CN104829712B - C, D types C.perfringens antitoxic serum and preparation method thereof - Google Patents
C, D types C.perfringens antitoxic serum and preparation method thereof Download PDFInfo
- Publication number
- CN104829712B CN104829712B CN201510202093.0A CN201510202093A CN104829712B CN 104829712 B CN104829712 B CN 104829712B CN 201510202093 A CN201510202093 A CN 201510202093A CN 104829712 B CN104829712 B CN 104829712B
- Authority
- CN
- China
- Prior art keywords
- perfringens
- type
- immunogene
- serum
- solution
- 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
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention relates to a kind of C, D type C.perfringens antitoxic serum and preparation method thereof；It is that c-type C.perfringens strain and D type C.perfringens strains are inoculated on blood plate culture medium and recovered respectively, reuse nutrient broth and increase bacterium Anaerobic culturel, respectively obtain c-type C.perfringens bacterium solution and D type C.perfringens bacterium solutions, toxin producing medium culture inactivation is recycled, to isolated after serum progress fundamental immunity and booster immunization.C, D type C.perfringens antitoxic serum prepared by the present invention, potency height, high specificity, has no toxic side effect；Available for animal C, D type C.perfringens disease it is immune, significant role can be played in the groove in the disease such as sheep struck, enterotoxemia caused by research pre-control control c-type, D types or the infection of C, D amphitypy C.perfringens.
Description
(1) technical field
The present invention relates to a kind of C, D type C.perfringens antitoxic serum and preparation method thereof.
(2) technical background
C.perfringens (C.perfringens) is also known as clostridieum welchii (C.welchii), be animal intestinal tract it is normal
Bacterium, however, when raising, environmental condition suddenly change when it can Infective, be a kind of conditioned pathogen.The bacterium can infect
Many animals, cause the complicated symptom such as necrotic enteritis and enterotoxemia or pathological change.C.perfringens is according to its point
The four kinds of main exotoxins (α, β, ε, ι) secreted are divided into five hypotypes of A, B, C, D, E.C-type C.perfringens principal causative toxin
Mainly α, β-exotoxin, it is the main pathogen of a variety of domestic animal intestines source toxaemia and " Sudden Death Syndrome ", as piglet red dysentery and sheep are sudden
Macaque.D types C.perfringens mainly produces two kinds of exotoxins of α, ε, can cause the animals such as lamb, sheep, goat, ox and squirrel
Enterotoxemia.The ground such as Heilungkiang, Qinghai, Yunnan, Ningxia has D type bacterium to cause the animal enterotoxemia such as ox, sheep in recent years
Report, heavy losses are caused to animal husbandry.The disease is classified as the two class infectious diseases of livestock and poultry by the Ministry of Agriculture.
In livestock breed aquatics, due to the use of antibiotic, in the case where selecting pressure, cause of disease constantly makes a variation.Perfringens
The Antibiotic Resistance of clostridium is more and more wider so that the antibiotic once played a great role increasingly incapability when in face of these diseases is
Power, vast raiser is usually allowed to suffer from great economic loss.Antiserum is also known as hyper-immune serum or disease-resistant serum, is a kind of
High-titer, specific antibody.
Using the pooled serum of two hypotypes of C, D, antibody enriches, and coverage rate is wide, can play good prevention in production
Effect.In China, C, D type C.perfringens antitoxic serum do not have the report for preparing and applying.So the present invention is invasive
Newly, and husbandry sector can be served.
(3) content of the invention
In order to solve the above problems, the invention provides a kind of C, D type C.perfringens antitoxic serum and its preparation
Method；C, D the type C.perfringens antitoxic serum prepared using the present invention, potency height, high specificity, is had no toxic side effect；
Available for animal C, D type C.perfringens disease it is immune, can be in research pre-control control c-type, D types or C, D amphitypy aerogenesis pod
The disease such as sheep struck, enterotoxemia plays significant role in the groove caused by the infection of film clostridium.
A kind of C, D type C.perfringens antitoxic serum, its preparation method comprise the following steps：
1. 3ul c-type C.perfringens strains and D type C.perfringens strains are inoculated in blood plate culture respectively
Recovered on base, reuse nutrient broth medium and increase bacterium Anaerobic culturel, respectively obtain c-type C.perfringens bacterium solution and the production of D types
Gas capsular clostridium bacterium solution.
2. 4ml c-type C.perfringens bacterium solutions and 3ml D type C.perfringens bacterium solutions are added to 100ml pH7.5
Toxin producing medium in, the shaken cultivation under anaerobic environment, 44 DEG C of culture 5h efficiently produce poison, then 4 DEG C of 4000g centrifugation 15min,
Again with filtration sterilization in the seitz filter in 0.22 μm of aperture, C, D type C.perfringens exotoxin solution are obtained, as C, D type
Second immunogene of C.perfringens antitoxic serum.
3. the formalin of its volume ratio 0.3% is added in C, D type C.perfringens exotoxin solution, fully vibration
Mix, 37 DEG C of inactivation 128h, during which every 5-6h shakings once, C, D the type C.perfringens toxoid for obtaining having inactivated are molten
Liquid.According to C, D type C.perfringens toxoid solution and white-oil adjuvant volume ratio 1:2 to C, D the type perfringens inactivated
The white-oil adjuvant emulsification of sterilizing is added in clostridium toxoid solution, obtains C, D type C.perfringens divalence white oil toxoid epidemic disease
Seedling, the first immunogene as C, D type C.perfringens antitoxic serum.
Prepared by 4.C, D type C.perfringens antitoxic serum is divided into fundamental immunity and booster immunization；It is first immune with first
The former healthy sheep healthy and strong to adult carries out fundamental immunity twice：Muscle branch injection C, D type C.perfringens divalence class poison
White oily vaccine；Reuse the second immunogene and carry out four booster immunizations：Muscle branch injects degerming C, D type C.perfringens
Exotoxin solution.Carried out by following immune programme for children：
Immune programme for children
5. vein sterile blood sampling after the immune 10d of last time, separates serum, obtains C, D type C.perfringens antitoxin
Serum.To C, D type C.perfringens antitoxic serum carry out safety examination, steriling test etc. (《People's Republic of China (PRC) beast
With biological products quality standard》Version in 2001).The antibody level of the serum product prepared is examined with Mouse neutralization test, finally
Minimum lethal dose (10ul) of 400 c-type C.perfringens exotoxins to mouse can be neutralized by measuring every 0.1ml serum, in energy
With 600 D type C.perfringens exotoxins to mouse minimum lethal dose (50ul).
Described blood plate medium component is：100 milliliters of distilled water, 3.7g bean powderes agar, 1g glucose and 5ml sheep
Blood；
Described nutrient broth medium composition：100 milliliters of distilled water, peptone 1g, powdered beef 0.3g and physiological saline
0.5g；
Described toxin producing medium composition is：Every 100mlPBS buffer solution 3g tryptones, 2g dextrin, the leaching of 2g yeast
Cream and 2.2gL- arginine；PBS composition is：1L deionized waters, 0.27g potassium dihydrogen phosphates (KH2PO4), 1.42g phosphoric acid
Disodium hydrogen (Na2HPO4), 8g sodium chloride (NaCl) and 0.2g potassium chloride (KCl)；
Serum 1mL is subcutaneously or intramuscularly injected in c-type or D type C.perfringens diseases Prevalent district, 1~5 days old lamb.It is right
Ill sick lamb, vein or intramuscular injection serum 3ml, if necessary after 4~5h, repeat injection once.Illness is grown up
5~10ml of sheep intravenous injection serum.
Beneficial effect
The serum titer is high, and prevention and control effect is good, can effectively prevent, control, treating and drawn by C, D type C.perfringens
The pulpy kidney disease of the sheep struck risen, lamb, calf and piglet diarrhea, cattle and sheep enterotoxemia and sheep, can be that aquaculture reduces loss,
Bring visual interests.
(4) illustrate
Fig. 1 C, D type C.perfringens antitoxic serums preparation
(5) embodiment
Detailed process prepared by the serum of embodiment 1
First, bacterium solution is prepared.By c-type C.perfringens strain (National Collection of Type
Cultures- Britain Culture Collection preservation preserving number:NCTC 3180) and D type C.perfringens
(National Collection of Type Cultures- Britain Culture Collection preservation preserving number:NCTC
8346) 37 DEG C of 36h recoveries on blood plate culture medium are inoculated in respectively, and one bacterium colony of picking increases bacterium training into nutrient broth respectively
Support, be 88%N in gas concentration2, 7%H2, 5%CO2Anaerobic environment under the conditions of 39 DEG C culture 13h, respectively obtain c-type aerogenesis
Capsular clostridium and D type C.perfringens, give over to inoculation toxin producing medium.
Secondly, immunogene is prepared.4mlC types C.perfringens and 3mlD type C.perfringens are added to 100ml
(per 100mlPBS buffer solution 3g peptones, 1.2g dextrin, 2g yeast extracts and 2.2gL- in pH7.5 toxin producing medium
Arginine) in, the shaken cultivation under anaerobic environment, 44 DEG C of culture 5h efficiently produce poison, and then 4 DEG C of 3800g centrifuge 15min again, then
With filtration sterilization in the seitz filter in 0.22 μm of aperture, C, D type C.perfringens exotoxin solution are obtained, is produced as C, D type
Second immunogene of gas capsular clostridium antitoxic serum.It is molten that exotoxin is added in C, D type C.perfringens exotoxin solution
The formaldehyde (concentration of formaldehyde 40%) of liquid volume ratio 0.3%, fully vibration mix, and 37 DEG C of 128h inactivations are put, during which at interval of 5-
6h shakes once, obtains C, D the type C.perfringens toxoid solution inactivated.By C, D type C.perfringens toxoid
Solution and white-oil adjuvant volume ratio 1:2 add the white-oil adjuvant emulsification of sterilizing in C, D type C.perfringens toxoid solution,
Obtain C, D type C.perfringens white oil toxoid bivalent vaccine, first as C, D type C.perfringens antitoxic serum
Immunogene.
Again, the preparation of C, D type C.perfringens antitoxic serum.C, D types C.perfringens antitoxic serum system
Immune and booster immunization based on back-up, fundamental immunity twice first is carried out to adult healthy, healthy and strong sheep with the first immunogene：
Muscle branch injects C, D type C.perfringens divalence white oil toxoid vaccine；It is immunized afterwards as booster immunization, booster immunization for 4 times
Use the second immunogene (muscle branch injection C, D type C.perfringens exotoxin solution).Carried out by following immune programme for children：
Immune programme for children
The preparation and inspection of serum product：Vein sterile blood sampling after the immune 10d of last time, separates serum, obtains C, D type
C.perfringens antitoxic serum.Safety examination, steriling test are carried out to C, D type C.perfringens antitoxic serum
(《People's Republic of China's veterinary biologicses quality standard》Version in 2001).The serum system prepared is examined with Mouse neutralization test
The antibody titer of product, 400 c-type clostridium perfringens toxoid minimum lethal doses can be neutralized by finally measuring every 0.1ml serum, in energy
With 600 D type clostridium perfringens toxoid minimum lethal doses.In C, D type C.perfringens disease Prevalent district, 1~5 age in days lamb
Under sheepskin or intramuscular injection serum 1mL is used to prevent, must to ill sick lamb vein or intramuscular injection 5~10ml of serum
When wanting after 4~5h, injection is repeated once.
The checking of the vaccine safety of embodiment 2. and immune effect.
Safety examination:16~20g healthy mices 5 are taken, the antiserum 0.5ml that respectively prepared by the intravenous injection present invention；
Body weight 250g-450g cavys 2 are used simultaneously, and antiserum 5ml prepared by the present invention, Continuous Observation 10 days is subcutaneously injected；It is local equal
Without adverse reactions such as granulomas, whole body is also no different paradoxical reaction.(reference《People's Republic of China's veterinary biologicses quality mark
It is accurate》)
The measure of absolute lethal dose:C-type and D type C.perfringens exotoxins solution prepared by the present invention is respectively with life
Manage after salt solution dilutes 5,10,15,20,30,40,50,60 times respectively to 8 groups of mouse peritoneal injection 0.5ml, every group 5, one dilute
The content of toxins of the whole dead maximum dilution multiples (60 be maximum dilution multiple) of 5 mouse of degree of releasing is the absolute lethal of mouse
Amount.It is 10ul (the i.e. outer poison of c-type C.perfringens to the dead amount of minimum of mouse finally to measure c-type C.perfringens exotoxin
When element dilutes 5,10,15,20,30,40,50 times, mouse peritoneal injection 0.5ml, mouse is all dead, thus toxin dilution 50
Measured again for c-type C.perfringens exotoxin is dead to the minimum of mouse, c-type C.perfringens exotoxin is contained by this dilution factor
10ul, thus c-type C.perfringens exotoxin is 10ul to the dead amount of minimum of mouse)；D type exotoxins are measured to mouse most
Small lethal dose be 50ul (i.e. 5,10 times of D types C.perfringens exotoxin dilution when, mouse peritoneal injection 0.5ml, mouse is complete
Portion is dead, thus toxin dilutes 10 times of minimum extremely amounts for D type C.perfringens exotoxins to mouse, D contained by this dilution factor
Type C.perfringens exotoxin is 50ul, thus D type C.perfringens exotoxins are 50ul to the dead amount of minimum of mouse).
Mouse neutralization test:C, D type C.perfringens antitoxic serum after last time booster immunization is taken to carry out small
Mouse neutralization test.By C, D type C.perfringens antitoxic serum of 0.2ml this experiment preparations respectively according to 1:50,1:100,
1:200,1:400,1:600,1:800,1:1000 dilution proportions respectively with outside the c-type C.perfringens of 2 times of minimum lethal doses
D type C.perfringens exotoxins (100ul) mixing of toxin (20ul) and 2 times of minimum lethal doses, and physiological saline is used respectively
Constant volume is put in 37 DEG C of incubator 2h to 1ml；The above-mentioned each mixed solution intraperitoneal injection of mice of dilution gradient of 0.5ml is taken respectively, is seen
Examine mouse survival situation.400 c-type exotoxin minimum lethal doses can be neutralized by measuring serum prepared by the 0.1ml present invention, can be neutralized
600 D type exotoxin minimum lethal doses.
Claims (1)
1. a kind of C, D type C.perfringens antitoxic serum, it is characterised in that be prepared by the following method to obtain：
1）3 μ L c-type C.perfringens strains and D type C.perfringens strains are inoculated on blood plate culture medium respectively
Increase bacterium Anaerobic culturel using nutrient broth medium after recovery, respectively obtain c-type C.perfringens bacterium solution and D type perfringens
Clostridium bacterium solution；
2）4 ml c-type C.perfringens bacterium solutions and 3 ml D type C.perfringens bacterium solutions are added to 100ml pH7.5's
In toxin producing medium, the shaken cultivation under anaerobic environment, 44 DEG C of 5 h of culture efficiently produce poison, then 4 DEG C of 4000 g centrifugation 15
Min, then with filtration sterilization in the seitz filter in 0.22 μm of aperture, obtain C, D type C.perfringens exotoxin solution, as C,
Second immunogene of D type C.perfringens antitoxic serums；
3）The formalin of its volume ratio 0.3% is added in C, D type C.perfringens exotoxin solution, fully vibration is mixed
It is even, 37 DEG C of 128 h of inactivation, during which every 5-6 h shakings once, obtain C, D the type C.perfringens toxoid inactivated
Solution；According to C, D type C.perfringens toxoid solution and white-oil adjuvant volume ratio 1:2 to C, D the type aerogenesis pod inactivated
The white-oil adjuvant emulsification of sterilizing is added in film clostridium toxoid solution, obtains C, D type C.perfringens divalence white oil toxoid
Vaccine, the first immunogene as C, D type C.perfringens antitoxic serum；
4）C, prepared by D types C.perfringens antitoxic serum is divided into fundamental immunity and booster immunization；First with the first immunogene pair
The healthy and strong healthy sheep of adult carries out fundamental immunity twice：Muscle branch injection C, D type C.perfringens divalence toxoid is white
Oily vaccine；Reuse the second immunogene and carry out four booster immunizations：Muscle branch injects the outer poison of degerming C, D type C.perfringens
Plain solution；Specific immune programme for children such as following table：
Immune programme for children
5）Vein sterile blood sampling after the immune 10d of last time, separates serum, obtains C, D type C.perfringens antitoxic serum；
Described blood plate medium component is：100 milliliters of distilled water, 3.7g bean powderes agar, 1g glucose and 5ml Sheep Bloods；
Described nutrient broth medium composition：100 milliliters of distilled water, peptone 1g, powdered beef 0.3g, physiological saline 0.5g；
Described toxin producing medium composition is：Per 100mlPBS buffer solution 3g tryptones, 2g dextrin, 2g yeast extracts and
2.2gL- arginine；PBS composition is：1L deionized waters, 0.27g potassium dihydrogen phosphates (KH2PO4), 1.42g phosphoric acid hydrogen two
Sodium (Na2HPO4), 8g sodium chloride (NaCl) and 0.2g potassium chloride (KCl).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510202093.0A CN104829712B (en) | 2015-04-23 | 2015-04-23 | C, D types C.perfringens antitoxic serum and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510202093.0A CN104829712B (en) | 2015-04-23 | 2015-04-23 | C, D types C.perfringens antitoxic serum and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104829712A CN104829712A (en) | 2015-08-12 |
| CN104829712B true CN104829712B (en) | 2018-03-27 |
Family
ID=53807943
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510202093.0A Active CN104829712B (en) | 2015-04-23 | 2015-04-23 | C, D types C.perfringens antitoxic serum and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104829712B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106177935A (en) * | 2016-08-19 | 2016-12-07 | 齐鲁动物保健品有限公司 | A kind of ruminant clostridial disease tetrad inactivated vaccine and preparation method thereof |
| CN107299070B (en) * | 2017-08-11 | 2020-09-04 | 中国兽医药品监察所 | D-type clostridium perfringens toxin for livestock and preparation method and special culture medium thereof |
| CN107789622A (en) * | 2017-10-18 | 2018-03-13 | 山东农业大学 | A kind of calf enterotoxemia C.perfringens β2The preparation method of toxin toxoid vaccine |
| CN107875377B (en) * | 2017-11-06 | 2020-01-03 | 山东农业大学 | Preparation method of clostridium perfringens type E toxoid vaccine |
| CN108892725A (en) * | 2018-04-18 | 2018-11-27 | 山东农业大学 | A kind of preparation method of E type C.perfringens antitoxic serum |
| CN109943507B (en) * | 2019-03-26 | 2021-08-17 | 中国兽医药品监察所 | Preparation method and application of veterinary A-type clostridium perfringens toxin |
| CN116445373B (en) * | 2023-06-14 | 2023-09-12 | 金宇保灵生物药品有限公司 | C clostridium perfringens toxigenic culture medium and preparation method and application thereof |
| CN117467001A (en) * | 2023-12-28 | 2024-01-30 | 金宇保灵生物药品有限公司 | Preparation method and application of clostridium perfringens antitoxin A serum |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1440810A (en) * | 2003-03-13 | 2003-09-10 | 山东农业大学 | Prepn of clostridium welchil toxoid vaccine and antitoxin serum |
| WO2008061950A1 (en) * | 2006-11-20 | 2008-05-29 | Intervet International B.V. | Use of clostridium perfringens type c bacterium for the manufacture of a vaccine |
| CN103432578A (en) * | 2013-09-12 | 2013-12-11 | 山东农业大学 | Preparation method of C-type clostridium perfringens toxin vaccine of piglet red dysentery |
| CN103463632A (en) * | 2013-09-05 | 2013-12-25 | 山东农业大学 | Preparation method for D type clostridium perfringen toxoid vaccine of cattle and sheep enterotoxaemia |
| CN103830747A (en) * | 2014-03-27 | 2014-06-04 | 山东农业大学 | Genetically engineered vaccine of epsilon toxin of clostridium perfringens and application thereof |
| CN103830722A (en) * | 2014-03-27 | 2014-06-04 | 山东农业大学 | Clostridium perfringens beta toxin genetic engineering vaccine and application thereof |
-
2015
- 2015-04-23 CN CN201510202093.0A patent/CN104829712B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1440810A (en) * | 2003-03-13 | 2003-09-10 | 山东农业大学 | Prepn of clostridium welchil toxoid vaccine and antitoxin serum |
| WO2008061950A1 (en) * | 2006-11-20 | 2008-05-29 | Intervet International B.V. | Use of clostridium perfringens type c bacterium for the manufacture of a vaccine |
| CN103463632A (en) * | 2013-09-05 | 2013-12-25 | 山东农业大学 | Preparation method for D type clostridium perfringen toxoid vaccine of cattle and sheep enterotoxaemia |
| CN103432578A (en) * | 2013-09-12 | 2013-12-11 | 山东农业大学 | Preparation method of C-type clostridium perfringens toxin vaccine of piglet red dysentery |
| CN103830747A (en) * | 2014-03-27 | 2014-06-04 | 山东农业大学 | Genetically engineered vaccine of epsilon toxin of clostridium perfringens and application thereof |
| CN103830722A (en) * | 2014-03-27 | 2014-06-04 | 山东农业大学 | Clostridium perfringens beta toxin genetic engineering vaccine and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104829712A (en) | 2015-08-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104829712B (en) | C, D types C.perfringens antitoxic serum and preparation method thereof | |
| CN103157100B (en) | hemophilus parasuis disease, swine streptococcosis bivalent inactivated vaccine and preparation method thereof | |
| CN102086447A (en) | Duck virus hepatitis strains and inactivated vaccine | |
| CN104623651B (en) | A kind of preparation method of the A type bacillus perfringens inactivated vaccine of rabbit clostridieum welchii disease | |
| CN103432578A (en) | Preparation method of C-type clostridium perfringens toxin vaccine of piglet red dysentery | |
| CN105112349B (en) | A kind of brucella melitensis molecular marker vaccine strain and its application | |
| CN108721616B (en) | A kind of avian pasteurella multocida capsular polysaccharide-protein conjugate vaccines and preparation method thereof | |
| CN106177935A (en) | A kind of ruminant clostridial disease tetrad inactivated vaccine and preparation method thereof | |
| CN103421832A (en) | Preparation and application of egg yolk antibody containing florfenicol drug resistance gene protein | |
| CN104096222B (en) | A kind of vaccine combination and its preparation method and application | |
| CN104163858B (en) | Pasteurella multocida acellular antigen, preparation method and applications thereof | |
| CN101708332A (en) | Rabbit triple inactivated vaccine, preparation method and application thereof | |
| CN104774796B (en) | Fowl enteropathogenic E. Coli inactivated vaccine and preparation method thereof | |
| CN104726413B (en) | The monoclonal antibody of one plant of Mycoplasma bovis nadh oxidase | |
| CN107653231B (en) | Duck-origin coronavirus low-virulent strain IBVDCV35 and application thereof | |
| CN103157101B (en) | Combined inactivate vaccine for haemophilus parasuis disease and streptococcus suis disease and preparation method for same | |
| CN107875377A (en) | A kind of preparation method of E types C.perfringens toxoid vaccine | |
| CN102688484B (en) | A kind of production method of chicken necrotizing enterocolitis (C type) inactivated vaccine | |
| CN103463632B (en) | The preparation method of the D type bacillus perfringens toxoid vaccine of cattle and sheep enterotoxemia | |
| CN103585622B (en) | The application of Vaccine strain for mycoplasma pneumonia of swine | |
| CN105713855A (en) | Strains, application of strains, vaccine and preparation method of vaccine | |
| CN108892725A (en) | A kind of preparation method of E type C.perfringens antitoxic serum | |
| SU1734762A1 (en) | Method for preparation of vaccine for prevention of dermatophytoses in pet and test animals | |
| CN104645324A (en) | Application of swine enzootic hyopneumoniae vaccine strain | |
| CN110172524A (en) | Chicken C.perfringens is formulated to the method for the drug resistance criterion of avilamycin |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |