RU2092187C1 - Inactivated vaccine for salmonellosis control in sheep - Google Patents

Abstract

FIELD: veterinary microbiology. SUBSTANCE: vaccine has the formalinized cell suspension of strains Salmonella abortusovis VIEV N 876, Salmonella typhimurium VIEV N 159, Salmonella dublin VIEV N 1449 in physiological solution, adjuvant - oily-lanolin mixture and immunostimulating agent - cell suspension Rodococcus equi VIEV N 2. Vaccine administrated at a single dose 1.0 ml makes the prolonged and stressed immunity and can be used for salmonellosis control in sheep. EFFECT: enhanced effectiveness of vaccine.

Description

 The invention relates to veterinary microbiology, in particular, to the production of an inactivated vaccine against salmonellosis of sheep.

 Sheep and goat salmonellosis (Paratyphoid, Paratyphoid abortion), an infectious disease caused by bacteria of the paratyphoid group, is characterized by abortion, septic events and the birth of unviable young animals. Abortions can occur in 25-30 of pregnant ewes. The damage is the sum of the costs of treatment, malnutrition of the young, the death of the aborted ewes. The disease is registered in all areas of the country with developed sheep breeding. When using effective vaccines, the number of abortions and the death of ewes is reduced tenfold from 25 to 2.5 or more.

 A live, avirulent vaccine against sheep salmonellosis is known, including the Salmonella antigen, serovar of sheep abortion from strain N 17 HC, which is diluted with saline before use.

 To obtain a vaccine, take the bacterial mass of live salmonella vaccine strain, concentrate, add cryoprotectants, pour into ampoules and freeze-dry.

 Sheep is vaccinated 1 month before the start of a random campaign in a dose of 1 ml once in farms unsuccessful for sheep salmonellosis. The duration of immunity is 6 months / 1 /.

 Also known is a concentrated, formolthiomersal vaccine against sheep salmonellosis, manufactured by the Armavir Biofactory. The vaccine contains antigens of 3 Salmonella serovars: sheep abortion, typhoid, and Dublin.

 The vaccine is used to prevent sheep salmonellosis in farms unsuccessful for this disease, it is administered twice: the first time 3-4 ml, the second time with an interval of 7-10 days 4-5 ml intramuscularly / 2 /.

 The objective of our research is to obtain an inactivated, areactogenic vaccine against salmonellosis of sheep, which provides the creation of intense, long-lasting immunity in the animal’s body, reducing the dose and frequency of administration.

 We isolated the serovars of Salmonella Abortion sheep N 876 and Dublin N 1149 from aborted fetuses of sheep used in the proposed vaccine as antigens in 1989.

 In addition, we isolated the strain Rodococcus equi N 2 from cattle from the economy of the Moscow region in 1986, which was used in the proposed vaccine as an immunostimulant, which was simultaneously filed.

 In addition to these strains, a previously known strain of Salmonella typhimurium N 159, obtained from the Collection of the Typical Culture Museum of the Bacteriology Laboratory of the All-Russian Scientific Research Institute of Experimental Veterinary Medicine, was used to obtain a vaccine against sheep salmonellosis. Ya.R. Kovalenko, Moscow, which has the following properties.

 Cultural and morphological signs. Gram-negative small rods, motile, forming convex colonies 1-3 mm in diameter, transparent or whitish. They grow well on ordinary nutrient media, on the BCH it forms a film and a parietal ring.

Physiological and biochemical properties. Aerob capable of growing under microaerophilic conditions. The optimum pH is 7.2-37 ^o C. Does not change milk, does not dilute gelatin, emits hydrogen sulfide, does not excrete indole, does not decompose urea, ferments glucose, mannitol, sorbitol, arabinose, maltose with the formation of acid, alctose, does not ferment sucrose, has enzymes lysine decarboxylase, ornithine dehydrolase. It is agglutinated by diagnostic salmonella serums 0-4, H i 1,2. Pathogenic for white mice. LD ₅₀ is equal to 10 • 10 ³ cells. The proposed vaccine includes antigens of 3 Salmonella serovars: abortion of sheep, mouse typhoid and dublin in physiological saline with 0.4-0.5 (v / v) formalin, adjuvant - oil-oil suspension of Rodococcus immunostimulant cells.

 The strain Salmonella abortusovis N 876 has the following properties.

 Cultural and morphological signs. Gram-negative small sticks, spores and capsules not forming, mobile. Small colonies with a diameter of up to 1 mm are formed on MPa, films and a parietal ring are not formed on MNS, the broth uniformly grows turbid.

Physiological and biochemical properties. Aerobes capable of growing under microaerobic conditions. The optimum pH is 7.2, t 37 ^o C. Do not change milk, do not dilute gelatin, emit hydrogen sulfide, do not emit indole, do not decompose urea, ferment glucose, mannitol with the formation of acid and very weak gas; lactose, sucrose, arabinose, sorbitol do not ferment, they have the enzyme lysine decarboxylase. Agglutinated with diagnostic salmonella serums 0-4, N-s, 1.6. Pathogenic for white mice. LD ₅₀ 47 • 10 ⁶ cells.

 The strain Salmonella dublin N 1449 has the following properties.

 Cultural and morphological properties. Gram-negative small sticks, spores and capsules do not form, are mobile, form convex colonies 1-3 mm in diameter, transparent or whitish. They grow well on ordinary nutrient media: on the BCH it forms a film and a parietal ring.

Physiological and biochemical properties. Aerobes capable of growing under microaerobic conditions, optimum pH 7.2, t 37 ^o C. Do not change milk, do not dilute gelatin, do not emit indole, emit hydrogen sulfide, do not decompose urea, ferment glucose, mannitol, sorbitol with the formation of acid and gas; lactose, sucrose, inositol, arabinose are not fermented, they have the enzyme lysine decarboxylase. Agglutinated with salmonella diagnostic sera: 0-9, H dr, (first phase). Pathogen for white mice LD ₅₀ is 7 • 10 ³ cells.

 The strain Rodococcus equi N 2 has the following properties.

Cultural and morphological characters. Gram-positive, motionless, rod-shaped and coccoid-shaped bacteria that form convex colonies up to 1 mm in diameter, grayish-yellow in color. Rodococcus equi N 2 is characterized by slow growth on simple nutrient media (3-4 days), but when 10 serum is enriched at 37 ^° C, good growth is observed after 24 hours. When the culture tubes are exposed to light, the colonies acquire a pigment of the color of salmon.

Physiological and biochemical properties. Aerob capable of growing under microaerobic conditions. The optimum pH is 7.2, t 37 ^o C. In relation to carbohydrates, amino acids, polyhydric alcohols is inactive. Ferment only glucose. It does not form indole and hydrogen sulfide, does not thin the gelatin, and does not lyse the red blood cells of animals. Has enzymes catalase and urease. Non-pathogenic when administered intraperitoneally to white mice weighing 8-20 g at a dose of 10 • 10 ⁹ microbial cells does not cause their death within 14 days.

 A method of preparing a vaccine is as follows.

 Example 1. For the preparation of 1 l of the vaccine take a mixture of antigens prepared in saline with 0.4-0.5 formalin. The concentration of cells in 1 ml of suspension is 100 billion.

Antigens Salmonella abortusovis N 876 200 ml
Salmonella typhimurium N 159 antigens 100 ml
Antigens Salmonella dublin N 149 100 ml
A suspension of Rodococcus equi N 2 cells at a physiological solution containing 150 ml of 100 ml in 1 ml
Light mineral oil and anhydrous lanolin in a ratio of 83-87: 17-13 - Else
After mixing all the components, the formalin concentration will be 0.2-0.25
A vaccine is used once in a dose of 1.0 ml for prophylactic and therapeutic purposes in sheep farms that are unsuccessful for sheep salmonellosis. The effectiveness of the vaccine in experimental conditions (using a triple infectious dose) was 80
Example 2. The vaccine is prepared analogously to example 1, but take a mixture of antigens prepared in physiological saline containing 0.4-0.5 formalin (1 ml of bacterial suspension contains 100 billion metric tons) in the following quantities:
Antigen Salm.abortusovis N 876 190 ml
Salm.typhimurium N 159 antigen 95 ml
Salm.dublin N 1449 antigen 95 ml
Suspension of Rodococcus equi N 2 cells on physiological a solution containing 150 ml of MT 95 ml in 1 ml
Oil-lanolin adjuvant in a ratio of 83-87: 17-13
Example 3. The vaccine is prepared analogously to example 1, but take a mixture of antigens prepared on a physiological solution containing 0.4-0.5 formalin (1 ml of antigen contains 100 billion metric tons)
Salmonella abortusovis N 876 antigen 210 ml
Sakm.typhimurium N 159 antigen 105 ml
Salmonella dublin N 1449 antigen 105 ml
Suspension of Rodococcus equi N 2 cells (concentration of 150 billion in 1 ml) 150 ml
Oil-lanolin mixture in a ratio of 83-87: 17-13
The components of the vaccine named in one of Examples 1, 2, 3 are placed in a homogenizer and, after 10-15 minutes of homogenization, the vaccine is packaged in bottles.

 As can be seen from the examples, the proposed vaccine is areactogenic, provides the creation in the animal’s body of long-lasting and intense immunity against sheep salmonellosis with a single dose of 1.0 ml.

 The vaccine in the indicated doses can be used for therapeutic purposes.

 The vaccine was tested with a positive result for 47 thousand sheep in the Alexander and Levokum districts of the Stavropol Territory in 1992-1994.

 The vaccine will find application in sheep farms that are unsuccessful in sheep salmonellosis.

Information sources:
1. Guidance on the use of a vaccine against salmonellosis of sheep from strain N 17 NS, 1991

 2. Polyvalent formol-thiomersal vaccine against sheep salmonellosis. In the reference book: Veterinary preparations, 1986 Kiev, Harvest, p.236.

Claims (1)

An inactivated sheep salmonellosis vaccine containing a formalinized suspension of cells of the strains from serovars Salmonella abortusovis, Salmonella typhimurium, Salmonella dublin in physiological saline and an adjuvant, characterized in that it additionally contains an immunostimulant in the form of a suspension of cells of the strain Rodococcus equi VI physiological 150 billion cubic meters / ml, from adjuvants - oil-oil mixture in the ratio of 83 87 17 13, from strains of serovar Salmonella abortusovis strain Salmonella abortusovis VIEV N876, from strains of serovar Salmonella typhimurium strain Salmonella typhimurium WIEV N159, from serovar strains Salmonella dublin strain Salmonella dublin WIEV N1449, in the following ratio of components, ml / l:
Formalized cell suspension of strain Salmonella abortusovis VIEV N876 with a concentration of 100 billion m.cl./ml 190 210
Formalized cell suspension of strain Salmonella typhimurium VIEV N159 with a concentration of 100 billion m.cl./ml 95 105
Formalized cell suspension of strain Salmonella dublin VIEV N1449 with a concentration of 100 billion m.cl./ml 95 100
Suspension of cells of the strain Rodococcus equi VIEV N2 with a concentration of 150 billion m.cl./ml 95 105
Oil-lanolin mixture in the ratio 83 87 17 13