The vaccine streptococcus and pasteurellosis nutria

 

(57) Abstract:

Usage: veterinary Microbiology, biotechnology, vaccine, prevention of Streptococcus and pasteurellosis nutria. The essence of the invention: manufacturing of vaccines against Streptococcus and pasteurellosis nutria separately cultivated strains Str. Zooepiolemicus VGNKI N K-DEPT, P. multocida VGNKI N 6011, 2394, 1015. Cultures of streptococci and pasteurellosis are combined in a ratio of 1:1 and add a 0.3% solution of formalin. After a period of inactivation and get the desired results control of the purity of the growth of bacterial mass contribute adjuvant - 3% solution of aluminium hydroxide at the rate of 15% by volume of the culture. The second adjuvant saponin added at the rate of 100 mg per 1 l of biomass. Adjuvants contribute with continuous agitation. Get saponin-formamidinesulfinic vaccine provides immunity formation in 3-5 days after vaccination, which persists for at least 12 months. Epizootic efficiency is 95% or more of all vaccinated animals. table 1.

The invention relates to veterinary Microbiology, namely, for construction of bacterial vaccines for the prevention of Streptococcus and pasteurellosis n what you registered as individually and in Association: dies young after precipitation (80%), females abortivum in the second half of pregnancy (up to 80%). Large costs are supplemented by the violation of the cultivation of animals, as well as through carrying out veterinary-sanitary measures in the elimination of these diseases.

Salmonellosis, streptococcosis and pasteurellosis nutria are factorial diseases of these animals. In this regard, specific prevention of the above diseases is required.

Prevention of salmonellosis is applied polyvalent vaccine against salmonellosis and colibacteriosis fur-bearing animals. The preparation is injected subcutaneously in the area of the inner thigh puppies nutria from 30 to 45 days of age, three doses of 0.25, 1.0, and 1.5 cm3with an interval of 8 to 10 days.

14 days after a course of prevention of Salmonella nutria vaccinates against streptococcosis. Intramuscularly formamidinesulfinic vaccine Streptococcus nutria twice in the thigh at doses of 1.0 and 1.5 cm3with an interval of 6 days regardless of age.

14 days after the last injection of a biological product against streptococcosis with the inner surface of the thighs, twice in doses of 0.5 cm3and 1.0 cm3every 10 to 12 days.

Course specific prophylaxis of especially dangerous infectious diseases for nutria in the application of monovaccines long-term (over 3 months), and therefore, despite their high specific activity, mortality of animals note from diseases that profilaktirujut last.

Therefore, the aim of the present invention is to construct the associated immunogenic vaccines against Streptococcus and pasteurellosis nutria to the existing system of prevention and Wellness activities to complement the associated vaccination of livestock and to achieve more effective epidemic control streptococcosis and pasteurellosis nutria.

A vaccine for the prevention of Streptococcus and pasteurellosis nutria were prepared on the basis of a strain of Streptococcus serogroup C N and pasteurellosis N 6011, 2394, 1015, who selected us from dead nutria and deposited in the collection of strains of microorganisms of the all-Russian State research Institute for control, standardisation and certification of veterinary preparations.

Example 1. For the manufacture of 1-series vaccine streptoco logicheskoi group C strain N K-DEPT and pasteurellosis N 6011, 2394, 1015. The contents of each ampoule separately sown in MPA with 1% glucose 3 4 Petri dishes according to the following procedure: 0.1 to 0.2 ml of the suspension culture of the strain was deposited on the surface of the agar and sterile glass spatula was distributing it over the surface of the agar. This spatula was performed on the surface of the agar consistently 3 more cups. The crops in MPA incubated 48 h at 37 37,5oC. at the same time doing the sowing on the BCH, MPB under paraffin oil and the environment Saburo 2 tubes of each medium and incubated under the same temperature conditions (environment Saburo 24 at 20oC) crops on MPB and environment Saburo 10 days. and on the BCH 3 days.

After a period of incubation of the culture with each nutrient medium separately tested for purity and character growth visual view and the view of smears stained by gram stain. Growth of cultures on nutrient media should be characteristic of pathogens Streptococcus and pasteurellosis.

From cultures grown on Petri dishes, select 2 3 colonies of Streptococcus and pasteurellosis separately and sown in BCH with the addition of 1% glucose and incubated at 37 37,5oC 18 22 PM Tested frequency growth view smears, gram stained, and sown in the BCH the ARS daily broth culture was perseval in preheated temperature 37-37,5oC BCH with glucose (1% ) in the 2.5-liter cylinders based 80-100 ml per 10 litres of Crops were cultivated at a temperature 37-37,5oC for 18-24 hours By this time the concentration of the bacterial mass should not be below 4109M. K. in 1 ml.

Make sure the frequency of streptococcal growth and pasteurellosis, biomass was poured into 20 liter bottles in a 1:1 ratio and added to 0.3% commercial formalin.

Inactivation of cultures should be performed within 48 hours at a temperature of 16-18oC, while shaking each cylinder up to 3-4 times within 60 minutes

After inactivation of selected samples of the cultures of each cylinder separately for completeness of inactivation. With this purpose, conducted crops in tubes on MPA, MPB, MPB under paraffin oil. Crops should not have growth over the 10 days of observation.

After a period of inactivation and get the desired results control of the purity of the growth (complete inactivation) in the bacterial mass was made sterile adjuvant 3% solution of aluminium hydroxide (GOST 18287-81) at the rate of 15% by volume of the culture. The second adjuvant saponin was added at the rate of 100 mg per 1 liter Adjuvants was made with continuous agitation bacterial mass.

Note: a 3% HP>C for 30 minutes

After 3 days. after making adjuvant have formed a series of vaccines, the pH of the finished vaccine should be around 7.0 to 7.2.

The filling of vaccines produce 20, 100, 200 ml sterile vials that cover aluminum caps, ensuring the tightness of the bottle contents.

Received the vaccine has the following composition about.

a) particulate antigen in the culture fluid containing streptococci and Pasteurella 18-22 time growth in concentration (4-5)109microbial cells of 1 cm383,7

b) glucose 1,0

C) formalin 0,3

d) aluminum hydroxide to 100

To determine the quality of saponin-formamidinesulfinic vaccine did sample from different locations of a series of 10 vials, from which 5 are used for testing, and 5 bottles kept in the archive within 12 months.

To determine the appearance, color, presence of any impurities, cereals, mold, nerazvivayuschihsya sediment vaccine vials were thoroughly shaken and viewed visually in transmitted light. At the same time the vials were checked for tightness and correct labeling.

The concentration of bodoro same accuracy class. For testing use 3 of the vaccine vial. The contents of each vial were tested separately. The pH of the vaccine is conducted according to instructions attached to the potentiometer. The vaccine should have a pH around 7.0 to 7.2.

To test for sterility used 5 vials of vaccine. The sowing was performed from each vial in two test tubes with MPA, MPB, MPB and environment Saburo. The crops is carried out in a volume of 0.2-0.3 ml of the vaccine. Nutrient crops and kept for 10 days, the growth of microorganisms is not mentioned.

The safety of the vaccine was conducted in Guinea pigs weighing 300-350 g and on white mice 16-18, Three vaccine vial was shaken and out of each vial of vaccine with sterility took 20-30 ml of the drug was transferred into a sterile vial, the overall sample was used to determine the safety and immunogenicity.

To determine the safety of the contents of the vial were thoroughly shaken and introduced five Guinea pigs subcutaneously in the dorsal area in a volume of 2 ml and ten white mice subcutaneously in the dorsal area in a volume of 0.5 ml Vaccine did not cause disease and death of Guinea pigs and white mice within ten days of observation.

To determine immunol the outer side of the thigh in the dose of 0.2 ml 14 days after immunization 40 vaccinated and 40 control white mice of the same weight were infected subcutaneously in the thigh podarowano ten times the lethal dose of vaccine strains of streptococci and pasteurellosis separately (20 mice). The observation period of 10 days.

The vaccine is considered immunogenic if it protects from disease and the deaths of at least 18 immunized mice in each experimental group, when all disease and the deaths of at least 38 of white mice in the control groups for 10 days.

We obtained a series of vaccines against Streptococcus and pasteurellosis nutria and known vaccine diplokokkovyh septicemia calves, lambs and piglets we have experienced in the production environment (see table). The analysis of the data presented in table N-1, significantly proves the efficacy of the vaccine series No. 1, in comparison with the known vaccine. The vaccine was administered intramuscularly in the thigh from the inside of 1.0 and 1.5 with an interval of 7-10 days. The vaccine series N 1 harmless, do not cause complications in vaccinated nutria, has a high activity and contributes to the prophylactics of clinical signs of streptococcosis and death of animals from disease to 85.4%

Vaccine n the current strain of Streptococcus zooepidemicus VGNKI N K-DEPT with a titer of 2.5 1093 109cells/ml in culture medium, a cell suspension of strains of Pasteurella multocida VGNKI N 6011, Pasteurella multocida VGNKI N 2394 and Pasteurella multocida VGNKI N 1015, taken in equal proportions, with a total titer of 2.5 1093 109cells/ml in culture medium, glucose, formalin, aluminum hydroxide and saponin in the following ratio of components, about.

A cell suspension of strain Streptococcus zooepidemicus VGNKI N K-DEPT in culture medium with a titer of 2.5 1093 10941,3 44,35

Suspension cells of strains of Pasteurella multocida VGNKI N 6011, Pasteurella multocida VGNKI N 2394 and Pasteurella multocida VGNKI N 1015, taken in equal proportions, with a total titer of 2.5 1093 109cells/ml in culture medium 41,3 44,35

Glucose 1,0 2,0

Formalin 0,3 0,4

Aluminium hydroxide 10 15

Saponin Ostalnoe

 

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