WO2008003734A1 - Combinaison de vaccins contre les streptocoques - Google Patents

Combinaison de vaccins contre les streptocoques Download PDF

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Publication number
WO2008003734A1
WO2008003734A1 PCT/EP2007/056789 EP2007056789W WO2008003734A1 WO 2008003734 A1 WO2008003734 A1 WO 2008003734A1 EP 2007056789 W EP2007056789 W EP 2007056789W WO 2008003734 A1 WO2008003734 A1 WO 2008003734A1
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WIPO (PCT)
Prior art keywords
water
streptococcus
cells
oil
vaccine
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PCT/EP2007/056789
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English (en)
Inventor
Luc Grisez
Chow Yong Ng
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Intervet International B.V.
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Publication date
Application filed by Intervet International B.V. filed Critical Intervet International B.V.
Priority to CN200780025577.6A priority Critical patent/CN101484184B/zh
Priority to BRPI0713274A priority patent/BRPI0713274B1/pt
Priority to MX2008016248A priority patent/MX2008016248A/es
Publication of WO2008003734A1 publication Critical patent/WO2008003734A1/fr

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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
    • A61K39/02—Bacterial antigens
    • A61K39/09—Lactobacillales, e.g. aerococcus, enterococcus, lactobacillus, lactococcus, streptococcus
    • A61K39/092—Streptococcus
    • 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
    • A61K2039/52—Bacterial cells; Fungal cells; Protozoal cells
    • A61K2039/521—Bacterial cells; Fungal cells; Protozoal cells inactivated (killed)
    • 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
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55566—Emulsions, e.g. Freund's adjuvant, MF59
    • 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

Definitions

  • the present invention relates to a combination vaccine for the protection of fish against streptococcal infection, and to methods for the preparation of such a combination vaccine.
  • Streptococcus Several species of the bacterial genus Streptococcus are currently known to cause infections in fish, more specifically in fish that are kept in aquaculture. Examples of such streptococcal species are Streptococcus iniae, S. difficile, S. agalactiae, S. dysgalactiae and S. phocae.
  • Streptococcus difficile is in fact a non-hemolytic Streptococcus agalactiae ⁇ Streptococcus agalactiae is a ⁇ -hemolytic bacterium).
  • Streptococcus iniae, Streptococcus difficile, Streptococcus agalactiae is used here.
  • Streptococcus iniae is frequently found in Tilapia, Rainbow trout, European sea bass and bream, Asian sea bass, Red drum, Rabbit fish, Japanese flounder, Yellowtail and hybrid striped bass.
  • the annual impact of Streptococcus iniae infection to aquaculture exceeds 100 million US-dollars.
  • Streptococcus difficile is frequently found in Asian sea bass, Tilapia, snapper and catfish.
  • Streptococcus agalactiae is currently mainly found to infect Tilapia.
  • Vaccines for combating Streptococcal infection in fish are known in the art. Many of the Streptococcus vaccines are based upon killed whole cells. Streptococcus agalactiae vaccines are i.a. described in the US-Patent Application US 2005/0208077. Streptococcus iniae vaccines are i.a. described in US-Patent US 6,379,677. Vaccines for combating Streptococcus iniae infection are also commercially available. An example is Norvax Strep Si, a vaccine against Streptococcus iniae infection and sold by Intervet Int. B.V.
  • a combination vaccine for the protection of fish against streptococcal infection comprises an immunogenic amount of Streptococcus difficile cells and an immunogenic amount of Streptococcus ini ⁇ e cells wherein the ratio of Streptococcus difficile cells to Streptococcus ini ⁇ e cells is more than or equal to (>) 40 to 10, this problem of suppression of the development of an anti-immune response to Streptococcus ini ⁇ e either occurs to an acceptable level, or is not seen at all.
  • the presence of an immunogenic amount of Streptococcus difficile cells and an immunogenic amount of Streptococcus ini ⁇ e cells is of course necessary to induce an immune response to each of the bacterial species.
  • the amount of Streptococcus difficile cells in the vaccine should however be at least four times the amount of Streptococcus ini ⁇ e cells.
  • a first embodiment of the present invention relates to a combination vaccine for the protection of fish against streptococcal infection that has as a characteristic that this combination vaccine comprises an immunogenic amount of Streptococcus difficile cells and an immunogenic amount of Streptococcus ini ⁇ e cells wherein the ratio of Streptococcus difficile cells to Streptococcus ini ⁇ e cells is > 40 to 10.
  • a combination vaccine wherein the amount of Streptococcus difficile cells in the vaccine is more than four times the amount of Streptococcus iniae cells gives even less suppression of the development of an anti-immune response against Streptococcus iniae.
  • a combination vaccine wherein the ratio of Streptococcus difficile cells to Streptococcus iniae cells is > 40 to 8 is preferred and ratios of > 40 to 6, > 40 to 4, > 40 to 2 and > 40 to 1 are in that order increasingly more preferred.
  • preferred forms of this embodiment relate to a combination vaccine according to the invention wherein that ratio is > 40 to 8, more preferably > 40 to 6, even more preferably > 40 to 4, still even more preferably > 40 to 2 and most preferably > 40 to 1.
  • Asian fish vaccines were, if possible at all, given as water-based vaccines. They comprised no oil phase.
  • water-in-oil emulsions water- in- oil-in-water emulsions or oil-in- water emulsions.
  • Such emulsion-vaccines have the advantage that they are more efficacious than their water-based counterparts.
  • emulsion vaccines allow for lower amounts of antigen to be used.
  • Another preferred form of this embodiment relates to a combination vaccine according to the invention wherein the vaccine is a water-in-oil vaccine.
  • Still another preferred form of this embodiment relates to a combination vaccine according to the invention wherein the vaccine is an oil-in-water vaccine.
  • the problem to be solved i.e. the problem that the presence of Streptococcus difficile in a combination vaccine is highly suppressive to the development of an anti-Streptococcus ini ⁇ e immune response, is encountered both in water-based vaccines and in emulsions on the basis of water and oil.
  • the solution as provided above is one solution to the problem.
  • an emulsion on the basis of water and oil is used for the vaccine, another solution was found to be feasible, possibly in combination with the solution given above.
  • a water-in-oil emulsion wherein the Streptococcus iniae cells and Streptococcus difficile cells are present in separate water droplets of the emulsion can relatively easy be made.
  • the standard procedure for making a basic water-in-oil emulsion is based upon the mixing of an amount of a single watery phase, a single oil phase and an amount of bacterial cells.
  • the watery phase can e.g. be just water, a water-based buffer and the like.
  • the oil phase can be a mineral or non-mineral oil (see below). Additionally, one or more emulsif ⁇ ers and/or detergents can be added.
  • the methods for making such emulsions, and the components for use in such emulsions are well-known in the art.
  • the present method differs from the standard method in that instead of a single watery phase two watery phases are used; one comprising Streptococcus iniae cells and the other one comprising Streptococcus difficile cells.
  • Each of the two watery phases is now emulsified with an amount of oil phase and after the separate water-in-oil emulsions are made, the two water-in-oil emulsions are mixed.
  • a variant to this method is e.g.
  • Streptococcus difficile cells are present in separate water droplets of the emulsion always fall within the scope of the invention, regardless the way they are made.
  • another embodiment of the present invention relates to a combination vaccine for the protection of fish against streptococcal infection
  • said vaccine is a water-in-oil vaccine comprising an immunogenic amount of Streptococcus iniae cells and Streptococcus difficile cells wherein the Streptococcus iniae cells and the Streptococcus difficile cells are present in separate water droplets of said water- in-oil emulsion.
  • Another possibility is, to make use of a water- in-oil- in- water emulsion, wherein the immunogenic amount of Streptococcus iniae cells and Streptococcus difficile cells are present in separate water droplets of said water- in-oil-in-water emulsion.
  • Such an emulsion comprises watery droplets, present inside small oil droplets, which in turn are surrounded by a watery phase.
  • another form of this embodiment relates to a combination vaccine for the protection of fish against streptococcal infection, wherein said vaccine is a water-in-oil-in- water vaccine comprising an immunogenic amount of Streptococcus iniae and Streptococcus difficile wherein the Streptococcus iniae component and the Streptococcus difficile component are present in separate water droplets of said water-in-oil-in- water emulsion.
  • Still another possibility to keep the Streptococcus iniae cells and the Streptococcus difficile cells separated in a water-in-oil-in- water emulsion is, to first make a water-in-oil emulsion wherein the watery phase comprises only the Streptococcus iniae cells or the Streptococcus difficile cells. In a next step, this water-in-oil emulsion can then be emulsified in a watery phase that comprises the other of the two bacterial components, i.e. the Streptococcus difficile cells or the Streptococcus iniae cells.
  • still another form of this embodiment relates to a combination vaccine for the protection of fish against streptococcal infection, wherein said vaccine is a water-in-oil-in- water vaccine comprising an immunogenic amount of Streptococcus iniae cells and Streptococcus difficile cells wherein the Streptococcus iniae cells are present in the oil- encapsulated water droplets of said water-in-oil-in- water emulsion and the Streptococcus difficile cells are present in the free watery phase of said water- in-oil- in-water emulsion.
  • said vaccine is a water-in-oil-in- water vaccine comprising an immunogenic amount of Streptococcus iniae cells and Streptococcus difficile cells wherein the Streptococcus iniae cells are present in the oil- encapsulated water droplets of said water-in-oil-in- water emulsion and the Streptococcus difficile cells are present in the free water
  • Still another form of this embodiment relates to a combination vaccine for the protection of fish against streptococcal infection, wherein said vaccine is a water-in-oil-in- water vaccine comprising an immunogenic amount of Streptococcus iniae cells and Streptococcus difficile cells wherein the
  • Streptococcus difficile cells are present in the oil-encapsulated water droplets of said water-in-oil- in- water emulsion and the Streptococcus iniae cells are present in the free watery phase of said water-in- oil-in-water emulsion.
  • Emulsions such as the water-in-oil and water-in-oil-in- water emulsions described above may have, after some time, a certain tendency towards sedimentation (w/o) or creaming (w/o/w) in the worst case followed by breaking. This phenomenon is i.a. depending on droplet size, time and temperature of storage, emulsif ⁇ ers used and so on.
  • another embodiment of the present invention relates to a water-in-oil or a water-in-oil- in- water combination vaccine for the protection of fish against streptococcal infection, wherein the vaccine comprises an immunogenic amount of Streptococcus difficile cells and an immunogenic amount of
  • Streptococcus iniae cells wherein the ratio of Streptococcus difficile cells to Streptococcus iniae cells is > 40 to 10, > 40 to 8, > 40 to 6, > 40 to 4, > 40 to 2 or > 40 to 1, in that order of preference, and wherein the Streptococcus iniae cells and the Streptococcus difficile cells are present in separate water droplets of said water-in-oil or water-in-oil-in- water emulsion.
  • Another form of this embodiment relates to a water- in- oil-in-water combination vaccine for the protection of fish against streptococcal infection, wherein the vaccine comprises an immunogenic amount of Streptococcus difficile cells and an immunogenic amount of Streptococcus iniae cells wherein the ratio of Streptococcus difficile cells to Streptococcus iniae cells is > 40 to 10, > 40 to 8, > 40 to 6, > 40 to 4, > 40 to 2 or > 40 to 1, in that order of preference, and wherein the combination vaccine for the protection of fish against streptococcal infection wherein the Streptococcus iniae cells are present in the oil-encapsulated watery phase droplets of said water- in-oil-in-water emulsion and the Streptococcus difficile cells are present in the free watery phase of said water-in-oil-in- water emulsion.
  • Still another form of this embodiment relates to a water-in-oil- in- water combination vaccine for the protection of fish against streptococcal infection
  • the vaccine comprises an immunogenic amount of Streptococcus difficile cells and an immunogenic amount of Streptococcus iniae cells wherein the ratio of Streptococcus difficile cells to Streptococcus iniae cells is > 40 to 10, > 40 to 8, > 40 to 6, > 40 to 4, > 40 to 2 or > 40 to 1, in that order of preference, and wherein the combination vaccine for the protection of fish against streptococcal infection wherein the Streptococcus difficile cells are present in the oil-encapsulated watery phase droplets of said water- in-oil-in-water emulsion and the Streptococcus iniae cells are present in the free watery phase of said water-in-oil-in-water emulsion.
  • Combination vaccines according to the present invention may in a preferred presentation also contain an immunostimulatory substance, a so-called adjuvant.
  • Adjuvants in general comprise substances that boost the immune response of the host in a non-specific manner.
  • a number of different adjuvants are known in the art. Examples of adjuvants frequently used in fish and shellfish farming are muramyldipeptides, lipopolysaccharides, several glucans and glycans and Carbopol( ⁇ ) (a homopolymer).
  • An extensive overview of adjuvants suitable for fish and shellfish vaccines is given in a review paper by Jan Raa (Reviews in Fisheries Science 4(3): 229-288 (1996)).
  • the vaccine may also comprise a so-called "vehicle".
  • a vehicle is a compound to which the bacterium adheres, without being covalently bound to it. Such vehicles are i.a. bio-microcapsules, micro-alginates, liposomes and macrosol
  • the vaccine may comprise one or more suitable surface-active compounds or emulsif ⁇ ers, e.g. Span or Tween.
  • Oil adjuvants suitable for use in water-in-oil, water-in-oil-in-water and oil-in-water emulsions are e.g. mineral oils or metabolisable oils.
  • Mineral oils are e.g. Bayol ® , Marcol ® and Drakeol ® .
  • Metabolisable oils are e.g. vegetable oils, such as peanut oil and soybean oil, animal oils such as the fish oils squalane and squalene, and tocopherol and its derivatives.
  • the amount of adjuvant added depends on the nature of the adjuvant itself, and information with respect to such amounts will be provided by the manufacturer.
  • the vaccine is mixed with stabilisers, e.g. to protect the bacteria from being degraded, to enhance the shelf-life of the vaccine, or to improve freeze-drying efficiency.
  • Useful stabilisers are i.a. SPGA, carbohydrates e.g. sorbitol, mannitol, trehalose, starch, sucrose, dextran or glucose, proteins such as albumin or casein or degradation products thereof, and buffers, such as alkali metal phosphates.
  • streptococcal vaccines can e.g. be administered by injection, usually as intraperitoneal injection (also referred to as IP injection). From a point of view of protection the IP -vaccination is the preferred vaccination, because it gives a good and long-lasting protection. This is especially true for inactivated vaccines.
  • Vaccines according to the invention basically comprise an immunogenic amount of a bacterium and a pharmaceutically acceptable carrier.
  • immunogenic as used herein is defined as the amount sufficient to induce an immune response in the target fish.
  • a pharmaceutically acceptable carrier can be as simple as water or a buffer, or an emulsion.
  • the amount of cells administered will depend on the Streptococcus species used, the presence of an adjuvant, the route of administration, the moment of administration, the age of the fish to be vaccinated, general health, water temperature and diet. When starting from commercially available vaccines, the manufacturer will provide this information.
  • vaccines for use according to the invention that are based upon bacterins can be given in general in a dosage of 10 3 to 10 10 , preferably 10 6 to 10 9 , more preferably between 10 8 and
  • Vaccines according to the invention that are based upon live attenuated bacteria can be given in a lower dose, due to the fact that the bacteria will continue replicating for a certain time after administration.
  • the vaccines for use according to the invention will preferably be vaccines that protect against streptococcal infection and additionally against one or more than one other fish-pathogenic virus or microorganism.
  • a preferred form of this embodiment relates to a vaccine wherein that vaccine comprises at least one other microorganism or virus that is pathogenic to fish, or one other antigen or genetic material encoding said other antigen, wherein said other antigen or genetic material is derived from a virus or microorganism pathogenic to fish.
  • Examples of commercially important fish pathogens in tropical and/or Mediterranean fish are Vibrio anguillarum, Photobacterium damselae subspecies piscicida, Tenacibaculum maritimum, Flavobacterium sp., Flexibacter sp., Lactococcus garviae, Edwardsiella tarda, E. ictaluri, Viral Necrosis virus, iridovirus, Spring Viremia of Carp virus and Koi Herpesvirus.
  • Examples of commercially important cold water fish pathogens are Vibrio anguillarum, Aeromonas salmonicidae, Vibrio salmonicidae, Moritella viscosa, Vibrio ordalii, Flavobacterium sp., Flexibacter sp., Streptococcus sp., Lactococcus garviae, Edwardsiella tarda, E.
  • ictaluri Piscirickettsia salmonis, Salmon Pancreatic Disease virus, Sleeping Disease virus, Viral Nervous Necrosis virus, Infectious Pancreatic Necrosis virus, Infectious haematopoietic necrosis virus, Infectious Salmon Anaemia virus, Viral Haemorrhagic Septicaemia virus and iridoviruses.
  • Salmonids are i.a. Lepeophtherius salmonis, Caligus elongatus, Cryptobia salmositica, Myxobolus cerebralis and Kudoa thyrsites.
  • a parasite infecting freshwater fish is e.g. Ichthyophthirius multifiliis.
  • Tilapia parasites are e.g. Dactylogyrus spp. and Trichodina spp. Marine fish may suffer i.a from the parasite Benedenia seriolae
  • the other microorganism or virus is selected from the following group of fish pathogens: Vibrio anguillarum, Photobacterium damselae subspecies piscicida, Tenacibaculum maritimum, Flavobacterium sp., Flexibacter sp., Lactococcus garviae, Edwardsiella tarda, E.
  • ictaluri Viral Necrosis virus, Spring Viremia of Carp virus, iridovirus, Infectious haematopoietic necrosis virus, Infectious Salmon Anaemia virus, Viral Haemorrhagic Septicaemia virus and Koi Herpesvirus, Aeromonas salmonicidae, Vibrio salmonicidae, Moritella viscosa, Vibrio ordalii, Piscirickettsia salmonis, Salmon Pancreatic Disease virus, Sleeping Disease virus, Viral Nervous Necrosis virus, Infectious Pancreatic Necrosis virus, iridoviruses, Lepeophtherius salmonis, Caligus elongatus, Cryptobia salmositica, Myxobolus cerebralis, Kudoa thyrsites, Ichthyophthirius multifiliis, Dactylogyrus spp., Trichodina spp.
  • the invention relates to methods for the preparation of a combination vaccine for the protection of fish against streptococcal infection
  • said vaccine is a water- in-oil vaccine or a water-in-oil-in- water vaccine comprising an immunogenic amount of Streptococcus iniae cells and Streptococcus difficile cells wherein the Streptococcus iniae cells and the Streptococcus difficile cells are present in separate water droplets of said water-in-oil or water-in-oil-in- water emulsion
  • the method comprises the steps of making a water-in-oil or water-in-oil- in- water emulsion comprising an immunogenic amount of Streptococcus iniae cells and a water-in-oil or water-in-oil-in-water emulsion comprising an immunogenic amount of Streptococcus difficile cells followed by the step of mixing the two emulsions.
  • the invention also relates to methods for the preparation of a combination vaccine for the protection of fish against streptococcal infection, wherein said vaccine is a water-in-oil-in-water vaccine comprising an immunogenic amount of Streptococcus iniae cells and Streptococcus difficile cells wherein the Streptococcus iniae cells are present in the oil- encapsulated water droplets of said water-in-oil-in-water emulsion and the Streptococcus difficile cells are present in the free watery phase of said water-in-oil- in-water emulsion, said method comprising the steps of making a water-in-oil emulsion wherein the watery phase comprises only the Streptococcus iniae cells or the Streptococcus difficile cells, followed by the step of emulsifying this water-in-oil emulsion in a watery phase that comprises the other of the two bacterial components, i.e. the Streptococcus difficile
  • the challenge strains used in this experiment were a S. difficile wild-type strain and S. iniae wild-type strain.
  • Vaccines All bacterial cultures used for the preparation of these vaccines were inactivated with formalin and were derived from S. difficile and S. iniae wild-type strains.
  • the commercially available ISA 736A VG was used as oil-adjuvant for vaccines preparations.
  • Table 1 showing the details on vaccines composition and concentration of each antigen used for the various formulations.
  • Grouping A total of 315 fish of similar size was used for this experiment and divided into 7 groups each consisting of 45 fish.
  • a challenge seed vial was taken from the ⁇ -5O 0 C freezer, thawed and its content inoculated into 100 0 ml of Tryptic Soy Broth
  • the culture was incubated at 32 0 C, placed on an orbital shaker with shaking speed set at 150 RPM After 23 hours incubation, the culture yielded an OD 660 of 0 804
  • a bacterial suspension with OD 660 of 0 715 was prepared and left at room temperature for 2 hours prior to use as challenge material Streptococcus difficile
  • a challenge seed vial was taken from the ⁇ -5O 0 C freezer, thawed and its content inoculated into 100 0 ml of Streptococcal Growth Medium
  • the culture was incubated at 32 0 C, placed on an orbital shaker with shaking speed set at 150 RPM After 24 hours incubation, the culture yielded an OD 660 value of 0 176
  • 0 9% NaCl portion of the culture was used to prepare a bacterial suspension with OD 660 of 0 133, followed by a further 100-fold dilution and the said suspension was used as challenge material
  • Table 3 showing average weight gain per fish over the post-vaccination period of 24 days.
  • Efficacy of vaccines The efficacy of the 6 bivalent vaccines were evaluated and expressed in terms of % RPS (Relative percentage survival). Data on % RPS for each challenge strain in each of the vaccinated group are shown in Table 4. Mortality recorded for all challenged fish were based on positive re-isolations of the challenged bacteria. Table 4 showing the % RPS for each challenge strain in each of the vaccinated groups and percentage mortality in the control groups challenged with a specific pathogen.
  • the figures 1 and 2 show the RPS values of each of the vaccines with respect to the results obtained after challenge with a specific bacterial strain at 24 days post- vaccination.
  • Figure 1 shows the efficacy of bivalent vaccines challenged with S. difficile.
  • Figure 2 shows the efficacy of bivalent vaccines challenged with S. iniae.
  • Figure 1 efficacy of bivalent vaccines challenged with S. difficile.
  • Figure 2 efficacy of bivalent vaccines challenged with S. iniae.

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Abstract

La présente invention concerne une combinaison de vaccins pour la protection des poissons contre une infection streptococcique, ainsi que des procédés pour la préparation d'une telle combinaison de vaccins.
PCT/EP2007/056789 2006-07-06 2007-07-05 Combinaison de vaccins contre les streptocoques WO2008003734A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
CN200780025577.6A CN101484184B (zh) 2006-07-06 2007-07-05 抗链球菌的联合疫苗
BRPI0713274A BRPI0713274B1 (pt) 2006-07-06 2007-07-05 combinação de vacinas para a proteção de peixes contra a infecção estreptocócica, e, método para a preparação de uma combinação de vacinas
MX2008016248A MX2008016248A (es) 2006-07-06 2007-07-05 Vacuna combinada estreptococica.

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US81925406P 2006-07-06 2006-07-06
US60/819,254 2006-07-06
EP06116740 2006-07-06
EP06116740.9 2006-07-06

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WO2008003734A1 true WO2008003734A1 (fr) 2008-01-10

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EC (1) ECSP099066A (fr)
HN (1) HN2009000005A (fr)
MX (1) MX2008016248A (fr)
WO (1) WO2008003734A1 (fr)

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WO2011122843A2 (fr) * 2010-04-02 2011-10-06 국립수산과학원 Vaccin inactivé combiné pour la prévention de la coccidiose des poissons et procédé pour préparer celui-ci
KR101180419B1 (ko) * 2009-07-16 2012-09-10 제주특별자치도(제주특별자치도해양수산연구원장) 어류의 세균성 질병에 대한 복합 불활화 백신
CN103550765A (zh) * 2013-11-11 2014-02-05 中国科学院海洋研究所 一种海豚链球菌dna疫苗及其应用
JP2015127666A (ja) * 2013-12-27 2015-07-09 大阪府 クドア・セプテンプンクタータの迅速検出法
WO2020069656A1 (fr) * 2018-10-05 2020-04-09 福又达生物科技股份有限公司 Procédé de détection de pathogène de poisson
CN116640867A (zh) * 2023-06-27 2023-08-25 四川农业大学 一种raa-lfd检测停乳链球菌的引物对和探针组合及其应用

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US20050208077A1 (en) * 2004-03-18 2005-09-22 Evans Joyce J Streptococcus agalactiae vaccine

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US20050208077A1 (en) * 2004-03-18 2005-09-22 Evans Joyce J Streptococcus agalactiae vaccine

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101180419B1 (ko) * 2009-07-16 2012-09-10 제주특별자치도(제주특별자치도해양수산연구원장) 어류의 세균성 질병에 대한 복합 불활화 백신
WO2011122843A2 (fr) * 2010-04-02 2011-10-06 국립수산과학원 Vaccin inactivé combiné pour la prévention de la coccidiose des poissons et procédé pour préparer celui-ci
WO2011122843A3 (fr) * 2010-04-02 2012-03-08 국립수산과학원 Vaccin inactivé combiné pour la prévention de la coccidiose des poissons et procédé pour préparer celui-ci
CN103550765A (zh) * 2013-11-11 2014-02-05 中国科学院海洋研究所 一种海豚链球菌dna疫苗及其应用
JP2015127666A (ja) * 2013-12-27 2015-07-09 大阪府 クドア・セプテンプンクタータの迅速検出法
WO2020069656A1 (fr) * 2018-10-05 2020-04-09 福又达生物科技股份有限公司 Procédé de détection de pathogène de poisson
CN116640867A (zh) * 2023-06-27 2023-08-25 四川农业大学 一种raa-lfd检测停乳链球菌的引物对和探针组合及其应用

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