Method of preparing immune multivalent serum against pneumoenteritis of young cattle of viral genesis

FIELD: medicine, veterinary medicine.

SUBSTANCE: invention relates to the field of veterinary medicine and biotechnology. The method includes selection of donors, selection of donor blood, holding at room temperature for clotting, separation of blood clot, serum treatment, placing in the refrigerator, checking on harmlessness and activity, packaging into sterile containers. At that a maternal and a closely related group of cattle are used as donors, who previously suffered from viral respiratory-intestinal infections, and the serum is pre-examined for antibodies presence to previously registered pneumoenteritis of viral genesis by raising the reaction of indirect hemagglutination. The blood is then incubated at room temperature for 1 hour and placed in a refrigerator at a temperature of 4-6C for 12-16 hours for complete retraction of blood clot. After checking of serum on harmlessness and activity the tanks are treated outside with 1% solution of "Zhavel-solid", uncorked in a sterile room, filtered, centrifuged and poured into sterile containers. Then the serum is irradiated with ultraviolet rays with a length of 254-256 nm during 1 hour and ozonised in a closed loop mode.

EFFECT: method does not use toxic substances, increases accuracy of determining activity of serum and accelerates duration of its preparation, and serum obtained with this method has a high specificity and sterility.

 

The technical field to which the invention relates.

The invention relates to veterinary medicine, in particular to methods for preparing multivalent immune serum against pneumoenteritis of young cattle (cattle) of viral origin and can be used for immune by using substitution immunotherapy of maternal antibodies and the creation of passive immunity in calves with severe lesions of the respiratory and intestinal infections of viral origin.

The level of technology

A method of obtaining immune serum, including immunization of animals, bleeding them for selection of serum, in which, to increase the output of the serum by the use of bone and muscle tissue of animals, after bleeding the animal carcass with remote internal organs and the skin is subjected to freeze-thawing, followed by grinding and the release of their blood. (see ed. St. SU # 533102, CL AC 35/16, AK 35/34, publ. 20.03.2000,)

The disadvantage of this method is the low efficiency of the immune serum.

A method of obtaining immune serum, including the introduction of antigen mixed with adjuvant-blockers in the popliteal lymph nodes and repeated immunization with antigen intramuscularly and intravenously, followed by separation of serum, that is, order to obtain monospecific serum to tahaawee acid staphylococci, as antigen used taikoubou acid staphylococci, which is injected with autologous blood treated with heparin (see ed. St. SU # 1775908, CL AC 39/395, publ. 15.08.1994 year).

The disadvantage of this method is the difficulty of obtaining the drug and the lack of effectiveness of the immune serum.

The closest in technical essence and the achieved positive effect and adopted by the authors for the prototype is a method for preparing immune multielemental serum against pneumoenteritis young cattle, consisting in the selection of donors, screening of blood donors, processing it and obtaining serum from blood with subsequent research on the safety and activity of extracted serum, the main stages of obtaining serum are: storage of blood at room temperature for 5-6 hours, storage of blood at 0-(+4) 48-72 hours, the suction serum in sterile containers, carbonizate phenol to a final concentration of 0.5%, sterilization by filtration through plate "SF", packaging, attirance, the control of sterility, safety, potency (see Methodological guidelines for the diagnosis and therapeutic interventions for viral respiratory diseases coarsely what about the cattle, The southern Department of agricultural Sciences, Institute of experimental veterinary medicine, Kharkov, 1979, p.41).

The disadvantages of this method are:

- application for conservation serum phenol solution, which according to its chemical characteristics, a highly toxic substance;

- when testing the serum on the activity of the application of several different types of serological reactions (RSC, RNVG, RZGA) with different timing and form of account credits;

selection of blood from donors of different age-sex groups (not related), containing antibodies that match only total virus-bacterial background of the economy, and virtually no specific related (maternal) antibodies optimally acceptable organism of the recipient.

Disclosure of inventions

The task of the invention is to develop a method of preparation of multivalent immune serum against pneumoenteritis young cattle viral Genesis with:

- high activity of specific antibodies;

- no toxic substances introduced into the body of the animal;

a precisely defined activity;

- accelerated period of preparation;

- accelerated period and the effectiveness of its actions in the body against opportunistic infections of viral origin;

- ease of preparation.

The technical result to the that can be obtained using the present invention, boils down to eliminating toxic substances, the accuracy of determining the activity of serum to accelerate the timing of its preparation, to improve the sensitivity in determining the presence and titer of antibodies to the disease.

The technical result is achieved by using the method of preparation of multivalent immune serum against pneumoenteritis young cattle viral origin, including the selection of donors, screening of donor blood, keeping at room temperature for coagulation, separation of blood clot, treatment serum, placing in the refrigerator, monitoring emissions and activity, filling sterile containers, at the same time as donors use a mother or closely related group of cattle, previously recovering from viral respiratory and intestinal infections, and previously studied blood and serum to identify or previously recorded pneumoenteritis viral origin by reaction of indirect haemagglutination, the blood is then incubated at room temperature for 1 hour and placed in a refrigerator at a temperature of 4-6C for 12-16 hours for full retraction blood clot, after testing capacity outside handle 1%solution "Zhavel-solid", uncork in a sterile room, centrifuged, Phil is trout and poured into sterile containers, then irradiated with ultraviolet rays with a length 254-256 nm for 1 hour and ozoniruyut blood in the closed loop.

Thus, the technical result is achieved due to the fact that in the method of preparation of multivalent immune serum as main components is used parent or sibling vysokomernoy serum from previously recovered viral respiratory and intestinal infections cows, and its components can be arbitrarily changed depending on what viral respiratory and intestinal infections dysfunctional one or another farm in the preparation of serum completely eliminated the use of chemical bactericidal drugs (such as thimerosal, phenol, etc.)than completely eliminates the toxicity of the drug, the use of the reaction of indirect haemagglutination, would allow, in the short term to determine the titer of antibodies immediately all respiratory gastrointestinal viral infection, which significantly reduces the time of preparation of serum, the presence of serum specific antibodies donor sibling groups, fast, optimal absorption of the drug by the body of the recipient.

The method of preparation of multivalent immune serum against pneumoenteritis of young cattle (cattle) viral Genesis is following who. The selection of the donor animals is carried out after a thorough epizootic survey of the economy and determine the cause of the disease of calves and the type of pathogen. Donors for the preparation of serum taken from mothers or closely related groups of animals, which contains the mother of the sick calves. The selection is conducted from the calculation of the titer of antibodies to the problematic disease. Having first studied blood and serum to identify or previously recorded pneumoenteritis viral origin by reaction of indirect haemagglutination, the blood is then incubated at room temperature for 1 hour and placed in a refrigerator at a temperature of 4-6C for 10-12 hours for full retraction blood clot, after retraction capacity outside handle 1%solution "Zhavel-solid", uncork in a sterile room, centrifuged, filtered and poured into sterile containers. Then passed through the apparatus ultraviolet irradiation and ozonation of blood "HOPE-ABOUT" with built-mercury-quartz lamps emitting ultraviolet short wave (80% 254 nm) in the closed loop. The procedure is performed with a pre-prepared sterile bottle (volume of 450 ml, filled with serum, 300 ml for replacement of air with oxygen while maintaining the sealed packaging), oxygen from the bottle through the Yu inline roller pump apparatus is pumped through the camera feed and returned into the vial. The method achieves a concentration of 1200 mg/l of ozone, with a strong bactericidal effect, for 20 minutes. Thus, sterility is achieved not only by the treatment of the serum with ozone, but additional ultraviolet radiation.

Furthermore, the residual ozone contained in the serum, with the introduction stimulates redox processes surrounding tissues than improves and accelerates its absorption in the body.

Examples of specific performance of the method of preparation of multivalent immune serum against pneumoenteritis of young cattle viral origin.

Example. The selection of the donor animals is carried out after a thorough epizootic survey of the economy and determine the cause of the disease of calves and type of the pathogen.

Donors for the preparation of serum taken from mothers or closely related groups of animals, which contains the mother of the sick calves. To do this, from clinically healthy cows receiving full medical examinations and the entire course of prevention research, pre-select the 10 ml of blood, and the serum examined by the method of reaction of indirect haemagglutination (rnga) detected or previously registered in the household pneumoenteritis viral origin.

Before the formation of the donor group of animals on achiev is Tatum studies is the selection of animals. The selection is conducted from the calculation of the titer of antibodies to the problematic disease. The higher the titer, and hence the higher the number in the serum, the more the animal is suitable for screening of donor blood. Calculation of required title, providing more or less stable immunity in the calf, rely on the presence of antibodies to a specific disease in a patient group of young and systemic immune background (to determine whether the circulation of these diseases on the farm at the moment). In this case, take into account not the amount of antibody titers in calves, and their presence, since most sick and weak calves humoral immunity is very low and the generation of antibodies may be small.

So, in the farms of the Stavropol territory has the highest percentage of calves identified antibody (descending) chlamydia to parainfluenza-3, respiratory syncytial infection and infectious bovine rhinotracheitis, to a lesser extent - to adenoviral infection. For previously registered in the farm viral diarrhea neither clinical signs nor antibodies in calves have been identified. Antibodies to the causative agent of viral diarrhea cattle in low titers were detected in several of the cows, which were also selected in the donor group.

Consequently, the mother of the donor are selected respectively to a specified list of diseases. Glastechnik viral diseases, which were previously registered in the data fields, but antibodies in calves is not detected, it was quite low titers of donor antibodies for therapeutic immune background.

A similar selection is carried out on any other problematic viral infections. When the selection in the group of clinically healthy, past the General clinical examination mothers donor animals were selected, each with high titers note at least two problematic diseases. After forming the group of donors conduct screening of donated blood within 800-1200 ml from one animal. Blood is taken with a sterile needle with a flexible polyurethane or silicone tube in sterile containers (vessels, banks) with a volume of 2000 cm3with a wide throat. The vessels fill no more than 2/3. Individuality selection does not adhere. The blood is allowed to stand for 1 hour at room temperature, then placed in a refrigerator at a temperature of 4-6C for 10-12 hours for full retraction blood clot, after retraction capacity outside handle 1%solution "Zhavel-solid", uncork in a sterile room, centrifuged, filtered and poured into sterile containers. Then passed through the apparatus ultraviolet irradiation and ozonation of blood "HOPE-ABOUT" with built-mercury-quartz lamps emitting ultrafiltation short range (80% - 254 nm) in the closed loop. The procedure is carried out with a pre-prepared sterile bottle (volume of 450 ml, filled with serum, 300 ml for replacement of air with oxygen while maintaining the sealed packaging), oxygen from the bottle using the built-in roller pump apparatus is pumped through the camera feed and return into the vial. The method achieves a concentration of 1200 mg/l of ozone, with a strong bactericidal effect, for 20 minutes. Then from each vial select 1-2 ml serum sterile syringe into a common tube. Selected national sample is examined for sterility in accordance with GOST 28085-89. At the same time put the bioassays on white mice to avoid toxicity. This same team of serum additionally investigated in rnga at all problematic viral infection. The overall title on every disease regularly recorded in the operating log. For immunization is used only sterile and non-toxic serum with optimal antibody titers. The use of multivalent immune serum for passive treatment-and-prophylactic immunization of young cattle, shown in the example.

Immune multivalent serum used for passive treatment-and-prophylactic immunization of calves with 3-4 weeks of age in therapeutic dose of 0.5-0.7 ml ha1 kg body weight of the calf. Injected subcutaneously once 1 time in 18-20 days to five to six months of age, depending on the form and severity of the disease, the General condition and weight of the animal, in prophylactic doses of 0.3-0.5 ml/kg, respectively. When complications of viral infection secondary bacterial flora multivalent serum used in combination with antibiotics after identifying bacterial PBA, determine its sensitivity with advanced poltically antibiotic. Serum prepared by the proposed method, used in some farms of the Stavropol region with a pronounced therapeutic effect, the systematic use it in to see b/C "Kuban" Kochubeevskogo area: after 3-4 injections of serum calves 1-2 months of age, mortality was reduced by 20%with the introduction of its patients 3-4-month-old calves - 30%, after regular use for 6 months to 60-70%by the end of the year the mortality of calves from viral respiratory and intestinal infections stopped altogether.

The present invention in comparison with the prototype and other known technical solution has the following advantages:

- complete elimination of contact with the drug in the organism of the recipient toxic substances;

- high activity of specific antibodies in the resulting serum;

- a single method and the accuracy of determining act is vnesti serum;

- faster and optimal absorption of the drug by the body of the recipient due to the oxygenation of body tissues and enhance metabolic processes in them on the injection of ozonated serum;

accelerated term and more effective therapeutic effect due to more elevated levels of antibodies against the causative agents of respiratory and intestinal infections of viral etiology;

- ease of preparation specified whey.

Method of preparation of multivalent immune serum against pneumoenteritis of young cattle viral origin, including the selection of donors, screening of donor blood, keeping at room temperature for coagulation, separation of blood clot, treatment serum, placing in the refrigerator, monitoring emissions and activity, packaged in sterile containers, characterized in that donors use maternal and closely related group of cattle, previously recovering from viral respiratory and intestinal infections, and pre-study serum for antibodies to the previously registered pneumoenteritis viral origin by setting the reaction of indirect haemagglutination, the blood is then incubated at room temperature for 1 h and placed in a refrigerator at temperature is 4-6C for 12-16 h for full retraction blood clot, after testing emissions and activity capacity outside handle 1%solution "Zhavel-solid", uncork in a sterile room, filtered, centrifuged and poured into sterile containers, then the whey is irradiated with ultraviolet rays with a length 254-256 nm for 1 h and ozoniruyut in the closed circuit.



 

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