Vaccine against newcastle disease of birds
(57) Abstract:The invention is intended for oral immunization of poultry against Newcastle disease. Vaccine "ARAWAK-NB includes extraembryonal fluid of chicken embryos containing culture vaccine strain La Sota". The vaccine also contains sucrose, peptone, casein, resin KB-4P-2, Aerosil AM-1-300 and additionally casein at a certain ratio of components. The vaccine produced by the method of contact of dehydration. The vaccine is applied poultry 20-270 days old oral with feed at a dose of 0.25 g/bird. The vaccine is harmless, reactogenic, highly effective for birds (protection against virus NB 80-100%), as well as environmentally friendly for staff. The invention increases the efficiency of the prevention of Newcastle disease. 7 table. The present invention relates to the field of biotechnology viral veterinary preparations, in particular to a vaccine against Newcastle disease, intended for oral immunization of chickens.Known vaccine against Newcastle disease, containing the antigen of the vaccine strain, sucrose or lactose, casein or skim milk, levamisol, magnesium sulfate and dithiothreitol effective to who by its complexity, that largely complicates the process. In addition, it is characterized by a significant loss of activity in manufacturing. The latter is due to the presence of phase transitions in the environment when drying freeze the way it was received. At the same time a multi-stage production, the use of complex equipment and scarce raw materials leads to a deterioration of technical and economic indicators.Also known vaccine against Newcastle disease, including extraembryonal fluid of chicken embryos containing culture vaccine strain La Sota, sucrose, peptone, resin KB-4P-2 and Aerosil AM-1-300. The vaccine used for aerosol immunization of poultry (2).The disadvantage of this vaccine is the impossibility of its use for oral immunization due to its low efficiency due to the inactivating action of gastric juice on the antigen. In addition, a significant content (9% by weight) in the composition of the vaccine Aerosil is environmentally unsafe for staff.The aim of the invention is the development of a highly effective vaccine against Newcastle disease for oral immunization birds obtained by the method of Cohn is P CLASS="ptx2">Extraembryonal fluid of chicken embryos containing culture vaccine strain La Sota" - 12,5-15,0
sucrose - 4,5-6,0
peptone - 3,0-4,0
casein - 0,7-2,0
resin KB-4P-2 - 72,0-77,3
Aerosil AM-1-300 - 1,0-2,0
The invention is illustrated by the following examples.Example 1.For the manufacture of vaccines use extraembryonal fluid of chicken embryos infected with strain La Sota virus NB at the age of 9-10 days. In an apparatus of the type RT-1 is placed without pre-treatment of 12.5 to 15.0%, by weight, the obtained antigen, which is dissolved by weight of 4.5-6.0% sucrose, 3,0 - 4,0% peptone, 0.7 to 2.0% casein. The components are mixed in the apparatus is performed at 8000 rpm for 1.5-2.0 minutes. For dehydration vaccinated suspension is mixed with ionoobmennoi resin KB-4P-2, on the surface of which is deposited particles Aerosil AM-1-300, taken by mass in the total number of 73,0-79.3 percent. The mixture of these components is carried out in a spherical drum when the speed of rotation on the roller machine 60-90 rpm for 5-7 minutesCook 3 composition of the proposed vaccine is given in table 1.The finished vaccine is a loose gray powder. The vaccine Packed in flavie tubes and running-metal bottle caps and plastic bags tightly sealed. Shelf life 12 months if stored in dark and dry place at temperature not above plus 4-6oC.Example 2.Study of biological activity of 3 formulations of vaccines "Arawak NB of the vaccine strain La Sota", prepared according to example 1 and stored for 3 and 8 months at a temperature of minus 8-12oC 2-6oC and 18 - 20oC. Biological activity determined by titration of 10-day-old rack and expressed in lg EID 50 g-1. The specificity of the response, the rack is determined by the setting reaction of haemagglutination. As a control have known vaccine containing antigen from strain La Sota" in the amount of 15% of the volume.The results of the study on the biological activity of the prepared compounds is proposed and known vaccines are given in table 2.The data are shown in table 2, suggests that the contact dehydration using a vaccine strain La Sota" has received 4 composition of the dry vaccine with biological activity from 9.0 to 9.2 lg EID 50 g-1.Storage of 3 formulations of the vaccine within 3 and 8 months at minus 8-12oC and 2-6oC practically does not lead to inactivation of the virus in vaccinati virus 0,5-0,7 lg EID 50 g-1. The best stabilizing effect was observed in sample No. 3 in terms of temperatures minus 8-12oC and 2-6oC. Thus, storage at 18-20oC for 8 months resulted in a decrease in its activity only at 1.0 lg.The introduction of the proposed vaccine casein and reducing the number of input data does not significantly affect the biological activity.Example 3.Study the stability of the virus NB of different strains in the cooking process and storage at a temperature of 2-6oC for 1-12 months.In use Newcastle disease virus (VSL) industrial strains "And(C)NR", "V-4", "Clone-30", "Bor-74 VGNKI" and "La Sota". Dry preparations prepare contact dehydrated as described in example 1 using the composition 3.The stability of the biological activity of the virus vaccine strains in the composition of the dry preparations are also determined by thermotest - largest reduction of the infectivity titer of the virus in the course of storage at a storage temperature of 37-38oC for 7 days.The research results are summarized in tables 3, 4.In table 3 the results of the research indicate that the practice of 8.3 (units "And(C)NR"") to 9.3 lg EID 50 cm3-1(g-1) (units "La Honeycomb"). The difference in the value of the credits finished products is not due to the sensitivity of the virus vaccine strain, and the titre original vaccinated suspensions. When stored at 2-6oC for 12 months shows that the most resistant to the storage temperature was the strains of "V-4", "Clone-30", "La Sota". The biological activity of these drugs for 5 months storage at 2-6oC remains practically at the original level. At the same time, the titer preparations of strains And(b)NR" and "Bor-74 VGNKI" during this period reduced by 1,03 and 0.6 lg, respectively.As can be seen from the table, the most sensitive thermotest were the strains of "Clone-30" and "Bor-74 VGNKI": their titles 7 days of storage at a temperature of 36-38oC decreased by 0.48 and 1.0 lg respectively. Virus vaccine strain La Sota in contact the drug was found to be highly resistant to such storage mode. The titer of it in these conditions, the retention has not changed.Example 4.Investigate the safety of the oral form of 3 formulations of vaccines "Arawak NB, prepared according to example 1 from strain La Sota" by feeding their Chicks 20-270-day age of the drug in 10-fold Cove toxicity and residual virulence in all formulations of the vaccine. The clinical status of the experimental Chicks did not differ from control - intact.Example 5.Evaluation of the immunogenicity of the N 2 vaccine "Arawak-NB, prepared according to example 1 from strain La Sota", stored at a temperature of 2-6oC for 3 months, with biological activity 3,h8,0g-1spend 100 chickens - broilers 14 days old. The dry preparation of the vaccine is mixed with feed based 150-350 mg/goal with 20 g of feed. The dose of the vaccine is fed within 2 days once a day (in the morning, at the rate of 10 g of feed 1 goal). The effectiveness of immunization control after 14 and 28 days for antibody titers detected in rtga and control of infection vaccinated chickens culture of a virulent strain of the T-53 virus NB at a dose of 3.0 lg LD50.The results of immunization of broiler chickens composition N 2 vaccine "Arawak-NB are presented in table 5.The data of table 5 indicate that oral immunization with a composition of N 2 dry vaccine "Arawak-NB dose of 250-300 mg/goal provides reliable protection of chickens from the control of infection. Most suitable for use is a dose of 250 mg/goal.Example 6.Assessing Imprimer 4, 40 chickens-broilers 14-day age of 20 animals in each experimental group prophylactic against Newcastle disease. To immunize birds use a dose of 0.25 g/bird. On the 28th day of chickens in both groups serological study set the antibody titers in rtga 4,0 log2, as well as their absolute resistance to control contamination of the culture of a virulent strain NB T-53".Example 7.Conduct a study of the immunogenicity of the N 3 vaccine "Arawak-NB, prepared according to example 1 from strain La Sota", stored for 4 months at a temperature of 2-6oC biological activity of 8.47 lg EID 50 g-1. Tests are conducted in a safe in Newcastle disease pricecostco on the population of 75.5 thousand heads. 3-4 days prior to vaccination and for 5 days after it excludes cottage bird antibiotics, sulfa and nitrofuranovye drugs. Vaccinate only healthy birds. In pricecostco vaccine "Arawak-NB evenly mixed with the feed in existing faucets. The vaccine is fed to poultry 20-270-day age two consecutive days in the morning feeding rate of 1 dose of 0.25 and 0.5 g/bird. The vaccine "Arawak-NB using 2 - and 3-fold, and once n is impressive. The vaccine is fed to the chickens, watching a uniform distribution of the feed in the feeders and its palatability. After vaccination the birds are fed and watered as usual. Results immunization control 7-10, 14-16, 60-70, 120-130, 270-300 days exploring rtha 25-40 blood serum samples from chickens of each group. The results obtained are presented in table 6.Note: birds are vaccinated "Arawak-NB after two vaccinations vaccine NB from strain Bor-74" at the age of 22 and 120 days.As can be seen from table 6, the immune response of chickens fed the vaccine at a dose of 250 g /thousand heads, formed after 14-16 days with an average geometric titer of 4.9 log2 and tension immunity 66,6% and reaches optimum values after 2 weeks of reimmunization (91,9% and 6.9 log2). The introduction of the vaccine to chickens at the age of 120-130 days leads to a slight increase of the geometric mean antibody titer to 7.2-7.8 for log2 and increases the effectiveness of immunization to 100%. To the end-of-life birds (18 months) the level of antibodies is reduced to 3.6 log2 when the degree of protection to 79.2%. Increasing doses of the vaccine twice as reliable improvement is. The use of "Arawak-NB" in background 2 times intranasal immunization of poultry vaccine "Bor-74 VGNKI" ispolzovanie livestock. In chickens of the control group immunized with vaccine "Bor-74", dynamics of formation of immunity does not differ from Arawak-NB, although the absolute values vary in the range of 1.0 to 2.0 log2.Example 8.On the industrial poultry flocks of chickens on livestock in 149,2 thousand, housed in 2 buildings, conduct the test composition N3 vaccine "Arawak-NB, prepared according to example 1. The vaccine "Arawak-NB mixed with fodder ask twice aged 36-44 days and 120 days at a dose of 0.25 g/bird after preliminary intranasal vaccine from strain Bor-74 VGNKI" aged 13-22 days according to the standard technique. Control chickens vaccinated by intranasal vaccine from strain Bor-74 VGNKI" in accordance with the instruction on its application. Results immunization control at 14, 21 and 28 days, exploring rtha 25-30 serum samples from chickens each experimental group. Efficiency of combined use of the N 3 vaccine "Arawak-NB and vaccine strain "Bor-74 VGNKI" are presented in table 7.As follows from table 7, the immune response of chickens to the introduction of vaccines "Arawak-NB reaches its maximum after 2-3 weeks after vaccination. The level of antibodies at this varies 4,92-5,12 log2 efficiency and ITRI of antihemagglutinin, several decrease, but do not reach beyond the level (84-86% of 4.44-4,9 log2, respectively). A similar pattern can be observed in chickens of the control group vaccinated with traditional intranasal method, which virtually leveled to 14 days after the second vaccination.Vaccine "Arawak-NB from strain La Sota", made by technology contact method of dewatering for oral administration, are harmless to birds, not causes of post-vaccination reactions, delayed onset of lay and dynamics of growth of the productivity of chickens; viacommunity (protection against virus NB 80-100%) and environmentally safe for staff. Integrated use for immunization vaccines "Arawak-NB with other vaccines can reduce costs and improve the effectiveness of efforts to combat Newcastle disease. Vaccine against Newcastle disease, including extraembryonal fluid of chicken embryos containing culture vaccine strain La Sota, sucrose, peptone, resin KB-4P-2 and Aerosil AM-1-300, characterized in that it additionally contains casein in the following ratio, wt.%:
Extraembryonal liquid chicken embryo is Zein - 0,7 - 2,0
Resin KB-4P-2 - 72,0 - 77,3
Aerosil AM-1-300 - 1,0 - 2,0
FIELD: veterinary medicine.
SUBSTANCE: vaccine has antigenic material produced from La-Sota strain reproduced in 9-10 days old SPF-hen embryos having infectious activity of at least 9.7 lg EID50/cm3 and hemagglutination activity equal to at least 1:512, protective medium and adjuvant of thymogen taken in effective proportions. The protective medium comprises 20% lactalbumin hydrolyzate solution and degreased milk in 1:5 proportion. The vaccine is produced by mixing its ingredients and with following preparation lyophilization. Ready vaccine is dry porous mass of light yellow color. The vaccine is applicable for carrying out specific prophylaxis. The vaccine is introduced as spray at a dose of 1 cm3/head.
EFFECT: high antigenic and immunogenic activity; no adverse side effects.
7 cl, 8 tbl