Method of obtaining hyperimmune serum against swine hemophilosis, streptococcosis, and pasteurellosis

FIELD: veterinary science.

SUBSTANCE: invention relates to veterinary science. The method includes preparing an antigen from inactivated and concentrated Pasterella multocida serotypes A, B and D, Haemophilus pleuropneumoniae serotypes I and II, Streptococcus suis serotype II (Castel strain) and administering initial immunisation and hyperimmunisation with increasing antigen doses followed by blood taking and end product recovery. Oxen are used as producers and hyperimmunisation is administered in 7 to 10 intramuscular injections with a total dose of 1800-2400 billion microbial bodies of each antigen at 2-3 days' intervals. Initial immunisation is administered hypodermically to different body areas two times with 10-15 billion microbial bodies injected at first and 20-25 billion microbial bodies injected in 14 days plus 10 cm3 aluminium hydroxide.

EFFECT: method is simple and time-saving and yields 3,2-3,5 times more serum.

2 tbl, 4 ex

 

The invention relates to biotechnology and can be used to obtain hyperimmune serum for the treatment and prevention of hemophilia, streptococcosis and pasteurellosis pigs. A known method of manufacturing a hyperimmune serum against hemophilia, streptococcosis and pasteurellosis pigs received from the producers of pigs (EN 2118169 AC 39/02, 39/09, 39/102 - prototype).

However, this method is uneconomical, since the lifetime of pigs is 5-6 months (depending on the length of the tail and the clinical condition of the producers). In addition, the process of immunization and krivovaty this species is extremely time-consuming. It is advisable to use as producers of oxen, the lifetime of which at least 5 years.

The technical result of the invention is to increase the yield of the target product and simplify the method, and the use of the drug at the same time against the three diseases.

This technical result of the proposed method is achieved by the fact that to create groundmounted the oxen (no more than once every five years) subcutaneously twice in different parts of the body is injected inactivated and concentrated gamefilthy, streptococcal and pasteurellosis antigens: first 10-15 billion microbial cells by adding 10 cm3Al(HE)3and after 14 days at 20-25 m is rd M.L.

Hyperimmunization oxen spend 7-10-intramuscular injection of increasing doses of inactivated and concentrated antigens purified from the culture fluid in a total dose of each antigen 1800-2400 billion microbial cells with an interval of 2-3 days.

In operation, after each blood collection (16-17 cm3blood per 1 kg of body weight) after 1-2 days the regular intramuscular injections of inactivated concentrated hemofilter, streptococcal and pasterilezom antigens in a dose of 400-500 billion MT each, and 8-10 days after this from the producers again take blood.

The method is as follows.

Example 1. Hyperimmunization oxen were carried out by 10 intramuscular injection of increasing doses of inactivated and concentrated hemofilter, streptococcal and pasterilezom antigens purified from the culture fluid in the total number of 2400 billion MT of each antigen with an interval of 2-3 days according to the scheme:

40-60-100-150-250-350-450 - 2-week break - 200-300-500 billion M.L.

The results of determination of the activity

The titers obtained serum was:

control (prototype)
to Pasteurella multocida1:128
the reaction passive1:5121:512
the haemagglutination (TPHA)D 1:1281:128
Haemophilus pleuropneumoniae 58701:3201:320
The reaction of agglutination (PA) GU-191:6401:640
Streptococcus suis Kasli
The reaction of agglutination (PA)1:2001:200

Example 2. In comparison with example 1 increased the number of injections and doses of antigens by hyperimmunization. Spend 11 injection in the total number of 3000 billion MT of each antigen according to the scheme: 40-60-100-150-250-350-450 - 2-week break - 200-300-500-600 billion M.L.

The results of determination of the activity

The titers obtained serum was:

Control (prototype)
to P.multocidaA 1:1281:128
TPHAIn1:5121:512
D1:1281:128
H.pleuropneumoniae 58701:3201:320
RASU-191:6401:640
Str. Suis (PA)1:2001:200

The increase in the number of injections is greater than 10 and the total number of antigens of more than 2400 billion M.T. did not lead to further increase in the antibody titre of the serum. Increasing the number of antigens and the number of injections is impractical because puts extra strain on the immune system of producers and not profitable from an economic point of view.

Example 3. Reduced the number of injections to 7 while reducing the total amount of injected antigens to 1800 billion MT:

40-60-100-200-300-500-600 billion M.L.

The titers obtained serum was:

Control (prototype)
to P.multocidaA1:1281:128
TPHAIn1:5121:512
D1:1281:128
H.pleuropneumoniae58701:3201:320
PASU-191:6401:640
Str. Suis (PA)1:2001:200

Reducing the number of injections to 7 while reducing the overall number of antigens to 1800 billion MT has not led to a decrease in the titer of antibodies in the serum.

Example 4. Reduced the number of injections to 6 while reducing the total amount of injected antigens to 1300 billion M.T. scheme:

40-60-150-250-350-450 billion M.L.

The titers obtained serum was:

Control (prototype)
to P.multocidaA1:641:128
TPHAIn1:2561:512
D1:641:128
H.pleuropneumonie58701:1601:320
PASU-191:3201:640
Str. Suis (PA)1:1001:200

Reducing the number of injections to 6 while reducing the total amount of injected antigens to 1300 billion MT of each antigen resulted in the decrease of antibody titers in the serum.

The proposed method is tested with positive results in laboratory and production environments.

The proposed method in contrast to the known has the following advantages:

1. The output increases syvorotkin of 3.2-3.5 times (table 2).

Table 2
View producerWeightQuantity of blood with 1
kg
Output
(kg)animal(ml)
(ml)
pig150-160131950-2080
oxen400-450166400-7200

2. More simple and less time-consuming.

The life of swine 5-6 months (depending on the length of the tail and the clinical condition of the producers). The process of immunization and krivovaty this species is extremely time-consuming. On the contrary, the lifetime of oxen at least 5 years.

Hence, the Grund - and hyperimmunization pigs is held every 5-6 months, while oxen - once in 5 years.

3. More economical that follows from points 1 and 2. In addition, this serum can use the I for the treatment and prevention of hemophilia, streptococcosis and pasteurellosis at the same time.

The way to obtain hyperimmune serum against hemophilia, streptococcosis and pasteurellosis pigs, comprising preparing the antigen of inactivated and concentrated cultures Pasterella multocida of serovariants a, b and D and Haemophilus pleuropneumoniae serovars I and II of Streptococcus suis serovar II (strain Kasli), grundmanis and hyperimmunization increasing doses of antigen and subsequent kremovatam and selection of the target product, characterized in that as producers use oxen, while hyperimmunization spend 7-10 times lower intramuscular injection of a total dose of each antigen 1800-2400 billion microbial cells with an interval of 2-3 days, and grundmanis carried out subcutaneously in different parts of the body twice, first by 10-15 billion microbial cells, and after 14 days at 20-25 billion microbial cells by adding 10 cm3aluminium hydroxide.



 

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5 tbl

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5 tbl

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4 tbl

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3 tbl

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6 tbl

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6 tbl, 4 ex

FIELD: molecular biology, veterinary.

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