WO2006068077A1 - Nouveau vaccin contre les maladies des poissons induites par une streptococcie - Google Patents

Nouveau vaccin contre les maladies des poissons induites par une streptococcie Download PDF

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Publication number
WO2006068077A1
WO2006068077A1 PCT/JP2005/023242 JP2005023242W WO2006068077A1 WO 2006068077 A1 WO2006068077 A1 WO 2006068077A1 JP 2005023242 W JP2005023242 W JP 2005023242W WO 2006068077 A1 WO2006068077 A1 WO 2006068077A1
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WIPO (PCT)
Prior art keywords
fish
vaccine
streptococcus
streptococci
culture
Prior art date
Application number
PCT/JP2005/023242
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English (en)
Japanese (ja)
Inventor
Terutoyo Yoshida
Masayoshi Asagi
Satoru Kurokawa
Original Assignee
Scientific Feed Laboratory Co., Ltd.
Meiji Seika Kaisha, Ltd.
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 Scientific Feed Laboratory Co., Ltd., Meiji Seika Kaisha, Ltd. filed Critical Scientific Feed Laboratory Co., Ltd.
Priority to JP2006548963A priority Critical patent/JPWO2006068077A1/ja
Publication of WO2006068077A1 publication Critical patent/WO2006068077A1/fr

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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
    • A61K39/09—Lactobacillales, e.g. aerococcus, enterococcus, lactobacillus, lactococcus, streptococcus
    • A61K39/092—Streptococcus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • 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
    • 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
    • C12R2001/46—Streptococcus ; Enterococcus; Lactococcus

Definitions

  • the present invention relates to a method for producing a vaccine effective for the prevention and treatment of new streptococcal disease in fish, a new fish disease vaccine preparation for new streptococcal disease, and a method for preventing or treating fish streptococcal disease. More specifically, a novel strain of the genus Streptococcus to which the causative bacteria of fish streptococci belong, a microorganism belonging to the genus Streptococcus and capable of producing a substance having a vaccine effect against a new type of streptococci of fish is used.
  • a method for producing a vaccine effect substance for preventing or treating streptococcal disease in fish, a vaccine preparation comprising the vaccine effect substance for preventing or treating fish streptococci obtained here, and a new type of fish using the novel strain The present invention relates to a vaccine and a vaccine composition for preventing and treating streptococcal disease.
  • Non-patent Document 2 Streptococcus dysgalactiae infection serotyped as a result of isolation of causative bacteria from this diseased fish and determination of general characteristics and 16S rDNA nucleotide sequence. This was confirmed and reported (Non-patent Document 2). Since then, it has been confirmed that it occurs from summer to autumn when the water temperature is high every year.
  • Patent Document 1 Japanese Patent No. 3543739
  • Non-Patent Document 1 Teixeira, M. et.al., International J. Nanole 'Ob' Systematic 'Bataterologi (International J. Systematic Bact erioU, 46 ⁇ 664—668, 1996 Year
  • Non-Patent Document 2 Japanese Society of Fisheries Science, Title # 1243 (A new streptococci isolated from a lesion of amberjack), April 1-5, 2003
  • Non-Patent Document 3 Annual Meeting of the Japanese Society of Fish Diseases, Title No. 207 (Study on Epidemiology of Lancefield Group C Streptococcus in Puri Fish), September 17-19, 2004 Disclosure of Invention
  • the present invention provides a preventive and therapeutic vaccine for a new type of streptococcal disease for fish caused by infection with Streptococcus dysgalactiae. Furthermore, the present invention provides a combination vaccine with a conventional vaccine against streptococcal disease and a combination vaccine with other fish disease vaccines.
  • Streptococcus microorganisms for example, a new strain of Streptococcus streptococcus disgalacticae subspecies disgalactia 07-12, have been used against new streptococci in fish.
  • the ability to produce substances with high and potent vaccine effects the culture has a strong vaccine effect against the new type of streptococcal disease of fish, and the culture is an excellent fish for streptococci
  • the present invention has been completed based on these findings.
  • Streptococcus disgalactea subspecies' disgalactea 07 12 was deposited as FERM BP-10442 on August 11, 2004 at the National Institute of Advanced Industrial Science and Technology, Japan Patent Biological Depositary. .
  • the present invention as a method for preventing and treating a new type of streptococci in fish caused by Streptococcus dysgalactiae, it has become possible to prevent and treat it by administration of a vaccine.
  • the present invention has made it possible to prevent and treat new streptococci even when drug-resistant bacteria emerge.
  • the vaccine according to the present invention does not interfere with the effect of mixing with a conventional Lactococcus garvieae vaccine, and other interferences such as an inactivated virus solution of irido virus (Mdo vims) are observed. It can be used as a multivalent vaccine by mixing with an inactivated vaccine solution.
  • FIG. 1 is a diagram showing the growth process of Streptococcus disgalactiae by medium.
  • FIG. 2 is a graph showing the effectiveness of a combination vaccine.
  • Streptococcus streptococcus' disgalactiae is a Lancefield C3 ⁇ 4 (Lancefield
  • Table 1 shows the bacteriological properties of the new streptococcus streptococcus disgalactiae subspecies' disgalactia 07-12 according to the present invention.
  • Lactose One D-Trehalose: +
  • the culture method of Streptococcus dysgalactia according to the present invention is not particularly limited.
  • a general streptococcal culture method may be used.
  • the medium used for the culture is not particularly limited as long as the bacterium can grow.
  • the broth medium Heart's infusion broth medium (
  • HI Infusion Broth
  • BHI Infusion Broth
  • THB Todd Hebbit Can be cultured in broth medium
  • THB blood agar medium
  • the pH during culture is 4.5-9.6.
  • the culture may be performed under any conditions such as stationary culture, shaking culture, and stirring culture, but good growth can be obtained by culturing with gentle stirring. In addition, there is no change in efficacy after 10 passages in THB medium or SCD medium.
  • the vaccine of the present invention can be produced by inactivating a culture broth of a microorganism belonging to the genus Streptococcus and capable of producing a substance having a vaccine effect against streptococcal disease in fish.
  • the method for inactivating the culture solution is not particularly limited, and a general method for inactivating bacteria can be used. Examples thereof include a method of chemically inactivating formalin, merzonin, ⁇ -pillar pillatatone, and the like. Alternatively, a physical inactivation method such as heat sterilization at 65 ° C for 30 minutes or 72 ° C for 15 minutes, or disruption of cells by ultrasonic treatment may be used.
  • a culture solution (culture) of Streptococcus disgalactia inactivated by the above method can be used.
  • the vaccine effect substance in the present invention may be one obtained by inactivating the bacterial cells isolated from the culture or the culture supernatant from which the bacterial cells have been separated and removed. More preferably, the inactivated culture solution of Streptococcus disgalactia 'Subspecies' Disgalactea 07-12, or the cells isolated from the culture, or the cells isolated What inactivated the removed culture supernatant is good. In particular, it is more preferable to use an inactivated culture supernatant from which bacterial cells have been separated and removed.
  • the route of administration is oral, immersion, intraperitoneal or intramuscular, and is not particularly limited, but is preferably intraperitoneal or intramuscular injection.
  • This vaccine can be administered prophylactically before the onset of symptoms, and can also be expected to be effective as a therapeutic agent if administered after onset.
  • the dose is 1 X 10 5 to 1 X 10 10 CFUZ tail, preferably 1 X 10 6 to 1 X 10 9 CFU / tail, more preferably 1 X 10 7 to 1 X as the number of bacteria before inactivation 10 8 CFU / tail.
  • Target fish for vaccine administration include yellowtail fish such as puri, amberjack, hiramasa, horse mackerel fish, etc. It will not specifically limit if it is a fish which suffers from a new type
  • the age of the fish at the time of administration is not particularly limited, as long as it has grown to a size that can be technically injected. In general, any fish with a body weight of 10g to 20g or more is acceptable. A single dose is sufficient, but it is more preferable to re-immunize after 3 to 12 months
  • the vaccine effect can be further enhanced by administration with a known adjuvant such as vegetable oil, mineral oil, aluminum gel or the like. Furthermore, the effect can be further enhanced by providing known immune enhancing agents such as vitamin E and peptide darican.
  • a pharmaceutically acceptable carrier is added to the vaccine composition of the present invention, and for example, it can be used in an appropriate dosage form such as an injection or liquid.
  • Pharmaceutically acceptable carriers include solvents, bases, stabilizers, preservatives, solubilizers, excipients, buffers and the like well known to those skilled in the art.
  • Streptococcus streptococcus ⁇ disgalactiae ⁇ subspecies' disgalactiae 07-12 (accession number:? 61 ⁇ 1 BP—10442) is commercially available BHI, SCD, and THB (Let ’s bettaton made by Dickinson ( Becton
  • BD cultured at 25 ° C for 24 hours, and the number of viable bacteria was measured by a conventional method.
  • the cells Regardless of which medium was used, the cells grew to 10 7 CFU / 0.1 mL or more. Increase in each medium The breeding process is shown in Fig. 1.
  • Formalin is added to 0.3 (vol / vol)% of the bacterial solution (cultured product) cultured at 25 ° C for 24 to 72 hours using the same THB or SCD medium as in Example 1, and inactivated at 25 ° C for 48 hours. did.
  • the inactivated bacterial solution was centrifuged at 5000 ⁇ G for 30 minutes to obtain bacterial cells of the sediment and culture supernatant.
  • the cells were resuspended in a 1/50 amount of phosphate buffered saline to obtain a concentrated bacterial solution, and the supernatant was concentrated by ultrafiltration to obtain concentrated supernatants of various concentrations.
  • Test vaccines of various compositions by mixing concentrated bacterial solutions and concentrated supernatants of various concentrations at different ratios, or by mixing with other vaccines, and further diluting with phosphate buffered saline. was made.
  • Test vaccines of various compositions prepared according to Example 2 were used.
  • a vaccine prepared from a bacterial solution cultured in fetal bovine serum (FBS) or a bacterial solution cultured in a THB medium supplemented with an appropriate amount of FBS was also used. Note that the bacterial solution obtained from all the media was cultured for 24 hours.
  • mice were attacked by intraperitoneal injection of Streptococcus' disgalactia 'subspices' desgalactia 07-12 (accession number: FERM BP-10442).
  • the number of attacking bacteria was 3.1 X 10 7 CFU / O. LmL / tail, and Table 2 was the number of attacking bacteria 4.9 X 10 7 CFU / O. LmL / tail).
  • the clinical symptoms were observed for 21 days and the findings were recorded.
  • the mortality rate in the immunization group was lower than the mortality rate in the attack control group (injected with phosphate buffered saline instead of the vaccine), confirming the effectiveness of the test vaccine.
  • the results of vaccine efficacy tests are shown in Tables 2 and 3.
  • Streptococcus dysgalactia subspecies' dysgalactia 07-12 vaccine prepared according to Example 2 Inactivated cells were mixed to 10 8 CFU / mL in 4-fold concentrated culture supernatant. Inactivated cells of Ratatococcus garbiae KG9408 (Lactococcus garviae K G9408) (Accession number: FERM BP-6749) were added to 10 9 CFU / mL to prepare a mixed vaccine. The bacterial solution was inactivated after 24 to 72 hours of incubation.
  • the test combination vaccine was injected into the peritoneal cavity of each test group (immune group) at 0.1 mL per fish.
  • 0.1 mL of phosphate buffered saline was injected instead of the vaccine. 14 days after immunization, intravascular injection of Streptococcus dysgalactia 07-12 culture solution into each of the immunized group and the control group (number of challenge bacteria: 1.6 X 10 7 CFU / 0.1)
  • Ratatococcus galbiae KG9408 number of attacking bacteria: 1.1 X 10 4 CFU / 0.1
  • the mortality rate of the new and old streptococcal mixed vaccine immunization group was lower than the mortality rate of the corresponding old and new streptococcal attack control group, and the vaccine effect was confirmed against both old and new streptococci.
  • the test results are shown in Fig. 2.
  • IP0D IP0D

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  • Health & Medical Sciences (AREA)
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  • Tropical Medicine & Parasitology (AREA)
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Abstract

La présente invention décrit un vaccin préparé par inactivation de la culture d'un micro-organisme appartenant au genre Streptococcus et produisant une substance qui induit un effet de type vaccin sur les streptococcies des poissons, par exemple, Streptococcus dysgalactiae subsp. dysgalactiae (FERM BP-10442). Ledit vaccin présente chez les poissons un excellent effet vis-à-vis de nouvelles streptococcies. Ledit vaccin se distingue des méthodes existantes employant des antibiotiques en ce qu'il permet le traitement prophylactique et thérapeutique de streptococcies sans risque d'apparition d'une souche résistant à un médicament.
PCT/JP2005/023242 2004-12-20 2005-12-19 Nouveau vaccin contre les maladies des poissons induites par une streptococcie WO2006068077A1 (fr)

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JP2006548963A JPWO2006068077A1 (ja) 2004-12-20 2005-12-19 新型レンサ球菌症用魚病ワクチン

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JP2004368300 2004-12-20

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007326794A (ja) * 2006-06-06 2007-12-20 Kyoritsu Seiyaku Kk 魚類ストレプトコッカス・ディスガラクティエを抗原とする不活化ワクチン

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2001342141A (ja) * 2000-03-29 2001-12-11 Gotoo Yoshoku Kenkyusho:Kk 水棲動物・陸棲動物用薬剤及び飼料
WO2001096381A2 (fr) * 2000-06-12 2001-12-20 University Of Saskatchewan Immunisation de bovins laitiers a l'aide de la proteine gapc contre l'infection a streptocoque
JP2002513398A (ja) * 1997-01-21 2002-05-08 ザ・テキサス・エイ・アンド・エム・ユニバーシテイ・システム フィブロネクチン結合タンパク質組成物および使用法
JP2003174886A (ja) * 2001-12-12 2003-06-24 Toray Ind Inc 新規タンパク質およびそのｄｎａ

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002513398A (ja) * 1997-01-21 2002-05-08 ザ・テキサス・エイ・アンド・エム・ユニバーシテイ・システム フィブロネクチン結合タンパク質組成物および使用法
JP2001342141A (ja) * 2000-03-29 2001-12-11 Gotoo Yoshoku Kenkyusho:Kk 水棲動物・陸棲動物用薬剤及び飼料
WO2001096381A2 (fr) * 2000-06-12 2001-12-20 University Of Saskatchewan Immunisation de bovins laitiers a l'aide de la proteine gapc contre l'infection a streptocoque
JP2003174886A (ja) * 2001-12-12 2003-06-24 Toray Ind Inc 新規タンパク質およびそのｄｎａ

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
MISAWA N. ET AL.: "Burizoku Gyorui ni Oite Hajimete Kakunin sareta Lancefield group C rensa Kyukin Kansensho", JAPANESE SOCIETY OF VETERIANY SCIENCE GAKUJUTSU SHUKAI KOEN YOSHISHU, vol. 138TH, August 2004 (2004-08-01), pages 123, XP003007379 *
NOMOTO R. ET AL.: "Burizoku Gyorui Oyobi Chikusan Yurai no Lancefield group C Rensa Kyukin no Seijo ni Kansuru Kenkyu", THE JAPANESE SOCIETY OF FISHERIES SCIENCE TAIKAI KOEN YOSHISHU, vol. 2005, April 2005 (2005-04-01), pages 71, XP003007380 *
NOMOTO R. ET AL.: "Lancefield group C Streptococcus dysgalactiae infection responsible for fish mortalities in Japan", JOURNAL OF FISH DISEASES, vol. 27, no. 12, 2 December 2004 (2004-12-02), pages 679 - 686, XP003007378 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007326794A (ja) * 2006-06-06 2007-12-20 Kyoritsu Seiyaku Kk 魚類ストレプトコッカス・ディスガラクティエを抗原とする不活化ワクチン

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