CN110013551A - A kind of fishing immersion immunity composite adjuvant and the preparation method and application thereof - Google Patents
A kind of fishing immersion immunity composite adjuvant and the preparation method and application thereof Download PDFInfo
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- CN110013551A CN110013551A CN201910421149.XA CN201910421149A CN110013551A CN 110013551 A CN110013551 A CN 110013551A CN 201910421149 A CN201910421149 A CN 201910421149A CN 110013551 A CN110013551 A CN 110013551A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
- A61K2039/552—Veterinary vaccine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
- A61K2039/55583—Polysaccharides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55588—Adjuvants of undefined constitution
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Abstract
The invention discloses a kind of less toxic, efficient, quick-acting, long-acting fishing immersion immunity composite adjuvants and the preparation method and application thereof, are related to aquaculture.Fishing immersion immunity composite adjuvant is made of following mass percent component: RH-40 25~35%, dehydrated alcohol 10~20%, astragalus polyose 10~15%, propolis 5~8%, ethyl acetate 3~6%, sodium ascorbate 1~3%, sodium chloride 1~2% and distilled water 11~55%.
Description
Technical field
The present invention relates to aquaculture, especially a kind of fishing immersion immunity composite adjuvant and the preparation method and application thereof.
Background technique
Free of contamination aquaculture vaccines can safely, effectively prevent aquiculture animal epidemic disease because of it, it has also become water in the world
The mainstream technology of control and prevention of disease is produced, the research and development of aquaculture vaccines develop towards " efficient, practical, low price, multiplicity " direction, impregnate mouth
The practicabilities technology such as clothes, multi-joint multivalence, regional differentiation immunization protocol will be applied onto production management.Wherein immersion immunity operation letter
List can realize that batch is injected, is small to fish body stimulation in the short time, more preferably to fingerling fry effect of inoculation, and immersion immunity has
The potentiality for exciting fish body mucosal immune response, for defending the cause of disease for entering fish body by mucous membrane tissue significant, promotion price
Value is high.The vaccine applied in aquaculture production at present is mostly inactivated vaccine, cannot be bred in animal body after inoculation, inoculation
Dosage is big, and protection period is short.And vaccine dosage is big during immersion immunity, and the immunity that fish body obtains is relatively weak, exempts from
Epidemic disease is at high cost, and the production for seriously constraining immersion immunity promotes and applies.Therefore it on the basis of constantly constructing new fish vaccine, grinds
Study carefully and how to improve the significant of immersion immunity effect.
The superiority and inferiority of fish vaccine immune effect and the selection of adjuvant, there are biggish relationships for application method.Immersion immunity exists
There is apparent superiority in terms of small dimension, mass disposal, can be significantly improved by the use of adjuvant and new operating technology
The immune protective rate of vaccine, therefore adjuvant has become the hot spot in fish immunology field with the research and development that vaccine uses, with
The application of new material new technology, more and more adjuvants and technology will enter immersion immunity, and improve immersion immunity effect.But mesh
Come to see, antigen submission in immersion immunity, also lacks science particular against former size, property, sensitive and tolerance etc.
Property systematic data supporting practical specific aim therefore is lacked to the screening for impregnating adjuvant and operating technology, constrain industry
Development.Less toxic, efficient, quick-acting, long-acting fish immunologic adjuvant is developed by way of multi-source, multipath, and safe, fast
Prompt, easy-operating vaccine has become fish disease prevention and cure, the development trend of especially great control and prevention of disease research using technology.
Summary of the invention
An object of the present invention is to provide a kind of less toxic, efficient, quick-acting, long-acting fishing immersion immunity composite adjuvant.
To achieve the above object, present invention provide the technical scheme that a kind of fishing immersion immunity composite adjuvant, by following
Mass percent component is made: RH-40 25~35%, dehydrated alcohol 10~20%, astragalus polyose 10~15%, propolis 5~
8%, ethyl acetate 3~6%, sodium ascorbate 1~3%, sodium chloride 1~2% and distilled water 11~55%.
Preferably, the fishing is made with immersion immunity composite adjuvant of following mass percent component: RH-40 30%, nothing
Water-ethanol 15%, astragalus polyose 12%, propolis 6%, ethyl acetate 5%, sodium ascorbate 2.5%, sodium chloride 1.5% and distillation
Water 25.5%.
The second object of the present invention is to provide the preparation method of above-mentioned fishing immersion immunity composite adjuvant.
To achieve the above object, the technical solution of offer of the invention is: propolis being completely dissolved in dehydrated alcohol, and is added
Enter RH-40 and ethyl acetate, be added dropwise after mixing content be 37.5% (mass ratio) astragalus polyose aqueous solution, ascorbic acid and
Sodium chloride to get homogeneous transparent immunity composite adjuvant.
The third object of the present invention is to provide the application of above-mentioned fishing immersion immunity composite adjuvant.
To achieve the above object, present invention provide the technical scheme that immune group is impregnated in 1.5% salt Aeration in the water
After 5min, by immunity composite adjuvant, 1:1~2 are mixed by volume with vaccine, are placed in water by the volume 1.5- after mixed in equal amounts
2% concentration uses, and concentration and dosage become larger with body size and increased, and is shown in Table 1.
1 vaccine of table uses concentration table corresponding with body size
, such as there is exception in the stable state that close observation security immunization measurement fish is wanted during mass immunization, should be timely
To corresponding batch exempted from fish density, in terms of take early stage security protection handle.It is usually chosen in fine, water
The suitable morning 8-10 point of temperature, 4-6 point in afternoon carry out.Fingerling hanging culture started dip operation after 2-3 hours again.
Soaking time should be tested in advance according to different cultivars sensibility.On the day before seed is impregnated, random sampling 20-
50 tails are tested, and using under concentration, fingerling quantity and aerobic condition as defined in the vaccine product specification, are observed defined
Whether there is abnormal response in soaking time, by extending soaking time or raising vaccine using concentration 50%~100% or adding
The ways such as big 20%~50% fry density determine whether immune operation is safe to this batch of fry under corresponding condition.
This product also can be used as injection adjuvant, and the dosage used with vaccine is 1:1-2, depending on vaccination particles size,
Grain is bigger, higher using the dosage of adjuvant.
In use process, the fish sensitive to dissolved oxygen, such as mandarin fish, bamboo shoot shell fish, it is used with caution.It is secondary after impregnating 7 days first times
Impregnate better effect.
The present invention has the advantage that compared with existing adjuvant
Fish are weak to antigen absorbability, and composite adjuvant of the present invention changes point in water body of vaccination particles
Cloth, stability is good, is not in layering and precipitating during transport and storage；It, can be with especially to the use of propolis adjuvant
The dosage for improving propolis to the full extent, protects vaccine antigen.
Composite adjuvant of the present invention is highly-safe, changes fish skin surface mucous characteristic, and antigen is promoted to absorb, also
Stress reaction caused by fish operate because of immersion etc. can be reduced, the immune international headache of seed scale is solved, epidemic disease can be reduced
50% or more seedling cost provides feasibility for industrial application of the vaccine in low value fish.
Composite adjuvant of the present invention can be used directly with vaccine, and viscosity is low, not need to emulsify, and simplify production technology,
Reduce operating cost.
Composite adjuvant of the present invention can promote the application of immersion immunity, reduce aquiculture animal morbidity, reduce
The use of antibiotic etc strengthens food hygiene and safety guarantee.
Specific embodiment
Illustrate the present invention With reference to embodiment: but it is understood that these specific embodiments are only intended to
Illustrate the present invention, rather than limiting the invention.Those skilled in the art completely can under the inspiration of the present invention, to this
The specific embodiment or technical characteristic of invention improve, but these technology technical solutions by improvement or replacement,
Still fall within protection scope of the present invention.
Embodiment 1
The operation place of the present embodiment are as follows: Jining City's aquatic products seed stock breeding station.
(1) cultivation grass carp is divided into 3 groups, every group of 300 tails are gone out by fish is exempted from the Aeromonas hydrophila containing adjuvant of the present invention
15min is impregnated in oxygenation in live vaccine solution, and control fish only impregnates 5min in 1.5% salt water.Immersion volume is 50L, and impregnates epidemic disease
Final bacterial concentration is not less than 4.2 × 10 in seedling mixed solution8CFU/mL。
(2) in net cage test, composite adjuvant and immersion immunity grass carp of aeromonas hydrophila inactivated vaccine, 2 weeks serum
Antibody titer, which reaches to reach within 160,6 weeks peak value 640,8 weeks, was reduced to 40 for 320,12 weeks.The results show that composite adjuvant immune group
Immunization time is longer, and serum antibody titer is also higher, peak is reached at 6 weeks, and composite adjuvant immersion immunity group is once impregnated
There is higher serum antibody titer within immune 8 weeks (about 2 months).
(3) go out the pool after cultivating 6 months.During grass carp cultivates, grows fine, disease problem do not occur, in breeding process
In, that because occurring, bad water colour, ammonia nitrogen, nitrite are not exceeded, pH value is high or change the reasons such as violent, and send out
Raw infected by Aeromonas hydrophila.
Embodiment 2
The operation place of the present embodiment are as follows: Guangdong Province's aquatic products vaccine Industry Technology Center.
(1) crucian is divided into 8 groups, every group of 120 tails.Immune group is containing respectively after 1.5% salt Aeration in the water impregnates 5min
15min is impregnated in inflation in 2% vaccine adjuvant (vaccine and adjuvant volume ratio 1:1 of the present invention) composite solution.After first immunisation 7d
Booster immunization is carried out with method, control group only impregnates 5min in 1.5% salt water.Every group of fish is randomly divided into 4 parallel groups after immune,
It is placed in indoor glass cylinder and inflates raising.Every group of immersion volume is 5L, in soaking vaccine solution bacterial antigens concentration be 4.6 ×
108CFU/mL。
(2) 20 days after secondary immersion, Aeromonas hydrophila SDLL2 plants of progress challenge viral dosage is used to the crucian for changing test.
Every group takes experiment fish to add up to 20 tails, injects the grass carp of health, every endnote penetrates 0.2 milliliter, and every group of 20 tails, water temperature is constant during test
At 28 DEG C or so, the incidence of test fish is observed day by day, is observed continuously 15.Correct evaluation test is wanted as a result, every test group
It at least needs with 10 tail grass carps, each test group of this test uses 20 tail grass carps, it is believed that the test is effective.
(3) secondary immersion immunity is shown, composite adjuvant makes to be exempted from fish relative protection ratio to reach 66.7%-88.0%, aobvious
Write the 50.5-60.2% higher than no adjuvant group.
Embodiment 3
The operation place of the present embodiment are as follows: Jining of Shandong Province city, deer low-lying area, Yutai County fry.
(1) in bighead, silver carp carrying seedling truck, 20,000 tails are transported, the long average 3cm of body, 22 DEG C of water temperature, is divided into two by about 600 jin
Water tank, each cabinet add water 300L, and 3 meters of the depth of water, the water surface is to 1 meter of tank lid, to accommodate increase oxygen.It uses during transportation
Oxygen cylinder continuously uninterrupted oxygenation.
(2) Aeromonas hydrophila, glimmering of adjuvant of the present invention is added according to volume 1:1 compatibility for the half an hour before pond-inputting, investment
Light pseudomonad inactivates bigeminy vaccine, and usage amount is every case 3L.Examine vaccine and adjuvant products without demulsification and phenomena such as layering
Afterwards, packaging is opened, 1.5kg salt is added, 5 times of water is added to dilute, in inlet tank of uniformly splashing, as oxygenation is more and more, foam is very
It is more.Therefore pond-inputting operation is quick as far as possible, avoids the long time stress.Temperature is higher, and soaking time is shorter.
(3) in pond culture, main silver carp of raising flowers, 800 tails/mu, specification is 750g/ tail.Collocation grass carp, crucian, silver carp, bream
Deng.Although having the influence of the extreme weathers such as arid, heavy rain, no morbidity medication in the whole process.Variegated carp grows fine, intermediate without dead
Fish feeds intake few.There are many surrounding pond dead fish, and especially 7.14 days front and back storms have nearby that occur turning over sugared anoxic existing in four mouthfuls of ponds
As this pond does not occur any exception, reduce the operation difficulty breeded fish because the water transfer of flower silver carp acts on.
Embodiment 4
The operation place of the present embodiment are as follows: Foshan Shuande town Le Liu.
(1) cement pit temporarily supports largemouth bass fingerling, and the octagon for the use of pond size being 2 meters of side length is increased using bottom micropore
Oxygen, largemouth bass seedling body weigh about 5g, and 1800 tails after discharging water, about 0.2 meter of the depth of water, make the shoal of fish concentrate on pool aottom center position, estimate pond
Middle water is about 200-250L or so, uses Aeromonas hydrophila, the Webster Aeromonas that adjuvant of the present invention is added according to 1:1 compatibility
Inactivate bigeminy vaccine, usage amount 2L.It 20 DEG C of water temperature, after impregnating 25 minutes, opens a sluice gate to be put into cultivating pool.Front and back is impregnated respectively to stop
It only feeds intake 1 day.
(2) cultivation pool area is 10 mu, and pond shape is in transmeridional rectangle, has independent culvert system, the depth of water is
2.0 meters or so, a small amount of crucian of intercropping and bighead.It is equipped with the production facilities such as corresponding aerator, feeder, water pump.Add within every 5-7 days
It water 1 time, changes within 15-20 days water 1 time, each quantity of exchanged water is 20cm or so, and keeping transparency is 35cm or so.It is installed on aerator
Dissolved oxygen controller sets the upper and lower limit of water dissolved oxygen according to cultivation needs, carries out Full-automatic oxygen-increasing, remains water dissolved oxygen
In 5mg/L or more.
(3) it is compared in the breeding process with other two same pond, this pond seed ingests vigorous, neat specification, packet
Including the fish of intercropping, all there is no bleeding, rotten body and Other diseases, and other two mouthfuls of ponds fin base portion position occur in early August
Rubescent and lower jaw is rubescent to anus area, and especially there are the bleeding striped vertical with fish body side line, chest in gill cover rear body two sides
Fin base portion is also usually rubescent obvious.There is also the phenomenons of flush and eye socket hyperemia for part fish body.Dissection inspection, body cavity
Inner product has ascites, and sky is without food and with congested phenomenon in enteron aisle.Pond on probation averagely reduces fish loss late than control pond
17.28%.
Claims (4)
1. a kind of fishing immersion immunity composite adjuvant, which is characterized in that be made of following mass percent component: RH-40 25~
35%, dehydrated alcohol 10~20%, astragalus polyose 10~15%, propolis 5~8%, ethyl acetate 3~6%, sodium ascorbate 1
~3%, sodium chloride 1~2% and distilled water 11~55%.
2. fishing according to claim 1 immersion immunity composite adjuvant, which is characterized in that by following mass percent component
It is made: RH-40 30%, dehydrated alcohol 15%, astragalus polyose 12%, propolis 6%, ethyl acetate 5%, sodium ascorbate
2.5%, sodium chloride 1.5% and distilled water 25.5%.
3. a kind of preparation method of fishing as claimed in claim 1 or 2 immersion immunity composite adjuvant, which is characterized in that by propolis
It is completely dissolved in dehydrated alcohol, and RH-40 and ethyl acetate is added, it is 37.5% astragalus polyose water that content is added dropwise after mixing
Solution, ascorbic acid and sodium chloride to get homogeneous transparent immunity composite adjuvant.
4. a kind of application of fishing as claimed in claim 1 or 2 immersion immunity composite adjuvant, which is characterized in that immune group exists
1.5% salt Aeration in the water impregnate 5min after, by immunity composite adjuvant, 1:1~2 are mixed by volume with vaccine, be placed in water by etc.
The concentration for measuring mixed volume 1.5-2% uses.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910421149.XA CN110013551A (en) | 2019-05-21 | 2019-05-21 | A kind of fishing immersion immunity composite adjuvant and the preparation method and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201910421149.XA CN110013551A (en) | 2019-05-21 | 2019-05-21 | A kind of fishing immersion immunity composite adjuvant and the preparation method and application thereof |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101690809A (en) * | 2009-09-30 | 2010-04-07 | 西北农林科技大学 | Propolis-astragalus polysaccharides compound nanometer emulsion adjuvant and preparation method and application thereof |
| CN102488755A (en) * | 2011-12-23 | 2012-06-13 | 天津康莱森生物科技集团有限公司 | Fishing Chinese herbal composition capable of inhibiting germs and preparation method thereof |
| CN104667274A (en) * | 2015-02-11 | 2015-06-03 | 中国水产科学研究院珠江水产研究所 | Adjuvant for improving immersion immunization effect of fishery vaccines and application of adjuvant |
-
2019
- 2019-05-21 CN CN201910421149.XA patent/CN110013551A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101690809A (en) * | 2009-09-30 | 2010-04-07 | 西北农林科技大学 | Propolis-astragalus polysaccharides compound nanometer emulsion adjuvant and preparation method and application thereof |
| CN102488755A (en) * | 2011-12-23 | 2012-06-13 | 天津康莱森生物科技集团有限公司 | Fishing Chinese herbal composition capable of inhibiting germs and preparation method thereof |
| CN104667274A (en) * | 2015-02-11 | 2015-06-03 | 中国水产科学研究院珠江水产研究所 | Adjuvant for improving immersion immunization effect of fishery vaccines and application of adjuvant |
Non-Patent Citations (1)
| Title |
|---|
| 李清: "《水生动物疾病与安全用药手册》", 30 April 2014 * |
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