CN101721696A - Trivalent oil emulsion inactivated vaccine for 4 type, 5 type and 13 type serum of haemophilus parasuis - Google Patents
Trivalent oil emulsion inactivated vaccine for 4 type, 5 type and 13 type serum of haemophilus parasuis Download PDFInfo
- Publication number
- CN101721696A CN101721696A CN200910263482A CN200910263482A CN101721696A CN 101721696 A CN101721696 A CN 101721696A CN 200910263482 A CN200910263482 A CN 200910263482A CN 200910263482 A CN200910263482 A CN 200910263482A CN 101721696 A CN101721696 A CN 101721696A
- Authority
- CN
- China
- Prior art keywords
- serum
- types
- type
- oil
- inactivated vaccine
- 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.)
- Pending
Links
Abstract
The invention relates to a trivalent inactivated oil emulsion vaccine for 4 type, 5 type and 13 type serum of haemophilus parasuis, which is prepared by the following steps: selecting the separated strains of three main popular types of serum of the haemophilus parasuis in China as antigens; after culturing and propagating for 24h with TSB broth, inactivating for 11-13h with 0.3% formaldehyde, and mixing the antigens of the three types of serum according to the proportion of 1:1:1; then adding oil phases of which the total volume is equal to the total volume of the antigens; and emulsifying to prepare the vaccine of the invention. The vaccine of the invention is used for preventing fibrinous polyserositis, arthritis and meningitis of pigs caused by the bacterial strains of the main types of serum of the current popular haemophilus parasuis.
Description
1. technical field the present invention relates to oil emulsion inactivated preparation method of the secondary haemophilus trivalent of a boar and the application in the prevention various diseases that secondary haemophilus causes by pig thereof.
2. secondary haemophilus (the Haemophilus parasuis of background technology pig, HPS) be the pathogen of pig Glasser ' s disease, a kind of commensalism bacterium of pig upper respiratory tract, it can be invaded body under given conditions and cause that with fiber disposition polyserositis, arthritis and meningitis be the systemic disease of feature.The secondary haemophilus of pig is a kind of non-hemolytic, non-mobility, NAD dependent form, the tiny bacillus of Gram-negative.Along with the development of world's pig industry, this disease has become a kind of important bacteria disease that influences pig industry in the global range.
In the breed of swinery, the prevention work of carrying out at ordinary times is particularly important.The secondary haemophilus of pig be a kind of in secondary disease, have multiple circulation way, cause the secondary haemophilus of pig to be settled in the intravital asynchronism(-nization) of piglet, the time of settling down is depended on and sow immunity situation, sow carry disease germs level, piglet maternal antibody level etc.There are some researches show; to the secondary haemophilus bacteria inactivation of sow inoculation pig Seedling; the piglet that the piglet that gives birth to after the sow immunity gives birth to than immune sow excessively pneumonia occurs and arthritic probability is little; and symptom is lighter relatively; weightening finish also will be higher than the latter, thereby has illustrated that maternal antibody has good protective action to piglet.But, increase along with age in days, the ablactational baby pig maternal antibody is decayed gradually so that the piglet that can not effectively prevent to carry disease germs infects the piglet that the secondary hemophilus infection of no pig is settled down, so managing to reduce the carry disease germs ratio of piglet is the effective ways that the secondary haemophilus of control pig impacts production, the immunity of piglet by effective vaccine just seemed most important.Solano (1998) shows that by piglet being carried out vaccination the immunoprotection time that can make piglet is longer.Studies confirm that immune commercially available vaccine successful story in a large number, the vaccine that uses mainly is the unit price and the bivalent inactivated vaccine of deactivation at present, can effectively control the generation of this eqpidemic disease by inoculating these vaccines.But because the serotype of the secondary haemophilus of pig own is more, whether commercially available vaccine has preferably that cross protection just becomes very real problem, so the commercially available vaccine of developing a kind of polyvalent serum type and containing unique bacterial strain is very crucial.
3. the secondary haemophilus inactivated vaccine of the existing pig of summary of the invention mostly is unit price and bivalence inactivated vaccine, respectively at serum 4 types, serum 5 types and serum 13 types.The invention provides a kind of tervalent inactivated vaccine, preventing the main serotype of the popular bacterial strain of the secondary haemophilus of current China pig simultaneously is the disease that serum 4 types, 5 types and 13 types cause.
4. technical scheme technical scheme of the present invention is, the secondary haemophilus serum of pig 4,5,13 type separated strains are cultivated respectively, adds the water that tween 80 is made oil emulsion after the deactivation; Utilize Marko's 52 lightweight paraffin oil and Si Ben-80, aluminium stearate to make oil phase.Water and oil phase equal-volume are mixed back emulsifying, thereby obtain the secondary haemophilus tervalence inactivated vaccine of pig.Its concrete preparation method is as follows:
1) cultivation of antibacterial: the secondary haemophilus serum of pig 4 types, 5 types and 13 types, three strain kind daughter bacterias are inoculated in the TSA solid medium respectively, cultivate in 37 ℃ of calorstats, the single bacterium colony of picking behind the 24h, access contains in the TSB fluid medium of NAD and calf serum, this bacterium liquid is added in 1% ratio behind the shaken cultivation 12-16h under 37 ℃, 180r/min condition in the TSB fluid medium of new preparation to breed in a large number again.Behind 37 ℃, 180r/min shaken cultivation 24h, collect bacterium liquid, with the total bacterial content of spectrophotometric determination antibacterial.
2) deactivation of antibacterial: add 0.3% formalin-inactivated by TSB fluid medium cumulative volume, in 37 ℃ of vibration deactivation 11-13h.
3) preparation of oil emulsion water: after the strain deactivation of three serotypes, the bacterial concentration of each serotype is all transferred to 1 * 10
10CFU/ml mixes by 1: 1: 1 volume ratio, presses cumulative volume 4% and adds the sterilization tween 80, and the limit edged stirs.
4) oil phase preparation: get 93 parts of Marko's 52 lightweight paraffin oil, add 1 part of aluminium stearate and stir till transparent, add 6 parts of Si Ben-80 subsequently, abundant mixing, 121 ℃ of high pressure steam sterilization 30min, it is standby to be cooled to room temperature.
5) preparation of oil-emulsion inactivated vaccine: the oil phase and the water equal-volume that prepare are mixed, and the limit edged stirs, and to dissolving fully, and carries out emulsifying in homogenizer.
The preservation condition of this trivalent oil-emulsion inactivated vaccine is: 2～8 ℃.
Advantage of the present invention is to prevent the disease that caused by the secondary haemophilus serum of pig 4,5,13 type bacterial strains, reaches 85% for the immunoprotection of homologous serotype bacterial strain.With respect to present commercially available vaccine, can single needle the more serotype of immunity, easy to use, and improved probability to other serotype cross protections.
The specific embodiment
In conjunction with implementing the present invention is described further and proves.
Embodiment 1
The secondary haemophilus serum of the pig that has prepared 4 types, 5 types, 13 type trivalent oil emulsion inactivated vaccines, 70 of Kunming mouses, sterile saline, specification is the 1ml syringe, vaccine strains.
Experimental procedure:
1,70 mices are divided into 30 of vaccinate groups, 30 of counteracting toxic substances groups, 10 of blank groups.
2, every cervical region subcutaneous injection of vaccine group mice vaccine 0.2ml, blank group injection equivalent sterile saline.
3, all mice all places raising under the same terms, behind the 14d, carries out the counteracting toxic substances protection test.
4, earlier with each vaccine strains expanding propagation 11-13h, then concentration is all transferred to 2 * 10
9CFU/ml.When doing the counteracting toxic substances protection test, vaccinate group mice is divided into 3 groups, 10 every group, difference lumbar injection SWUN0662, SWUN0661, SWUN0642 bacterial strain bacterium liquid, every injected in mice bacterium liquid 1ml.The counteracting toxic substances group also is divided into three groups, and 10 every group, the counteracting toxic substances mode of counteracting toxic substances group is with the vaccine injection group.Behind the counteracting toxic substances, note observing the clinical symptoms of mice, and record death time and mortality rate.
Experimental result: the counteracting toxic substances matched group is in 24h, and mortality rate is all more than 70%, and the counteracting toxic substances protective rate of vaccine immunity group is all about 80%.
Embodiment 2
The secondary haemophilus trivalent of the pig that has prepared inactivated vaccine, 20 of Kunming white mice, sterile saline, 1ml syringe experimental procedure:
1, mice is divided into two groups, one group is the vaccinate group, and another group is the blank group of injecting normal saline.
2, every vaccinate 0.2ml of the mice of vaccinate group, matched group injection equivalent normal saline.All mices all place under the same terms and raise.
3, observe spirit of mice and situation such as search for food in the 28d of injection back.
Experimental result: in the 28d, lethargy does not appear in mice, undesirable conditions such as feed intake reduction.Show that this vaccine has higher safety for mice.
Claims (6)
1. the secondary haemophilus serum of pig 4 types, 5 types, 13 type trivalent oil emulsion inactivated vaccines is characterized in that separated strain by main popular serotype 4,5,13 types of the secondary haemophilus of the current pig of China is mixed in proportion to make the trivalent oil emulsion inactivated vaccine after deactivation.The composition of contained adjuvant has mineral oil, surfactant.
2. the used separated strain of trivalent oil emulsion inactivated vaccine according to claim 1 is the secondary haemophilus serum of pig 4,5,13 types, and strain number is SWUN0662, SWUN0661, SWUN0642, all separates from Guangdong.
3. trivalent oil emulsion inactivated vaccine according to claim 1, wherein said antigenic preparation method is, the secondary haemophilus serum of pig 4,5,13 type separated strains are inoculated in 1% the 0.02%NAD solution and the TSB meat soup of 5% newborn calf serum, 37 ℃, 180r/min shaken cultivation 24h, the adding final concentration is 0.3% formalin-inactivated 11-13h, and the culture concentration with three kinds of bacterial strains all is concentrated into 1 * 10 at last
10CFU/ml.
4. trivalent oil emulsion inactivated vaccine according to claim 1, wherein said mineral oil are Marko's 52 lightweight paraffin oil.
5. trivalent oil emulsion inactivated vaccine according to claim 1, wherein said surfactant are Si Ben-80, tween 80, aluminium stearate.
6. trivalent oil emulsion inactivated vaccine according to claim 1, the preparation method of wherein said oil-emulsion inactivated vaccine, this method may further comprise the steps:
1) cultivation of antibacterial: the secondary haemophilus serum of pig 4 types, 5 types and 13 types, three strain kind daughter bacterias are inoculated in the TSA solid medium respectively, cultivate in 37 ℃ of calorstats, the single bacterium colony of picking behind the 24h, access contains in the TSB fluid medium of NAD and calf serum, this bacterium liquid is added in 1% ratio behind the shaken cultivation 12-16h under 37 ℃, 180r/min condition in the TSB fluid medium of new preparation to breed in a large number again.Behind 37 ℃, 180r/min shaken cultivation 24h, collect bacterium liquid, use the spectrophotometric determination bacterial concentration.
2) deactivation of antibacterial: press TSB fluid medium cumulative volume 0.3% and slowly add formaldehyde, in 37 ℃ of vibration deactivation 11-13h.
3) preparation of oil emulsion water: after the strain deactivation of three serotypes, the bacterial concentration of each serotype is all transferred to 1 * 10
10CFU/ml, the bacterium liquid that will adjust then after the concentration mixes by 1: 1: 1 volume ratio, slowly adds the sterilization tween 80 and stirs by 4% of cumulative volume.
4) oil phase preparation: get 93 parts of Marko's 52 lightweight paraffin oil, add 1 part of aluminium stearate, the limit edged stirs, and till transparent, adds 6 parts of Si Ben-80 again, abundant mixing, and 121 ℃ of high pressure steam sterilization 30min, it is standby to be cooled to room temperature.
5) preparation of oil-emulsion inactivated vaccine: the oil phase and the water equal-volume that prepare are mixed emulsifying in homogenizer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910263482A CN101721696A (en) | 2009-12-18 | 2009-12-18 | Trivalent oil emulsion inactivated vaccine for 4 type, 5 type and 13 type serum of haemophilus parasuis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910263482A CN101721696A (en) | 2009-12-18 | 2009-12-18 | Trivalent oil emulsion inactivated vaccine for 4 type, 5 type and 13 type serum of haemophilus parasuis |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101721696A true CN101721696A (en) | 2010-06-09 |
Family
ID=42443644
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910263482A Pending CN101721696A (en) | 2009-12-18 | 2009-12-18 | Trivalent oil emulsion inactivated vaccine for 4 type, 5 type and 13 type serum of haemophilus parasuis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101721696A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102499982A (en) * | 2011-12-21 | 2012-06-20 | 青岛易邦生物工程有限公司 | Method for producing trivalent inactivated vaccine against Haemophilus parasuis infection |
| CN102908615A (en) * | 2011-08-01 | 2013-02-06 | 普莱柯生物工程股份有限公司 | Novel haemophilus parasuis disease trivalent inactivated vaccine and preparation method thereof |
| US20150098968A1 (en) * | 2013-10-04 | 2015-04-09 | Merial Limited | Haemophilus parasuis vaccine serovar type four |
| CN105112342A (en) * | 2015-09-17 | 2015-12-02 | 龙岩学院 | Haemophilus parasuis strain |
| WO2016119078A1 (en) * | 2015-01-29 | 2016-08-04 | 山东省农业科学院畜牧兽医研究所 | Combined use of haemophilus parasuis lc strain and lz-20100109 strain |
| CN109806389A (en) * | 2019-02-22 | 2019-05-28 | 河南省农业科学院畜牧兽医研究所 | A kind of trivalent inactivated vaccine against Haemophilus parasuis infection and its application |
| CN114805554A (en) * | 2022-04-13 | 2022-07-29 | 西南民族大学 | Refined yolk antibody injection for resisting streptococcus suis and haemophilus parasuis and preparation method thereof |
-
2009
- 2009-12-18 CN CN200910263482A patent/CN101721696A/en active Pending
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102908615A (en) * | 2011-08-01 | 2013-02-06 | 普莱柯生物工程股份有限公司 | Novel haemophilus parasuis disease trivalent inactivated vaccine and preparation method thereof |
| CN102908615B (en) * | 2011-08-01 | 2014-11-19 | 普莱柯生物工程股份有限公司 | Novel haemophilus parasuis disease trivalent inactivated vaccine and preparation method thereof |
| CN102499982B (en) * | 2011-12-21 | 2013-07-17 | 青岛易邦生物工程有限公司 | Method for producing trivalent inactivated vaccine against Haemophilus parasuis infection |
| CN102499982A (en) * | 2011-12-21 | 2012-06-20 | 青岛易邦生物工程有限公司 | Method for producing trivalent inactivated vaccine against Haemophilus parasuis infection |
| JP2016533353A (en) * | 2013-10-04 | 2016-10-27 | メリアル インコーポレイテッド | Hemophilus paraswiss vaccine serotype 4 |
| US20150098968A1 (en) * | 2013-10-04 | 2015-04-09 | Merial Limited | Haemophilus parasuis vaccine serovar type four |
| US9504740B2 (en) * | 2013-10-04 | 2016-11-29 | Merial, Inc. | Haemophilus parasuis vaccine serovar type four |
| WO2016119078A1 (en) * | 2015-01-29 | 2016-08-04 | 山东省农业科学院畜牧兽医研究所 | Combined use of haemophilus parasuis lc strain and lz-20100109 strain |
| CN105112342A (en) * | 2015-09-17 | 2015-12-02 | 龙岩学院 | Haemophilus parasuis strain |
| CN109806389A (en) * | 2019-02-22 | 2019-05-28 | 河南省农业科学院畜牧兽医研究所 | A kind of trivalent inactivated vaccine against Haemophilus parasuis infection and its application |
| CN109806389B (en) * | 2019-02-22 | 2022-03-22 | 河南省农业科学院畜牧兽医研究所 | Haemophilus parasuis trivalent inactivated vaccine and application thereof |
| CN114805554A (en) * | 2022-04-13 | 2022-07-29 | 西南民族大学 | Refined yolk antibody injection for resisting streptococcus suis and haemophilus parasuis and preparation method thereof |
| CN114805554B (en) * | 2022-04-13 | 2023-08-18 | 西南民族大学 | Refined egg yolk antibody injection for resisting streptococcus suis and haemophilus parasuis and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101721696A (en) | Trivalent oil emulsion inactivated vaccine for 4 type, 5 type and 13 type serum of haemophilus parasuis | |
| CN103263666B (en) | Porcine circovirus 2 type, porcine mycoplasmal pneumonia bivalent inactivated vaccine and preparation method thereof | |
| CN103157100B (en) | hemophilus parasuis disease, swine streptococcosis bivalent inactivated vaccine and preparation method thereof | |
| CN103083655A (en) | Vaccine composition for preventing and treating porcine circovirus type 2, haemophilus parasuis and mycoplasma hyopneumoniae infection and preparation method thereof | |
| CN103948918A (en) | Method for preparing trivalent inactivated vaccine for preventing mink hemorrhagic pneumonia | |
| JP2012521391A (en) | Vaccines to protect against various serotypes of Streptococcus bacterium | |
| CN105112349B (en) | A kind of brucella melitensis molecular marker vaccine strain and its application | |
| CN107513506A (en) | Mycoplasma hyopneumoniae, vaccine combination and its application | |
| CN105039233B (en) | A kind of B. abortus molecular marker vaccine strain and its application | |
| US9593303B2 (en) | Method for continuously culturing Ehrlichia and Neorickettsia risticii | |
| CN103602637B (en) | Vaccine strain for mycoplasma pneumonia of swine | |
| WO2016119078A1 (en) | Combined use of haemophilus parasuis lc strain and lz-20100109 strain | |
| CN104774796B (en) | Fowl enteropathogenic E. Coli inactivated vaccine and preparation method thereof | |
| CN105754905A (en) | Pseudomonas aeruginosa of minks and application of pseudomonas aeruginosa | |
| CN104056263B (en) | A kind of prevention and treatment are by vaccine combination and the preparation method and application of the respiratory disease of atrophic rhinitis secondary | |
| CN103157101B (en) | Combined inactivate vaccine for haemophilus parasuis disease and streptococcus suis disease and preparation method for same | |
| CN103656634B (en) | Resisting porcine circovirus and porcine contagious pleuropneumonia infect vaccine combination and preparation | |
| CN103585622B (en) | The application of Vaccine strain for mycoplasma pneumonia of swine | |
| CN106139137A (en) | A kind of vaccine combination and application thereof | |
| CN102309753A (en) | Riemerella anatipestifer serum 1 type, 2 type and 4 type trivalent oil emulsion inactivated vaccine | |
| CN110862938B (en) | Porcine pathogenic escherichia coli strain and inactivated vaccine thereof | |
| KR101210082B1 (en) | Vaccine composition for swine polyserositis and manufacturing method thereof | |
| CN103182077B (en) | Application of serum 4 type haemophilus parasuis vaccine strain | |
| KR101209964B1 (en) | Vaccine composition for swine polyserositis and manufacturing method thereof | |
| CN103566364B (en) | Porcine contagious pleuropneumonia, Streptococcus suis bivalent inactivated vaccine and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20100609 |