CS248586B1 - Inactivated vaccine against ovine listeriosis - Google Patents

Abstract

The solution relates to inactivated vaccine substances useful in immunoprophylaxis listeriosis of sheep, which in the vaccinees creates a state of specific protection against clinically manifest form of the disease. The vaccine contains bacterial bodies of 5 strains of Listeria monocytogenes in the same volume ratio. In immunized sheep, it significantly protects against intravenous infection with virulent with a brute of this kind of microorganism induces an increase in immunized rabbits Specific agglutinating antibodies. Nachadza use in animal mass production and contributes to a substantial reduction in incidence of this disease in sheep farming all category and thus to increase the economy efficiency in sheep farming.

Description

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SNIRC JULIUS MVDr. CSc., NITRA, BALASCAK JURAJ RVDr. CSc., PBTROVANY, GUBRAN DUSAN MVDr., NITRA,

KOVAC JOZEF MVDr., FIELD KBSOV

Inactivated sheep listeriosis vaccine

The invention relates to an inactivated vaccine useful in the immunoprophylaxis of sheep listeriosis, which in a vaccine creates a state of specific protection against a clinically manifest form of the disease. The vaccine contains bacterial bodies from 5 strains of Listeria monocytogenes in the same volume ratio.

In immunized sheep it significantly protects against intravenous infection with virulent horseradish of this kind of microorganism, in immunized rabbits it induces the rise of specific agglutination antibodies. It is used in large-scale livestock production and contributes to a substantial reduction in the incidence of this disease in sheep breeding of all categories and thereby to increase the economic efficiency in sheep breeding.

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The invention relates to an inactivated vaccine from heated virulent strains of Listeria monooytogenes isolated from brain tissue ovia affected by listeric meningitis, which causes a vaccine condition of the vaccine in the vaccine against a clinically manifest form of the disease and which can be used for immunotherapy.

Listeriosis, as an infectious non-contagious palsy of humans and animals caused by Listeria monooytogenes, has become one of the main causes of economic losses in ovieo in recent years. The occurrence of this disease does not only create a serious animal health problem, but also a serious problem in the field of general health.

Of the different categories of livestock, sheep are the most affected. In these animals, the disease causes serious, economic loss due to high mortality, abortion rates, growth retardation in intensive fattening lambs, increased costs of epidemiological measures and therapeutic interventions,

In general, various types of antibiotics and sulfonamide or various combinations thereof are used in the therapy of the disease. SCHELD, T.I., FLETCHER, D.D., FINK, F.N., SANDE, M.A. : Response to therapy in an experimental rabbit model of meningitis due to Listeria monooytogenes, (J.Infect.Dis., Θ4θ, 1979, pp. 287-291) determined in paraffinic model that the most effective therapy for listeric meningitis is the parenteral administration of ampicillin antibiotics. and gentamycin · However, antULotic therapy can only be successful after a certain clinical stage of unless there are manifestly produced nerve symptoms of the disease. In addition, the use of antibiotics generates a positive selection pressure for the presence of resistant strains of this microorganism.

The high costs of eradicating this infectious disease reduce the economic efficiency of breeding ovieo. In this situation, the introduction of a spe- cific immunogen would appear to be the most effective measure in the fight against listeriosis, which would produce sufficient immunization in vaccinated animals.

248 588 protection against clinically manifest form of listeriosis. At present, a specific immunogen is not commercially produced in our country, and very few are very few experts in the field of immuno-prophylaxis of this disease in ova.

In 1981, KEMPSKI, W. Report on the damping of listeria by using inactivated auto-prophylactic saccharides / Zycie listeriozy owiec przy uzyciu autoszpzepionek wraps. (The Problems of Listeriosis, Prooeedings of the Seventh International Symposium, Sofia, 1979, pp. 344-338), was used for Speoifia's prophylaxis! of this disease, the inactivated vacoin, which contained, as a standard, strains of serotypes I and IV into filled with auto-hormone strains isolated from holdings with an acute disease. In addition to the use of inactivated auto-hormone vacoin in the immunoprophylaxis of this disease, a live attenuated vacoin was prepared from strains of Listeria monooytogenes serotypes 1 and 4b / IVANCV, I., DRAGANOV, M., DIKOVA, Ts .: Study on the aotive immunoprophylaxis of sheep lister. Prohlems of listeriosis, Prooeedings of the sewenth International Symposium, Sofia, 1979, pp.324-333 /,

Accordingly, there is a need for the introduction of a suitable vacoin for immunoprophylaxis of listeriosis ovieo. The application of the vacoin satisfies practical breeding conditions and can be used for protective vaccination in endemiocyte infested listeriosis recurring as well as in the susceptibility of simple cliniocy manifest disease. The vaccine can be used as an emergency at the outbreak of the disease when recommending the following measures: sick individuals with clinically manifest symptoms of listeric encephalitis are resisted, in addition, Speo-Oxygen therapeutics are treated and concurrently vaccinations and revacoinations are performed.

The bag prepared according to the invention can find a broad practical application in animal production in animal husbandry, where it will substantially contribute to reducing the occurrence of listeriosis and thus improve economic efficiency in animal husbandry.

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The vaccine according to the invention comprises 4 to 6% of a mixture of inactivated bacterial bodies of Listeria monocytogenes of five strains in 1 ml in equal volume, emulsified in an adjuvant which is an aluminum hydroxide gel, or a mixture of adjuvant oils.

Our experiments have shown that it is possible to produce an inactivated vaccine against sheep listeriosis from a bacterial culture prepared by culturing virulent strains of Listeria monooytogenes, which have been isolated from the brain of lambs and sheep affected by acute meningo-enoephaloritis. These strains were deposited in the collection of microorganisms of the University of J. E. Purkyne in Brno, cl. Peace Defenders 10. The strains were assigned the following deposit numbers: CCM 3847, CCM 3848, CCM 3849, CCM 3850, CCM 3851. The strains were tested for their serotyping and virulence. Characteristics of individual strains:

CCM 3847 serotype 1 / 2a, ΙΕ ^ θ for mouse 1, OSOx10 ⁸ #

CCM 3848 serotype 4b, 1Ι) ^ θ for mouse 2,797x10?, ·

CCM 3849 serotype 1 / 2a, ΙΒ ^ θ P ^ o my ^ 4,743x10 ^;

CCM 3850 serotype 4a, 11> _5θ for mouse 5,633 ^χ 1θ ^ »

CCM 3θ51 serotype 1 / 2a, LD ^ q for mouse 2,649x10 ?.

The minimum infectious dose to sheep after intravenous administration was 3.3 x ^{10 10,} and the maximum infectious dose of 2.7 x ¹⁰ 6 colonies formed germs.

The vaccine prepared by said strains has a good specific immunostimulatory effect in sheep against challenge intravenous infection! with a virulent strain of Listeria rnonocytogenes,

Immunoprophylactically, the vaccine can be used in all age categories of sheep and an optimal immunization scheme has been developed for its practical application, respecting the current state of knowledge in the field of etiopathogenesis of this disease. Use of the vaccine in sheep breeding will significantly reduce the incidence of clinically manifest forms of listeriosis. as a result, the economic efficiency of breeding this kind of livestock will be increased.

Example

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Listeria monooytogenes strains CCM 3847, CGM 3848, CCM 3849, CCM 3θ5θ, CCM 3851 are propagated in sterile soybean medium, a solution of amino acids obtained by enzymatic hydrolysis of delipidized soybean, enriched with 1 volume of 40% glucose solution and a sputter. Each strain of this microorganism is propagated separately. After 18 h culture at 37 ° C ^{, a} sample of each strain ^is tested to determine the optical density, pH of the bacterial mass and sterility. The resulting density must be at least $ 30 and the saw should not drop below 7 »2 - 7.4.

The bacterial mass obtained is inactivated by the addition of 10 µl formaldehyde so that the resulting concentration of formaldehyde does not exceed a maximum of 1.6 mg / ml. The inactivated bacterial cultures are subsequently leveled using equal volumes of bacterial cultures of the individual vaccine strains. The resulting concentration of inactivated microorganisms in the equalized bactorial meat is 4 to 6 x 10 6.

The mixed bacterin thus prepared is bound to an adjuvant, which may be Al (OR ₎ or a mixture of adjuvant oils. When using Al / OH / gel, this is added to the final concentration of 800 g / 100 ml in inactivated bacterin. When using adjuvant oils, these are emulsified in a ratio by adding 1 part adjuvant oils to 4 parts of inactivated bacterin.

The vaccine is satisfactory if it is free from bacterial and fungal contamination, is harmless if, after subcutaneous administration, 0.3 ml of vacoins do not die within 8 days after administration of either vaccinated mice and after subcutaneous administration with a rabbit of 2 x 1.5 ml in the range 14 days, elicits a specific agglutination antibody titer ranging from 1:80 to 1: 120.

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The X-value of the inactivated vaccine determined by the Mortality Index after infection! is 0.11 in ovine immunized, as opposed to 1.705 in non-immunized sheep in laboratory conditions. The efficacy of the inactivated vaccine in ovieo farms with a staionic incidence of listeriosis was determined on the basis of the proportion of decline in mortality with the diagnosis of listeriosis in vaccinated animals versus vaccinated animals. Average **

Claims (3)

PRBDMET OF THE INVENTION 248 588 An inactivated vaccine against listeriosis ovieo characterized in that it contains 4 to 6% of the mixture-inactivated balcterial body of Listeria monooytogenes of CCM 3847, CCM 3848, CCM 3849, CCM 3850, CCM 3851 in 1 ml in 1 ml. ratio, emulsified in adjuvant · 2. The inactivated vaccine of claim 1, wherein the adjuvant is an aluminum hydroxide gel or a mixture of adjuvant oils.