CN108553640A - A kind of chicken coccidiasis attenuated live vaccines and its preparation method and application - Google Patents
A kind of chicken coccidiasis attenuated live vaccines and its preparation method and application Download PDFInfo
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- 229960005486 vaccine Drugs 0.000 title claims abstract description 67
- 241000287828 Gallus gallus Species 0.000 title claims abstract description 65
- 230000002238 attenuated effect Effects 0.000 title claims abstract description 24
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 241000499566 Eimeria brunetti Species 0.000 claims abstract description 40
- 241000179199 Eimeria mitis Species 0.000 claims abstract description 37
- 239000007788 liquid Substances 0.000 claims abstract description 6
- 239000000825 pharmaceutical preparation Substances 0.000 claims abstract description 5
- 230000036039 immunity Effects 0.000 claims description 19
- 241000223924 Eimeria Species 0.000 claims description 6
- 201000010099 disease Diseases 0.000 claims description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 5
- 230000003053 immunization Effects 0.000 claims description 5
- 239000004744 fabric Substances 0.000 claims description 3
- 238000002649 immunization Methods 0.000 claims description 3
- 230000005847 immunogenicity Effects 0.000 abstract description 4
- 206010059866 Drug resistance Diseases 0.000 abstract description 3
- 239000000203 mixture Substances 0.000 abstract description 3
- 239000003640 drug residue Substances 0.000 abstract description 2
- 238000003912 environmental pollution Methods 0.000 abstract description 2
- 238000004321 preservation Methods 0.000 abstract description 2
- 235000013330 chicken meat Nutrition 0.000 description 54
- 230000037396 body weight Effects 0.000 description 20
- 239000002775 capsule Substances 0.000 description 13
- 241000224483 Coccidia Species 0.000 description 10
- 235000019786 weight gain Nutrition 0.000 description 7
- 210000003250 oocyst Anatomy 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 5
- 238000009360 aquaculture Methods 0.000 description 3
- 244000144974 aquaculture Species 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000001900 immune effect Effects 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- 230000007918 pathogenicity Effects 0.000 description 3
- 235000021053 average weight gain Nutrition 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000003224 coccidiostatic agent Substances 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000008004 immune attack Effects 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 235000010894 Artemisia argyi Nutrition 0.000 description 1
- 208000003322 Coinfection Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 241000499563 Eimeria necatrix Species 0.000 description 1
- 241000223932 Eimeria tenella Species 0.000 description 1
- 206010053759 Growth retardation Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 244000030166 artemisia Species 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 208000027503 bloody stool Diseases 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 210000005069 ears Anatomy 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 231100000001 growth retardation Toxicity 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 208000035861 hematochezia Diseases 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 208000020442 loss of weight Diseases 0.000 description 1
- 238000003305 oral gavage Methods 0.000 description 1
- 210000004681 ovum Anatomy 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 235000013594 poultry meat Nutrition 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000000384 rearing effect Effects 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000004584 weight gain Effects 0.000 description 1
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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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P33/00—Antiparasitic agents
- A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
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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
- A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
-
- 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
- A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
- A61K2039/541—Mucosal route
- A61K2039/542—Mucosal route oral/gastrointestinal
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Abstract
The invention discloses a kind of chicken coccidiasis attenuated live vaccines and its preparation method and application, the vaccine includes：100~500/part of E.brunetti, 100~500/part of Eimeria mitis, the vaccine are stored in preservation liquid and prepare by causing weak worm strain to cause weak worm strain to be uniformly mixed with the precocity of Eimeria mitis the precocity of E.brunetti；The vaccine composition is scientific and reasonable, at low cost, using rear drug residue free, does not generate drug resistance, not can cause environmental pollution, and immunogenicity is good, safe to use, can be applied to prepare the pharmaceutical preparation for preventing chicken coccidiasis.
Description
Technical field
The present invention relates to coccidial vaccine and its application field, more particularly to a kind of chicken coccidiasis attenuated live vaccines and its preparation
Methods and applications.
Background technology
Chicken coccidiasis is as caused by one or more Eimerias parasitize chicken intestinal mucosal epithelial cells with intestines
Inflammation, diarrhea, growth retardation, feed conversion rate decline and the dead epidemic disease being characterized.In 7 universally acknowledged breeder Amys
In ear ball worm, E.brunetti belongs to the worm kind of medium pathogenicity, and Eimeria mitis is that pathogenicity is relatively weak
Worm kind.From Epidemic status it is found that 7 kinds of coccidias are found in larger regional extent, but the prevalence rate of various coccidias
There are larger differences.E.brunetti can be found all over the world, but the report countries in the world in relation to prevalence rate
Each department are not quite similar, and there is prevalence certain regionality, most literature to report the pathogenic main table of E.brunetti
It is now morbidity and loss of weight rather than death, the economic loss caused by poultry husbandry is larger in terms of chicken weightening, feed conversion rate.With
There are higher prevalences in some areas for slow Eimeria, individually infect or can all make Broiler Chicken with other coccidia mixed infections
Decline again, feed conversion rate reduces.
Prevent chicken coccidiasis at present and relies primarily on coccidiostat and coccidia work egg capsule vaccine.The abuse of coccidiostat is led
The aggravation of coccidia drug resistance and food medicament residue are caused, human health is endangered, pollutes environment.The anti-of chicken coccidiasis is carried out using vaccine
It is a main trend to control, and the chicken coccidiasis vaccine listed now is mainly for the Eimeria tenella for causing serious bloody stool or death
Worm, Eimeria Necatrix, huge Eimeria and heap-type Eimeria carry out immunoprophylaxis, and temporarily draw without effective be directed to
Play the coccidial vaccine that chicken is slow-growing, weightening is poor.As it can be seen that exploitation is for causing, chicken is slow-growing, the coccidia epidemic disease for the difference that increases weight
Seedling is necessary.
As it can be seen that the prior art could be improved and improve.
Invention content
Place in view of above-mentioned deficiencies of the prior art, the purpose of the present invention is to provide a kind of chicken coccidiasis attenuated live vaccines
And its preparation method and application, it is intended to it solves in the prior art without for by E.brunetti and Eimeria mitis
The problem of coccidial vaccine that caused chicken is slow-growing, weightening is poor.
In order to achieve the above object, this invention takes following technical schemes：
A kind of chicken coccidiasis attenuated live vaccines, including：100~500/part of E.brunetti, Eimeria mitis
100~500/parts.
In the chicken coccidiasis attenuated live vaccines, including：250/part of E.brunetti, Eimeria mitis
250/parts.
In the chicken coccidiasis attenuated live vaccines, the E.brunetti is that the precocity cause of E.brunetti is weak
Worm strain, the Eimeria mitis are that the precocity of Eimeria mitis causes weak worm strain.
In the chicken coccidiasis attenuated live vaccines, the immunization ways of the vaccine include oral immunity and eye drip.
A kind of preparation method of chicken coccidiasis attenuated live vaccines, including：The precocity of E.brunetti is caused into weak worm strain
It causes weak worm strain to be uniformly mixed with the precocity of Eimeria mitis, is stored in and preserves in liquid.
A kind of application of chicken coccidiasis attenuated live vaccines, the vaccine are applied to prepare the pharmaceutical preparation for preventing chicken coccidiasis
In.
Advantageous effect：
The present invention provides a kind of chicken coccidiasis attenuated live vaccines and its preparation method and application, the vaccine composition science is closed
Reason, it is at low cost, using rear drug residue free, drug resistance is not generated, do not can cause environmental pollution, immunogenicity is good, uses peace
Entirely, 1 × 10 is highly resistant to after chicken is immune5A E.brunetti and 1 × 105The infection of a Eimeria mitis, can
Applied to the pharmaceutical preparation for preventing chicken coccidiasis is prepared, having filled up shortage in the prior art, chicken is slow-growing, increases for causing
The defect of the coccidial vaccine of the method for double differences improves the economic benefit of chicken aquaculture.
Specific implementation mode
The present invention provides a kind of chicken coccidiasis attenuated live vaccines and its preparation method and application, for make the purpose of the present invention,
Technical solution and effect are clearer, clear, and the present invention is described in more detail by the following examples.It should be appreciated that herein
Described specific examples are only used to explain the present invention, is not intended to limit the present invention.
The present invention provides a kind of chicken coccidiasis attenuated live vaccines, including：100~500/part of E.brunetti,
100~500/part of Eimeria mitis；E.brunetti and Eimeria mitis mainly cause chicken morbidity,
Rate of body weight gain reduces, and to reduce the conversion ratio of feed, reduces the economic benefit of chicken aquaculture；The vaccine immunogenicity of above-mentioned composition
It is good, it is safe to use, 1 × 10 can be reasonably resistant in the dosage range5A E.brunetti and 1 × 105A gentle Chinese mugwort
The infection of U.S. ear ball worm, prevent the chicken caused by E.brunetti, Eimeria mitis is slow-growing, under rate of body weight gain
The phenomenon that drop.When the immunizing dose of E.brunetti, Eimeria mitis is less than above range, can not generate effectively
Immune effect；When the immunizing dose of E.brunetti, Eimeria mitis is more than above range, immune response
In strong, the upgrowth situation of chicken may be influenced.
Preferably, the vaccine includes：250/part of E.brunetti, 250/head of Eimeria mitis
Part；Under the dosage, the immune effect that chicken obtains is best, and the influence to the upgrowth situation of chicken is minimum, rate of body weight gain highest.
Preferably, the E.brunetti is that the precocity of E.brunetti causes weak worm strain, the gentle Amy
Ear ball worm is that the precocity of Eimeria mitis causes weak worm strain.Precocious worm strain is weaker relative to original worm strain pathogenicity, but energy
Its immunogenicity is enough kept, therefore immune effect of vaccine has fully guaranteed, while use is upper more safe, avoids vaccine
Cause potentially to dissipate poison threat during use and cause excessive damage to the enteron aisle of chicken, so that improving aquaculture cost.
Specifically, the immunization ways of the vaccine include oral immunity and eye drip；More specifically, the oral immunity includes
Spice is immune, drinking-water is immunized, gavages.
A kind of preparation method of chicken coccidiasis attenuated live vaccines, including：The precocity of E.brunetti is caused into weak worm strain
It causes weak worm strain to be uniformly mixed with the precocity of Eimeria mitis, is stored in and preserves in liquid, preserved in 2~8 DEG C of environment；Tool
Body, the preservation liquid includes 1% toluene-sodium-sulfonchloramide solution.
A kind of application of chicken coccidiasis attenuated live vaccines, the vaccine are applied to prepare the pharmaceutical preparation for preventing chicken coccidiasis
In.
The preparation of 1 chicken coccidiasis attenuated live vaccines of embodiment
E.brunetti, Eimeria mitis Sporulated Oocysts are counted respectively with blood counting chamber, obtain its concentration,
The required quantity of each component coccidian oocyst is calculated according to the proportioning of table 1, is respectively put into different clean containers, is added and protects
Liquid storage is simultaneously stirred, and is obtained the mixing worm strain egg capsule solution of corresponding number of components, is placed in 2~8 DEG C of environment and saves backup.
1 chicken coccidiasis attenuated live vaccines of table at being grouped as
| E.brunetti（A/head part） | Eimeria mitis（A/head part） | |
| Proportioning 1 | 100 | 100 |
| Proportioning 2 | 250 | 250 |
| Proportioning 3 | 500 | 100 |
| Proportioning 4 | 100 | 500 |
Embodiment 2
Take 3~7 age in days SPF chickens（Finger is grown in barrier system or isolator, and no international, especially domestic popular main chicken passes
The chicken group for cause of disease of catching an illness）110, it is divided into 11 groups, every group 10, weighs.Wherein, the 1st, 2 group is to match being immunized for 1 vaccine
Worm group is attacked, the 3rd, 4 group is to match the immune of 2 vaccines to attack worm group, and the 5th, 6 group is attacked worm group for proportioning the immune of 3 vaccines,
7th, 8 group is to match the immune of 4 vaccines to attack worm group, and the 9th, 10 group is attacked worm group to be not immune；11st group is not attacked to be not immune
Worm group.
By the isolated rearing under the same conditions respectively of 11 groups of SPF chickens, after administered by oral gavage is 21 days immune, each group chicken weighs and attacks
Worm, the 1st, 3,5,7,9 group is orally attacked E.brunetti malicious Sporulated Oocysts 1 × 10 by force5A/only；2nd, 4,6,8,10 group
It is oral to attack Eimeria mitis malicious Sporulated Oocysts 1 × 10 by force5A/only, it observes 7, records each group clinical symptoms.It receives respectively
Take the 2nd, 4,6,8,10 group attack excrement and 1st of the worm after 120~144 hours, 3,5,7,9 groups attack excrement of the worm after 144~168 hours
Just, OPG is calculated separately（Gram excrement egg sac number）With egg capsule slip.Worm is attacked to weigh the weight of each group chicken after 7 days and calculate phase
To rate of body weight gain, and using body weight increase rate, egg capsule slip and survival rate as test index.
Wherein,
OPG=a × 100, a are the egg sac number being calculated under the tally microscope of Maxwell；
Egg capsule slip（%）=（It is not immune attack worm group OPG-and be immunized attack worm group OPG）÷ is not immune to attack worm group OPG × 100%；
Attack the quantity of the weight ÷ experiment chickens of counterpoise before worm=attack before worm；
Attack the quantity of the weight ÷ experiment chickens of counterpoise after worm=attack after worm；
Average weight gain=increasing weight after attacking worm-increases weight before attacking worm；
Average body weight increase rate=each immune group attacks worm group or the not immune average weight gain ÷ for attacking worm group is not immunized and does not attack the flat of worm group
Weightening × 100%.
2 each group chicken of table attacks the weightening result before and after worm
Wherein, weightening data of the worm after 7 days are attacked in the weightening data after " a " indicates 21 days immune in table 2, " b " expression；" cloth " indicates
E.brunetti, " and " indicate Eimeria mitis.
Before as shown in table 2, attacking worm, use the other average body weight increase rate of each group of the vaccine immunity of proportioning 1~4 for 95.7%,
94.6%, 92.9%, 93.4%, each immune group SPF chickens do not occur death.After attacking worm, using the difference of the vaccine immunity of proportioning 1
Attack E.brunetti, two groups of other body weight increase rates of Eimeria mitis are 64.0%, 77.2%；Likewise, using
Two groups of other body weight increase rates of the vaccine immunity of proportioning 2 are respectively 85.4%, 91.0%；Using the two of the vaccine immunity of proportioning 3
The other body weight increase rate of group is respectively 87.3%, 79.1%；Using two groups of other body weight increase rate difference of the vaccine immunity of proportioning 4
It is 66.5%, 94.6%.
After attacking worm, two groups of E.brunetti, Eimeria mitis other ovum are attacked using the vaccine immunity for matching 1
Capsule slip is respectively 89.3%, 85.2%；It is respectively 95.4%, 94.1% using two groups of immune other egg capsule slips of proportioning 2；
It is respectively 97.5%, 87.7% using two groups of immune other egg capsule slips of proportioning 3；Using two groups of the vaccine immunity of proportioning 4
Other egg capsule slip is respectively 88.2%, 97.6%.
3 each group chicken of table attacks OPG values and egg capsule yield result after worm
Wherein, " cloth " indicates E.brunetti in table 3, " and " indicate Eimeria mitis.
In conjunction with the testing result of table 2 and 3, it is clear that, in proportioning 1~4, after attacking worm, attack E.brunetti unavoidably
The body weight increase rate that epidemic disease attacks worm group is only 42.8%, and the not immune body weight increase rate for attacking worm group for attacking Eimeria mitis is only
66.2%, illustrate the high dose infection of both coccidias so that chicken is slow-growing, weightening is poor, and each immune group chicken is opposite
Rate of body weight gain is apparently higher than corresponding be not immunized and attacks worm group.Specifically, other the relative weight gain of group of the vaccine immunity using proportioning 1
Not immune to attack worm group high by 21.2% than corresponding for rate（Attack E.brunetti）With 11.0%（Attack Eimeria mitis）；It adopts
With the other body weight increase rate of group of the vaccine immunity of proportioning 2, not immune to attack worm group high by 42.6% than corresponding（Attack Bu Shi Amy ears
Coccidia）With 24.8%（Attack Eimeria mitis）；Using the other body weight increase rate of group for the vaccine immunity for matching 3 than corresponding
Not immune to attack worm group high by 44.5%（Attack E.brunetti）With 12.9%（Attack Eimeria mitis）；Using proportioning 4
Not immune to attack worm group high by 23.7% than corresponding for the other body weight increase rate of group of vaccine immunity（Attack E.brunetti）With
28.4%（Attack Eimeria mitis）；And it each immune attacks after worm group attacks E.brunetti, Eimeria mitis
Egg capsule slip is above 85.2%, it was demonstrated that proportioning 1~4 has achieved the effect that be immunized well, and the shadow of the weightening to chicken
Sound is low, is highly resistant to the attack of both coccidia high doses of E.brunetti, Eimeria mitis.
Further, before attacking worm after being immunized, the group body weight increase rate using the vaccine immunity of proportioning 1 and proportioning 2 is apparent
The group of the vaccine immunity matched better than other 2 kinds；After attacking worm, using the other body weight increase rate of group of the vaccine immunity of proportioning 2
It is better than the group of the vaccine immunity using proportioning 1,3,4 with egg capsule slip；Therefore, during combined immune and the vaccine after worm is attacked
Influence to the rate of body weight gain of chicken and the egg capsule slip after worm is attacked, applicant show that the optimum proportioning of the vaccine is proportioning
2 scheme contains E.brunetti Precocious Strains Sporulated Oocysts 250, Eimeria mitis in i.e. every part vaccine
It is most slight to the influence of chicken during vaccine reaction when Precocious Strains Sporulated Oocysts 250, it attacks after worm and can provide relatively strong
Immune protective effect.
It, can according to the technique and scheme of the present invention and its hair it is understood that for those of ordinary skills
Bright design is subject to equivalent substitution or change, and all these changes or replacement should all belong to the guarantor of appended claims of the invention
Protect range.
Claims (6)
1. a kind of chicken coccidiasis attenuated live vaccines, which is characterized in that including：100~500/part of E.brunetti, and
100~500/part of slow Eimeria.
2. chicken coccidiasis attenuated live vaccines according to claim 1, which is characterized in that including：E.brunetti 250
A/head part, 250/part of Eimeria mitis.
3. chicken coccidiasis attenuated live vaccines according to claim 1, which is characterized in that the E.brunetti is cloth
It is that the precocity of Eimeria mitis causes weak worm strain that the precocity of family name's Eimeria, which causes weak worm strain, the Eimeria mitis,.
4. chicken coccidiasis attenuated live vaccines according to claim 1, which is characterized in that the immunization ways of the vaccine include
Oral immunity and eye drip.
5. the preparation method of the chicken coccidiasis attenuated live vaccines described in a kind of any one of claim 1-4, which is characterized in that packet
It includes：It causes weak worm strain to cause weak worm strain to be uniformly mixed with the precocity of Eimeria mitis the precocity of E.brunetti, preserves
In preserving liquid.
6. a kind of application of chicken coccidiasis attenuated live vaccines according to any one of claims 1-4, which is characterized in that the epidemic disease
Seedling is applied to prepare in the pharmaceutical preparation for preventing chicken coccidiasis.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109646674A (en) * | 2019-01-25 | 2019-04-19 | 中国农业大学 | A kind of coccidia disease live-vaccine and its application |
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