Salmonella vaccine manufacture method

FIELD: medicine.

SUBSTANCE: invention belongs to vaccine manufacture for infectious diseases prophylaxis. Salmonella vaccine manufacture includes detoxication of exo- and endotoxins in suspention of autoclaved salmonells consecuently with two detoxicants - 0.2% formalin solution at 40±1 C° during 6-7 days and 0.5±0.1% aethonium solution at 41±1 C° during 7-8 days, followed by sterilisation at 1.0 atm during 20 minutes. Derived anatoxins are sorbed on aluminium hydroxide and sterilised at 1 atm during 20 minutes.

EFFECT: anatoxin-vaccine with enhanced effectivity and safety is used for specific prevention of salmonellosis in piglets, calves, carnivorous and poultry.

3 ex

 

The invention relates to Microbiology, in particular to the manufacture of a toxoid vaccine for the prevention of salmonellosis calves, pigs, carnivores and birds.

Known methods for producing inactivated bacterial and viral vaccines, anatoxin based on the use of 0.8±0.2% solution of formalin (sparrows Microbiology and immunology. - M.: Medicine, 1999. - S-268).

Getting Salmonella vaccine includes the cultivation of Salmonella in a nutrient medium, sterilization, detoxification of toxins 0,8±0,2% formalin solution, sorption on aluminum hydroxide and the introduction of thimerosal. Because of the carcinogenic properties of formaldehyde, thimerosal and incomplete detoxification of enterotoxins with some success have been tested peroxide, tyrosine, antibiotics, phenol, RIP, photo-oxidation, however, in the case of inactivated Salmonella and colibacteriosis vaccines they were ineffective.

The prototype was taken the way of getting Salmonella vaccine, which includes the cultivation of Salmonella in a nutrient medium, sterilization and detoxification of toxins 0,8±0,2% formalin solution, sorption on aluminum hydroxide with subsequent submission of the preservative is thimerosal (Size J.F. ñ, B. Û.F., Kirillov L.V. Veterinary medicines. - M.: Kolos, 1981. - S-227).

The disadvantage of this method is the detoxification of 0.8 is 0,2% formalin solution, since formaldehyde has carcinogenic properties, resulting in the use of it in such concentration unsafe. The vaccine also introduces a toxic preservative is thimerosal.

One of the ways to solve the problem is a decrease in the concentration of formalin. But this reduces its effectiveness, which leads to incomplete detoxification Salmonella vaccine.

Learn some detoxication revealed the effectiveness bischetvertichnoe ammonium compounds, in particular, etoniya (1,2-ethylene-bis-(N-dimethylcarbamate) ammonium dichloride). Safety and effectiveness etoniya confirmed by applying it in humane medicine as ointments, pastes and mortars with dermatitis, keratitis, stomatitis for the accelerated healing of wounds, etc.

The aim of the invention is to improve the efficiency and safety of Salmonella toxoid vaccine.

This goal is achieved through irreversible and complete detoxification complex Salmonella toxins consistently two detoxication, the reduction of the concentration of formalin to 0.2% by making the second detox etoniya in a concentration of 0.5±0.1% for a final detoxification at 40±1°C for 6-7 days, the removal of thimerosal.

While the initial concentration of formaldehyde is reduced to 0.003 to 0.05%, and etoniya, from 0.5% to 0.2% to 0.3% due to binding of the s protein molecules of toxins and microorganisms.

In the patent and scientific and technical literature technical solutions are not found, similar to the claimed using etoniya, indicating the novelty of the proposed solution.

Using 0,5±0,1% etoniya decreasing the concentration of formalin from 0.8±0.2% to 0.2% in the proposed method allows to achieve stable detoxification and polymerization of the whole complex of Salmonella toxins and get harmless and effective Salmonella toxoid vaccine.

Installed complete and irreversible detoxify all types of Salmonella toxins, including enterotoxins, by using the first stage of detoxification of 0.2% solution of formalin instead of 0.8±0.2% and in the second stage 0,5±0,1% etoniya with not only detox, but preservative, bactericidal and fungicidal properties allowed to offer sustainable manufacturing technology effective and harmless Salmonella toxoid vaccines without mercury-containing preservative is thimerosal.

The invention is illustrated by the following examples.

Example 1. Getting Salmonella toxoid vaccines (CAB). To the resulting suspension autoclaved Salmonella with a concentration of 60±10 billion/ml of microorganisms and soluble toxins with a concentration of 1.2±0.2 mg/ml was added formaldehyde and 0.2% concentrations detoxification at 40±1°C for 7 days, and then added atony 0.5% concentration to complete detoxification at 41±1°C for 7 days all kinds of Salmonella toxins, including enterotoxins, which formalin is not able to detoxify. After sorption of the toxoid vaccine on aluminum hydroxide (3 mg/cm3conducted packing of the drug of 20.0 and 100.0 ml flasks and sterilized at 1.0 ATM for 20 minutes.

The resulting toxoid vaccine is tested for safety and effectiveness.

Example 2. Test Salmonella toxoid-vaccine emissions was carried out on 10 white mice, 20-60 days old calves and piglets, 12 broilers and 6 pigeons by subcutaneous injection to mice in a volume of 0.3 ml, calves and piglets 6-7 ml, chicken and pigeons to 0.5 ml Under observation for 12 days, all animals remained clinically healthy, without visible changes in the injection.

Example 3. Test Salmonella toxoid-vaccine activity was performed using a 14 goals piglets, 12 calves 20-3-day-old and 8 pigeons by intramuscular injection of toxoid vaccines pigs and calves at a dose of 4-5 ml, pigeons at a dose of 1 ml twice with an interval of 7 days. Within 15 days after the second vaccination, all vaccinated animals and unvaccinated three pigs, three cows, four pigeons entered the living virulent is ultura S.cholerosius daily growing in the BCH at a dose of 200-300 thousand microbial phone Observation of the animals was carried out for 20 days. It was found that vaccinated calves, piglets and six of eight pigeons are not sickened by Salmonella, and all unvaccinated control calves, pigs and pigeons are sick in the period of 7-12 days after infection with virulent Salmonella.

All of diseased calves and piglets assigned to specific treatment with immune serum with antibiotics and saline solutions.

Infected with Salmonella pigeons died within 10-15 days.

The way of getting Salmonella vaccines, including detoxification Exo - and endotoxins in suspension with autoclaved Salmonella, sorption obtained anatoxina on aluminum hydroxide, characterized in that the detoxification of toxins consistently spend two detoxication - 0,2%formalin solution at 40±1°C for 6-7 days and 0,5±0,1%solution etoniya at 41±1°C for 7-8 days and after adsorption on aluminum hydroxide target product is sterilized at 1.0 ATM for 20 minutes



 

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Composition // 2389481

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4 ex

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