Serum against cattle diseases caused by viruses of infectious rhinotracheatis, paraflu, rota, corona and mucosa diarrhea-disease, polyspecific, hyperimmune, method of prevention and treatment of cattle diseases caused by viruses of infectious rhinotracheitis, parainfluenza, rota, corona and mucosa diarrhea-disease

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions relates to field of veterinary virology. Serum contains antibodies in titres: to rotavirus, virus of mucosa diarrhea-disease - not less than 1:1600 (in EIA), virus of infectious rhinotracheitis - not less than 1:16 (in NR), coronavirus and parainfluenz-3 virus not less than 1:152 (in IHAR). Serum is obtained by hyperiimunisation of oxen-producers with polyvalent viral antibody, containing mixture of culture viral suspensions of cattle rotavirus with titre 7.0-8.0 Ig TCD 50/ml, cattle coronavirus with titre 6.0-7.0 Ig TCD 50/ml, virus of cattle infectious rhinotracheitis with titre 7.5-8.0 Ig TCD 50/ml, parainfluenza-3 virus with titre 7.5-8 Ig TCD 50/ml, virus of mucosa diarrhea-disease with titre 4.5-9.5 Ig TCD 50/ml. With preventive purpose serum is introduced to a calf not later than 2-3 hours after birth and re-introduced after 24 hours, subcutaneously into several points in dose 1.0 cm3 per 1 kg of animal weight. In case if disease symptoms appear, serum is introduced to a calf in therapeutic doses 1.5 cm3 per 1 kg of weight, in combination with antibiotics subcutaneously in several points 1 time per day 2-3 days running.

EFFECT: group of inventions provides reliable passive immunity in extremely short terms and reduces the number of vaccinations.

2 cl

 

The present invention relates to the field of veterinary Virology, in particular to the manufacture of specific hyperimmune serum against disease of cattle caused by a virus of infectious bovine rhinotracheitis, parainfluenza, Rota, corona and diarrhoea diseases of the mucous membranes, and is intended for a specific immunization and passive prophylaxis of mixed forms of infectious enteritis and enterocolitis of newborn calves.

In the structure of morbidity of newborn calves gastrointestinal disease of viral etiology, clinically manifested by diarrhea, occupy a special place and are one of the main causes of death of young animals.

This group of diseases causing farms enormous economic damage resulting from the high mortality of the patients, the high cost of treatment, reduction in weight and holding of General and specific measures.

Without effective protection of newborn animals from infectious diseases is not possible to obtain healthy young and biologically valuable animal products.

Available in the literature data of recent years and the accumulated knowledge and experience testify that the mass enteritis and enterocolitis of newborn calves with clinical signs of diarrhea are caused once the ranks etiological factors: if the basis for the emergence of non-communicable diseases are functional disorders of the gastrointestinal tract of the newborn, closely related to metabolic disorders in cows-mothers due to inadequate feeding and violations of the rules of detention, for infectious diarrhoea huge role Rota-, corona-, herpes viruses (infectious bovine rhinotracheitis virus and so on), and to a lesser extent parovirus and virus diarrhea - diseases of the mucous membranes of cattle etc.

The infectious process is compounded by the fact that these agents act on the naked body of a newborn in combination with each other (virus + virus virus + bacteria and so on), causing a mixed infection, leading to more severe disease is often fatal.

Diarrhea of newborn calves of mixed etiology characterized by stationarity, severe and prolonged form of currents. Newborn calves infected in infected maternity wards in the first minutes of life with the first breath contaminated air and the first breath of infected milk, and then in 2-5 days, leading to their death against the background of progressive developing irreversible pathological changes.

The emergence and development of infectious process caused by the high concentration of livestock, latent persistence of pathogens in the body of cows and heifers, but cause acute disease in newborn calves.

Most rasprostranen the m and effective way to protect newborn calves from agents, involved in the pathogenesis of mixed infection is to increase the specific resistance of the body by soldering colostrum from vaccinated cows-mothers.

Development in recent years in many countries, including in the Russian Federation, helped to create mono-, bivalent and associated vaccines, causing the effect of immunity in calves to enteritis. Antibodies obtained from the colostrum of cows, protect calves from disease up until they will not develop their own mechanisms of immunity. For various reasons, some calves after birth does not receive the necessary amount of this natural protection, which is associated with insufficient intake of immunoglobulins from the colostrum of mothers and insufficient receive colostrum in the first hours after birth or violation of the mechanism of absorption of immunoglobulins in the colostrum intestine of newborn animals.

Therefore, only antibodies, entered calves oral, protect them from diseases, provide full protection from the development of clinically apparent disease and stop the excretion of viruses and bacteria.

Given this situation, with the specific purpose of passive prophylaxis and treatment in different countries practiced oral administration of hyperimmune serum or immunoglobulins obtained from hyperimmunizing animal-producers, gamma-globulin isolated from the colostrum of the first divania or from the blood serum of cattle.

The well-known "Method of obtaining adhesive and antitoxic serum against escherichiosis farm animals", which is the neutralization of pathogenic action of Escherichia animals of different species (1).

However, these dyes do not provide a therapeutic effect when mixed forms of infectious diarrhea of newborn calves.

As a rule, in 75-85% of cases are co-infected, and monozygote does not have specific activity against Rota-, corona-, herpes - major causative agents of this disease.

Also known poly-specific hyperimmune serum against Rota-, corona-, herpes viruses, and E. coli (K-99, And 20) for local protection and immunotherapy mixed forms of diarrhea of newborn calves.

The serum contains antibodies in the credits: rotavirus is not less than 1:3200 (ELISA), the coronavirus is not less than 1:512 (rnga), herpesvirus - 1:512 (PH), Escherichia having adhesin To 99 and 20 is not less than 1:1600 (RA), heat-stable enterotoxin of E. coli is not less than 1:4 (RDP).

Received hyperimmunization oxen-producers polyvalent viral antigen containing a mixture of culture viral suspensions, and inactivated antigens of E. coli, containing their somatic, adhesive (K-99, And 20) antigens in physiological solution, toxoids in the cultural medium (broth of Hottinger) in the following ratio of their components in two different immunogenic (vol.%):

1. Culture viral suspension for intravenous immunization:

Rotavirus cattle with title 107,0-107,830-35
TCC 50/ml
Coronavirus cattle with title 105,0-105,830-35
TCC 50/ml
Herpes virus of cattle with a titer of 107,6-10830-35
TCC 50/ml

2. Exericise antigens for subcutaneous immunization:

Somatic and adhesive antigen-9923-27
the concentration of 10-15 billion M.K. 1 cm3
Somatic and adhesive antigens And 2023-27
the concentration of 10-15 billion M.K. 1 cm3
The toxoid To-99 - 20 E. coli45-52
Formalin0,4-0,5

For the manufacture of viral antigens to hyperimmunization oxen-producers conduct separate breeding and cultivation of viruses, and the Rota-, corona - and herpes virus suspension containing at least 30-35 mg/ml of a specific protein are pooled in equal proportions and used to hyperimmunization oxen-producers.

The method of obtaining exericising antigens is as follows: get toxoid and somatic adhesive antigens, which are then mixed in equal volume ratios and used to hyperimmunization animals.

For hyperimmunization use polietilene consisting of Rota-, corona-, herpes viruses and Escherichia coli containing adhesive antigens To 99 and 20.

A mixture of antigens Rota-, corona-, herpes viruses injected four times in increasing amounts. For desensitization of the body of the immunized animals for 1-1 .5 h before intravenous them subcutaneously injected a mixture of viral antigens in a tenfold lower dose.

Esherihioznae antigen is injected subcutaneously in the neck region for the first time with incomplete hell is Vantomme's adjuvant, later, without adjuvant. Test blood sample is carried out from the jugular vein on the 20th day after the last injection of antigens. The resulting serum from each producer using serological reactions is examined for the presence of antibodies. Producers in serum which antibody titers are low, the antigen is administered repeatedly at doses according to a recent introduction, and the blood sample is not produced.

Production krovavaya permitted in the presence of antibodies in the serum producers to rotovirus not below 1:3200 in IFA; coronavirus - not less than 1:512 and rtha; herpesvirus - not less than 1:256 in PH; Escherichia - not less than 1:1600 in RA and, provided that the temperature in animals should not exceed 39,5°C, and after the preliminary exposure to them on a starvation diet for 12 h with unlimited watering. On the 20th day after blood collection producers enter polianthes in doses according to a recent introduction.

Evaluating the effectiveness of poly-specific hyperimmune serum is carried out in the acute experience in newborn calves.

For therapeutic purposes the serum used for sick calves intravenously in severe intoxication at a dose of 60-80 cm3at low - dose 40-60 cm3. If necessary, the serum is injected again after 24 hours subcutaneously at a dose of 50 cm3.

Prophylactic serum administered orally at a dose of 60-80 ml per reception is on the first portion of colostrum (the first 1-1 .5 hours life of the calf) and the re-introduction not later than 24-30 hours after the first in the same dose (2). (The prototype.)

Known associated vaccine against rotavirus, coronavirus calves and viral diarrhea is a disease of the mucous membranes of cattle, in which we used the following strains: strain RM (rotavirus) strain 2-CL-2 (cov), and the strain VK-V1 # 36 (virus diarrhea - diseases of the mucous membranes). All virus strains were isolated from disadvantaged households. The Russian Federation and adapted for reproduction in a variety of cell cultures (3).

The objective of the research was to develop technology for the production of poly-specific hyperimmune serum against disease of cattle caused by a virus of infectious bovine rhinotracheitis, parainfluenza, Rota-, corona - and diarrhea - diseases of the mucous membranes, providing passive immunity in the shortest time possible, and emergency treatment for developing disease, as well as reducing the number of inoculations with hyperimmunization and treatment.

This has been achieved thanks to the fact that hyperimmunization oxen-producers spend a culture suspension of the following composition with the introduction of the bulls:

Rotavirus cattle with a title
7,0-8,0 lg TCD 50/ml30-40
Coronavirus cattle with a title
6,0-7,0 lg TCD 50/ml30-40
The virus of infectious bovine rhinotracheitis
cattle with a title
6,5-7,0 lg TCD 50/ml20-30
AI virus-3 cattle with a title
7,5-8,0 lg TCD 50/ml20-30
Virus diarrhea - diseases of the mucous membranes with a title
4,5-6,5 lg TCD 50/ml30-40

Serum contains ready-made antibodies against the virus of infectious bovine rhinotracheitis, parainfluenza, Rota, corona and diarrhoea diseases of the mucous membranes and has a preventive effect and therapeutic effect, creates passive immunity that persists for up to three months.

Serum obtained from the blood of oxen (calves cattle or deer), which hyperimmunizing commercial vaccines against infectious bovine rhinotracheitis, parainfluenza, Rota, corona virus and diarrhea is a disease of the mucous.

Biopreparat, to make the serum,use the following commercial vaccine:

1. Vaccine against parainfluenza-3, infectious rhinotracheitis and bovine viral diarrhea - diseases of the mucous membranes of cattle emulsion inactivated production FGI ARRIAH.

2. Vaccine Rota-, coronavirus infections and viral diarrhea is a disease of the mucous cattle emulsion inactivated production ARRIAH.

3. Vaccine associated inactivated against rotavirus, coronavirus enteritis of calves and viral diarrhea in cattle "REDECOR-Supreme VIEW" production VIEW.

As producers use a healthy, have good fatness of herds (calves for fattening or reindeer) 2-5 years old, weight not lower than 300-400 kg

All incoming animals are subject to quarantine and screening for infectious diseases, according to the current regulations on the procedure for preparation and sanitary treatment of animals used for production of biologics.

Feeding oxen-producers must be balanced on the main parameters.

After the end of the quarantine period (30 days), if the animal meets the requirements, proceed to hyperimmunization according to the developed scheme.

Components for immunization
Scheme hyperimmunization
Immunization daysThe dose of antigensRoute of administration
1234
1 day1. Vaccine Rota-, coronavirus infections and viral diarrhea is a disease of the mucous cattle emulsion inactivated production ARRIAH10 ml from two sides (20 ml)Subcutaneously in the inguinal crease
2. Vaccine against parainfluenza-3, infectious rhinotracheitis and bovine viral diarrhea - diseases of the mucous membranes of cattle emulsion inactivated production ARRIAH10 ml from two sides (20 ml)Subcutaneously in the field predtopochnyh lymph nodes
3. (instead of 1) Vaccine associated inactivated against rotavirus, coronavirus enteritis of calves and viral diarrhea in cattle "REDECOR-Supreme VIEW" production VIEW.(instead of 1.) 10 ml from two sides (20 ml)Subcutaneously in the inguinal crease
14th day1. The vaccine is Rotel Rota-, coronavirus infections and viral diarrhea is a disease of the mucous cattle emulsion inactivated production ARRIAHas in day 1as in day 1
2. Vaccine against parainfluenza-3, infectious rhinotracheitis and bovine viral diarrhea - diseases of the mucous membranes of cattle emulsion inactivated production ARRIAH
3. Vaccine associated inactivated against rotavirus, coronavirus enteritis of calves and viral diarrhea in cattle "REDECOR-Supreme VIEW" production VIEW.
28-day1. Vaccine Rota-, coronavirus infections and viral diarrhea is a disease of the mucous cattle emulsion inactivated production ARRIAHas in day 1Subcutaneous
2. Vaccine against parainfluenza-3, infectious rhinotracheitis and bovine viral diarrhea - diseases of the mucous membranes of cattle emulsion inactivated production ARRIAH
42nd day1. Vaccine Rota-, coronavirus infections and viral diarrhea is a disease of the mucous cattle emulsion inactivated carried the STV ARRIAH as in day 1Subcutaneous
2. Vaccine against parainfluenza-3, infectious rhinotracheitis and bovine viral diarrhea - diseases of the mucous membranes of cattle emulsion inactivated production ARRIAH
3. Vaccine associated inactivated against rotavirus, coronavirus enteritis of calves and viral diarrhea in cattle "REDECOR-Supreme VIEW" production VIEW

During operation, pay special attention to the health of the producers. Daily clinical examination with measurement of body temperature of animals.

Trial krovavaya spend from the jugular vein into sterile test tubes. The resulting serum from each producer using serological reactions is examined for the presence of antibodies to the virus IRT - PH, rotavirus and virus diarrhoea diseases of mucous in ELISA, coronavirus and the AI virus-3 in RTG.

Production krovavaya permitted in the presence of antibodies in the serum producers to rotavirus and virus diarrhoea is a disease of the mucous in the ELISA is not lower than 1:1600, the virus RTIs in PH not lower than 1:16 and to the coronavirus and the AI virus-3 in RTG - not less than 1:512.

The body temperature of animals in the production Krasovskii should not exceed 39,5°C. Animals have to undergo pre is WriteLine on the starvation diet for 12 hours with unlimited watering.

Producers in the serum which antibody titers below, the blood sample is not produced.

The first krovavaya from producers by volume should not exceed 0.6 l of blood per 100 kg weight of the animal, and in the subsequent 1.6 liters per 100 kg of mass.

Every year doing a month long break in the exploitation of animals. On the 30th day of rest producers enter polianthes in doses according to the subsequent introduction.

Producers operate within 3-5 years, after which they implement in the prescribed manner.

Blood processing and obtaining serum.

To prevent blood from clotting used anticoagulant - 10%solution of sodium citrate (C6H5Na3O7·5H2O, M.V. 348,17), which is prepared by 5-7%sodium chloride solution and sterilized by toclafane.

To separate the formed elements of the blood obtained was centrifuged blood is subjected to separation (separator type AC-2ZH). At the initial stage of separation (within 10-15 minutes) the formed elements of blood are not completely separated from the plasma, and the first portion flows out of the separator with erythrocytes. Such a plasma is collected separately and sephirot again.

The resulting plasma defibrinated in sterile reactor, adding 30%solution of calcium chloride based 1.3 cm3to 1 liter of plasma.

Serum, conserve the comfort of a 5%solution GC phenol to a final concentration of 0.5% and with the aid of the vacuum pump is pumped into sterile containers, where stand for 2 months at 4-10°C.

After settling the serum is filtered and sterilized through a plate with the calculation of the load 5 l 1 plate.

Each batch of serum should the manufacturer take control according to THE current (or a HUNDRED).

Define the appearance of color, the presence of impurities, mold, etc.

Definition of sterility performed according to GOST 28065.

To check on the safety of a mixture of serum samples from 5 vials injected subcutaneously in a volume of 5.0 cm35 Guinea pigs. During 10 days of observation, the animals need to stay healthy.

Activity polyspecific sera with antigens of rotavirus and virus diarrhoea is a disease of the mucous determined in ELISA, virus IFF - PH, and against coronavirus virus and PG-3 - response inhibition of haemagglutination (rtga).

On each bottle with serum paste labels indicating: manufacturer, its trademark, the name of the serum, the number of its in a bottle, batch numbers and control, the date of manufacture, shelf life, storage conditions, and denote THE (one HUNDRED).

The order of application

The proposed serum used for the treatment and prophylaxis of acute gastrointestinal and respiratory sableman the th calves (birth to 9 months of age), caused by parainfluenza virus, infectious bovine rhinotracheitis, diarrhea, diseases of the mucous membranes, rotavirus and coronavirus cattle.

For the prevention of gastrointestinal and respiratory diseases calf serum no later than 2-3 hours after birth and again after 24 hours. The serum is injected subcutaneously at multiple points of the calculation 1.0 cm3on 1 kg of weight of animal.

In the case of the onset of symptoms of the digestive or respiratory calf injected the serum into a therapeutic dose (1.5 cm3on 1 kg of body weight) in combination with antibiotics subcutaneously at multiple points once a day for 2-3 days in a row.

Contraindications to the use of serum no.

Serum was tested with positive results in 20 regions of the Russian Federation from 2002 to 2010,

Serum will find wide application in all regions of the Russian Federation in animal husbandry for the prevention and treatment of gastrointestinal and respiratory diseases caused by Rota and corona viruses, virus diarrhea - diseases of the mucous membranes, viruses PG-3 and RTIs. The drug will dramatically reduce the above-mentioned diseases and thereby to preserve the health of the youngster, and get a successful breeding stock.

Sources of information

1. Patent RU NO. 2043772, AK, 39/42; AC 39/40, 1995

2. Patent RU NO. 2145504, AK 39/42, AK 39/40,1999

3. The dissertation on competition of a scientific degree of the candidate of vet. Sciences Belova NB Antigenic and protective properties associated vaccine against rotavirus, coronavirus enteritis of calves and viral diarrhea is a disease of the mucous membranes of cattle. M., 2005, p.40-45.

1. Serum against disease of cattle caused by a virus of infectious bovine rhinotracheitis (RTI), parainfluenza (PG-3), Rota, corona and diarrhoea diseases of the mucous membranes, poly-specific hyperimmune, characterized in that it contains antibodies in the credits: rotavirus, a virus diarrhoea is a disease of the mucous - not less than 1:1600 (ELISA), virus RTI - not less than 1:16 (PH), coronavirus and the AI virus-C not lower than 1:512 (rtga)obtained by hyperimmunization oxen-producers polyvalent viral antigen containing a mixture of culture viral suspensions rotavirus cattle with a title 7,0-8,0 lg TCD 50/ml, coronavirus cattle with a title 6,0-7,0 lg TCD 50/ml of the virus of infectious bovine rhinotracheitis in cattle with the title of 7.5 to 8.0 lg TCD 50/ml virus, PG-C with a title of 7.5-8 lg TCD 50/ml, virus diarrhea - diseases of the mucous membranes - with a titer of 4.5 to 9.5 lg TCD 50/ml

2. Method of prevention and treatment of diseases of cattle, caused by the virus of infectious bovine rhinotracheitis, parainfluenza, Rota, corona and diarrhoea diseases of the mucous membranes, characterized in that for the profilaktiki gastrointestinal and respiratory diseases calf injected serum, described in paragraph 1, not later than 2-3 hours after birth and again after 24 h, subcutaneously at multiple points of the calculation 1.0 cm31 kg weight of the animal, and in the case of the onset of symptoms calf injected the serum therapeutic doses of 1.5 cm3on 1 kg of weight in combination with antibiotics subcutaneously at multiple points 1 time per day 2-3 days in a row.



 

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22 cl, 11 ex

FIELD: medicine.

SUBSTANCE: method of preparing diagnostic agglutinating serum for pathogenic Yersinia strains involves hyperimmunisation of rabbits with antigen representing 0.4-0.6% formalin inactivated antigen (pYV+) of Yersinia enterocolitica My 03R strain into auricular cranial vein. Immunisation of rabbits with said antigen is fourfold in dosage as follows: 290-310 million kl/ml, 490-510 million kl/ml, 0.99-1.1 billion kl/ml and 1.9-2.1 billion kl/ml, respectively with dosage interval 6-7 days. Further the producer is examined for immunogenic properties. Serum separated from the sampled blood is preserved.

EFFECT: method ensures preparation of high-quality agglutinating serum used in yersiniosis diagnostics in animals.

3 ex

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

FIELD: veterinary science.

SUBSTANCE: invention refers to veterinary microbiology and biotechnology can be used for development of specific colibacillosis diagnostic aids. According to the invention the method covers producing antibody erythrocytic colibacillosis diagnosticum and provides carrier processing with antibodies to Escherichia antigens being adhesive antigens extracted from Escherichia cultures with phosphate-carbamide buffer 1.8-2.0 M of pH 7.2-7.4 at temperature 40-45°C during 25-30 min. The culture fluid containing Escherichia cell culture and phosphate-carbamide buffer are taken in mass ratio 1:0.4-0.6 respectively. The supernatant from the extraction is heated up at 65-68°C within 25-30 min. Gamma globulin fractions are used as serum antibodies for carrier processing.

EFFECT: method allows for high-quality end product due to improved sensitivity and specificity.

3 ex

FIELD: veterinary science.

SUBSTANCE: invention refers to veterinary microbiology and biotechnology, can be used for development of specific diagnostic aids. According to the invention the method covers producing antibody erythrocytic pasteurellosis diagnosticum by carrier processing with serum antibodies of Pasteurella antigens being capsular antigens extracted from Pasteurella cultures with 2.0-2.5% sodium chloride brine at 40-42°C during 30-40 min, with centrifuging the extract, separating the supernatant thereafter warmed up at 65-70°C during 15-30 min followed with sterilisation filtration. The serum antibodies for making diagnosticum are gamma globulin fraction or G or M immunoglobulins.

EFFECT: application of the method allows for high-quality end product due to improved sensitivity and specificity.

2 cl, 9 ex

FIELD: medicine microbiology.

SUBSTANCE: claimed method includes administering before contamination to the animal subsequently in right pope then after 14 days in left pope 0.2 ml of non-complete Freund's adjuvant with equal volume of physiological salt solution. Subsequent administering after 14 days in right pope up to 10 LD50 of Bacillus anthracis 81/1 spore dredge doesn't cause animal death for long period (monitoring time is 35 days).

EFFECT: immune serum for screening of anthrax diagnosis preparation.

1 dwg, 1 tbl, 8 ex

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