RU2220743C2 - Method for manufacturing associated vaccine against salmonellosis, pasteurellosis and enterococcal infection in piglets - Google Patents

Abstract

FIELD: biotechnology, veterinary microbiology. SUBSTANCE: the present method deals with inoculation and separate controlled cultivation of vaccine strains of salmonella, pasteurella and streptococci followed by inactivation of these cultures with formaldehyde, adsorption and mixing adsorbed cultures. Moreover, as vaccine salmonella strain one should apply S. cholerae-suis strain N 370, its inactivation is carried out with formaldehyde at 43-48 C for 6-8 d, moreover, final formaldehyde concentration in the culture corresponds to 0.02-0.08%. The present innovation enables to increase immunogenic activity of associated vaccine. EFFECT: higher efficiency. 1 tbl

Description

 The invention relates to biotechnology and veterinary medicine and can be used in the industrial production of the associated vaccine against salmonellosis, pasteurellosis and enterococcal infection of piglets for the prevention in animals of infectious diseases caused by salmonella, pasteurella and streptococcus.

 The existing method of manufacturing a vaccine against salmonellosis, pasteurellosis and enterococcal infection of piglets, as well as inactivated vaccines against salmonellosis of calves and piglets, has several disadvantages, which include an excessive concentration of formaldehyde in inactivated cultures of Salmonella cholerae-suis, Salmonella dublin and Salmonella typhimurium - 0.1 % (or 0.4% in terms of formalin), which negatively affects the immunogenicity of the drugs, and a long inactivation time of 15-16 days [1, 2, 3].

 The aim of the invention is the creation of a highly immunogenic associated vaccine that provides reliable protection against salmonellosis, pasteurellosis and enterococcal infection of piglets.

 This goal is achieved using a gentle regime of inactivation of microorganisms, which allows to halve the time of inactivation of Salmonella and three times lower the final concentration of formaldehyde in an inactivated culture of Salmonella compared to the prototype [1].

 The method is as follows. Sowing and separate controlled cultivation of vaccine strains of pasteurell, Salmonella and streptococci is carried out in accordance with the method [4, 5, 6].

Inactivation of streptococcal cultures is carried out with formaldehyde at a temperature of 45.5 ^o C for 24 hours, and the final concentration of formaldehyde in the culture is 0.05% [6].

Inactivation of pasteurell cultures is carried out with formaldehyde at a temperature of 44.5-45.5 ^o C for 4-6 hours, and the final concentration of formaldehyde in the culture is 0.02% [7].

The resulting bacterial mass of the production strain of S. cholerae-suis 370 with a bacterial concentration of 4 • 10 ⁹ cells / cm ^{3 is} pumped into a previously prepared sterile reactor (or glass bottle) and fractional formaldehyde is added with constant stirring to a final concentration of 0.02-0 , 08%, the pH was adjusted to 7.1-7.2 pH units by adding a 10% sodium hydroxide solution, stirred for 15-20 minutes and kept at a temperature of 43-48 ^o C for 6-8 days.

 The culture of pasteurell, salmonella and streptococcus after inactivation is adsorbed on an adjuvant and mixed according to the instructions [1].

 The technological parameters of the manufacture of the associated vaccine against salmonellosis, pasteurellosis and enterococcal infection of piglets, indicated in the examples, are shown in the table.

Example 1
A method of manufacturing a vaccine with minimum parameter values
The process of inactivation of the production strain of Salmonella is carried out with formaldehyde at a temperature of 43 ^o C for 6 days, and the final concentration of formaldehyde in the culture is 0.02%.

Example 2
A method of manufacturing a vaccine with optimal values of the parameters
The process of inactivation of the production strain of Salmonella is carried out with formaldehyde at a temperature of 45.5 ^o C for 7 days, and the final concentration of formaldehyde in the culture is 0.05%.

Example 3
A method of manufacturing a vaccine with maximum values of the parameters
The process of inactivation of the production strain of Salmonella is carried out with formaldehyde at a temperature of 48 ^o C for 8 days, and the final concentration of formaldehyde in the culture is 0.08%.

Example 4
A method of manufacturing a vaccine with parameter values below the minimum
The process of inactivation of the production strain of Salmonella is carried out with formaldehyde at a temperature of 40.5 ^o C for 5 days, and the final concentration of formaldehyde in the culture is 0.01%.

Example 5
A method of manufacturing a vaccine with parameter values above the maximum
The process of inactivation of a production strain of Salmonella is carried out with formaldehyde at a temperature of 50.5 ^o C for 9 days, and the final concentration of formaldehyde in the culture is 0.110%.

 The vaccine prepared according to examples 1-3 has a high immunogenic activity and meets the requirements of technological conditions, while the vaccine prepared according to example 4 does not completely inactivate Salmonella endotoxins, and the vaccine prepared according to example 5 has increased reactogenicity.

Sources of information
1. Instructions for the manufacture and control of a vaccine against salmonellosis, pasteurellosis and enterococcal infection of piglets associated, approved July 16, 1990 (prototype).

 2. Instructions for the manufacture and control of a concentrated formolvac vaccine against salmonellosis (paratyphoid) calves, approved May 14, 1990

 3. Instructions for the manufacture and control of the vaccine against paratyphoid piglets, approved on November 11, 1988.

 4. RF patent 1566533 dated April 7, 1993, A 61 K 39/102 "Method for the manufacture of a vaccine against pasteurellosis of birds."

 5. RF patent 2129016 of April 20, 1999, A 61 K 39/112 "Method for the manufacture of a vaccine against salmonellosis of farm animals."

 6. Application for the grant of a patent 2000128386/13 (030343) "Vaccine associated with aluminum hydroxide against infectious pneumonia of pigs of bacterial etiology and method for its manufacture" dated November 15, 2000

 7. Auth. St. RF 1792708 dated October 8, 1992, A 61 K 9/102 "Method for the production of sera for the treatment and prevention of pasteurellosis of farm animals, mainly pigs, fur animals and rabbits."

Claims (1)

A method of manufacturing an associated vaccine against salmonellosis, pasteurellosis and enterococcal infection of piglets, comprising seeding and separate controlled cultivation of vaccine strains of salmonella, pasteurella and streptococci, followed by inactivation of cultures with formaldehyde, adsorption on adjuvant and mixing of adsorbed cultures, characterized in that the vaccine strain is in quality use strain S. cholerae-suis No. 370, the inactivation of which is carried out with formaldehyde at 43-48 ° C for 6-8 days, with the final concentration formaldehyde radio culture is 0.02-0.08%.

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