WO2000005341A1 - Anti-flexiblebacter maritimus vaccine (fm-95) for the prevention of the flexibacteriosis marina disease in turbot and salmonidae fish, and process for producing said vaccine - Google Patents

Anti-flexiblebacter maritimus vaccine (fm-95) for the prevention of the flexibacteriosis marina disease in turbot and salmonidae fish, and process for producing said vaccine Download PDF

Info

Publication number
WO2000005341A1
WO2000005341A1 PCT/ES1999/000232 ES9900232W WO0005341A1 WO 2000005341 A1 WO2000005341 A1 WO 2000005341A1 ES 9900232 W ES9900232 W ES 9900232W WO 0005341 A1 WO0005341 A1 WO 0005341A1
Authority
WO
WIPO (PCT)
Prior art keywords
vaccine
turbot
maritimus
flexibacteriosis
marina
Prior art date
Application number
PCT/ES1999/000232
Other languages
Spanish (es)
French (fr)
Inventor
Ysabel Santos Rodriguez
Francisco Pazos Alvarez
Original Assignee
Universidade De Santiago De Compostela
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Universidade De Santiago De Compostela filed Critical Universidade De Santiago De Compostela
Publication of WO2000005341A1 publication Critical patent/WO2000005341A1/en

Links

Classifications

    • 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
    • A61K39/0216—Bacteriodetes, e.g. Bacteroides, Ornithobacter, Porphyromonas
    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/36—Adaptation or attenuation of cells
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales

Definitions

  • Anti-Flexibacter maritimus vaccine for the prevention of "marine flexibacteriosis” disease in turbot and salmonid fish, and obtaining procedure.
  • Anti-Flexibacter maritimus (FM-95) vaccine for the prevention of "marine flexibacteriosis” disease in turbot and salmonid fish grown in seawater, and obtaining procedure.
  • the vaccine is composed of formolized cells of a strain of Flexibacter maritimus isolated from turbot. This vaccine confers a high degree of protection (greater than 90%) against the strains of Flexibacter maritimus (serotype O2) causing mortalities in cultured fish.
  • the strain of Flexibacter maritimus used in this vaccine is the isolated LPV 1.7 (serotype O2) and is deposited in the Spanish Type Culture Collection with the reference CECT 5045.
  • This strain has the typical characteristics of the species, being Gram-negative filamentous bacilli , positive oxidase and catalase, nitrate reducers, glucan-galactosamine producers and unable to produce hydrogen sulfide. They are also negative for the tests of indole, methyl red and Voges-Proskauer.
  • the species Flexibacter maritimus constitutes a homogeneous group from the biochemical and physiological point of view. However, the existence of serological and genetic variability has been detected Santos et al. 1998. ICES Leafletts.
  • LPS lipopolysaccharides
  • Incubation is performed at 25 ° C for 72 hours. At the end of the incubation period, formalin is added at a final concentration of 0.35% to kill the bacteria, and it is kept under stirring for three more hours, after which it is transferred to 4
  • the culture is centrifuged (10,000 rpm for 30 min.), The supernatant is discarded, and the cell precipitate is resuspended in phosphate buffered saline (PBS, sodium chloride, 8 g / L; potassium chloride 0.3 g / L; 0.73 g / L sodium phosphate; 0.2 g / L monopotassium phosphate; pH 7.4) with formalin at a final concentration of 0.35%.
  • PBS phosphate buffered saline
  • the vaccine mixture is adjusted to an optical density of 1.1 (A58O: Absorbance58 ⁇ ) -
  • the resulting vaccine solution contains approximately 10 ⁇ 0 cells / mL.
  • Sterility control is carried out by sowing the vaccine mixture in FMM agar plates, in tryptone soy agar plates (TSA) with NaCl at a final concentration of 1% (TSA-1), and in thioglycolate tubes, incubating for 72 hours at 25 and 37 ° C.
  • TSA tryptone soy agar plates
  • TSA-1 tryptone soy agar plates
  • TSA-1 tryptone soy agar plates
  • TSA-1 tryptone soy agar plates
  • thioglycolate tubes incubating for 72 hours at 25 and 37 ° C.
  • the specificity control is carried out by agglutination on slides, using the antiserum obtained in rabbit against strain CECT 5045 and as an antigen the whole cells used in the manufacture of the vaccine.
  • the vaccine should be kept at 4 ° C until use.
  • This vaccine should be administered by intraperitoneal injection.
  • fish are inoculated with 0.1-0.2 mL / fish (depending on the weight of the fish) of undiluted vaccine mixture.
  • the strain of Flexibacter maritimus used in this vaccine is called CECT 5045, and has been isolated from turbot grown in Galicia.
  • the efficacy of this vaccine was evaluated in turbot both on a laboratory scale and on an industrial scale.
  • the power expressed as Relative Survival Percentage (RSP) has been greater than 90%.

Abstract

Process for producing the anti-Flexibacter maritimus FM-95 vaccine for the prevention of the flexibacteriosis marina disease produced by the Flexibacter maritimus bacterium in turbot and salmonidae fish bred in sea water. The process is characterized in that the vaccine comprises the bacterial cells which have been inactivated with formol of the strain of Flexibacter maritimus isolated from turbot (strain deposited in the Spanish collection of type cultures with the reference CECT 5045). The vaccine can be administered by injection and confers to the fish efficient protection against flexibacteriosis marina.

Description

TÍTULO TITLE
Vacuna anti-Flexibacter maritimus (FM-95) para la prevención de la enfermedad "flexibacteriosis marina" en rodaballo y peces salmónidos, y procedimiento de obtención.Anti-Flexibacter maritimus vaccine (FM-95) for the prevention of "marine flexibacteriosis" disease in turbot and salmonid fish, and obtaining procedure.
DESCRIPCIÓNDESCRIPTION
Vacuna anti-Flexibacter maritimus (FM-95) para la prevención de la enfermedad "flexibacteriosis marina" en rodaballo y peces salmónidos cultivados en agua de mar, y procedimiento de obtención. La vacuna está compuesta por células formolizadas de una cepa de Flexibacter maritimus aislada de rodaballo. Esta vacuna confiere un elevado grado de protección (mayor al 90 %) frente a las cepas de Flexibacter maritimus (serotipo O2) causantes de mortalidades en peces cultivados.Anti-Flexibacter maritimus (FM-95) vaccine for the prevention of "marine flexibacteriosis" disease in turbot and salmonid fish grown in seawater, and obtaining procedure. The vaccine is composed of formolized cells of a strain of Flexibacter maritimus isolated from turbot. This vaccine confers a high degree of protection (greater than 90%) against the strains of Flexibacter maritimus (serotype O2) causing mortalities in cultured fish.
La flexibacteriosis marina, causada por la especie Flexibacter maritimus es una de las enfermedades bacterianas más importantes que afectan a peces marinos a nivel mundial. Esta enfermedad fue diagnosticada por primera vez en Galicia en el año 1991 (Pazos et al. 1993. FHS/AFS Newslett. 21 (3): 1-2) en rodaballos y salmones cultivados, convirtiéndose desde entonces en uno de los principales problemas para la piscicultura marina de nuestra área y causando importantes pérdidas económicas. El aislamiento del microorganismo fue posible gracias al empleo del medio para Flexibacter maritimus (FMM) diseñado en nuestro laboratorio (Pazos et al. 1996. Journal of Fish Diseases, 19: 193-197). El tratamiento de la flexibacteriosis marina con antibióticos no ofrece resultados satisfactorios y en la actualidad no existen vacunas comerciales para prevenir la enfermedad (Pazos et al. 1997. International Workshop on Aquaculture Application of Controlled Drug and Vaccine Delivery. Villa Manin di Pasariano. Italia, p. 49).Marine flexibacteriosis, caused by the species Flexibacter maritimus is one of the most important bacterial diseases that affect marine fish worldwide. This disease was first diagnosed in Galicia in 1991 (Pazos et al. 1993. FHS / AFS Newslett. 21 (3): 1-2) in turbot and farmed salmon, since then becoming one of the main problems for Marine fish farming in our area and causing significant economic losses. Isolation of the microorganism was made possible by the use of the medium for Flexibacter maritimus (FMM) designed in our laboratory (Pazos et al. 1996. Journal of Fish Diseases, 19: 193-197). The treatment of marine flexibacteriosis with antibiotics does not offer satisfactory results and there are currently no commercial vaccines to prevent the disease (Pazos et al. 1997. International Workshop on Aquaculture Application of Controlled Drug and Vaccine Delivery. Villa Manin di Pasariano. Italy, p. 49).
La cepa de Flexibacter maritimus empleada en esta vacuna es el aislado LPV 1.7 (serotipo O2) y está depositada en la Colección Española de Cultivos Tipo con la referencia CECT 5045. Esta cepa presenta las características típicas de la especie, siendo bacilos filamentosos Gram-negativos, oxidasa y catalasa positivos, reductores de nitratos, productores de glucano-galactosamina e incapaces de producir ácido sulfhídrico. Son además negativos para las pruebas del indol, rojo de metilo y Voges-Proskauer. La especie Flexibacter maritimus constituye un grupo homogéneo desde el punto de vista bioquímico y fisiológico. Sin embargo, se ha detectado la existencia de variabilidad serológica y genética Santos et al. 1998. ICES Leafletts. En prensa). Las proteínas y lipopolisacáridos (LPS) de la envoltura celular de esta especie son inmunogénicos para rodaballo y peces salmónidos (Pazos et al. 1997. VII International Conference on Diseases of Fish and Shellfish. European Association of Fish Pathologists. Edimburgo, Reino Unido, p. 73).The strain of Flexibacter maritimus used in this vaccine is the isolated LPV 1.7 (serotype O2) and is deposited in the Spanish Type Culture Collection with the reference CECT 5045. This strain has the typical characteristics of the species, being Gram-negative filamentous bacilli , positive oxidase and catalase, nitrate reducers, glucan-galactosamine producers and unable to produce hydrogen sulfide. They are also negative for the tests of indole, methyl red and Voges-Proskauer. The species Flexibacter maritimus constitutes a homogeneous group from the biochemical and physiological point of view. However, the existence of serological and genetic variability has been detected Santos et al. 1998. ICES Leafletts. In press). The proteins and lipopolysaccharides (LPS) of the cell envelope of this species are immunogenic for turbot and fish Salmonids (Pazos et al. 1997. VII International Conference on Diseases of Fish and Shellfish. European Association of Fish Pathologists. Edinburgh, United Kingdom, p. 73).
PREPARACIÓN A partir del preinóculo bacteriano en fase logarítmica de crecimiento se inoculan matraces de dos litros conteniendo un litro de medio de cultivo caldo FMM (peptona 5 g/L; extracto de levadura 0,5 g/L; acetato sódico 0,01 g/L; agua de mar 1 L; pH 7,3 ± 0,1).PREPARATION From the bacterial pre-circle in the logarithmic growth phase, two-liter flasks are inoculated containing one liter of FMM broth culture medium (5 g / L peptone; 0.5 g / L yeast extract; 0.01 g / sodium acetate L; 1 L seawater; pH 7.3 ± 0.1).
La incubación se realiza a 25 °C durante 72 horas. Finalizado el período de incubación, se añade formol a una concentración final de 0,35 % para matar las bacterias, y se mantiene durante tres horas más en agitación, al cabo de las cuales se transfiere a 4Incubation is performed at 25 ° C for 72 hours. At the end of the incubation period, formalin is added at a final concentration of 0.35% to kill the bacteria, and it is kept under stirring for three more hours, after which it is transferred to 4
°C. Después de 12 horas se centrifuga el cultivo (10.000 rpm durante 30 min.), se desecha el sobrenadante, y el precipitado celular se resuspende en tampón fosfato salino (PBS, cloruro sódico, 8 g/L; cloruro potásico 0,3 g/L; fosfato sódico 0,73 g/L; fosfato monopotásico 0,2 g/L; pH 7,4) con formol a una concentración final del 0,35 %. La mezcla vacunal se ajusta a una densidad óptica de 1,1 (A58O: Absorbancia58θ)- La solución vacunal resultante contiene aproximadamente 10^0 células/mL.° C. After 12 hours the culture is centrifuged (10,000 rpm for 30 min.), The supernatant is discarded, and the cell precipitate is resuspended in phosphate buffered saline (PBS, sodium chloride, 8 g / L; potassium chloride 0.3 g / L; 0.73 g / L sodium phosphate; 0.2 g / L monopotassium phosphate; pH 7.4) with formalin at a final concentration of 0.35%. The vaccine mixture is adjusted to an optical density of 1.1 (A58O: Absorbance58θ) - The resulting vaccine solution contains approximately 10 ^ 0 cells / mL.
El control de esterilidad se lleva a cabo sembrando la mezcla vacunal en placas de agar FMM, en placas de agar triptona de soja (TSA) con NaCl a una concentración final del 1 % (TSA-1), y en tubos de tioglicolato, incubando durante 72 horas a 25 y 37 °C. El control de especificidad se realiza mediante aglutinación en portaobjetos, utilizando el antisuero obtenido en conejo frente a la cepa CECT 5045 y como antígeno las células enteras empleadas en la fabricación de la vacuna.Sterility control is carried out by sowing the vaccine mixture in FMM agar plates, in tryptone soy agar plates (TSA) with NaCl at a final concentration of 1% (TSA-1), and in thioglycolate tubes, incubating for 72 hours at 25 and 37 ° C. The specificity control is carried out by agglutination on slides, using the antiserum obtained in rabbit against strain CECT 5045 and as an antigen the whole cells used in the manufacture of the vaccine.
La vacuna debe ser conservada a 4 °C hasta su utilización.The vaccine should be kept at 4 ° C until use.
MODO DE ADMINISTRACIÓNADMINISTRATION MODE
Esta vacuna debe administrarse por inyección intraperitoneal. Para ello, se inoculan los peces con 0,1 - 0,2 mL/pez (dependiendo del peso del pez) de mezcla vacunal sin diluir.This vaccine should be administered by intraperitoneal injection. For this, fish are inoculated with 0.1-0.2 mL / fish (depending on the weight of the fish) of undiluted vaccine mixture.
La cepa de Flexibacter maritimus empleada en esta vacuna se denomina CECT 5045, y ha sido aislada a partir de rodaballo cultivado en Galicia.The strain of Flexibacter maritimus used in this vaccine is called CECT 5045, and has been isolated from turbot grown in Galicia.
La eficacia de esta vacuna fue evaluada en rodaballo tanto a escala de laboratorio como a escala industrial. La potencia, expresada como Porcentaje de Supervivencia Relativa (RSP) ha sido superior al 90 %. The efficacy of this vaccine was evaluated in turbot both on a laboratory scale and on an industrial scale. The power, expressed as Relative Survival Percentage (RSP) has been greater than 90%.

Claims

REIVINDICACIONES
1.- Cepa de Flexibacter maritimus LPV 1.7, aislada de rodaballo cultivado en Galicia, depositada en la Colección Española de Cultivos Tipo (CECT) con el número de referencia CECT 5045. 1.- Flexibacter maritimus LPV 1.7 strain, isolated from turbot grown in Galicia, deposited in the Spanish Type Culture Collection (CECT) with the reference number CECT 5045.
2.- Vacuna anti-Flexibacter maritimus (FM-95) para la prevención de la enfermedad flexibacteriosis marina en rodaballo y peces salmónidos cultivados en agua de mar, compuesta por células bacterianas de la cepa LPV 1.7 de este microorganismo inactivadas con formol.2.- Anti-Flexibacter maritimus vaccine (FM-95) for the prevention of marine flexibacteriosis disease in turbot and salmon fish grown in seawater, composed of bacterial cells of the LPV 1.7 strain of this microorganism inactivated with formalin.
3.- Procedimiento para la obtención de la vacuna de la reivindicación 2, caracterizado por el cultivo de las células bacterianas en caldo para Flexibacter maritimus3. Method for obtaining the vaccine of claim 2, characterized by the culture of the bacterial cells in broth for Flexibacter maritimus
(FMM) y su inactivación por formol a una concentración final del 0,35 % durante 3 horas a 25 °C, y porque el cultivo inactivado se centrifuga y el precipitado celular se resuspende en tampón fosfato a una densidad óptica de 1,1 (Absorbancia580)-(FMM) and its inactivation by formalin at a final concentration of 0.35% for 3 hours at 25 ° C, and because the inactivated culture is centrifuged and the cell precipitate is resuspended in phosphate buffer at an optical density of 1.1 ( Absorbance580) -
4.- Procedimiento, según la reivindicación 3, para la obtención de la vacuna FM-95 anti-Flexibacter maritimus, caracterizado porque la concentración de células bacterianas en la preparación vacunal es de 10 células/mL. 4. Method according to claim 3 for obtaining the FM-95 anti-Flexibacter maritimus vaccine, characterized in that the concentration of bacterial cells in the vaccine preparation is 10 cells / mL.
PCT/ES1999/000232 1998-07-22 1999-07-21 Anti-flexiblebacter maritimus vaccine (fm-95) for the prevention of the flexibacteriosis marina disease in turbot and salmonidae fish, and process for producing said vaccine WO2000005341A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
ES009801549A ES2139549B1 (en) 1998-07-22 1998-07-22 ANTI-FLEXIBACTER MARITIMUS VACCINE (FM-95) FOR THE PREVENTION OF THE DISEASE "MARINE FLEXIBACTERIOSIS" IN TURBINE AND SALMONID FISH, AND PROCEDURE FOR OBTAINING.
ESP9801549 1998-07-22

Publications (1)

Publication Number Publication Date
WO2000005341A1 true WO2000005341A1 (en) 2000-02-03

Family

ID=8304597

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/ES1999/000232 WO2000005341A1 (en) 1998-07-22 1999-07-21 Anti-flexiblebacter maritimus vaccine (fm-95) for the prevention of the flexibacteriosis marina disease in turbot and salmonidae fish, and process for producing said vaccine

Country Status (2)

Country Link
ES (1) ES2139549B1 (en)
WO (1) WO2000005341A1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008074797A (en) * 2006-09-22 2008-04-03 Univ Kinki Vaccine for fish gliding bacterial disease

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2939719A1 (en) * 2021-10-26 2023-04-26 Univ Santiago Compostela Immunogenic composition for the prevention of marine tenacibaculosis caused by Tenacibaculum maritimum and Tenacibaculum soleae in fish, procedure for obtaining and use (Machine-translation by Google Translate, not legally binding)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08214788A (en) * 1995-02-08 1996-08-27 Sanwa Denpun Kogyo Kk Feed for fished and shellfishes containing specific antigen protein or peptide containing antigenic determinant of the same protein

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08214788A (en) * 1995-02-08 1996-08-27 Sanwa Denpun Kogyo Kk Feed for fished and shellfishes containing specific antigen protein or peptide containing antigenic determinant of the same protein

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
ALSINA M. AND BLANCH A.R.: "First isolation of Flexibacter maritimus from culturated turbot (Scophthalmus maximus)", BULL. EUR. ASS. FISH. PATHOL., vol. 13(5), 1993, pages 157 - 159 *
BERNARDET D.F., Immunization with bacterial antigens; Flavobacterium and Flexibacter infections. En: Fish Vaccinology, GUDDING R. et al. (Eds), Dev. Biol. Stand. Bases, Karger, 1997, Vol. 90, pages 179-188. *
BERNARDET J.F. ET AL: "Comparative study on Flexibacter maritimus strains isolated form farmed sea bass (Dicentrarchus labrax) in France", FISH PATHOLOGY, vol. 29(2), 1994, pages 105 - 111 *
CHEN M.F. ET AL: "Isolation and Characterization of Flexibacter maritimus from marine fishes of California", JOURNAL OF AQUATIC ANIMAL HEALTH, vol. 7, 1995, pages 318 - 326 *
DATABASE WPI Week 9644, Derwent World Patents Index; AN 1996-436855 *
HANDLINGER J. ET AL: "The pathology of Flexibacter maritimus in aquaculture species in Tasmania, Australia", JOURNAL OF FISH DISEASES, vol. 20, 1997, pages 159 - 168 *
MAGARINOS B. ET AL: "Response of Pasteurella piscida and Flexibacter maritimus to skin mucus of marine fish", DISEASES OF AQUATIC ORGANISMS, vol. 21, 1995, pages 103 - 108 *
PAZOS F. ET AL: "Evaluation of media for the successful culture of Flexibacter maritimus", JOURNAL OF FROM DISEASES, vol. 19, 1996, pages 193 - 197 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008074797A (en) * 2006-09-22 2008-04-03 Univ Kinki Vaccine for fish gliding bacterial disease

Also Published As

Publication number Publication date
ES2139549B1 (en) 2000-10-01
ES2139549A1 (en) 2000-02-01

Similar Documents

Publication Publication Date Title
Aronson Spontaneous tuberculosis in salt water fish
Gutierrez et al. Protective properties of egg yolk IgY containing anti‐Edwardsiella tarda antibody against paracolo disease in the Japanese eel, Anguilla japonica Temminck & Schlegel
EP0555366A1 (en) Bacterial attenuation method and vaccine
CN112029696B (en) Edwardsiella piscicola derived from turbot and application thereof
EP3071226B1 (en) Fish vaccine
US20150132341A1 (en) Fish vaccine
US7707970B2 (en) Vaccine against salmonid rickettsial septicaemia based on Arthrobacter cells
CN104560851A (en) Aeromonas salmonicida live vaccine preparation as well as preparation method and application of lyophilized vaccine product
JP3601602B2 (en) Non-toxic microorganisms and their use: Salmonella
US20160296615A1 (en) Fish vaccine
WO2000005341A1 (en) Anti-flexiblebacter maritimus vaccine (fm-95) for the prevention of the flexibacteriosis marina disease in turbot and salmonidae fish, and process for producing said vaccine
Raj et al. Characterization and infectivity evaluation of Vibrio harveyi causing white patch disease among captive reared seahorses, Hippocampus kuda
Dack et al. Further studies on Bacterium necrophorum isolated from cases of chronic ulcerative colitis
US8758774B2 (en) Bivalent vaccine for marine fish and method for making the same
WO1999055835A1 (en) Anti-vibrio anguillarum vaccine (gava-3) for the prevention of the vibriosis disease in the turbot and salmonidae, and preparation process
ES2321787B1 (en) PROCEDURE FOR OBTAINING ANTI-EDWARDSIELLA TARDA VACCINE (EDT) FOR THE PREVENTION OF EDWARDSIELOSIS DISEASE OF THE ROLL.
Nayak et al. Original Research Article Characterization, pathogenicity, antibiotic sensitivity and immune response of Flavobacterium columnare isolated from Cirrhinus mrigala and Carassius auratus
Wollstein Studies on the phenomenon of d'Herelle with Bacillus dysenteriae
JPH09505085A (en) Pasteurellae antigens and related vaccines
JPH05336950A (en) Cultivation for preparing oral live typhoid vaccine
CN117305191B (en) Drug-resistant Edwardsiella tarda for killing turbot
Smith Anaerobic pneumococcus
Meyer et al. A Disease in Wild Rats Caused by Pasteurella muricida, n. sp.: Plague Studies. 2
JP3543739B2 (en) Fish disease vaccine
ES2224864B2 (en) PROCEDURE FOR OBTAINING THE ANTI-LACTOCOCCUS GARVIEAE ORAL VACCINE FOR THE PREVENTION OF THE LACTOCOCOSIS DISEASE OF THE TRUCK.

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): CA NO US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
NENP Non-entry into the national phase

Ref country code: CA