CN
- China
- Prior art keywords
- layer
- centrifugal
- water
- egg capsule
- chicken
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241000287828 Gallus gallus Species 0.000 title claims abstract description 43
- 241000223924 Eimeria Species 0.000 title claims abstract description 19
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 238000000034 method Methods 0.000 title abstract description 44
- 210000003250 oocyst Anatomy 0.000 title abstract description 12
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 claims abstract description 20
- 238000012545 processing Methods 0.000 claims abstract description 5
- 238000012216 screening Methods 0.000 claims abstract description 5
- 239000002775 capsule Substances 0.000 claims description 47
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 38
- 210000003608 fece Anatomy 0.000 claims description 32
- 239000007788 liquid Substances 0.000 claims description 29
- 239000010871 livestock manure Substances 0.000 claims description 25
- 238000002156 mixing Methods 0.000 claims description 17
- 239000000126 substance Substances 0.000 claims description 10
- 239000006228 supernatant Substances 0.000 claims description 8
- 239000002689 soil Substances 0.000 claims description 6
- 238000005119 centrifugation Methods 0.000 claims description 4
- 239000004744 fabric Substances 0.000 claims description 4
- 210000002784 stomach Anatomy 0.000 claims description 4
- 238000011010 flushing procedure Methods 0.000 claims description 3
- 238000010790 dilution Methods 0.000 claims description 2
- 239000012895 dilution Substances 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 15
- 229960005486 vaccine Drugs 0.000 abstract description 14
- 238000000926 separation method Methods 0.000 abstract description 6
- 238000000151 deposition Methods 0.000 abstract description 3
- 239000000427 antigen Substances 0.000 abstract description 2
- 102000036639 antigens Human genes 0.000 abstract description 2
- 108091007433 antigens Proteins 0.000 abstract description 2
- 238000000746 purification Methods 0.000 abstract description 2
- 230000002550 fecal effect Effects 0.000 abstract 1
- 235000013330 chicken meat Nutrition 0.000 description 36
- 208000003495 Coccidiosis Diseases 0.000 description 14
- 206010023076 Isosporiasis Diseases 0.000 description 14
- 229920006395 saturated elastomer Polymers 0.000 description 14
- 229930006000 Sucrose Natural products 0.000 description 12
- 239000005720 sucrose Substances 0.000 description 12
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 11
- 239000003814 drug Substances 0.000 description 10
- 238000005516 engineering process Methods 0.000 description 10
- 239000000243 solution Substances 0.000 description 10
- 241000224483 Coccidia Species 0.000 description 9
- 229940079593 drug Drugs 0.000 description 9
- 239000012535 impurity Substances 0.000 description 9
- 230000001165 anti-coccidial effect Effects 0.000 description 8
- 239000002253 acid Substances 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 238000000432 density-gradient centrifugation Methods 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 230000002265 prevention Effects 0.000 description 4
- 241000223934 Eimeria maxima Species 0.000 description 3
- 241000223932 Eimeria tenella Species 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 244000144977 poultry Species 0.000 description 3
- 235000013594 poultry meat Nutrition 0.000 description 3
- 239000002893 slag Substances 0.000 description 3
- 241000251468 Actinopterygii Species 0.000 description 2
- 229930191564 Monensin Natural products 0.000 description 2
- GAOZTHIDHYLHMS-UHFFFAOYSA-N Monensin A Natural products O1C(CC)(C2C(CC(O2)C2C(CC(C)C(O)(CO)O2)C)C)CCC1C(O1)(C)CCC21CC(O)C(C)C(C(C)C(OC)C(C)C(O)=O)O2 GAOZTHIDHYLHMS-UHFFFAOYSA-N 0.000 description 2
- UKHWDRMMMYWSFL-UHFFFAOYSA-N Nicarbazin Chemical compound CC=1C=C(C)NC(=O)N=1.C1=CC([N+](=O)[O-])=CC=C1NC(=O)NC1=CC=C([N+]([O-])=O)C=C1 UKHWDRMMMYWSFL-UHFFFAOYSA-N 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 230000008021 deposition Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000011031 large-scale manufacturing process Methods 0.000 description 2
- 229960005358 monensin Drugs 0.000 description 2
- GAOZTHIDHYLHMS-KEOBGNEYSA-N monensin A Chemical compound C([C@@](O1)(C)[C@H]2CC[C@@](O2)(CC)[C@H]2[C@H](C[C@@H](O2)[C@@H]2[C@H](C[C@@H](C)[C@](O)(CO)O2)C)C)C[C@@]21C[C@H](O)[C@@H](C)[C@@H]([C@@H](C)[C@@H](OC)[C@H](C)C(O)=O)O2 GAOZTHIDHYLHMS-KEOBGNEYSA-N 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000009987 spinning Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- MOOFYEJFXBSZGE-QJUDHZBZSA-N 1,2-bis[(z)-(4-chlorophenyl)methylideneamino]guanidine Chemical compound C=1C=C(Cl)C=CC=1\C=N/N=C(/N)N\N=C/C1=CC=C(Cl)C=C1 MOOFYEJFXBSZGE-QJUDHZBZSA-N 0.000 description 1
- ZSZFUDFOPOMEET-UHFFFAOYSA-N 2-(4-chlorophenyl)-2-[2,6-dichloro-4-(3,5-dioxo-1,2,4-triazin-2-yl)phenyl]acetonitrile Chemical compound C1=CC(Cl)=CC=C1C(C#N)C1=C(Cl)C=C(N2C(NC(=O)C=N2)=O)C=C1Cl ZSZFUDFOPOMEET-UHFFFAOYSA-N 0.000 description 1
- 208000019901 Anxiety disease Diseases 0.000 description 1
- 206010003694 Atrophy Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- ZEFNOZRLAWVAQF-UHFFFAOYSA-N Dinitolmide Chemical compound CC1=C(C(N)=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O ZEFNOZRLAWVAQF-UHFFFAOYSA-N 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 241000223931 Eimeria acervulina Species 0.000 description 1
- 241000179199 Eimeria mitis Species 0.000 description 1
- 241000499563 Eimeria necatrix Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- KQXDHUJYNAXLNZ-XQSDOZFQSA-N Salinomycin Chemical compound O1[C@@H]([C@@H](CC)C(O)=O)CC[C@H](C)[C@@H]1[C@@H](C)[C@H](O)[C@H](C)C(=O)[C@H](CC)[C@@H]1[C@@H](C)C[C@@H](C)[C@@]2(C=C[C@@H](O)[C@@]3(O[C@@](C)(CC3)[C@@H]3O[C@@H](C)[C@@](O)(CC)CC3)O2)O1 KQXDHUJYNAXLNZ-XQSDOZFQSA-N 0.000 description 1
- 239000004189 Salinomycin Substances 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000036506 anxiety Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000037444 atrophy Effects 0.000 description 1
- 238000009361 aviculture Methods 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000013872 defecation Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 229960000248 diclazuril Drugs 0.000 description 1
- 229960003934 dinitolmide Drugs 0.000 description 1
- 239000003640 drug residue Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 231100000225 lethality Toxicity 0.000 description 1
- 230000002101 lytic effect Effects 0.000 description 1
- RWVUEZAROXKXRT-VQLSFVLHSA-N maduramicin Chemical compound O1[C@@H](C)[C@H](OC)[C@@H](OC)C[C@H]1O[C@@H]1[C@H]([C@@]2(C)O[C@H](CC2)[C@@]2(C)O[C@]3(O[C@@H]([C@H](C)[C@@H](O)C3)[C@@H](C)[C@H]3[C@@H]([C@@H](OC)[C@H](C)[C@@](O)(CC(O)=O)O3)OC)CC2)O[C@@H]([C@@H]2[C@H](C[C@@H](C)[C@@](C)(O)O2)C)C1 RWVUEZAROXKXRT-VQLSFVLHSA-N 0.000 description 1
- 229950006915 maduramicin Drugs 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 210000004681 ovum Anatomy 0.000 description 1
- 230000024241 parasitism Effects 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 230000008447 perception Effects 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 229960004591 robenidine Drugs 0.000 description 1
- 229960001548 salinomycin Drugs 0.000 description 1
- 235000019378 salinomycin Nutrition 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- GOLXNESZZPUPJE-UHFFFAOYSA-N spiromesifen Chemical compound CC1=CC(C)=CC(C)=C1C(C(O1)=O)=C(OC(=O)CC(C)(C)C)C11CCCC1 GOLXNESZZPUPJE-UHFFFAOYSA-N 0.000 description 1
- 125000000185 sucrose group Chemical group 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 229940031418 trivalent vaccine Drugs 0.000 description 1
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses a method for batch preparation of chicken Eimeria coccidium oocysts, and particularly relates to a separation and purification method for producing oocysts (antigens) of chicken Eimeria coccidium vaccines. The method for preparing the chicken Eimeria coccidium oocysts comprises the following process steps: fecal sample processing, continuous screening, saturated brine float centrifuging, oocyst centrifugal depositing and oocyst purifying. According to the method, the industrial preparation of the chicken Eimeria coccidium oocysts can be realized, the production efficiency can be improved, and the cost can be reduced.
Description
Technical field
A kind of chicken Eimeria egg capsule batch preparation relates to the separation purification method of the egg capsule (antigen) of producing chicken Eimeria vaccine.
Background technology
Sick about the chicken Eimeria
Eimeria (Eimeria) coccidia (Coccidium) is the one type of obligatory parasitism protozoon that parasitizes the chicken enteron aisle, causes coccidiosis of chicken (Coccidiosis).According to statistics; The chicken coccidia that the whole world has been reported has 15 kinds; Wherein, Have 5 kinds to have significantly pathogenicly, they are: Eimeria tenella (E.tenella), poison Eimeria (E.necatrix), Eimeria maxima (E.maxima), heap shape Eimeria (E.acervulina) and gentle Eimeria (E.mitis).In the pathogenic coccidia of 5 kinds of tools, there are 3 kinds to chick tool hazardness, they are: Eimeria tenella, Eimeria maxima and heap shape Eimeria.
Coccidiosis is a kind of ubiquitous infectious eqpidemic disease, and every in the world have the place of chicken just to have the chicken coccidia, and the generation of coccidiosis is just arranged, and the outburst of this disease usually appears in the mass-producing chicken house incident suitable place that is coccidiosis of chicken.Coccidiosis of chicken is that a kind of infection is rapid, propagates soon and extensively, and the high lytic infection of pathogenicity rate, lethality rate is sick, is preventing and treating improperly under the situation, and infection rate and mortality ratio all can be up to more than 50%.Therefore, coccidiosis of chicken causes serious economy loss to aviculture, and the annual loss that causes because of this disease in the whole world is up to tens billion of yuans, and the financial loss that China causes because of this disease every year is no less than 10,000,000,000 yuan.In today that poultry husbandry intensification degree improves constantly, coccidiosis of chicken is an indispensable link in the prevention and control of poultry husbandry eqpidemic disease.
The control of coccidiosis of chicken
From the eighties in last century, the prevention and control of coccidiosis of chicken have obtained major progress, mainly have benefited from the appearance of a large amount of anticoccidial drugs, and obtain gratifying effect.Yet; Mostly a large amount of anticoccidial drugs is chemical classes product (like Robenidine, nicarbazine, dinitolmide, diclazuril, nicarbazine and sulfa drugs) and antibiotics product (like monensin, Salinomycin., draw husky rhzomorph, that draws rhzomorph, Maduramicin, Hainan mycin), and the life-time service of these products must cause the appearance of resistance and drug residue phenomenon.And the use of various anticoccidial drugs is the administration of companion material, in fact the nineties in last century anti-medicine phenomenon very general, the development pressure pcl of new drug increases in this case, the prevention and control difficulty of the enterprise of raising chickens strengthens day by day.
Got into since 21 century; Pharmaceutical chemicals and antibiotic use are restricted in the global range; The food safety system receives increasing concern; Developed country has forbidden the use of chemical anticoccidial drug basically, and the use of antibiotics anticoccidial drug also is subject to significant restrictions, and has prohibited this series products in the U.S., Japan and the European Community and has appeared in the feed.The application that does not add restriction in China owing to various anticoccidial drugs, resistance generally produces, and the product that has (like monensin) has not had clinical use value.The food safety policy of China comes into effect in recent years; Human consumer's self health perception strengthens day by day simultaneously; The atrophy day by day of the market of anticoccidial drug; And China also will implement forbid or limit the marketing policy of using anticoccidial drug in the recent period, and coccidiosis of chicken prevents and treats situation anxiety suddenly like this.
For effective control coccidiosis of chicken, guarantee the sound development of poultry husbandry, press for the new method of preventing and treating of seeking, this wherein the vaccine inoculation prevention be one of approach that has future.Yet traditional coccidian oocyst stripping technique is not suitable for industrialization production, need be applicable to the technical matters of large-scale production in a hurry.
Existing egg capsule stripping technique
Chicken coccidial oocyst must separate just from chicken manure; Existing stripping technique has multiple, but can reduce 2 types, and one type is floating method; Another kind of is density gradient centrifugation, wherein floating saturated brine floating method, sucrose solution floating method, saturated sal epsom floating method and the saturated chromatosulfuric acid floating method of comprising; It is sucrose density gradient centrifugation that gradient centrifugation has only a kind of.
Floating method
Because the density (1.103～1.064) of coccidian oocyst is between the density of water and saturated brine (sucrose solution, saturated sal epsom and saturated chromatosulfuric acid); When the egg capsule solution that will contain impurity was placed on that whizzer is centrifugal in the water, egg capsule and a spot of material sank than water-mass density is big together, because buoyancy; Than the little impurity come-up of water-mass density; After centrifugal, abandon supernatant, removed the impurity littler than water-mass density; When the egg capsule solution that will contain impurity is placed in the saturated brine (sucrose solution, saturated sal epsom and saturated chromatosulfuric acid) when centrifugal; Because buoyancy, egg capsule and a spot of material be than the little come-up together of saturated brine density, centrifugal after; Abandon deposition, removed the impurity bigger than saturated brine density.Subsequently, isolating egg capsule is handled with Youxiaolin, removes the bur on bacterium and egg capsule surface.
Saturated brine floating method operation steps:
The 1st step: get infected chicken fresh excreta 10g, add water and be mixed into liquid manure for 10 times;
The 2nd step: liquid manure filters through 60 order sub-sieves, keeps filtrating;
The 3rd step: filtrating is supplied the water yield and is filtered through 200 order sub-sieves, keeps filtrating;
The 4th step: it is centrifugal through 2000rpm * 10min to filtrate, and abandons supernatant;
The 5th step: throw out adds saturated brine, and 2000rpm * 10min is centrifugal;
The 6th step: repeated for the 5th step;
The 7th step: get supernatant liquid and add water, 3000rpm * 15min is centrifugal, and throw out is egg capsule;
The 8th step: throw out adds water, the centrifugal flush away salinity of 3000rpm * 15min.
Sucrose solution floating method operation steps:
The 1st step: get infected chicken fresh excreta 10g, add water and be mixed into liquid manure for 2～5 times;
The 2nd step: liquid manure filters through 60 order sub-sieves, keeps filtrating;
The 3rd step: filtrating is supplied the water yield and is filtered through 100 or 200 order sub-sieves, keeps filtrating;
The 4th step: filtrating adds equivalent sucrose solution (sheatherShi liquid) and mixes, and is centrifugal through 3000rpm * 3min;
The 5th step: fish for above-mentioned centrifuge tube surface supernatant liquid in another tool water receptacle with 20～25 order dredgers;
The 6th step: 3000rpm * 3min is centrifugal, the flush away sugar soln;
The 7th step: repeated for the 6th step, throw out is egg capsule.
Saturated sal epsom floating method operation steps:
The 1st step: get infected chicken fresh excreta 10g, add water and be mixed into liquid manure for 2～5 times;
The 2nd step: liquid manure filters through 60 order sub-sieves, keeps filtrating;
The 3rd step: filtrating is supplied the water yield and is filtered through 100 or 200 order sub-sieves, keeps filtrating;
The 4th step: filtrating adds the saturated Adlerika of equivalent and mixes, and is centrifugal through 3000rpm * 3min;
The 5th step: fish for above-mentioned centrifuge tube surface supernatant liquid in another tool water receptacle with 20～25 order dredgers;
The 6th step: 3000rpm * 3min is centrifugal, the saturated Adlerika of flush away;
The 7th step: repeated for the 6th step, throw out is egg capsule.
Saturated chromatosulfuric acid floating method operation steps:
The 1st step: get infected chicken fresh excreta 10g, add water and be mixed into liquid manure for 10 times;
The 2nd step: 1500rpm * 5min is centrifugal, keeps throw out;
The 3rd step: add 4～5 times of amount chromatosulfuric acid liquid in the throw out, abundant mixing under the condition of ice bath, it is centrifugal to carry out 1500rpm * 5min at once;
The 4th step: with suction pipe sucking-off supernatant, add 20 times of frozen water, 1500rpm * 5min is centrifugal again;
The 5th step: repeat centrifugal 2 times of 1500rpm * 5min, throw out is egg capsule.
Sucrose density gradient centrifugation
Ultimate principle
Between 1.103～1.064, process density gradient according to the density of coccidian oocyst, add,, collect the egg capsule layer between two gradients through centrifugal with the isolating egg capsule of saturated brine floating method with sucrose, with density greatly and the little impurity of density separate with egg capsule.
Operation steps:
The 1st step: press the saturated brine floating method and handle ight soil, separate worm's ovum;
The 2nd step: sucrose solution gradient preparation (pressing Suo Xun, Yang Xiaoye, 2005);
The 3rd step: get the isolating concentrated egg capsule liquid 5ml of saturated brine floating method, place on the liquid level the centrifugal 30min of 1500g;
The 4th step: collect each gradient layer of inspection and deposition respectively, have each layer of egg capsule to collect respectively, with saline water or tap water centrifuge washing, collecting precipitation thing;
The 5th step: throw out is resuspended in the 5ml water, once more gradient centrifugation, repeated collection and washing process.
The analysis of existing manufacturing technology:
The saturated brine floating method is the most frequently used coccidian oocyst stripping technique in laboratory, and this technology is simple relatively, is easy to grasp favorable repeatability, reliable results.But be used for industrialization production major defect is arranged.To handle the 300kg chicken manure day is that the industrialization production of split run production capacity is example, and the liquid manure amount after the processing can reach 3300 liters, and after 60 orders and 200 mesh sieves divided, 90% liquid manure became filtrating (excrement slag＜10%), and whole filtrating reaches 2970 liters.
The whizzer of present domestic maximum capacity is 4 * 500ml (rotating speed is not less than 3000rpm), once centrifugally can only handle 2000ml, handle 2970 liters of filtratings, and a bench centrifuge needs non-stop run 1485 times; Like 1 bench centrifuge darg 10 times; Then need 148 super-magnum centrifuges, this does not also comprise follow-up a large amount of spinning and purifying, therefore; This technology obviously can not be used for large-scale production, and the vaccine of producing through this kind method uses owing to too high also being inappropriate in the production of cost.
With chicken coccidia pentavalent vaccine is example, and 3 fryer day in age week defecation are by 100 grams, and daily output 300kg ight soil needs 3000 chickens; Each plants 600, and every chicken (by 7 days) in one-period ovulates, and capsule is about can to prepare 10000 plumage vaccines, and each cycle can be produced 6,000,000 plumage vaccines; Annual by production in 300 days; Totally 42 production cycles, total can be produced vaccine 2.52 hundred million plumages, and this is the output of a medium-scale vaccine factory.
In view of this, the industrialized manufacturing technique of vaccine to chicken coccidiosis can not prolong uses conventional art, and formulating new technical process is the only way of industrialization.
The defective of sucrose solution floating method though the liquid manure amount reduces, needs the sucrose solution of adding and filtrating equivalent basically with the saturated salt water law; Still need a large amount of centrifugal treating repeatedly; Not only need a large amount of whizzers, and centrifugal treating is time-consuming too much, this method can not be used for industrialization production.
Saturated sal epsom floating method is except that having above-mentioned defective; This fado is used for the laboratory and separates Tyzzeria perniciosa (Tyzzeria perniciosa) and Wenyonella philiplevinei (Wenyonella philiplevine) egg capsule, seldom is used for the separation of chicken pathogenic Eimeria egg capsule.
Saturated chromatosulfuric acid floating method not only needs a large amount of centrifugal treating, also needs condition of ice bath, and this does not obviously meet the working condition of vaccine to chicken coccidiosis, and cost is too high.
The sucrose density gradient centrifugation process is complicated, condition preparation difficulty, and sepn process has comprised the saturated brine floating method, and weak point is no longer tired to be stated.
Summary of the invention
The purpose of this invention is to provide the method that industrialization prepares chicken Eimeria egg capsule.
Method of the present invention is:
A, excrement appearance are handled: get the infected chicken fresh excreta; Add the water dilution mixing of 3 times of its weight, behind the adding stomach en-thorough mixing, leave standstill the water that adds 7 times of excrement weight after 10 minutes again; Mixing makes into liquid manure, and wherein pepsic add-on is a 0.05mg/ gram ight soil;
B, screening processing continuously: liquid manure is handled through multi-layer type vibrosieve filter, promptly at the uniform velocity adds liquid manure from charging opening, is filtered by the 1st layer to the 4th layer continuous sub-sieve; Add the water flushing simultaneously; After take off the 4th layer of screen cloth, water washes out all trapped substances, to the graduated containers for future use of tool;
Multi-layer type vibrosieve filter: or claim the multi-layer type vibrating screen classifier, establish 4 layers altogether, the 1st layer of 60 order, the 2nd layer of 200 order, the 3rd layer of 325 order, the 4th layer of 1250 order;
C, saturated brine are floating centrifugal: it is floating that trapped substance adds the abundant mixing of saturated brine of 5 times of volumes, and 2500rpm * 10min is centrifugal;
D, egg capsule centrifugation: the swimmer on sucking-off centrifuge tube surface adds to and is less than 10 times of water mixings of its volume, and 2500rpm * 10min is centrifugal;
E, egg capsule purify: remove supernatant, add water to centrifuge tube and be full of mixing, 2500rpm * 10min is centrifugal, and throw out is egg capsule.
The characteristics of present technique:
(1) simplified technology: present technique comes down to the technique improvement to the saturated salt water law, and whole egg capsule sepn process has reduced by three unit, the simplified working procedures of Technology 37.5%.
(2) be applicable to industrialization production: with the 300kg chicken manure is example; 3300 liters of liquid manures are through screening processing continuously; 60 orders, 200 orders and 325 mesh sieves have been held back the excrement slag bigger than egg capsule, and all flow through 1250 mesh sieves less than the excrement slag and the water of egg capsule, the trapped substance that resides in 1250 mesh sieves have only egg capsule with the identical impurity of egg capsule size; Its total amount is about 2% of liquid manure total amount, promptly 66 liters.With above-mentioned large vol whizzer centrifugal treating, 1 bench centrifuge needs 33 times, if use 6 bench centrifuges, then the work of every machine split run is 5.5 times.Meet the matched and the post setting of medium-sized vaccine factory, if will enlarge output, only needing increases by 6 bench centrifuges, can reach and produce the above pentavalent coccidial vaccine of 500,000,000 plumages per year, as the production trivalent vaccine then output can reach more than 700,000,000 plumages.
(3) excrement appearance adds a small amount of stomach en-(0.05mg/ restrains ight soil) in handling, and its role is to destroy intestinal tissue, foreign organic matter in the ight soil, helps screening and filters, and simultaneously, also can remove the adhesion that reaches egg capsule and foreign material between the egg capsule.
(4) shortened egg capsule and separated the time spent, compared, saved disengaging time more than 50% with conventional art.Reduced centrifugal number of times, traditional stripping technique is after filtering through 200 mesh sieves, and egg capsule and water and bulky stool appearance impurity concrete must pass through centrifugal treating and just can make egg capsule and water sepn together, and then it is floating to carry out salt solution.And new technology has been saved centrifugal before floating, and reason is directly to have accomplished 3 separation: 1. big than egg capsule foreign material; 2. than little foreign material of egg capsule and 3. water.
For industrialization production, as continue to use traditional technology, the spinning of this operation needs equally a large amount of repeatedly centrifugal repeatedly, is impracticable, and new technology has been eliminated " bottleneck " of original technology fully.
(5) significantly reduce the usage quantity of saturated brine, reduced cost.Because sieve apparatus has increased by 325 order removal of impurities continuously, has reduced the swimmer total amount, can reduce the usage quantity about 30% of saturated brine.
(6) improved the separation rate of egg capsule,, increased contacting of filtrate and screen cloth because sieve apparatus can produce the ternary motion continuously, can more effective filtration egg capsule, make the separation rate raising 10% of egg capsule, finally can increase the output 10% of vaccine.
Embodiment
Get infected chicken fresh excreta 20kg, add 60 liters of abundant mixings of water, add stomach en-1000mg, mixing leaves standstill 10min; Add 140 premium on currency afterwards again, be mixed into liquid manure; About 220 liters of liquid manure total amount; Start continuous sieve apparatus; Liquid manure from charging opening at the uniform velocity (10-15 liter/minute) add, add 20 liters of entry after liquid manure adds again, keep the sieve apparatus running simultaneously; To wash the not filtration egg capsule that is retained in the 1st, 2,3 layer of screen cloth as far as possible; Take off the 4th layer of sub-sieve afterwards, water flushing trapped substance is in heaps, is collected in the graduated container of tool subsequent use; (V: V) the saturated brine mixing is floating, is sub-packed in the 500ml centrifuge tube, and 2500rpm * 10min is centrifugal to add 5 times by the trapped substance amount; The swimmer on sucking-off centrifuge tube surface adds at least 10 times of water mixings, and 2500rpm * 10min is centrifugal; It is centrifugal that taking precipitate adds enough water (filling it up with centrifuge tube) 2500rpm * 10min, and throw out is egg capsule.
Claims (1)
1. a chicken Eimeria (Eimeria coccidia) egg capsule batch preparation is characterized in that:
A, excrement appearance are handled: get the infected chicken fresh excreta; Add the water dilution mixing of 3 times of its weight, behind the adding stomach en-thorough mixing, leave standstill the water that adds 7 times of excrement weight after 10 minutes again; Mixing makes into liquid manure, and wherein pepsic add-on is a 0.05mg/ gram ight soil;
B, screening processing continuously: liquid manure is handled through multi-layer type vibrosieve filter, promptly at the uniform velocity adds liquid manure from charging opening, is filtered by the 1st layer to the 4th layer continuous sub-sieve; Add the water flushing simultaneously; After take off the 4th layer of screen cloth, water washes out all trapped substances, to the graduated containers for future use of tool;
Multi-layer type vibrosieve filter: or claim the multi-layer type vibrating screen classifier, establish 4 layers altogether, the 1st layer of 60 order, the 2nd layer of 200 order, the 3rd layer of 325 order, the 4th layer of 1250 order;
C, saturated brine are floating centrifugal: it is floating that trapped substance adds the abundant mixing of saturated brine of 5 times of volumes, and 2500rpm * 10min is centrifugal;
D, egg capsule centrifugation: the swimmer on sucking-off centrifuge tube surface adds to and is less than 10 times of water mixings of its volume, and 2500rpm * 10min is centrifugal;
E, egg capsule purify: remove supernatant, add water to centrifuge tube and be full of mixing, 2500rpm * 10min is centrifugal, and throw out is egg capsule.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100292716A CN102174408B (en) | 2011-01-10 | 2011-01-10 | Method for batch preparation of chicken Eimeria coccidium oocysts |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100292716A CN102174408B (en) | 2011-01-10 | 2011-01-10 | Method for batch preparation of chicken Eimeria coccidium oocysts |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102174408A CN102174408A (en) | 2011-09-07 |
| CN102174408B true CN102174408B (en) | 2012-08-22 |
Family
ID=44517666
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011100292716A Active CN102174408B (en) | 2011-01-10 | 2011-01-10 | Method for batch preparation of chicken Eimeria coccidium oocysts |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102174408B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102726617A (en) * | 2012-07-05 | 2012-10-17 | 烟台青大生物制药有限公司 | Feed additive composed of cellulose and sodium bicarbonate, feed and applications of feed additive and feed, as well as chicken coccidiosis live oocyst vaccine |
| CN103251938B (en) * | 2013-04-15 | 2015-07-15 | 内蒙古神元生物工程股份有限公司 | Industrial production process of eimeria coccidiosis attenuated vaccine |
| CN103881913A (en) * | 2014-03-07 | 2014-06-25 | 广东省农业科学院动物卫生研究所 | Industria production method for separating and purifying chicken eimeria coccidium oocyst |
| CN111154653B (en) * | 2020-01-16 | 2020-11-20 | 中国农业科学院兰州兽医研究所 | Method for purifying insect eggs through iodixanol density gradient centrifugation |
| CN111733079B (en) * | 2020-07-08 | 2021-09-07 | 佛山市正典生物技术有限公司 | Separation composition, preparation method thereof and chicken coccidian oocyst separation method using same |
| CN113528347B (en) * | 2021-06-28 | 2023-04-18 | 佛山市正典生物技术有限公司 | Method for improving sorting success rate of fluorescent Eimeria oocysts and application |
| CN114381375A (en) * | 2021-12-21 | 2022-04-22 | 佛山市正典生物技术有限公司 | Treatment method and application of chicken Eimeria maxima excrement mucus |
| CN114380474B (en) * | 2021-12-21 | 2022-10-21 | 佛山市正典生物技术有限公司 | Excrement treating agent and method and application thereof for treating excrement containing chicken cecum core |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1837348A (en) * | 2006-04-07 | 2006-09-27 | 佛山市正典生物技术有限公司 | Culture solution and preservation solution for coccidian oocyst |
| CN1946837B (en) * | 2004-03-31 | 2010-06-23 | 恩布里克斯公司 | Method of purifying oocysts using enzymatic digestion of fecal debris |
-
2011
- 2011-01-10 CN CN2011100292716A patent/CN102174408B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1946837B (en) * | 2004-03-31 | 2010-06-23 | 恩布里克斯公司 | Method of purifying oocysts using enzymatic digestion of fecal debris |
| CN1837348A (en) * | 2006-04-07 | 2006-09-27 | 佛山市正典生物技术有限公司 | Culture solution and preservation solution for coccidian oocyst |
Non-Patent Citations (2)
| Title |
|---|
| 朱民.5种分离纯化粪便中隐孢子虫卵囊方法的比较.《上海预防医学杂志》.中国期刊全文数据库,2006,第18卷(第4期),全文. * |
| 黄兵.鸡球虫的分离与保存.《中国寄生虫学与寄生虫病杂志》.中国期刊全文数据库,2006,第24卷全文. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102174408A (en) | 2011-09-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102174408B (en) | Method for batch preparation of chicken Eimeria coccidium oocysts | |
| CN208292897U (en) | Totally-enclosed CBD automatically extracts production line | |
| CN105601762B (en) | A kind of method for extracting Hijiki polysaccharide | |
| CN101638637A (en) | Kit for processing human marrow, cord blood and peripheral blood cells and cell processing method | |
| CN105175564B (en) | The industrialization extraction separation method and separator of ganoderma spore polysaccharide | |
| CN103881913A (en) | Industria production method for separating and purifying chicken eimeria coccidium oocyst | |
| CN106318896A (en) | Method for preparing and purifying cedarwood protoplast | |
| CN106216294A (en) | A kind of Chinese crude drug cleaning drying device | |
| CN109370977A (en) | A method of isolating and purifying rice protoplast | |
| CN112877215A (en) | Separation and purification method and inoculation method of Eimeria tenella sporozoites | |
| CN107955084B (en) | Reaction tank deslagging and separating system special for extracting chlorella polysaccharide | |
| CN106689002A (en) | Green healthy controllable industrial prawn culture method | |
| CN102702377A (en) | Termitomyces albuminosus polysaccharide separation and purification method and injection thereof | |
| CN102492780A (en) | Method for removing impurities from sorgo | |
| CN207608551U (en) | A kind of separator of coral polyp and zooxanthellae | |
| CN109355174A (en) | A kind of multi-level sieve of microalgae filtering | |
| CN209376483U (en) | A kind of feed coefficient or native pool circulating water culture system | |
| CN108034691A (en) | A kind of method of the abundance of microorganism in accurate counting invertebrate hemolymph | |
| CN103251938B (en) | Industrial production process of eimeria coccidiosis attenuated vaccine | |
| CN105968166B (en) | A process for producing diosgenin by combining dry crushing-enzymolysis fermentation-counter current alcohol extraction techniques and a counter current alcohol extraction device therefore | |
| CN204981708U (en) | Separator that industrialization of glossy ganoderma spore polysaccharide was drawed | |
| CN201756486U (en) | Device used for coagulating enriching cryptosporidium oocysts and giardia Lamblia cysts | |
| CN1270720C (en) | Yolk lecithin for injection and method for making same | |
| RU2678122C1 (en) | Method for producing reproductive products of mussels mytilus galloprovincialis lam | |
| CN113528347B (en) | Method for improving sorting success rate of fluorescent Eimeria oocysts and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| PP01 | Preservation of patent right |
Effective date of registration: 20160901 Granted publication date: 20120822 |
|
| RINS | Preservation of patent right or utility model and its discharge | ||
| PD01 | Discharge of preservation of patent |
Date of cancellation: 20170301 Granted publication date: 20120822 |
|
| PD01 | Discharge of preservation of patent | ||
| PP01 | Preservation of patent right |
Effective date of registration: 20170303 Granted publication date: 20120822 |
|
| PP01 | Preservation of patent right | ||
| PD01 | Discharge of preservation of patent |
Date of cancellation: 20170903 Granted publication date: 20120822 |
|
| PD01 | Discharge of preservation of patent | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Method for batch preparation of chicken Eimeria coccidium oocysts Effective date of registration: 20180703 Granted publication date: 20120822 Pledgee: Zhang Yongping Pledgor: Inner Mongolia Shenyuan Biomedical Co.,Ltd. Registration number: 2018990000521 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20220803 Address after: 010000 Business Incubation Park of Wuchuan County, Hohhot City, Inner Mongolia Autonomous Region 302-303 (West Building of Agricultural Bank of Wuchuan Kezhen) Patentee after: Inner Mongolia Yuanben Biomedical Technology Co.,Ltd. Address before: 010010 Ruyi Road, Ruyi economic and Technological Development Zone, the Inner Mongolia Autonomous Region, Hohhot Patentee before: Inner Mongolia Shenyuan Biomedical Co.,Ltd. |
|
| TR01 | Transfer of patent right |