Inactivated emulsion vaccine against salmonellosis piglets

 

(57) Abstract:

The invention is intended for the specific prevention of animal diseases. Inactivated emulsion vaccine against salmonellosis pigs contains a mixture of virulent antigens, immunogenic strains of Salmonella serovars cholerasuis, Salmonella typhimurium physiological solution, adjuvant and preservative - 0,4 - 0,5% (vol./about.) formalin. As the antigen of serovar Salmonella cholerasuis it contains a strain of Salmonella cholerasuis N 8. As the antigen of serovar Salmonella typhimurium strains Salmonella typhimurium N 159. As immunostimulant using suspension cells Rodococcus equi N 2 physiological solution. Adjuvant - makelanrinne mixture. The vaccine will create a busy long immunity of piglets at a reduced dose and frequency of administration.

The invention relates to the field of veterinary Microbiology, in particular the production of inactivated vaccines against salmonellosis piglets.

Salmonellosis swine is an infectious disease caused by bacteria belonging to the group of paratyphoid C. acute over the age of 45 days, the pigs refuse to eat, depressed, lying down, the body temperature increases lubochna-intestinal tract, gradually encroaching exhaustion. When delayed treatment of piglets die in 2-7 days in 70-80% of cases. Damage is the sum of the cost of treatment, mortality of young animals. The disease is registered in all regions of the country. When using effective vaccines the death of piglets decreases.

Famous dried live vaccine from strain TC-177. It is an inactivated broth culture of the attenuated strain of Salmonella choleraesuis TC-177, dried by sublimation. The vaccine is used for prophylactic purpose in disadvantaged for salmonellosis swine farms where the agent is installed Salmonella choleraesuis. Instill in all clinically healthy animals with 2 weeks of age. The vaccine is injected subcutaneously twice.

Now apply formulatio. It is made from three strains of Salmonella: Salmonella choleraesuis - 50%, Salmonella typhimurium - 25%, Salmonella dublin - 25%. The vaccine is used for prophylactic purpose in farms, troubled by salmonellosis piglets. If the farm has no sick animals, vaccinated all piglets piglets and weaners aged 20-30 days of age. The vaccine is administered twice under the skin of 4-5 ml with an interval of 5-8 days. The piglets older than 2 months of age enter Pacala 3 ml, after 5-8 days - 4-5 ml and 10-20 days after the second vaccination with 5 ml.

The objective of our research - getting killed, reactogenic vaccine against Salmonella pigs, providing busy creating long-term immunity in the animal body, reducing the dose, frequency of administration.

We have isolated serovar Salmonella cholera CMS N 8, selected from the fallen pig used in the vaccine as an antigen.

In addition, we used as immunostimulant strain Rodococcus equi N 2, isolated from cattle from the farms of the Moscow region in 1986.

In addition to these strains to obtain a vaccine against salmonellosis piglets was used formerly known strain of Salmonella murine typhus N 159, obtained from the Museum collection type culture laboratory of Bacteriology of the all-Russian research Institute of experimental veterinary medicine named. J. P. Kovalenko, Moscow, which has the following properties:

Cultural and morphological characteristics.

Gram-negative, small, slightly curved rods, motile, forming convex colonies 1-3 mm diameter, transparent or whitish color. Khoroshevskoe properties.

Aerobe, able to grow in microaerobic conditions. The optimum pH of 7.2, temperature - 37oC.

Does not change the milk, not liquefies gelatin, produces hydrogen sulfide, does not emit indole, does not decompose urea; sprayway glucose, mannitol, sorbitol, arabinose, Inositol, maltose with the formation of acid; lactose, sucrose is not sprayway; has enzymes: isindeterminate, Interdigitated. Agglutinated salmonellezne sera - O - 1, 4, 5, 12; H - i; 1,2.

Pathogenic for white mice, LD50equal 5 103cells intraperitoneal infection.

The proposed vaccine comprises antigens 2 Salmonella serovars: cholera CMS and typhimurium in physiological solution with 0.4 - 0.5% formalin, beta - Maslyakova mixture containing a suspension of cells rhodococci (immunostimulant).

The strain of Salmonella cholera CMS N 8 has the following properties:

Cultural and morphological characteristics.

Motile gram-negative rods with rounded ends, spores and capsules do not form.

At MPA - the colonies of small, translucent, convex, with aging become muddy. On BCH does not form a film and parietal rings, broth evenly turbid.

Does not change the milk, not liquefies gelatin, does not emit indole, does not decompose urea; breaks down glucose with the formation of acid and gas), mannitol, xylose, lactose, sucrose, Inositol, arabinose, dulcet not sprayway; has enzymes: isindeterminate, Interdigitated.

Agglutinated Salmonella sera: O - 6,7, H - c; 1,5.

Pathogenic for white mice, LD50equal to 16 cells intraperitoneal infection.

Strain Rodococcus equi N 2 has the following properties:

Cultural and morphological characteristics.

Gram-positive, stationary rod-shaped and coccoid forms of bacteria that form a convex colonies up to 1 mm in diameter, grayish-yellow color.

For rhodococcal N 2 characterized by slow growth on simple nutrient media (4-9 days), but when the enrichment of 10% serum at 37oC shows good growth after 24 hours. During curing tubes with culture in the light - colonies acquire the pigment color of salmon.

Physiological and biochemical properties.

Aerobe capable to grow in microaerobic conditions. The optimum pH of 7.2 to 7.4, temperature 30oC.

In respect of carbohydrates, Amin who calls for gelatin, not analyzes the red blood cells of animals. Has enzymes: catalase, urease. Non - intraperitoneal injection white mice, weighing 18-20 g dose of 10 to 109microbial bodies, does not cause their death within 14 days.

The method of preparation of the vaccine consists of the following.

Example 1. To prepare 1 liter of vaccine take a mixture of antigens prepared with physiological solution with 0.4 - 0.5% formalin. The concentration of cells in 1 ml suspension 40 billion

Antigens:

Salmonella choleraesuis - 300 ml

Salmonella typhimurium - 100 ml

Suspension cells Rodococcus equi physiological solution containing 1 ml of 200 billion microbial cells 100.

Light mineral oil and lanolin anhydrous ratio 83-87:17-13 - rest.

After mixing all the components, the concentration of formalin is 0,2 - 0,25%.

Apply the vaccine is a single dose of 1.0 ml prophylactically in pig farms, troubled by salmonellosis piglets.

The effectiveness of the use of the vaccine under experimental conditions (using a five-infecting dose) amounted to 100%.

Example 2. The vaccine is prepared analogously to example 1, but take a mixture of antigens, prigot the rd microbial cells) in the following amounts:

Antigens:

Salmonella choleraesuis - 315 ml

Salmonella typhimurium - 105 ml

A suspension of microbial cells rhodococci (concentration in 1 ml 200 billion) - 105 ml.

Maslyakova mixture in the ratio 83-87:17-13 - rest.

Example 3. The vaccine is prepared analogously to example 1, but take a mixture of antigens prepared with physiological solution containing 0.4 - 0.5% formalin (1 ml contains 40 billion microbial cells).

Antigens:

Salmonella choleraesuis - 285 ml

Salmonella typhimurium - 95 ml

Suspension cells rhodococci physiological solution containing 1 ml of 200 billion microbial cells - 95 ml.

Maslyanichnoy adjuvant in a ratio 83-87:17-13 - rest.

Named in one of the examples 1, 2, 3 components of the vaccine are placed in a homogenizer and after 10-15 minutes of homogenization vaccine Packed in bottles.

As can be seen from the examples, the proposed vaccine is reactogenic, provides for the creation in the animal body long and active immunity against Salmonella infection of pigs with a single dose of 1.0 ml.

The vaccine has been tested by us on 70 pigs with positive results in 1995-1996,

The vaccine will be used in pig salmonellas piglets comprising a mixture of virulent antigens, immunogenic strains of serovars Salmonella choleraesuis, Salmonella tuphimurium physiological solution, adjuvant, preservative 0,4 - 0,5% (V/V) formalin, characterized in that as antigens of serovar Salmonella choleraesuis contains a strain of Salmonella choleraesuis 8 as the antigen of serovar Salmonella typhimurium contains a strain of Salmonella typhimurium 159, as immunostimulant contains a suspension of cells Rodococeus equi 2 maslyanino mixture, the components included in the vaccine, take the following value is based on l: antigens on the basis of physiological solution with the addition of 0.4 - 0.5% formalin and concentration of microbial cells in 1 ml of 40 billion: antigen Salmonella choleraesuis 8 285 - 315 ml antigen of Salmonella typhimurium 159 95 - 105 ml of re-suspended cells Rodococcus equi 2 physiological solution containing 200 billion microbial cells in 1 ml of 95 - 105 ml, adjuvant - Maslyakova mixture in the ratio of 83 - 87 : 17 - 13 - rest.

 

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