RU2470663C1 - Swine salmonellosis vaccine, method for preparing, method for preventing swine salmonellosis - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to veterinary microbiology and concerns a swine salmonellosis vaccine, a method for preparing it and a method for preventing swine salmonellosis. The described method for preparing involves separate culture of the strains S.choleraesuis No. 370 and S.typhimurium No. 415 in a biogenerator on Hottinger's broth pH 7.2-7.5 and amine nitrogen 100-120 mg % at air supply 1 l/min per 1 l of a nutrient medium and agitation rate 100 rpm for 10 hours, biomass lysis with hydroxylamine, deposition of the dissolved antigens, setting out, centrifugation, removal of a supernatant, mixing up of the lysates 1:1, addition of edible pectin, lyophilisation to produce a finished vaccine. The dry antigen lysate 1 mg contains 2.5-3.5×10 microbial cells. A method for preventing swine salmonellosis consists in the oral introduction of the vaccine into piglets from 30th-35th to 45th-54th day of life in a dose of 400-600 bln microbial cells of the corpuscular antigen daily for 10 days with the veterinary probiotic Lactobifadol 10-12 g head/day.

EFFECT: group of inventions enables preserving seed stock, producing healthy crop; it may be used in salmonellosis unfavourable pig farms.

3 cl, 6 tbl, 5 ex

Description

The invention relates to the field of veterinary microbiology, in particular to obtaining a vaccine against pig salmonellosis, a method for its manufacture and prevention of salmonellosis.

Salmonellosis of productive mammals of animals and birds is not only a significant problem of veterinary medicine from an economic point of view, but also an important social problem, since it poses a threat and often causes massive outbreaks of foodborne toxicoinfections of salmonella etiology in humans. Despite the considerable amount of work devoted to the control and prevention of this disease, it is still widespread in livestock farms and causes significant economic damage to the industry. According to recent veterinary reporting data (2003-2009, FGU Center for Veterinary Medicine), salmonellosis in the Russian Federation is invariably one of the top ten in the structure of infectious diseases of cattle, small cattle, pigs and poultry. During this period, the proportion of salmonellosis in the structure of infectious diseases of cattle was 2.3-3.8%, small cattle - 2.5-9.9%, pigs - 3.4-14.1%, poultry (salmonellosis + typhoid pullorosis) - 1.4-2.02%.

The most important element of a comprehensive system for combating salmonella infection is vaccination.

Currently, a method of vaccination against salmonellosis of pigs is known, comprising administering a dry complex lysate antigen of the broth culture of S. choleraesuis No. 370 and S. typhimurium No. 415 strains, with 1 mg of dry lysate antigen corresponding to 2.5-3.5 × 10 ⁹ microbial cells of the corpuscular antigen, taken in equal proportions.

For prophylaxis, antigens are administered orally to piglets orally in a mixture with mixed feed for 10 days daily in doses equivalent to 200.400 and 600 billion microbial cells, respectively, and the optimal dose was 400-600 billion microbial cells per head.

Oral immunization of the test antigen was performed against the background of the probiotic drug Lactobifadol (1).

Also known is the lysate antigen (vaccine) from the broth culture of strains S. choleraesuis No. 370 and S. typhimurium No. 415, taken in equal proportions destroyed by hydroxylamine.

As the immunizing principle of the lysate antigen, in accordance with the strains used, are O-antigenic complexes 6 and 7, H-antigenic complexes of the 1st phase C and 2nd phase 1.5; and O-antigenic complexes 1, 4, 12, H-antigenic complexes of the 1st phase I and 2nd phase 1, 2, respectively. 1 mg of dry hydroxylamine antigen contains components of Salmonella cells in an amount equivalent to 2.5-3.5 × 109 m.k. (2). Prototype.

The objective of our research was to obtain a vaccine against salmonellosis of pigs, which ensures the creation of long-lasting and intense immunity in the body when administered orally.

We managed to achieve the set goal due to the fact that:

- as antigens contains strains of S. choleraesuis No. 370 and S. typhimurium No. 415, taken in equal proportions, while 1 mg of dry lysate antigen corresponds to 2.5-3.5 × 10 ⁹ microbial cells.

- Salmonella biomass of strains S. choleraesuis No. 370 and S. typhimurium No. 415 are separately cultured in a bioreactor on Hottinger broth pH 7.2-7.5 and an amine nitrogen index of 100-120 mg% with an air supply of 1 l / min per 1 liter of nutrient medium and stirring speed of 100 rpm for 10 hours, add hydroxylamine (NaH ₂ OH · HCl) at the rate of 0.065 mg x / h per 1 billion microbial cells with constant stirring, incubated for 4 days at a temperature of 38 ... 39 ° C, adjust the pH of the lysate to 7.4 with 10% NaOH, mix the lysates 1: 1, then the dissolved antigens are precipitated by sequential addition leniem Na ₂ HPO ₄ and CaCl ₂ - 1 mL lysate loss 4mg CaHPO _4, settled lysate with precipitate during the day, then centrifuged 2500-3000 rev / min for 20 minutes, the supernatant was removed and added pectin based food 1% of the resulting lysate, freeze-dried and receive a vaccine.

- a method for the prevention of salmonellosis in pigs is the oral administration of the vaccine to pigs aged 30-35 to 45-54 days of life at a dose equivalent to 400-600 billion microbial cells of the corpuscular antigen daily for 10 days with a veterinary probiotic Lactobifadol at a dose of 10-12 g per head per day.

The essence of the proposal is as follows:

The salmonellosis vaccine is:

antigen from a mixture of broth cultures of strains S. choleraesuis No. 370 and S. typhimurium No. 415, taken in equal proportions and destroyed by hydroxylamine, with 1 mg of dry lysate antigen corresponding to 2.5-3.5 × 10 ⁹ microbial cells.

The manufacturing method is as follows:

- conduct separate cultivation of strains of S. choleraesuis No. 370 and S. typhimurium No. 415 in a bioreactor on Hottinger broth pH 7.2-7.5 and an amine nitrogen index of 100-120 mg% with an air supply of 1 l / min per 1 liter of nutrient medium and a stirring speed of 100 rpm for 10 hours;

- the dosage of biomass is determined using the optical turbidity standard of Freight One named after Tarasevich (sowing dose was 100 million m.k. / cm ³ nutrient medium);

- then add hydroxylamine (NaH ₂ OH · HCl) at the rate of 0.065 mg x / h per 1 billion microbial cells with constant stirring;

- incubated for 4 days at a temperature of 38 ... 39 ° C;

- adjust the pH of the lysate to 7.4 with 10% NaOH, mix the lysates in a ratio of 1: 1;

- then the precipitation of the dissolved antigens is carried out by the sequential addition of Na ₂ HPO ₄ and CaCl ₂ (per 1 ml of lysate precipitation of 4 mg CaHPO ₄ );

- defend the lysate with sediment during the day, and then centrifuged (2500-3000 rpm for 20 minutes);

- remove the supernatant and add food pectin at the rate of 1% to the resulting lysate;

- freeze-dried and get the finished vaccine;

- after they carry out control on sterility and harmlessness.

The resulting vaccine is used prophylactically on piglets at a dose equivalent to 400-600 billion microbial cells of the corpuscular antigen daily for 10 days with the probiotic Lactobifadol at a dose of 10-12 g per head / day.

The proposed vaccine is harmless and provides the creation in the body of an animal of intense immunity lasting up to 6 months. The most optimal for immunization of piglets is the age from 30-35 to 45-54 days of life.

Example 1

The reactogenicity of salmonella lysate antigens with various methods of their administration was tested on outbred mice with a live weight of 16-18 g and the LD50 value was determined by intraperitoneal, subcutaneous and oral administration of the above antigens. Of 360 mice, 36 groups of 10 animals each were formed. Corpuscular antigens were administered to each group in doses of 2.5 × 10 ⁹ , 5 × 10 ⁹ , 10 × 10 ⁹ , 20 × 10 ⁹ , 40 × 10 ⁹ and 80 × 10 ⁹ microbial cells per head, and lysed antigens - in doses equivalent to these values in a volume of 0.5 cm ³ .

Table 1 Comparative reactogenicity of salmonella corpuscular and lysate antigens with different methods of their administration to white mice Route of administration Salmonella strains Types of Antigen Magnitude LD ₅₀ , mic. class intraperitoneally S.choleraesuis No. 370 corpuscular 6.58 × 10 ⁹ lysed 23.72 × 10 ⁹ S.typhimurium No. 415 corpuscular 6.18 × 10 ⁹ lysed 25.11 × 10 ⁹ subcutaneously S.choleraesuis No. 370 corpuscular 11.61 × 10 ⁹ lysed 60.99 × 10 ⁹ S.typhimurium No. 415 corpuscular 17.70 × 10 ⁹ lysed 79.90 × 10 ⁹ oral S.choleraesuis No. 370 corpuscular 24.07 × 10 ⁹ lysed 128.46 × 10 ⁹ S.typhimurium No. 415 corpuscular 19.92 × 10 ⁹ lysed 121.99 × 10 ⁹

So the reactivity of S. choleraesuis No. 370 lysate antigens, expressed in LD ₅₀ values, was 3.6 times less than the reactivity of corpuscular antigens when administered intraperitoneally, by 5.3 times (p≤0.05) for subcutaneous and oral administered. Similar data were obtained for S. typhimurium antigens No. 415. Lysate antigens were less reactogenic compared with corpuscular antigens 4.1 times (p≤0.05) with intraperitoneal administration, 4.5 times (p≤0.05) with subcutaneous and 6.1 times (p≤≤ 0.05) when administered orally.

Thus, it was found that salmonella antigens obtained by lysis of bacterial cells of S. choleraesuis No. 370 and S.typhimurium No. 415 strains with hydroxylamine are less reactogenic both in the intraperitoneal and in the subcutaneous and oral methods of their application to white outbred mice as compared to corpuscular ( whole cell) antigens. This provides the opportunity to use them to immunize animals at higher doses than corpuscular antigens, without the risk of side effects.

Example 2

In the experiment, lysate antigens from Salmonella serovar S.typhimurium strain 415 were used. White outbred mice with a live weight of 16-18 g were taken for immunization, which were previously checked for the absence of Salmonella corresponding serovar in their intestines, and in the feces and blood - specific antibodies to them (in RNAT and RIGA with erythrocyte diagnosticum). 6 groups of 20 animals each were formed from mice. Animal groups No. 1, 2, and 3 were immunized for 5 consecutive days with a daily dose of antigen equivalent to 20 billion microbial cells without a protector, with 1% pectin and sorbed on bran, respectively. Pectin and bran were used as protectors to prevent the destruction of antigens in the acidic environment of the stomach of animals. Mice of groups 4, 5, and 6 received similar antigens, but for 10 consecutive days and at a daily dose equivalent to 10 billion microbial cells. Thus, the total dose of antigens obtained by mice in all groups was equal and amounted to 100 billion microns. class on the head. Antigen without protector and with 1% pectin was introduced through a probe, and sorbed on bran was mixed into dry food.

On the 5th, 10th, 20th, 25th and 30th day after the last injection of drugs from each group, 3 mice were killed. The contents of the large intestine and blood serum from them were examined in RIGA with a erythrocyte diagnosticum for the presence of coproantibodies and humoral antibodies to S. typhimurium. Antibody titers were expressed as natural logarithms with base 2.

Table 2 presents the average titers of humoral antibodies detected in the blood serum of mice.

table 2 The dynamics of humoral antibodies to S. typhimurium in the blood serum of vaccinated mice Types of Antigens Immunization schedule The duration of the study (day after immunization) and the average titers n = 3) of humoral antibodies (log ₂ ) 5 10 fifteen twenty 25 thirty No protectors 5 days 2 U 2 one one one 10 days four 2 3 3 2 one With 1% pectin 5 days 1.7 2 1.7 one one one 10 days 3 3 2.7 2 2 1.3 Bran sorbed 5 days 2.7 2 3 four 2.7 2 10 days 2.7 2 3 3.3 3 3.3 The control - - - - - -

The data obtained by studying the titers of antibodies to S. typhimurium in the feces of mice are presented in table 3.

Table 3 Dynamics of coproantibodies to S. typhimurium in oral vaccinated mice Types of Antigens Immunization schedule The duration of the study (day after immunization) and the average titers (n = 3) of secretory antibodies (log ₂ ) 5 10 fifteen twenty 25 thirty No protectors 5 days 5 10 eleven 8 7 5 10 days eleven 10.7 8 8 6 4.7 With 1% pectin 5 days 5 7 7 7.3 7 6 10 days 7.3 6.3 5.7 6 6.3 6 Bran sorbed 5 days four 7.3 8.3 6.3 6 5.7 10 days 4.3 8 6.7 6 5 4.7 The control - - - - - -

A comparison of the levels of humoral and coproantibodies suggests that coproantibodies are found in significantly higher titers. The peak level of coproantibodies (15-20 days of observation) was on average higher than the peak level of humoral antibodies by 4-5 dilutions or 16-32 times. By the end of the month of observation, this indicator was 3-4 dilutions or 8-16 times.

Thus, it was found that the scheme of oral immunization, providing for multiple (five- or 10-fold) administration of the antigen provides a sufficiently long contact with lymphoid tissue of the intestine. The antibody response in this case to antigens with and without protectors practically does not differ either in magnitude or duration in the tested time frame. Long periods of oral immunization are also more expedient from the point of view that group immunization under production conditions with this scheme will allow for a more accurate and uniform distribution of the vaccine preparation in feed, i.e. more accurate dosing, and therefore a more uniform immune background in the vaccinated livestock.

Example 3

The study of the immunogenicity of lysate antigens during oral immunization is shown in the direct infection of mice.

In order to study the immunogenicity of the test lysate antigens during oral immunization of animals, 15 groups of white outbred mice were formed with a live weight of 18-20 g. Animal groups No. 1-5 were immunized with a preparation consisting of mixed in equal amounts of lysed Salmonella cells of S. choleraesuis strains No. 370 and S.typhimurium No. 415 with the addition of a probiotic veterinary preparation Lactobifadol, group No. 6-10, mice immunized without a probiotic and group No. 11-15 an intact control. Immunization was carried out for 10 consecutive days in a daily dose equivalent to 10 billion microbial cells of Salmonella (the total dose of antigens is equivalent to 100 billion micro cells per head). Infection of mice was carried out intraperitoneally in doses multiple of 10 virulent S. choleraesuis strain No. 370 at 0.5 ml per head. Observation of animals was carried out for 10 days.

Table 4 Results of direct infection of orally vaccinated mice Infectious doses Immune animals Control (non-immune) With probiotic Without probiotic number of goals fell number of goals fell number of goals fell goals % goals % goals % 5 × 10 ¹ 8 0 0 9 0 0 8 3 37.5 5 × 10 ² 10 one 10 9 2 22.2 8 6 75 5 × l0 ³ 10 3 thirty 8 3 37.5 8 8 one hundred 5 × 10 ⁴ 10 6 60 10 6 60 8 8 one hundred 5 × 10 ⁵ 9 7 77.8 7 6 85.7 eleven eleven one hundred LD ₅₀ 1.8 × 10 ⁵ 1.1 × 10 ⁵ 2.6 × 10 ²

In the direct infection experiment, it was shown that the LD ₅₀ in immunized animals was significantly different from control non-immune mice. Thus, immunization for 10 consecutive days in a daily dose equivalent to 10 billion salmonella microbial cells provides 700 times increased resistance to infection with S. choleraesuis strain No. 370 in mice with a probiotic and 400 times without a probiotic compared with non-immunized animals . Therefore, immunization within 10 days causes the formation of specific immunity to salmonellosis.

Example 4

For experience in one of the salmonella-free farms, 7 groups of piglets of 8 goals each were selected, selected according to the principle of analogues at the age of 14-16 days. Pigs of groups 1, 3, and 5 received the test vaccine mixed with animal feed for 10 days daily in doses equivalent to 200, 400, and 600 billion microbial cells of the corpuscular antigen, respectively, from 45 to 54 days of life. Animal groups No. 2, 4 and 6 were immunized according to a similar scheme, but additionally from 15 to 45 days of life and during the entire period of immunization they received 10 g of probiotic Lactobifadol mixed with mixed feed per head / day. Pigs of group No. 7 served as an intact control.

On the 7th, 14th, 21st, 31st and 41st days after the termination of immunization, samples of feces and blood were taken from piglets. The contents of the large intestine and blood serum were examined in RIGA with erythrocyte diagnostics for the presence of humoral and coproantibodies to S. typhimurium and S. choleraesuis.

Table 5 The dynamics of humoral and coproantibodies in piglets Study time, days Doses of antigen, piglet groups and antibody titers (M ± m; log ₂ ) 200 billion 400 billion 600 billion The control 1 g 2 g 3 g 4 g 5 g 6 g 7 g Humoral antibodies to S.typhimurium 7 2.0 ± 0.9 1.8 ± 0.3 2.2 ± 0.4 2.6 ± 0.6 2.6 ± 0.3 2.8 ± 0.3 0 fourteen 3.3 ± 0.3 4.0 ± 0.06 3.6 ± 0.3 4,5 ± 0,03 4.0 ± 0.7 5.5 ± 0.5 0 21 2.5 ± 0.7 3.0 ± 0.05 3.3 ± 0.5 3.5 ± 0.2 3.6 ± 0.7 4.0 ± 0.05 0 31 2.0 ± 0.9 2.5 ± 0.3 1.8 ± 0.6 3.0 ± 0.2 2.3 ± 0.3 3.6 ± 0.1 0 41 0 2.0 ± 0.09 0 2.0 ± 0.06 0 2.4 ± 0.5 0 Humoral antibodies to S. choleraesuis 7 2.3 ± 0.3 2.3 ± 0.3 2.3 ± 0.3 2.6 ± 0.3 3.0 ± 0.6 3.3 ± 0.3 0 fourteen 3.3 ± 0.3 3.7 ± 0.3 4.0 ± 0.6 4.7 ± 0.3 4.0 ± 0.9 5.3 ± 0.3 0 21 3.3 ± 0.3 3.5 ± 0.2 3.5 ± 0.9 3.5 ± 0.3 3.5 ± 0.5 4,5 ± 0,2 1.0 ± 0.5 31 1.8 ± 0.6 2.0 ± 0.03 2.6 ± 0.7 2.6 ± 0.3 2.6 ± 1.0 3.0 ± 0.3 1.0 ± 0.3 41 0 2.0 ± 0.3 1.0 ± 0.6 2.0 ± 0.2 1.0 ± 0.7 2.8 ± 0.3 1.0 ± 0.3 Co-counterpart to S. typhimurium 7 5.6 ± 0.3 5.5 ± 0.7 4.3 ± 0.3 7.0 ± 0.6 5.6 ± 0.7 7.6 ± 0.3 0 fourteen 7.3 ± 0.7 8.3 ± 0.7 7.6 ± 0.3 8.6 ± 0.3 9.3 ± 0.3 10.6 ± 0.3 0 21 7.3 ± 0.9 8.5 ± 0.3 7.6 ± 1.0 9.0 ± 0.3 8.0 ± 0.3 9.5 ± 0.3 2.0 ± 0.3 31 5.0 ± 0.6 5.6 ± 0.3 5.3 ± 0.9 6.6 ± 0.3 6.6 ± 0.3 7.0 ± 0.6 2.0 ± 0.2 41 3.5 ± 1.5 4.0 ± 0.7 3.0 ± 0.6 4.0 ± 0.3 4.0 ± 0.6 5.0 ± 1.0 3.0 ± 0.3 Co-counterpart S.choleraesuis 7 5.0 ± 0.6 7.0 ± 1.0 5.3 ± 0.5 6.6 ± 0.3 7.0 ± 0.6 8.0 ± 0.3 0 fourteen 7.3 ± 0.6 8.3 ± 0.2 7.3 ± 0.9 9.5 ± 0.3 9.0 ± 0.6 10.3 ± 0.3 0 21 7.0 ± 1.0 7.5 ± 0.2 7.0 ± 0.5 9.3 ± 0.3 7.6 ± 0.3 10.0 ± 0.2 2.0 ± 0.5 31 4.6 ± 0.6 5.5 ± 0.3 5.0 ± 0.5 6.0 ± 1.0 6.0 ± 1.0 6.3 ± 0.3 2.0 ± 0.2 41 3.0 ± 0.9 4.0 ± 0.3 3.0 ± 1.0 3.0 ± 0.5 4.0 ± 0.7 5.0 ± 0.3 3.5 ± 0.5

Table 5 presents the average titers of humoral antibodies detected in the blood serum of piglets. From the data of the table it is seen that all three tested doses of antigen provided the synthesis of specific antibodies to salmonella of both serovars. Antibody titers reached their maximum by 14 days after the cessation of immunization, remained at that level for 5-7 days and then decreased. The intensity of the humoral immune response was dose-dependent in nature, and antibody titers increased with increasing dose of antigen.

The introduction of Lactobifadol into the treatment regimen of piglets provided a more intensive antibody genesis due to the immunomodulating effect of the probiotic. This effect was observed at all three doses tested. Smaller values of m values in groups 2, 4 and 6, which is especially pronounced at the peak of antibody titers, indicate a smaller scatter in the absolute values of antibody titers or a more uniform specific immune response in these piglet groups.

The results shown in table 5 on the dynamics of coproantibodies indicate that the nature and dynamics of the antibody response from the intestinal lymphoid tissue is generally similar to that in its evaluation by humoral antibodies. The titers of coproantibodies reached their peak values by 14-21 days after the last antigen delivery, after which their level began to gradually decrease. A comparison of the levels of humoral and coproantibodies indicates that coproantibodies were found in much larger quantities. The peak level of coproantibodies (14-21 days of observation) for S. typhimurium was on average 4-5 dilutions or 16-32 times higher than humoral ones, and for S. choleraesuis - 3-6 dilutions or 8-64 times. By the end of the observation period (41 days after immunization), humoral antibodies to S. typhimurium could not be detected, while their level to S. choleraesuis was 2-2.8 log ₂ . The level of coproantibodies by this term was 3-5 log ₂ for both serovariants of the pathogen.

Example 5

For the experiment in one of the salmonella-deficient farms, 302 deep-sows were selected, which were inoculated with the test antigens intramuscularly, twice for 25-28 and 15-18 days before farrowing. All piglets obtained from sows were divided into two groups. Group No. 1 (882 goals) from the 30th to the 35th day of life received the test vaccine orally in a mixture with animal feed. Group No. 2 (918 goals) was vaccinated with the same vaccines intramuscularly, twice at the age of 30 and 43 days. When weaning from sows and transferring to the growing area from piglets, similar groups (sectors) were formed: No. 1 - 644 heads; No. 2 - 638 goals. At the same time, a control sector of 643 heads from piglets obtained from sows vaccinated three times with the PPD vaccine was completed. The control pigs themselves against salmonellosis were not vaccinated.

The animals were observed before they were transferred to the feeding site, taking into account the safety and average live weight of one head transferred for fattening in each group. Before immunization, 20 days after the termination of oral immunization and 10 days after the second intramuscular injection of the test salmonella antigens in 10 piglets in each group, blood was taken for serological studies.

Studies have shown that the proposed vaccine in tested doses is harmless to piglets 30-35 and 45-54 days of age. Oral immunization with salmonella lysate antigens by the proposed method provides long-term contact with intestinal lymphoid tissue and a pronounced specific immune response of the body. The optimal dose is a daily dose equivalent to 400-600 billion per head of corpuscular antigen.

Under production conditions, oral immunization increased the safety of piglets by 6%, and the live weight of piglets transferred for fattening by 1.4 kg compared to intramuscularly vaccinated animals.

The proposed vaccine was tested by us with a positive result in 2006-2011. in accordance with the Russian Scientific and Technical Program for Fundamental and Priority Applied Research on the Scientific Support for the Development of the Agro-Industrial Complex (AIC), assignment 02.04.01 on the basis of the Microbiology Laboratory with the Museum of Typical Cultures of the All-Russian Scientific Research Institute of Experimental Veterinary Medicine (VIEV) named after Ya.R. Kovalenko (RAAS); CJSC Skorodnyanskoye, Gubkinsky District, Belgorod Region; Peasant farm “Spinu” of the Mozhaisk district of the Moscow region on a livestock of 620 mice, 2797 pigs.

The vaccine will find application in farms in the country that are unsuccessful for pig salmonellosis, will allow to maintain the breeding stock and get a healthy offspring.

Information sources

1. J. "Veterinary medicine." - 2011. - No. 1. - S. 34-36.

2. J. "Veterinary medicine and feeding" (Vetkorm). - 2009. - No. 5. - S.4-5.

Claims (3)

1. The vaccine against salmonellosis of pigs, characterized in that the antigens contain strains of S. choleraesuis No. 370 and S. typhimurium No. 415 taken in equal proportions, while 1 mg of dry lysate antigen corresponds to 2.5-3.5 10 ⁹ microbial cells. 2. A method of manufacturing a vaccine against pig salmonellosis, characterized according to claim 1, characterized in that the biomass of Salmonella strains S. choleraesuis No. 370 and S. typhimurium No. 415 separately cultivated in a bioreactor on Hottinger broth pH 7.2-7.5 and indicator amine nitrogen 100-120 mg% with an air supply of 1 l / min per 1 liter of nutrient medium and a stirring speed of 100 rpm for 10 hours, add hydroxylamine (NH ₂ OH · HCl) at the rate of 0.065 mg per 1 billion microbial cells with constant stirring, incubated for 4 days at a temperature of 38 ... 39 ° C, adjust the pH of the lysate d 7.4 using 10% NaOH, the lysates were mixed 1: 1, then dissolved antigens precipitated by sequential addition of Na ₂ HPO ₄ and CaCl ₂ - 1 mL lysate loss 4mg SaNRO _4, settled lysate with precipitate during the day, then centrifuged 2500-3000 rpm for 20 minutes, remove the supernatant and add food pectin at the rate of 1% to the resulting lysate, freeze-dry and get the vaccine. 3. A method for the prevention of salmonellosis in pigs is the oral administration of the vaccine, characterized according to claim 1, to piglets aged 30-35 to 45-54 days of life in a dose equivalent to 400-600 billion microbial cells of the corpuscular antigen, daily for 10 days with a veterinary probiotic Lactobifadol in a dose of 10-12 g per head per day.