RU2802251C1 - Vaccine composition for the prevention of enzootic abortion in sheep - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to a vaccine composition containing the Chlamydia VNITIBP-21 antigen and a kit for the prevention of enzootic abortion in sheep.

EFFECT: invention can be used to prevent enzootic abortion in sheep.

3 cl, 4 tbl, 9 ex

Description

The present invention relates to the field of microbiology, veterinary medicine, biotechnology, pharmacy, in particular to a new strain of Chlamydia abortus "Chlamydia VNITIBP-21" used for the manufacture of inactivated vaccines and the production of specific diagnostic sera and globulins and as an antigen for the detection of antibodies to chlamydia in farm animals . The claimed invention can be used in the pharmaceutical industry, agriculture, as well as in research activities.

State of the art

Chlamydia abortus is a Gram-negative bacterium that is an obligate intracellular pathogen of eukaryotic cells. This bacterium has two membranes and no flagella. When released into the environment through vaginal secretions, the bacteria remain alive and can easily spread to infect other organisms.

Chlamydia abortus is found in cows, goats, sheep and pigs, where it causes abortion later in pregnancy by colonizing the placenta.

Chlamydia abortus is the cause of enzootic abortion in sheep (BOA). This pathogen with zoonotic potential is recognized as a major cause of reproductive failure in small ruminants and other domestic animals [1-2].

The disease has a significant economic impact as it is the most important cause of loss of offspring in sheep in parts of Europe, North America, Africa, Europe and other parts of the world. Chlamydia abortus poses a potential risk to pregnant women, as several cases of abortions have been reported following contact with lambing sheep or goats infected with Chlamydia abortus [3-5].

The prior art known strain of Chlamydia psittaci used for the manufacture of vaccines against chlamydia cats [6]. The disadvantage of this strain is that the live vaccine made from it has residual reactogenicity, and as a result, not very high immunogenicity.

The Chlamydia psittaci K-1 DEP strain is known in the Russian Federation for the manufacture of a vaccine against chlamydia in cats and fur animals [7].

Another strain of Chlamydia abortis bovis is known from the prior art, using which an associated vaccine was made for the prevention of respiratory diseases in cattle, containing viruses of infectious rhinotracheitis and diarrhea, chlamydia antigen from the strain of Chlamydia abortis bovis "K-8 VIEV" in titer 10 ^{6, 5} - ^7.5 ELD ₅₀ /ml, formalin and adjuvant based on mineral oil and lanolin [8].

An analogue of the claimed technical solution was chosen strain of Chlamydia abortis bovis.

The disadvantage of the selected analogue is its unsuitability for use in the manufacture of diagnostics and for the production of effective vaccines against reproductive pathologies and other reproductive disorders in sheep caused by Chlamydia abortis in the Russian Federation due to differences in the antigenic structure and immunobiological properties.

So far, isolated isolates of the epizootic strain of Chlamydia abortus, a homologous strain for the manufacture of diagnostic and vaccine preparations, are unknown.

Creation and production of a new virulent strain of Chlamydia abortus (chlamydia), isolated in the Russian Federation and having stable cultural properties and high biological, immunogenic antigenic activity and used for the industrial production of therapeutic and diagnostic drugs, which will improve strategies to combat this problem, including more effective diagnosis and prevention measures.

Disclosure of the essence of the invention

The technical task is to obtain a strain of Chlamydia abortus (chlamydia) with stable infectious, antigenic and immunogenic activity, which can be used for the manufacture of immunological therapeutic and prophylactic drugs, immunoglobulins and as an antigen for detecting antibodies to chlamydia in farm animals.

The technical result consists in obtaining a chlamydia strain with stable infectious, antigenic and immunogenic activity, which can be used for the manufacture of inactivated vaccines for the prevention of enzootic abortion in sheep (OEA); to obtain specific diagnostic sera, immunoglobulins, as an antigen for the detection of antibodies to chlamydia in farm animals.

Used terms and definitions.

Membrane filtration is a pressure separation process that uses membranes to separate components in a liquid solution or suspension based on their size and differences in charge. Ultrafiltration is a type of membrane filtration in which hydrostatic pressure forces liquid against a semi-permeable membrane. Solvent and low molecular weight solutes pass through the membrane and are called permeate, while suspended solids and high molecular weight solutes are retained (retentate) [9-10].

Tangential Filtration - Fluid is pumped tangentially along the membrane surface. The applied pressure serves to force some of the liquid through the membrane to the filtrate side. In tangential filtration, the retained components do not accumulate on the membrane surface, instead they are carried away by the tangential flow.

The main applications of tangential filtration are concentration, diafiltration (desalting and buffer replacement) and fractionation of large biomolecules from small ones. Cross-flow filtration units are also used to clarify and remove cells or cell debris from a cell culture or fermentation medium.

Tangential ultrafiltration is described in [11].

Diafiltration is the filtration of a product by membrane filter media with the supply of a wash liquid to the product, as a result of which the concentration of the product components that have passed through the filters decreases, i.e. these substances are washed out without the inevitable increase in the concentration of the components of the product that did not pass through the filters or its concentration. As a wash liquid, wash liquids foreign to the product, separately supplied water or a solvent, the product's own permeate taken, for example, from a connected diafiltration stage, or a mixture of both [12-13] are used.

The Chlamydia abortus strain "Chlamydia VNITIBP-21" was isolated in 2021 from the internal tissues of an aborted sheep fetus in a livestock farm in the village of Turma, Medvedevsky district of the Republic of Mari El, where cases of abortions, stillbirths and the appearance of non-viable fetuses in sheep were observed. The strain is adapted to propagation in chick embryos (EC) and continuous cell culture lines, has passed 25 passages during sequential cultivation on EC yolk membranes.

In 2021, the strain of Chlamydia abortus "Chlamydia VNITIBP-21" was deposited in the collection of microorganisms of the Federal State Budgetary Scientific Institution "Federal Scientific Center - All-Russian Research Institute of Experimental Veterinary Medicine named after K.I. Scriabin and Ya.R. Kovalenko (Federal State Budgetary Scientific Institution of the Federal Scientific Center of the VIEV RAS), registration number B-1371 (Certificate of deposit No. 29 dated November 26, 2021 Chlamydia abortus "VNITIBP-21").

The strain of Chlamydia abortus "Chlamydia VNITIBP-21" is characterized by the following features and properties.

Morphological features. Elementary bodies of a rounded shape with a diameter of 0.2-0.3 microns, reticular - 0.5-0.7 microns, stained according to Stemp, Machiavello and Romanovsky-Giemsa. Stemp stain gives elementary bodies of chlamydia a red-violet color against a blue background of the preparation; according to Machiavello, chlamydia stain bright red, cells of pathological material - light blue, cell nuclei - dark blue; according to Romanovsky-Giemsa, elementary inclusions of chlamydia acquired a red-violet color, and the reticular bodies of the affected cell turned blue, the cell nuclei turned burgundy-violet and their cytoplasm turned blue.

The strain of Chlamydia abortus "Chlamydia VNITIBP-21" is a gram-negative bacteria, spherical in shape, does not form spores, no flagella and no capsule. The cell wall consists of a 2-layer membrane. They have glycolipids, the main method of staining according to Romanovsky-Giemsa. Bacteria have 2 forms of existence: elementary bodies (inactive infectious particles, outside the cell); reticular bodies (inside cells, vegetative form).

biological properties. The strain of Chlamydia abortus "Chlamydia VNITIBP-21" multiplies well in the yolk membranes of 6-7-day-old chicken embryos or in continuous McCoy cell culture. The titer of chlamydia when cultivated on these biological substrates at a temperature of 36-37°C for 40-48 hours reaches a maximum of 6.0±0.01 lg TCID ₅₀ /ml. During reproduction in a transplanted cell culture, it accumulates at the 25th passage at the level of 5.5±0.5 lg TCID ₅ o/ml. In McCoy cell culture, the strain does not cause a cytopathic effect. The strain is passaged in the body of white mice with intranasal, intraperitoneal and intracerebral routes of infection.

To the claimed strain of Chlamydia abortus "Chlamydia VNITIBP-21" susceptible animals (naturally susceptible and laboratory) are sheep, goats, rabbits, cows, pigs, horses.

antigenic properties. When administered intranasally in the body of rabbits or mice, the strain of Chlamydia abortus "Chlamydia VN1TIBP-21" induces the formation of antibodies in a titer from 1:256 to 1:512 in the complement binding reaction (hereinafter referred to as RSK). There are species-specific and group-specific antigens common to all chlamydia. When the pathogen is stained according to Machiavello and Romanovsky-Giemsa, the morphology is typical for Chlamydia abortus. In RSK with specific diagnostic sera of sheep abortion at a dilution of 1:40, the activity (titer) of the antigen is at least 1:64; in a serum dilution of 1:80, the activity (titer) of the antigen is at least 1:128.

The claimed strain retains stable infectious and antigenic activity. It is stable in antigenic and genetic characteristics when passaged on McCoy cell culture for 20 passages (observation period), in yolk sacs for 6-7 days of developing EC for 15 passages (observation period).

The immunogenicity of the claimed strain was tested on white mice (non-linear laboratory mice ICR (CD-1)). Accommodation, maintenance and care of laboratory rodents used for scientific purposes were carried out in accordance with GOST 33216-2014 “Guidelines for the maintenance and care of laboratory animals. Rules for keeping and caring for laboratory rodents and rabbits” [14].

Young animals, healthy and equal in weight and age, were divided into nine groups of 7-10 mice in each, where from the first to the third groups, the inactivated Chlamydia abortus "Chlamydia VNITIBP-21" antigen with a protein content of 20-40 was administered twice with an interval of 3 weeks. mg/ml, other groups (from the fourth to the sixth) were injected twice with an interval of 3 weeks, Chlamydia abortis antigen "Chlamydia VNITIBP-21" with a protein content of 40-60 mg/ml, the following groups (from the seventh to the ninth) twice with an interval of 3 weeks introduced Chlamydia abortus "Chlamydia VNITIBP-21" with a protein content of 60-80 mg/ml. Were formed three control groups of 7-10 from non-immunized animals.

Resistance index>2.5. The antigen produced from the Chlamydia abortus strain "Chlamydia VNITIBP-21" in the inactivated adsorbed emulsified vaccine ensures the formation of intense and long-term immunity in vaccinated animals.

In experimental experiments on white mice, it was found that double immunization with a vaccine based on the Chlamydia abortus strain "Chlamydia VNITIBP-21" protected white mice from the development of chlamydia infection after artificial (experimental) infection by 93-95% with the known strain of Chlamydia abortis bovis "K-8 VIEV" and 90-93% from the strain of Chlamydia psittaci "K-8-K VIEV".

pathogenic properties. The Chlamydia abortus strain "Chlamydia VNITIBP-21" is pathogenic for white mice, chicken embryos and sheep.

After infection with the culture of the strain in sick animals, the phenomena of anorexia, rhinitis, conjunctivitis, paralysis of the limbs, as well as abortions, the birth of dead babies are noted. The complete death of white mice (100%) is set for 6-10 days.

7

The strain of Chlamydia abortus "Chlamydia VNITIBP-21" is resistant to streptomycin; sensitive to the action of tetracycline antibiotics, to a lesser extent penicillin (see table 2).

The strain of Chlamydia abortus "Chlamydia VNITIBP-21" retains a stable infectious and antigenic activity for 25 passages.

The strain of Chlamydia abortus "Chlamydia VNITIBP-21" retains its biological properties for one year at a temperature of minus 70°C, in lyophilized form - for at least 3 years.

The strain of Chlamydia abortus "Chlamydia VNITIBP-21" is free from bacterial, fungal, microplasma and viral contaminants.

The strain of Chlamydia abortus "Chlamydia VNITIBP-21" is used for the manufacture (production) of immunological drugs: vaccines, diagnostic sera.

In vaccine compositions based on the strain Chlamydia abortus ((Chlamydia VNITIBP-21), oil adjuvant compositions of the water-in-oil or water-oil-water or oil-in-water type can be used [15-16].

The possibility of carrying out the claimed invention is disclosed in the following examples.

Example 1. Cultivation and isolation of the strain Chlamydia abortus ((Chlamydia VNITIBP-21" using chicken embryos.

The starting material for strain isolation was a 20-25% suspension of ground tissues of an aborted sheep fetus in 0.01-0.02 M phosphate buffer solution with a pH of 7.2-7.4.

The suspension of pounded tissues is obtained by rubbing the internal organs of an aborted sheep fetus to a homogeneous state using any materials, instruments and/or devices known to a person skilled in the art.

To isolate the Chlamydia abortus strain ((Chlamydia VNITIBP-21), chicken embryos of 6-8 days of age were used, the membranes of their yolk sacs were inoculated with a tissue suspension (0.2 ml) of the internal organs of aborted fetuses according to the procedure known to a specialist from the prior art. Chlamydia isolates propagated in the epithelial cells of the yolk membranes of chicken embryos, causing their death on the 5-10th day after infection, reaching an infectious titer of up to 10 ⁶⁵ ELD ₅₀ /0.3 ml Purification and concentration of the chlamydia pathogen was carried out by tangential ultrafiltration.Concentrated antigen Chlamydia abortus "Chlamydia VNITIBP-21" has a compliment binding titer of 1:128-1:256. Further, inactivated by boiling for 15-20 minutes, the concentrated antigen extraction material is treated with ether-acetone extraction known to a person skilled in the art.

Several isolates of Chlamydia Abortus (chlamydia) were isolated from the internal organs of an aborted sheep fetus, one of the most productive, immunogenic and antigenic was given the name-strain of Chlamydia Abortus "Chlamydia VNITIBP-21".

Example 2. Determination of sensitivity to antibiotics of the Chlamydia abortus strain "Chlamydia VNITIBP-21".

Used a known method for determining sensitivity to antibiotics-diffusion method using standard paper discs with antibiotics.

The claimed strain of Chlamydia Abortus "Chlamydia VN1T1BP-21" is the most resistant to the action of streptomycin; most sensitive to the action of antibiotics of the tetracycline series, to a lesser extent to penicillin from the antibiotics of the penicillin series.

Example 3. Confirmation of the immunogenic properties of the Chlamydia abortus strain "Chlamydia VNITIBP-21"

To confirm the immunogenic properties of the Chlamydia abortus strain "Chlamydia VNITIBP-21" was compared with the strains of Chlamydia abortis bovis "K-8 VIEV" and Chlamydia psittaci "K-8-K VIEV".

Confirmation of the immunogenic properties of the claimed strain was carried out on white non-linear laboratory mice by intraperitoneal double injection of the inactivated antigen at a dose of 0.4-0.5 ml. Healthy young laboratory mice of the same weight and age were selected (accommodation, maintenance and care according to GOST 33216-2014). The selected mice were divided into nine groups of at least 7-10, where the first three groups of mice were injected with Chlamydia abortus antigen ((Chlamydia VNITIBP-21" with a protein content of 20-40 mg / ml), the next three groups of mice were injected with Chlamydia abortis antigen "Chlamydia VNITIBP- 21" with a protein content of 40-60 mg/ml, the following three groups of mice were injected with Chlamydia abortus "Chlamydia VNITIBP-21" with a protein content of 60-80 mg/ml. Three control groups (same in age and weight) were formed by 7- 10 of non-immunized animals After 21-28 days after the last immunization, experimental and control mice are infected intranasally with a 10% suspension of the yolk membranes of chicken embryos infected with a strain of chlamydia.

One control group A, experimental groups of mice 1st, 4th and 7th were infected with a strain of Chlamydia abortus "Chlamydia VNITIBP-21" with an infectious activity of 10 ⁵⁰ -10 ⁵⁵ ELD ₅₀ /0.3 ml at a dose of 0.05 ml . Control group B, experimental groups of mice 2nd, 5th and 8th were infected with Chlamydia abortis bovis "K-8 VIEV" with infectious activity 10 ^5.0 -10 ^5.5 ELD ₅₀ /0.3 ml at a dose of 0 .05 ml. Control group C, experimental groups of mice 3rd, 6th and 9th were infected with Chlamydia psittaci "K-8-K VIEV" with infectious activity 10 ^5.0 -10 ⁵ , ⁵ ELD ₅₀ /0.3 ml per dose 0.05 ml. In control groups A, B and C, unvaccinated mice showed 100% death after infection. The results of this experiment are presented in table 3.

Conclusion. As a result of the studies, it was proved that a double intraperitoneal injection of inactivated chlamydia strain Chlamydia abortus "Chlamydia VNITIBP-21" protects 98.5-99.5%) of white mice from intranasal infection with this strain, 93-95%) from infection with a known strain Chlamydia abortis bovis "K-8 VIEV" and 90-93% of infection with a strain of Chlamydia psittaci "K-8-K VIEV" (see table 3).

Example 4. Confirmation of the antigenic properties of the Chlamydia abortus strain "Chlamydia VNITIBP-21".

The antigenic activity of the strain Chlamydia abortus "Chlamydia VNITIBP-21" is confirmed in the complement fixation reaction by comparing the antibody titers in hyperimmune sera of guinea pigs obtained against chlamydia strains: Chlamydia abortus "Chlamydia VNITIBP-21", Chlamydia abortis bovis "K-8 VIEV", Chlamydia psittaci "K-8-K VIEV" (double immunization with an interval of 3 weeks at a dose of 0.9-1.0 ml), as well as antigens from these strains. Characteristics of the antigenicity of the specified strain in comparison with other known strains are presented in table 4.

These data confirm that the antigenic activity in the study with homologous serum of the Chlamydia abortus strain "Chlamydia VNITIBP-21" is two times higher than the known strain of Chlamydia abortis bovis "K-8 VIEV" and the strain of Chlamydia psittaci "K-8-K VIEV".

Example 5. Obtaining a vaccine from an inactivated cultural raw material from the Chlamydia abortus strain "Chlamydia VNITIBP-21" using the roller method of cultivation.

The inactivated emulsion vaccine against chlamydia is prepared from the mat brood of the Chlamydia abortus strain "Chlamydia VNITIBP-21" (see example 1), which is used to infect a transplanted McCoy cell culture of 2-3 days of age with a concentration of more than 100,000 cells/ml and obtain seed brood . Grown chlamydia meet the following requirements: the infectivity titer after reproduction in a monolayer of McCoy cells is not less than 5.0 lg TCID50/ml, pH 7.2-7.4 and in the absence of contamination.

A monolayer of McCoy cells is dissociated by trypsinization (mixture of trypsin and versene) from confluent cultures and then transferred to DMEM medium containing 5% pestivirus-free fetal calf serum (DMEM medium), with a final concentration of about 400,000 cells per 1 ml . Fractions of this suspension are mixed in 100 ml aliquots with 20 ml aliquots of fractions of the mat brood of the strain Chlamydia abortus "Chlamydia VNITIBP-21" and the final mixture, after preliminary holding for 1-2 hours at a temperature of 37°C for the adsorption of chlamydia, is transferred to roller vessels. Add nutrient medium DMEM with 5% fetal serum and antibiotic (gentamicin at a concentration of 50 μg/ml or its analogues) and then incubated at 37°C for 76-92 hours in an atmosphere of CO ₂ .

After this last incubation, the roller vessels are subjected to three consecutive freeze/thaw cycles. The resulting material is freed from cellular debris by centrifugation at 1000 rpm for 20 minutes. The suspension, cleared of detritus, should have the appearance of a clear liquid of a pinkish-cherry color. The resulting material is monitored for infectious activity.

The purified suspension is subjected to inactivation. Inactivation is carried out with formalin in the amount of 0.14-0.16% and for 120-122 hours at a temperature of 37°C.

The emulsion vaccine is prepared by mixing the cultured inactivated raw material - Chlamydia abortus strain "Chlamydia VNITIBP-21" and an oil adjuvant in the form of a water-oil-water emulsion.

The oil adjuvant-the rest is added to the inactivated raw material thus obtained, and thoroughly mixed, and the vaccine is obtained in the following ratio of components, wt %:

Cultural inactivated mixture, including the strain of chlamydia Chlamydia abortus 50.00 "Chlamydia VNITIBP-21" (antigen)

Oil adjuvant rest.

The vaccine thus prepared is a pale pink emulsion.

The resulting vaccine is packaged under sterile conditions in glass vials of 2.0, sealed, and controlled in accordance with the requirements for sterility, harmlessness and antigenic activity. Vials containing the resulting vaccine are packed into kits, which include instructions for use.

The vaccine is checked for sterility by inoculation on the following nutrient media: MPB, MPA, MPBB, Sabouraud medium and thioglycol medium. In all cases, the growth of microflora is not observed.

The vaccine retains its biological properties for 18 months if stored in a dry, dark room at a temperature of 4-8°C.

Example 6. Confirmation of the safety of the claimed vaccine.

The claimed vaccine is administered intramuscularly in a double dose of the optimal dose of five guinea pigs selected in accordance with GOST 33216-2014. Within 14 days, the change in the behavior of the animals was observed.

Conclusion. All animals are alive and healthy, the behavior corresponds to the physiological norm.

Example 7. Determination of the antigenic activity of the vaccine.

To assess the antigenic activity, the vaccine is administered seronegative to chlamydial infection to four sheep of 3-5 months of age of the Romanov breed in compliance with the rules of asepsis and antisepsis intramuscularly in a volume of 2.0 ml. Before immunization and 21 days after vaccination, blood is taken from animals and serum is examined for the presence of specific antibodies in ELISA using the diagnostic kit "Indirect enzyme immunoassay for the detection of antibodies directed against Chlamydophila abortus in ruminants, pigs and horses" ("GO Screen® Chlamydophila abortus Indirect Multi-species") manufactured by ID.Vet., France, according to the manufacturer's instructions. Accounting for the results of the presence of antibodies to chlamydia in the laboratory is carried out by assessing the percentage of S / P (test sample / positive ratio), using a wavelength of 405 nm in spectrophotometry. S/P is calculated according to the formula specified in the manufacturer's instructions. The result of the analysis of samples with a percentage of S/P:

- less than or equal to 50% is considered negative;

- between 50% and 60% is considered uncertain;

- greater than or equal to 60% is considered positive.

The results of the studies indicate that before immunization, no antibodies to chlamydia were detected in the blood sera of sheep, and 21 days after vaccination, specific antibodies were detected in all 4 samples - the S / P ratio significantly exceeded 60% (from 78 to 84).

Thus, the resulting inactivated emulsion vaccine from the Chlamydia abortus strain "Chlamydia VNITIBP-21" is a sterile, harmless preparation and has sufficient antigenic activity. A single injection of the vaccine provides a high level of antibodies in the blood serum of vaccinated animals.

Example 8. Determination of the immunogenic activity of the vaccine in laboratory mice.

Form 2 groups of white mice with a live weight of 18-20 g, 20 animals each. White mice of the 1st group are vaccinated subcutaneously at a dose of 0.2 ml with an inactivated vaccine against chlamydia from the Chlamydia abortus strain "Chlamydia VNITIBP-21", prepared according to example 5.

White mice of the 2nd group are not vaccinated (control). On the 21st day after vaccination, laboratory animals of groups 1 and 2 are infected subcutaneously at a dose of 0.3 ml with a culture of a titrated control strain of Chlamydia abortus with a titer of 10 ⁶ ' ⁵ EID50 / 0.3 ml. Do not establish the death of white mice in the 1st experimental group and establish the complete death of white mice (100%) in the 2nd control group.

Example 9 Determination of the immunogenic activity of a vaccine in target animals.

The high immunogenic activity of the proposed vaccine was also confirmed in an acute experiment on susceptible animals, pregnant ewes. In the experiment, 60 pregnant ewes were selected from a farm safe for salmonellosis, chlamydia, campylobacteriosis and leptospirosis. The chlamydia vaccine prevented 100% of ewes from aborting after being infected with a field isolate. In non-vaccinated groups (7 animals), 5 out of 7 animals aborted from chlamydia, 2 non-viable offspring were obtained from 2 animals.

Thus, vaccination of ewes with a chlamydia vaccine provides 100% immunological protection against these diseases.

Example 10. Determination of the immunogenic activity of the vaccine in the last month of the expiration date (month 18).

Form 2 groups of white mice with a live weight of 18-20 g, 20 animals each. White mice of the 1st group are vaccinated subcutaneously at a dose of 0.2 ml of the declared inactivated vaccine after its storage, in the last month of the vaccine expiration date (see example 5).

White mice of the 2nd group are not vaccinated (control). On the 21st day after vaccination, laboratory animals of groups 1 and 2 are infected subcutaneously at a dose of 0.3 ml with a culture of a titrated control strain of Chlamydia abortus with a titer of 10 ⁶ ' ⁵ EID ₅₀ / 0.3 ml. Do not establish the death of white mice in the 1st experimental group and establish the complete death of white mice (100%) in the 2nd control group.

Conclusion. The vaccine retains immunogenic activity at 18 months of storage.

All the above examples confirm the implementation of the technical solution.

The presented examples do not limit the scope of the claims of the present invention and serve only for the purpose of illustrating and disclosing the claimed technical solution.

Industrial Applicability

The set technical task, namely, obtaining a strain of Chlamydia abortus (chlamydia), which has a stable infectious, antigenic and immunogenic activity, which can be used for the manufacture of immunological immunoglobulin preparations and as an antigen for the detection of antibodies to chlamydia in farm animals, has been achieved, which is confirmed the examples given.

The practical application of the claimed technical solution makes it possible to use it for the manufacture of inactivated vaccines and the production of specific diagnostic sera and globulins and as an antigen for the detection of antibodies to chlamydia in farm animals.

List of literary sources

1. Halbert G. Diseases of sheep. Can Vet J. 2008;49(7):702;

2. Longbottom, D.; Coulter, L.J. Animal chlamydioses and zoonotic implications. J. Comp. Pathol. 2003, 128, 217-244;

3. Rodolakis A., Mohamad K.Y. Zoonotic potential of Chlamydophila. Vet. microbiol. Jan 2010; 140(3-4):382-391;

4. Pospischil A., R. Thoma R., Hilbe M., Grest P., Gebbers F.-O. Abortion in woman caused by caprine Chlamydophila abortus (Chlamydia psittaci serovar 1), 2002. Swiss Medical Weekly, vol. 132, no. 5-6, pp.64-66;

5. Walder G., Hotzel H., Brezinka C. et al., "An unusual cause of sepsis during pregnancy: recognizing infection with Chlamydophila abortus," Obstetrics & Gynecology, 2005. vol. 106, no. 5, pp. 1215-1217;

6Mitzel Jr. et al. Vaccination Against Feline Pneumonitis, Am. J. Vet. Res., 1977, 38(9), pp.1361-1364;

7. Patent 2122579 CHLAMYDIA PSITTACI strain used for the manufacture of an inactivated vaccine against chlamydia in cats and fur animals. Application No. 97104467. Application date: 1997.03.21. Published: 1998.11.27;

8. Auto. USSR N 1789219, class. A61K 39/118, publ. 1993;

9. Brock T. Membrane filtration. M.: Mir; 1987. 462 p.;

10. Anthony Bennett, "Metman technology: Developments in ultrafiltration technologies"// Filtration + Separation V. 49, Issue 6, November-December 2012, Pages 28-33, doi.org/10.1016/S0015-1882(12)70287- 2

11. Shiloach J, Martin N, Moes H. Tangential flow filtration. Adv Biotechnol Process (1988), 8: 97-125; Van Reis R, Zydney A. Membrane separations in biotechnology. Curr Opin Biotechnol (2001), 12:208-211);

12. Madsen R.F., Design of sanitary and sterile UF- and diafiltration plants, Separation and Purification Technology, 22-23 (2001) p.79-87;

13. Patent No. 2338433 of the Russian Federation Method for product diafiltration and device for implementing the method. Published: 20.11.2008 Bull. No. 32);

14. GOST 33216-2014 Guidelines for the maintenance and care of laboratory animals. Rules for keeping and caring for laboratory rodents and rabbits;

15. Garcon N., Leroux Roels G., Cheng W.F. (2011). Vaccine adjuvants. Perspectives in Vaccinology. 1, 89-113;

16. Edwige Boogie On, Elodie Gonsalves, Alexander Shevtsov, Polina Mezonnas, Stepan Remyga, Oleg Goryushev, Sebastian Deville, Nicolas Bertaud and Juliette Ben ArusA New Adjuvant Combined with Inactivated Influenza Enhances Specific CD8 T Cell Response in Mice and Decreases Symptoms in Swine Upon challenge. Article in Viral Immunology(2015) 28(9):524-31. doi: 10.1089/vim.2014.0149;

17. Patent 2085213 Vaccine for the prevention and treatment of chlamydia in farm animals and fur animals. Application No. 95 95108959. Application date: 1995.06.05. Published: 1997.07.27.

Claims (4)

1. Vaccine composition for the prevention of enzootic abortion in sheep, containing a cultural inactivated mixture, including the antigen of the Chlamydia abortus strain "Chlamydia VNITIBP-21" and an oil adjuvant at a ratio of components, wt.%: culture inactivated mixture, including the antigen of a strain of chlamydia Chlamydia abortus "Chlamydia VNITIBP-21" 50.00 oil adjuvant rest 2. A set for the prevention of enzootic abortion in sheep, including a sealed vial containing the vaccine composition according to claim 1, instructions for use. 3. The kit according to claim 2, further characterized in that the vaccine composition according to claim 1 is sterile.