WO2002072139A1 - Vaccin aviaire efficace contre le virus de la maladie de la bursite infectieuse - Google Patents

Vaccin aviaire efficace contre le virus de la maladie de la bursite infectieuse Download PDF

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WO2002072139A1
WO2002072139A1 PCT/US2002/002589 US0202589W WO02072139A1 WO 2002072139 A1 WO2002072139 A1 WO 2002072139A1 US 0202589 W US0202589 W US 0202589W WO 02072139 A1 WO02072139 A1 WO 02072139A1
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vaccine
virus
ibdv
attenuated
cells
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PCT/US2002/002589
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Jagdev M. Sharma
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Regents Of The University Of Minnesota
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Priority to US10/471,266 priority Critical patent/US20040146530A1/en
Publication of WO2002072139A1 publication Critical patent/WO2002072139A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • 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
    • A61K39/12—Viral antigens
    • 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
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/525—Virus
    • A61K2039/5254—Virus avirulent or attenuated
    • 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
    • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
    • 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
    • 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
    • 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
    • A61K2039/70—Multivalent vaccine
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2720/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsRNA viruses
    • C12N2720/00011—Details
    • C12N2720/10011—Birnaviridae
    • C12N2720/10034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2720/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsRNA viruses
    • C12N2720/00011—Details
    • C12N2720/10011—Birnaviridae
    • C12N2720/10061—Methods of inactivation or attenuation
    • C12N2720/10064—Methods of inactivation or attenuation by serial passage

Definitions

  • Marek's disease virus MDV
  • infectious bursal disease virus IBDV
  • Newcastle disease virus NDV
  • infectious bronchitis virus IBN
  • infectious laryngotracheitis virus ILTN
  • avian encephalomyelitis AEV
  • chick anemia virus CAV
  • fowlpox virus FPV
  • avian influenza virus AIV
  • reovirus avian leukosis yirus
  • AMV reticuloendotheliosis virus
  • REV avian adenovirus and hemorrhagic enteritis virus
  • HEV hemorrhagic enteritis virus
  • IBDV is a member of the Birnaviridae family whose genome consists of two segments of double-stranded R ⁇ A. It causes an acute, highly contagious disease in young chickens. IBDV causes mortality in chickens and induces immunosuppression. Immunosuppressed chickens become more susceptible to Marek's disease, coccidiosis, inclusion body hepatitis, hemorrhagic aplastic anemia, gangrenous dermatitis, infectious chicken anemia agent, salmonellosis and colibacillosis.
  • the main target cells for IBD viral replication are the actively dividing B lymphocytes.
  • the virus infection leads to the destruction of lymphoid cells in the bursa of Fabricius and, to a lesser degree, in other lymphoid organs such as cecal tonsils and spleen.
  • the bursa of Fabricius is a unique, primary lymphoid organ in avian species, where B lymphocytes maturate and differentiate.
  • the bursa is the principal reservoir of virus replication, and peak virus titers in the bursa can be detected between three to five days after IBDV infection.
  • productive viral replication is often associated with necrosis, apoptosis of lymphoid cells, inflammatory change, atrophy, and hemorrhages.
  • the virus In the birds that survive the acute phase of the disease, the virus is cleared, the bursal follicles become repopulated with B cells and immune competence is reestablished.
  • the mechanisms that limit virus replication and promote recovery are not known and may involve virus-specific immune responses.
  • Chicken is the only avian species known to be susceptible to clinical disease and characteristic lesions caused by IBDV. Turkeys, ducks and ostriches are susceptible to infection with IBDV although, thus far, there have been no published reports on the susceptibility of these species to clinical disease. Most commercial chickens get exposed to IBDV early in life. In unprotected flocks, the virus causes mortality and immunosuppression. Although mortality can be quite significant at times, the major economic concern for the poultry industry is the ability of IBDV to cause immunosuppression. Immunosuppressed flocks perform poorly and show reduced economic return. Both broiler and layer flocks are vulnerable to the immunosuppressive effects of the virus and must be protected by vaccination.
  • IBDV vaccines used in commercial flocks are selected by the ability of the vaccines to induce vigorous antibody responses.
  • IBD is one of the major economically important diseases of poultry industry world- wide. The disease causes recurring economic loss.
  • IBDV vaccines are sold yearly worldwide.
  • IBD outbreaks continue to occur.
  • the present invention provides a poultry vaccine that is an attenuated infectious bursal disease virus (IBDV), wherein the vaccine is effective in inducing immunological protection against IBDV infection and is non-pathogenic.
  • IBDV infectious bursal disease virus
  • the term "attenuated” is defined as being rendered less virulent in a non- chemically-induced or genetically engineered manner. In particular, in this specification "attenuated” means passaged in tissue culture.
  • non- pathogenic is used herein to mean non- virulent or unable to induce illness.
  • cell-adapted is defined herein to mean virus that is transferred or passaged from one culture of cells to an next culture of cells.
  • the present invention provides a poultry vaccine that contains a biological agent or microbial component that is effective in inducing improved protection against IBDV by stimulating a strong cellular response in addition to a strong humoral immune response as compared to traditional IBDV vaccines.
  • the live biological agent of the vaccine may be a virus.
  • the virus may be an attenuated virus, a recombinant virus or a virus that has been altered by chemical, physical or molecular means.
  • the terms "traditional” or "conventional” are used to refer to currently available IBDV vaccines.
  • the new IBDV vaccine of the present invention is different from conventional vaccines because the new vaccine is selected on its ability to generate anti-IBDV cellular immunity.
  • the conventional vaccines are selected on their ability to generate humoral immunity without an assessment of their ability to generate cellular immunity.
  • IBDV vaccines are generally classified by those skilled in the art as being “mild,” “intermediate” or “hot.” These terms refer to the residual virulence of the vaccine virus. Thus, a “mild” vaccine would retain a minimal, if any, virulence, and a “hot” vaccine would be highly virulent. In general, however, a mild vaccine would not generate a protective immune response.
  • Conventional intermediate and hot vaccines are known to cause variable, often extensive, bursal necrosis and associated immunosuppression. The conventional vaccines induce poor protection in chicks bearing anti-IBDV antibodies such as newly hatched progeny of IBDV- exposed hens. The new vaccine causes reduced bursal damage and fewer immunosuppressive side effects, and is more effective in the presence of preexisting antibody (such as maternal immunity) in providing protective immunity to the animal than conventional vaccines.
  • the vaccines of the present invention is effective in such avian species as chicken, turkey, duck or ostrich, in particular chicken.
  • the vaccine is IBDV that has been attenuated in macrophage cells.
  • the vaccine may further contain more than one strain of IBDV, or may contain additional biological agents or microbial components effective against a second disease to form a multivalent vaccine.
  • the vaccine may contain a mixture of "mild” or "intermediate” virus in addition to "hot” virus. This mixture may be in the ratio of, including but not limited to, 1 :99, 25:75, 50:50, 80:20, 90:10, 95:5, 99:1 or 99.9:0.1 intermediate or mild to hot virus.
  • the vaccine may contain one or more adjuvants.
  • the present invention also provides a method of protecting poultry by administering to the poultry an immunologically protective amount of a vaccine of the present invention.
  • immunologically protective means that the vaccine is effective in inducing an immune response.
  • An immunological response to a composition or vaccine is the development in the host of a cellular and/or antibody-mediated immune response to the polypeptide or vaccine of interest. Usually, such a response consists of the subject producing antibodies, B cell, helper T cells, suppressor T cells, and/or cytotoxic T cells directed specifically to an antigen or antigens included in the composition or vaccine of interest.
  • the present invention also provides a method of making an attenuated IBDV vaccine.
  • the vaccine is prepared by passaging IBDV in macrophage cells for at least 2, 5, 10 or 13 passages.
  • the IBDV may be passaged for about 30, 20, 16 or 15 passages.
  • the IBDV may be passaged for 10 to 16 passages, or 13 to 15 passages.
  • the cells used in the tissue culture passaging may be poultry macrophage cells, such as those from chickens.
  • the cells may be the MQ-NCSU cell line.
  • IBDV vaccine that stimulates the chick's own cellular immune system (T cells) in addition to humoral immune system would be more effective, unique, and highly desirable, leading to a more efficacious vaccination process.
  • All previously available IBDV vaccines were selected on their ability to generate a strong humoral response.
  • the present research has shown that cellular immunity is also critical for protection against IBDV.
  • a vaccine that generates a strong cellular immunity as well as humoral immunity is needed.
  • the protective efficacy of such a vaccine especially if the vaccine is effective in the face of maternal antibody, would be much better than the presently existing vaccines.
  • bursectomized chickens allow the replication of IBDV in a number of non-bursal tissues.
  • Several serial passages of the virus have been made in bursectomized chickens. This virus is valuable as a vaccine because it has reduced ability to destroy the bursa and thus has reduced immunosuppressive side effects.
  • cytokine genes include types I and II interferons.
  • the efficacy of these plasmid in inducing protective immunity in chickens has been investigated. Cytokine containing recombinant IBDV initiated a better immune response than IBDV lacking such gene inserts.
  • the present invention provides a poultry vaccine that contains a biological agent or microbial component that is effective in stimulating an improved protective cellular and humoral immune response to IBDV as compared to traditional IBDV vaccines.
  • the vaccine is effective in such avian species as chicken, turkey, duck or ostrich, in particular chicken.
  • the live biological agent of the vaccine may be a virus.
  • the virus may be an attenuated virus, a recombinant virus or a virus that has been altered by chemical, physical or molecular means.
  • the vaccine may further contain more than one strain of IBDV or additional biological agents or microbial components effective against other diseases to form a multivalent vaccine.
  • the vaccine may contain one or more adjuvants.
  • the present invention also provides a method of protecting poultry by administering a vaccine that is effective in inducing cellular and humoral immunity and that contains a biological agent or microbial component that is effective in stimulating a protective cellular and humoral immune response to IBDV.
  • the vaccine of the present invention can be administered via conventional modes of administration or in ovo. Methods of in ovo immunization are set forth, for example, in U.S. Patent No. 6,048,535. Vaccination can be performed at any age. For in ovo vaccination, vaccination would be done in the last quarter of embryonal development but may be done at any time during embryonation.
  • the vaccines according to the invention can, for example, be administered intramuscularly, subcutaneously, orally, intraocularly, intratracheally, intranasally, ovo, in drinking water, in the form of sprays or by contact spread.
  • chickens are given the first vaccine in ovo or at one day of age. Subsequent vaccinations are done according to need. Breeder chickens can be vaccinated before and during the lay cycle (several inoculations).
  • Vaccines are often formulated and inoculated with various adjuvants.
  • the adjuvants aid in attaining a more durable and higher level of immunity using small amounts of antigen or fewer doses than if the immunogen were administered alone.
  • the mechanism of adjuvant action is complex, and may involve the stimulation of cytokine production, phagocytosis and other activities of the reticuloendothelial system as well as a delayed release and degradation of the antigen.
  • Suitable adjuvants include but are not limited to surfactants, e.g., hexadecyl amine, octadecylamine, lysolecithin, dimethyldioctadecylammonium bromide, N,N- dioctadecyl-N'-N-bis(2-hydroxyethyl-propane di-amine), methoxyhexadecyl- glycerol, and pluronic polyols; polanions, e.g., pyran, dextran sulfate, poly IC, polyacrylic acid, carbopol; peptides, e.g., muramyl dipeptide, aimethylglycine, tuftsin, oil emulsions, alum, and mixtures thereof.
  • surfactants e.g., hexadecyl amine, octadecylamine, lysolecithin
  • the immunogenic product may be incorporated into liposomes for use in a vaccine formulation, or may be conjugated to proteins such as keyhole limpet hemocyanin (KLH) or human serum albumin (HSA) or other polymers.
  • KLH keyhole limpet hemocyanin
  • HSA human serum albumin
  • Vaccines containing live virus can be prepared and marketed in the form of a suspension, or lyophilized.
  • Lyophilized vaccines can preferably contain one or more stabilizers. Suitable stabilizers are, for example, SPGA (Bovarnik (1950): J. Bacteriology 59; 509), carbohydrates (such as sorbitol, mannitol, starch, sucrose, dextran or glucose), proteins (such as albumin or casein), or degradation products thereof, protein-containing materials (such as bovine serum or skimmed milk) and buffers (such as alkali metal phosphates).
  • the aim of inactivation of IBD viruses is to eliminate both reproductive ability and virulence of the viruses. In general, this can be achieved by chemical or physical means.
  • Chemical inactivation can be effected by treating the viruses with, for example, enzymes, formaldehyde, ⁇ -propiolactone, ethylene-imine or a derivative thereof, an organic solvent (such as a halogenated hydrocarbon) and/or a detergent (such as TW ⁇ N, TRITON X, sodium desoxy-cholate, sulphobetain or cetyl trimethylammonium salts).
  • an organic solvent such as a halogenated hydrocarbon
  • a detergent such as TW ⁇ N, TRITON X, sodium desoxy-cholate, sulphobetain or cetyl trimethylammonium salts.
  • inactivating substance is neutralized afterwards; material inactivated with formaldehyde can, for example, be neutralized with thiosulphate.
  • Physical inactivation can preferably be carried out by subjecting the viruses to energy-rich radiation, such as UV light, X-radiation or ⁇ -radiation. If desired, the pH can be brought back to a value of about 7 after treatment.
  • energy-rich radiation such as UV light, X-radiation or ⁇ -radiation.
  • the pH can be brought back to a value of about 7 after treatment.
  • TW ⁇ N and SPAN may also be added to the inactivated virus material.
  • the invention also includes combination vaccines, where the IBDV material according to the present invention is combined with one or more additional homologous or heterologous biological agents effective against various diseases.
  • the vaccines according to the invention are suitable for protecting poultry (such as chickens and turkeys) against IBD (Gumboro's disease).
  • IBD Tuboro's disease
  • Example 1 Development of an IBDV vaccine that causes reduced damage to the bursa yet replicates in the bird and induces strong immunity.
  • the cloned or uncloned viruses were tested for (1) induction of cellular or humoral immunity; (2) protective efficacy following in ovo or post-hatch administration;
  • a virulent or intermediate isolate of IBDV is chemically mutated using established procedures. Viral mutant are screened and the mutant(s) that have lost virulence but have retained immunogenicity are identified. A virulent or intermediate isolate of IBDV is genetically cloned and site-directed mutations are induced in the genome. The mutant virus has enhanced immunogenic potential.
  • Example 2 Reduction of pathogenicitv and enhancement of immunogenicity of intermediate IBDV vaccine.
  • the effects of concurrent in ovo or post-hatch administration of cytokine- inducing chemical or compounds and intermediate IBDV are examined.
  • a commercially available inhibitor of iNOS gene is also examined. This gene codes for nitric oxide. Recent data have show that nitric oxide is responsible for much of the pathological lesions induced by intermediate IBDV and the use of the inhibitor prevents lesions.
  • Example 3 Preparation of a recombinant vaccine vectored in a bacterial vector.
  • Viral genes in concert with cytokine genes are incorporated in bacterial plasmids.
  • the recombinant plasmids are used for immunizing chickens.
  • the viral gene plus cytokine gene combinations that produce the most desirable immune response are used as vaccines. These selected combinations are used to construct live recombinant vaccines in viral vectors.
  • a "mixed" or combination vaccine is an alternative approach to protecting maternal antibody-bearing chicks against IBD.
  • the efficacy of a mixture containing a minimum infectious dose of a high virulence virus and a high dose of a low virulence virus as an effective vaccine in maternal antibody positive birds was examined. The idea was that in chicks that have high antibody levels, the high virulence virus was likely to break through the antibody and initiate immunity. In chicks with low or negligible antibody levels, although the high virulence virus would home to the bursa and initiate infection, the presence of an overwhelming dose of the low virulence virus would reduce the pathogenic effects of the high virulence virus.
  • the preliminary data showed that less that 10 embryo lethal dose 50 (ELD 50 ) of IM-IBDV (high virulence virus) can induce bursal lesions in SPF chickens. This dose is being tried in association with 10 6 tissue culture infectious dose 50 (TCID 50 ) of a mild strain of IBDV.
  • ELD 50 embryo lethal dose 50
  • TCID 50 tissue culture infectious dose 50
  • the vaccine may contain a mixture of "mild” or "intermediate” virus in addition to "hot” virus. This mixture is in the ratio of, but not limited to, 1 :99, 25:75, 50:50, 80:20, 90:10, 95:5, 99: 1 , or 99.9:0.1 intermediate or mild to hot virus.
  • Example 5 Attenuated Vaccines
  • Other research groups have attempted to attenuate/adapt IBDV to macrophage cell lines, but were unsuccessful. (See, Abdel-Alim, G and Y.M. Saif. 2001. Imunogenicity and antigenicity of very virulent strains of infectious bursal disease virus. Avian Diseases 45:92-101.)
  • the present inventor was able to successfully adapt virulent, intermeidate and mild IBDV to replicate in a chicken macrophage cell line.
  • the virulent strain, Irwin Moulthrop (IM) of IBDV was attenuated in chicken macrophage cell line (MQ-NCSU).
  • the adapted virus, designated IM-IBDV-Mac caused lytic infection in cells and replicated to high titers.
  • IM-IBDV-Mac also replicated in chicken embryo fibroblasts and vero cells, where it induced typical cytopathic effects.
  • the adaptation of IM-IBDV to macrophages resulted in the loss of the ability of the virus to cause mortality in embryonated eggs or chickens but retained its immunogenic potential.
  • the data indicate that the replication of microphage-adapted virus in chicken embryo fibroblast (CEF) cells began at the fourteenth passage (PI 4). P14 or later passages, but not earlier passages, replicated vigorously in CEF. The replication in CEF became more vigorous with increasing passages in microphage cells.
  • CPE cytopathic effect
  • Table 1 Summary of characteristics of IM-IBDV adapted to replicate in MQ- NCSU cells
  • Passage # The number of times IM-IBDV was serially passed in macrophage cells (MQ-NCSU cells). The serial passages were made by freeze/thawing cells and medium from the previous passage, centrifuging the mixture to clear debris, and inoculating a new batch of growing microphage cells.
  • Egg mort. The ability of the virus to cause embryo mortality when inoculated inlO- to 12-day-old embryonated chicken eggs. Generally, virulent and egg-adapted viruses tend to cause high egg mortality.
  • Chick mort. The ability of the virus to cause mortality when inoculated in hatched chickens. SPF chickens were tested at 2-4 weeks of age. Gross bursa: Chickens exposed to the virus had grossly detectable bursal lesion typical of IBDV. Virulent viruses cause extensive gross bursal lesions. Micro. Bursa: The ability of the virus to cause microscopic bursal lesions characteristic of IBDV. The ability of a virus to cause microscopic lesions is generally associated with the extent of viral replication in the bursa (B cells). Virulent viruses replicate extensively in the bursa; attenuated viruses replicate less extensively, depending upon the degree of attenuation.
  • Antigen B S Th Sections of bursa (B), spleen (S) and thymus (Th) were examined by immunohistochemistry for the presence of virus antigen. +/- means that the presence of viral antigen was equivocal. Presence of viral antigen in the spleen and thymus may indicate that the virus is capable of extra-bursal replication and may induce immunity without causing excessive bursal destruction.
  • Antibody Chickens were examined for antibody levels two weeks after inoculation with the virus. Antibody levels are shown as very high (++++) to undetectable (-). Highly immunogenic (protective) viruses induce high levels of antibody. Protection: Birds were exposed to the virus and two weeks later, challenged with virulent virus (PO of IM-IBDV). "+" indicates protection against mortality and lesion induction by the virulent virus.

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Abstract

L'invention concerne un vaccin pour les volailles. Ce vaccin contient un agent biologique ou un composant microbien efficace pour stimuler une réaction immunitaire humorale et protectrice des cellules contre le virus de la maladie de la bursite infectieuse (IBDV). L'invention concerne des procédés de fabrication du vaccin et de vaccination des volailles par l'administration dudit vaccin.
PCT/US2002/002589 2001-03-09 2002-01-30 Vaccin aviaire efficace contre le virus de la maladie de la bursite infectieuse WO2002072139A1 (fr)

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US27486101P 2001-03-09 2001-03-09
US60/274,861 2001-03-09

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005049077A2 (fr) * 2003-11-17 2005-06-02 Regents Of The University Of Minnesota Vaccin aviaire efficace contre le virus de la bursite infectieuse aviaire

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3548055A (en) * 1969-05-14 1970-12-15 Research Corp Infectious bursal disease vaccine
US4530831A (en) * 1982-11-02 1985-07-23 Akzo N.V. Infectious Bursal Disease vaccine
US4824668A (en) * 1987-03-09 1989-04-25 Sterwin Laboratories Inc. Attenuated infectious bursal disease virus strain and vaccine therefrom

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3548055A (en) * 1969-05-14 1970-12-15 Research Corp Infectious bursal disease vaccine
US4530831A (en) * 1982-11-02 1985-07-23 Akzo N.V. Infectious Bursal Disease vaccine
US4824668A (en) * 1987-03-09 1989-04-25 Sterwin Laboratories Inc. Attenuated infectious bursal disease virus strain and vaccine therefrom

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005049077A2 (fr) * 2003-11-17 2005-06-02 Regents Of The University Of Minnesota Vaccin aviaire efficace contre le virus de la bursite infectieuse aviaire
WO2005049077A3 (fr) * 2003-11-17 2005-11-24 Univ Minnesota Vaccin aviaire efficace contre le virus de la bursite infectieuse aviaire

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