The remedy for the prevention of infectious laryngotracheitis birds

 

(57) Abstract:

The invention relates to veterinary pharmacology. The remedy for the prevention of infectious laryngotracheitis birds contains viral vaccine from strain "VNIIBT" and bioprotector - liposomal vesicles of soy lecithin is taken in the amount of 40 to 65 mg per 1 ml of the vaccine. Vesicles may contain soy lecithin, and/or alpha-tocopherol, and/or cholesterol. 14 days after a single immunization with the specified tool immune response to 4 - 7% higher compared to the prototype. 2 C.p. f-crystals.

The invention relates to medicine, in particular to the drugs used in veterinary medicine, and can be used for specific protection against infectious laryngotracheitis birds.

Infectious laryngotracheitis (ILT) is an acute contagious disease of birds - is a cause of high mortality rates, we recover individuals reduced daily gain and egg production. The complex responses to TLI one of the main tools is vaccination with killed and live VirusWall.

Known (Babkin C. F. and other Veterinary: Rep. mivd. thematic. Sciences. 3b. Kiev, 1975, vol.42, S. 14-19) inactivated emulsion vaccine by GCI, however in the case of poultry farms (hundreds of thousands) intramuscular vaccination too time-consuming.

Also known (Dutko Y. S., Shevchenko, A. A., Izotova M. N., Kisner A. T. veterinary medicine, 1991, N3, S. 32-34) dry virusvaktsinu "NT" strain "SNIIP", which use the group method, aerosol or oral application, or individual method (kloachnye). Dry vaccine diluted in distilled water. However, virusvaktsinu "NT" does not provide a sufficiently rapid engraftment and accumulation of virus in the mucosa of the larynx and trachea of birds.

The closest to the essence and the achieved therapeutic effect to the proposed tool is to prevent the TLI, including VirusWall from strain "VNIIBT" and bioprotector (Malosco centuries, Sokolov, C. D., Bull C. F., Cabanova L. P., saddle centuries Poultry, 1983, No. 12, S. 21-23). Virusvaktsinu against TLI birds from strain "VNIIBT" made from vaccinated extraembryonal fluid and chorioallantoic membranes of chicken embryos. As bioprotector known means contains peptone, food gelatin and sucrose, taken in the ratio 2:5:5, or with peptone sucrose, or lactose with peptone and skimmed cow's milk.

what creates the greatest protection against infection inexpensive and less time consuming. This tool provides rapid engraftment and accumulation of virus in the mucosa of the larynx and trachea, which gives better protection from diseases TLI, prevents acute outbreaks and clinical manifestations TLI.

However, the first introduction of this tool is manifested increased reactogenicity, accompanied by an increased mortality of Chicks, especially broilers.

The objective of the present invention is to reduce reactogenicity birds means to prevent the TLI.

This task is solved in that the means for prevention TLI birds, including VirusWall from strain "VNIIBT" and bioprotector, as bioprotector use of liposomal vesicles of soy lecithin is taken in the amount of 40-65 mg / 1 ml VirusWall.

Liposomal vesicles containing lipid membrane soy lecithin or soy lecithin by addition of 1-2% alpha-tocopherol, or soy lecithin, cholesterol and alpha-tocopherol, taken in the ratio (in g) 100:(30-35):(1-2).

Viral vaccine derived from 9-day-old chick embryos infected with a virus TLI birds strain "VNIIBT", sootvetstvenno in the invention, was in the range of 5.3 lg EID50- 7,5 lg EID50(EID - embryo infective dose).

Obtaining the claimed means described in the examples.

Example 1. In 10 ml of chloroform was dissolved 3 g of soya lecithin and dried 1 l round-bottom flask on a rotary evaporator to remove chloroform. Then formed in the flask film of lipids add 65 ml of viral masses, 28 ml of a 5% sorbitol and 7 ml of phosphate buffer (pH 7.6) and received a suspension resuspended at room temperature until a homogeneous suspension, which in volume of 3 ml is poured into ampoules. 1 ml of vaccine accounts for 46.2 mg of lipids.

The lyophilization is carried out after the preliminary freezing at -30 (-35)oC and further evaporation of the water solvent.

Example 2. In 10 ml of chloroform was dissolved 4 g of soya lecithin and 0.04 g of alpha-tocopherol. Next, as in example 1.

1 ml of the vaccine have to 62.2 mg of lipids.

Example 3. In 10 ml of chloroform was dissolved 2 g of soybean lecithin, 0,67 g cholesterol, 0.02 g of alpha - tocopherol and dried 1 l round-bottom flask on a rotary evaporator to remove chloroform. Then formed in the flask film of lipids add 65 ml wire temperature to obtain a homogeneous suspension, which volume of 3 ml is poured into ampoules. 1 ml of the vaccine makes up 41.4 mg of lipids.

Ligalizaziy carried out after pre-freezing and subsequent evaporation of the water solvent.

Example 4. The film of lipids obtained as in example 3, add 65 ml of viral masses, 28 ml of 5% sucrose and 7 ml of phosphate buffer (pH 7.6) and received a suspension resuspending and poured into vials as in example 1. 1 ml of the vaccine makes up 41.4 mg of lipids.

Example 5. In 10 ml of chloroform was dissolved 3 g of soybean lecithin, 0.06 g of DL-alpha-tocopherol and dried 1 l round-bottom flask on a rotary evaporator to remove chloroform. Then formed in the flask film of lipids add 120 ml of 5% sucrose and 30 ml of phosphate buffer (pH 7.6) and received a suspension resuspending, pour in a bottle of 250 ml and after pre-freezing at -30(-35)oC and evaporation of the water solvent lyophilizer.

Then into the vial containing the lyophilized liposomes add 65 ml of viral masses, 28 ml of 5% sucrose and 7 ml of phosphate buffer (pH 7.6) and received a suspension resuspending for 2-3 minutes at room temperature.

The resulting homogeneous suspension according to the norms of THE 9384-001 - 00495674-96. Solubility, residual moisture and sterility it meets SPECIFICATIONS.

The titer of the virus in all examples is equal 5,32-between 6.08 lg EID50/ml.

To determine thermal stability of the inventive tool and vaccine prototype kept at a 37.5oC for 7 days. The titer of the virus in the proposed drug decreased from of 5.82 lg EID50/ml to 5,39 lg EID50/ml; during this same time, the titer of the virus vaccine prototype decreased from 5.81 to 4,55 lg EID50/ml.

Test was conducted immunogenic properties of the proposed drug (vaccine with liposomes) in comparison with the means of the prototype. Vaccination was carried out in 2 groups of 20 chickens, and 20 chickens constituted the control group.

After a single immunization aerosol method claimed vehicle was not observed post-vaccination reactions in chickens. After a single immunization vaccine prototype 3 of 20 Chicks from the third to the eighth day was observed depression and loss of appetite.

After a single immunization claimed by means of the cellular immune response (E-ROCK) to the 9-th day after immunization was higher by 3.5% compared with the prototype; on day 14 after immunization, the percentage of phagocytosis was higher by 4-7%, phagocytic number 0 is of immunogenic properties of the proposed drug in comparison with the prototype 20 chickens after a single vaccination of the inventive agent and vaccine prototype, and 20 chickens of the control group were infected with field virus TLI (strain "24A") at a dose of 1000 EID500.2 ml intratrahealno. Observations were conducted within 10 days of the fallen were investigated to establish the cause of death.

Clinical signs of ILT in infected people appeared on the 4th - 5th day and was expressed in the depression and decreased activity when food intake. On 6-7 day in single patients had cough, neck stretch when inhaling.

In the group vaccinated with the claimed means, ill 2 of 20 chickens; in the group vaccinated with the vaccine prototype, ill 5 chickens, one of the number of cases fell in the control (non-vaccinated) group at 2-4 day ill 18 chickens, Palo with clinical signs of ILT - 3 chickens. At autopsy of dead chickens found hyperemia and edema of the larynx, hemorrhages on the mucous membrane of the larynx, strands and blood clots.

Was also held twice vaccination claimed means 34670 chicken egg breed at the age of 41-42 days (1 vaccination) and 54-55 days (II vaccination) aerosol method. Dose, aspirated Chicks at the first vaccination was 3 EID50when the second - 7,5 EID50. Post-vaccination reactions in chickens were not observed. Vaccination is not Pauli rnga).

1. The remedy for the prevention of infectious laryngotracheitis birds, including viral vaccine from strain "VNIIBT" and bioprotector, characterized in that as bioprotector tool contains liposomal vesicles of soy lecithin is taken in the amount of 40 to 65 mg per 1 ml of VirusWall.

2. Means under item 1, wherein the liposomal vesicles contain soy lecithin and alpha-tocopherol, taken in the ratio of 100 : (1 - 2).

3. Means under item 1, wherein the liposomal vesicles contain soy lecithin, cholesterol and alpha-tocopherol, taken in the ratio 100 : (30 - 35) : (1 - 2).

 

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