CN109663125A - A kind of chicken intestinal diorder salmonella inactivated vaccine and its application - Google Patents
A kind of chicken intestinal diorder salmonella inactivated vaccine and its application Download PDFInfo
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Abstract
The present invention relates to microorganisms technical field, in particular to a kind of fowl salmonella vaccine further relates to the application of the salmonella.A kind of chicken intestinal diorder salmonella inactivated vaccine, the vaccine composition includes chicken intestinal diorder Salmonella bacterium solution and adjuvant, wherein the adjuvant is made of the raw material of following parts by weight: 50 parts of aqueous oil adjuvant, 1.5-3.5 parts of Arlacel-80,1.5-2.5 parts of Tween-80,0.1 part of small peptide, 1.5-2.5 parts of propolis.The salmonella inactivated vaccine of offer of the invention is with strong points, and to local popular Salmonella strains good immune effect, preparation process is easy to operate, and cost is relatively low, easily stored, suitable for there is the scale farm in laboratory.
Description
Technical field
The present invention relates to microorganisms technical field, in particular to a kind of fowl salmonella vaccine further relates to the sramana
The application of Salmonella.
Background technique
As aquaculture industry of China is to intensive, large-scale development, the disease incidence of livestock and poultry is risen year by year, and new disease is not
Disconnected to increase, the common disease for especially endangering fowl poultry safety has the trend spread rapidly, becomes the weight of puzzlement aquaculture industry of China
Want problem.Up to the present, poultry salmonella infection is commonplace, mostly dysentery characterized by white mucous stool type and enteritis type, and antibiotic is anti-sand control
The door most effective means of Salmonella, but as drug resistance of the salmonella to antibiotic gradually rises, the clinical effectiveness of antibiotic by
Gradually decline, to affect antibiotic to the prevention and treatment effect of salmonella infection.To improve curative effect, clinical antibiotics make
It is higher than with dosage and instructs dosage, so as to cause antibiotic residue problem in livestock products, therefore clinically prevents and treats salmonella infection
It is badly in need of new antibiotic alternative or product.
The Development Techniques of vaccine have developed very mature, and the common adjuvant of vaccine mainly has aluminium glue, propolis, white oil, Chinese medicine
Extract etc..Salmonella inactivated vaccine can be by kind of a fowl maternal antibody, in chick early stage as a kind of prevention and control salmonella
The means of infection, can also be inoculated with salmonella inactivated vaccine by early immune, and excitation chick generates antibody and protects body not
By salmonella infection.Therefore the popularization and application of salmonella inactivated vaccine and industrialization prospect are very wide.
The serotype and immunogenicity of Salmonella strains decide the specificity and response immune response of vaccine.Screening is faced
The serological type strain of bed often hair salmonella infection, and the Biological indicators such as its safety and immunogenicity are detected, it is to develop the epidemic disease
The element task of seedling.Clinically multiple salmonella infection bacterial strain serotype is mostly enteritis type, white diarrhea, typhoid fever type etc., root
According to relevant regulations, white diarrhea needs control techniques, cannot research and develop vaccine, and typhoid fever type clinic is relatively fewer, therefore for clinical more
The chicken intestinal diorder salmonella infection of hair develops corresponding vaccine gesture necessary.In the past 20 years with immunology, molecular biology skill
The development of art, the development and application of bacterial vaccine are more rapid, extensive.
Summary of the invention
In order to solve the problems, such as that, without the prevention Bacterium enteritidis infection of effective inactivated vaccine in existing poultry husbandry, the present invention mentions
A kind of Salmonella gallinarum inactivated vaccine composition is supplied.
It is a further object of the present invention to provide the applications of above-mentioned chicken intestinal diorder salmonella inactivated vaccine.
The present invention is achieved by the following measures:
Vaccine composition described in a kind of chicken intestinal diorder salmonella inactivated vaccine includes chicken intestinal diorder Salmonella bacterium solution and adjuvant, wherein institute
Adjuvant is stated to be made of the raw material of following parts by weight: 50 parts of aqueous oil adjuvant, 1.5-3.5 parts of Arlacel-80, Tween-80 1.5-
2.5 parts, 0.1 part of small peptide, 1.5-2.5 parts of propolis.
Preferably, the Salmonella bacterium solution the preparation method comprises the following steps: choose infection Bacterium enteritidis broiler chicken, isolate sand
Door Salmonella；Salmonella is purified, salmonella is expanded culture, inactivates, it is dense that the salmonella of inactivation is diluted to its
Degree is greater than 5 × 109CFU/mL, 4 DEG C save backup.
The preparation method of above-mentioned chicken intestinal diorder salmonella inactivated vaccine, using following steps:
(1) match liquefaction phase adjuvant: by aqueous oil adjuvant, it is entirely molten that Arlacel-80 is heated to solids, is uniformly mixed to obtain oily phase adjuvant；
(2) prepare water phase: by adjuvant Tween-80, propolis, small peptide is added in bacterium solution, is uniformly mixed to obtain water phase；
(3) the water phase 1:1 in mass ratio emulsification of the oily phase adjuvant of step (1) preparation and step (2) preparation uniformly can be obtained
Inactivate salmonella vaccine.
Application of the above-mentioned chicken intestinal diorder salmonella inactivated vaccine in preparation prevention or treatment chicken intestinal diorder drug.
Beneficial effect
The salmonella inactivated vaccine of offer of the invention is with strong points, to local popular Salmonella strains immune effect
Good, preparation process is easy to operate, and cost is relatively low, easily stored, suitable for there is the scale farm in laboratory.
Specific embodiment
In order to better understand the present invention, it further illustrates combined with specific embodiments below.
Aqueous oil adjuvant used in following embodiment and comparative example is bought from Beijing Bo Aolong Immune Technology Corp..
Embodiment 1:
A kind of chicken intestinal diorder salmonella inactivated vaccine, is obtained by following steps:
Adjuvant is made of the raw material of following parts by weight: 50 parts of aqueous oil adjuvant, and 1.5 parts of Arlacel-80,2.5 parts of Tween-80,
0.1 part of small peptide, 1.5 parts of propolis.
Salmonella is expanded culture, is inactivated, 4 DEG C save backup；With liquefaction phase adjuvant: weighing water by weight ratio
Property oil adjuvant, Arlacel-80, it is entirely molten to be heated to solids, is uniformly mixed；It prepares water phase: the salmonella of inactivation is diluted to it
Concentration is greater than 5 × 109CFU/mL, then by adjuvant Tween-80, propolis, small peptide are added in bacterium solution, are uniformly mixed.By it is oily mutually and
Inactivation salmonella vaccine uniformly can be obtained by 1:1 emulsification in water phase.
Embodiment 2:
A kind of chicken intestinal diorder salmonella inactivated vaccine, is obtained by following steps:
Adjuvant is made of the raw material of following parts by weight: 50 parts of aqueous oil adjuvant, 3 parts of Arlacel-80,2.0 parts of Tween-80, small
0.1 part of peptide, 1.5 parts of propolis.
Salmonella is expanded culture, is inactivated, 4 DEG C save backup；With liquefaction phase adjuvant: weighing water by weight ratio
Property 50 parts of oil adjuvant, Arlacel-80, it is entirely molten to be heated to solids, is uniformly mixed；It prepares water phase: the salmonella of inactivation is diluted
It is greater than 5 × 10 to its concentration9CFU/mL, then adjuvant Tween-80, propolis, small peptide are added in bacterium solution, it is uniformly mixed.It will be oily
Mutually is emulsified by 1:1 and inactivation salmonella vaccine uniformly can be obtained with water phase.
Embodiment 3:
A kind of chicken intestinal diorder salmonella inactivated vaccine, is obtained by following steps:
Adjuvant is made of the raw material of following parts by weight: 50 parts of aqueous oil adjuvant, and 3.5 parts of Arlacel-80,1.5 parts of Tween-80,
0.1 part of small peptide, 2.5 parts of propolis.
Salmonella is expanded culture, is inactivated, 4 DEG C save backup；With liquefaction phase adjuvant: weighing water by weight ratio
Property 50 parts of oil adjuvant, Arlacel-80, it is entirely molten to be heated to solids, is uniformly mixed；It prepares water phase: the salmonella of inactivation is diluted
It is greater than 5 × 10 to its concentration9CFU/mL, then adjuvant Tween-80, propolis, small peptide are added in bacterium solution, it is uniformly mixed.It will be oily
Mutually is emulsified by 1:1 and inactivation salmonella vaccine uniformly can be obtained with water phase.
Comparative example 1
A kind of chicken intestinal diorder salmonella inactivated vaccine, is obtained by following steps:
Adjuvant is made of the raw material of following parts by weight: 50 parts of aqueous oil adjuvant, and 3 parts of Arlacel-80,2.0 parts of Tween-80, bee
1.5 parts of glue.
Salmonella is expanded culture, is inactivated, 4 DEG C save backup；With liquefaction phase adjuvant: weighing water by weight ratio
Property 50 parts of oil adjuvant, Arlacel-80, it is entirely molten to be heated to solids, is uniformly mixed；It prepares water phase: the salmonella of inactivation is diluted
It is greater than 5 × 10 to its concentration9CFU/mL, then adjuvant Tween-80, propolis are added in bacterium solution, it is uniformly mixed.By it is oily mutually and
Inactivation salmonella vaccine uniformly can be obtained by 1:1 emulsification in water phase.
Comparative example 2
A kind of chicken intestinal diorder salmonella inactivated vaccine, is obtained by following steps:
Adjuvant is made of the raw material of following parts by weight: 50 parts of oil adjuvant, 3 parts of Arlacel-80, and 2.0 parts of Tween-80, propolis 1.5
Part.
Salmonella is expanded culture, is inactivated, 4 DEG C save backup；With liquefaction phase adjuvant: weighing oil by weight ratio
50 parts of adjuvant, Arlacel-80, it is entirely molten to be heated to solids, is uniformly mixed；It prepares water phase: the salmonella of inactivation is diluted to it
Concentration is greater than 5 × 109CFU/mL, then adjuvant Tween-80, propolis are added in bacterium solution, it is uniformly mixed.By oily phase and water phase
Inactivation salmonella vaccine uniformly can be obtained by 1:1 emulsification.
Effect test
Illustrate performance characteristics of the invention by experimental data:
1. testing program
Healthy 3 Japanese instar chickling 120 is selected, is randomly divided into 6 groups: A group, 20, the chicken intestinal diorder Salmonella prepared according to embodiment 1
Bacterium inactivated vaccine, only, neck subcutaneous injection is immune by 0.5ml/；B group, 20, the chicken intestinal diorder salmonella prepared according to embodiment 2
Inactivated vaccine, only, neck subcutaneous injection is immune by 0.5ml/；C group, is gone out according to chicken intestinal diorder salmonella prepared by embodiment 3 by 20
Live vaccine, only, neck subcutaneous injection is immune by 0.5ml/；D group, is inactivated according to chicken intestinal diorder salmonella prepared by comparative example 1 by 20
Vaccine, only, neck subcutaneous injection is immune by 0.5ml/；E group, inactivates epidemic disease according to chicken intestinal diorder salmonella prepared by comparative example 2 by 20
Seedling, only, neck subcutaneous injection is immune by 0.5ml/；F group, 20, blank assay control is not immune only to attack poison.
2. evaluation index
2.1 clinical symptoms periodically observe and record the clinical symptoms of chicken, such as appetite, breathing, the state of mind daily.
2.2 pathologic findings dissect dead chicken, observe pathological change.
2.3 efficacy determinations are cured, are improved, is effective, is dead.
7,14,21d after 2.4 serological method A, B, C, D, E groups are immune adopt together with control F group and pass through slide agglutination
Anti- enteritis type Salmonella gallinarum antibody agglutination titer in hematometry serum.
After 2.5 Immunization method vaccine immunity 21d, each 10 chickens of A, B, C, D, E, F group, chest muscle injection are taken respectively
Corresponding vaccine manufacture salmonella strain 1,000,000,000/only；Observation 10 days, all chickens of dissect.
3 results
3.1 clinical symptoms
After healthy chick is immune, mental symptom, feeding situation have no significant change.Morbidity chicken mainly shows after infection salmonella
For spirit is depressed, loose random, loss of appetite, drinking-water increases, anus is stained with color loose stool, there are the phenomena of mortality in severe infections person.
3.2 pathological change
Morbidity chick dissect is as it can be seen that liver enlargement, unsharp border have a tiny canescence necrosis point, and intestinal mucosa is congested, bleeding,
There is white downright bad point on surface, and meropia phleboedesis is big.
The measurement of 3.3 antibody titers (is measured) using slide agglutination
It takes a blood sample within 21 days after vaccine immunity test chicken, the anti-enteritis type Salmonella gallinarum antibody of chicken serum is measured using slide agglutination method
Agglutination titer, testing result are shown in Table 1.
Antibodies toward salmonella testing result after table 1 is immune
3.4 Immunization methods
Poison is attacked within 21 days after 5 kinds of vaccine immunities, respectively the Salmonella gallinarum bacterium solution of intramuscular injection one minimum morbidity dosage, observation note
Record test chicken condition of morbidity death.Test result is shown in Table 2.
2 three kinds of vaccine immune chickens of table attack malicious salmonella S2 plants of immuning effect test results
Poison is attacked to Salmonella gallinarum according to the vaccine that embodiment 1, embodiment 2, embodiment 3, comparative example 1, comparative example 2 are prepared respectively
Immune protective rate (according to clinical symptoms) be respectively 6/10,6/10,7/10,4/10,2/10 protection, control chicken 10/10 fall ill；
From the point of view of dead and clinical symptoms, immune protective rate is respectively 6/10,6/10,7/10,4/10,6/10 protection, compares chicken 10/
10 morbidities, 4/10 death.These results suggest that using vaccine prepared by the present invention (according to embodiment 1, embodiment 2, embodiment 3)
It is immune, there is good effect to local chicken intestinal diorder salmonella infection is controlled, is superior to prepare according to comparative example 1 and comparative example 2
Vaccine.
Claims (4)
1. a kind of chicken intestinal diorder salmonella inactivated vaccine, which is characterized in that the vaccine composition includes chicken intestinal diorder salmonella
Liquid and adjuvant, wherein the adjuvant is made of the raw material of following parts by weight: 50 parts of aqueous oil adjuvant, Arlacel-80 1.5-3.5
Part, 1.5-2.5 parts of Tween-80,0.1 part of small peptide, 1.5-2.5 parts of propolis.
2. chicken intestinal diorder salmonella inactivated vaccine according to claim 1, which is characterized in that the system of the Salmonella bacterium solution
Preparation Method are as follows: the broiler chicken for choosing infection Bacterium enteritidis isolates salmonella；Salmonella is purified, by salmonella
It expands culture, inactivates, the salmonella of inactivation is diluted to its concentration greater than 5 × 109CFU/mL, 4 DEG C save backup.
3. a kind of preparation method of chicken intestinal diorder salmonella inactivated vaccine of any of claims 1 or 2, which is characterized in that use
Following steps:
(1) match liquefaction phase adjuvant: by aqueous oil adjuvant, it is entirely molten that Arlacel-80 is heated to solids, is uniformly mixed to obtain oily phase adjuvant；
(2) prepare water phase: by adjuvant Tween-80, propolis, small peptide is added in bacterium solution, is uniformly mixed to obtain water phase；
(3) the water phase 1:1 in mass ratio emulsification of the oily phase adjuvant of step (1) preparation and step (2) preparation uniformly can be obtained
Inactivate salmonella vaccine.
4. chicken intestinal diorder salmonella inactivated vaccine according to claim 1-3 is in preparation prevention or treatment chicken intestinal diorder
Application in drug.
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| CN105899228A (en) * | 2013-10-11 | 2016-08-24 | 加利西亚健康服务(Sergas) | Live attenuated vaccines |
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