RU2420571C1 - Vaccine and method of goose parvovirus infection vaccination - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: offered is a vaccine containing dimerethylenimine inactivated goose parvovirus antigen of Goosa Parvovirus Strain, VGNKI No.71, clone 6 of the activity not less 6.5-7.5 lg "ТЦД"₅₀/cm³ mixed with the oil adjuvant Montanide ISA-70, taken in the ratio 1:1-1:4; the dimerethylenimine concentration is not less than 0.1%. A method of goose parvovirus infection vaccination includes the intramuscular introduction of the vaccine preparation to a bird of a parental flock in a single dose 2.5·10⁴-2.5·10⁵ "ТЦЦ"₅₀/cm³ 30 days prior to the expected oviposition.

EFFECT: vaccine preparation is harmless for the birds, exhibits the evident antigenic and immunogenic activity, invokes an intense long immune response in geese for the entire reproductive period; the single application of the inactivated preparation is more technological and saving, reduces immune system stresses and loads in the birds.

2 cl, 2 tbl, 2 ex

Description

The invention relates to the field of veterinary virology, in particular to the production of vaccines for the specific prophylaxis of parvovirus infection of geese (geese enteritis virus), as well as to methods of treating and preventing this disease.

Parvovirus infection of geese (viral enteritis of geese) is an acute contagious disease of young geese and musky ducks, which characterizes damage to the gastrointestinal tract, liver, heart, pancreas and other parenchymal organs, and is associated with high mortality (from 30 to 100%). The causative agent is parvovirus, which belongs to the family Parvoviridae.

The disease is widespread in European countries (Germany, France, England, Hungary, Czechoslovakia) and on other continents. In the Russian Federation, it is registered in many farms in various zones.

The most effective way to combat PVI geese is vaccination along with the use of citrate blood and blood serum of geese convalescents and hyperimmune serums obtained on various biological models.

A known vaccine against geese viral enteritis and a method for the prevention of geese viral enteritis (1).

The vaccine contains culture fluid, Goosa parvovirus strain VGNKI No. 71, clone 6 with biological activity 10 ^6.0 -10 ⁷ TCD ₅₀ and a protective drying environment at a ratio of culture fluid and protective environment 1: 1-1.0: 1.5. A method for the prevention of the disease involves the introduction of the proposed vaccine to goslings of 1-2 days of age at a dose of 50-70 thousand TCD ₅₀ in 0.50-0.75 cm ³ or to adult geese twice, 50-30 days before the laying of a dose of 75- 100 thousand TCD ₅₀ in 0.75-1.00 cm ³ .

However, this vaccine, even when applied twice, does not create prolonged intense immunity in birds for the entire reproductive period.

A well-known inactivated vaccine against geese viral enteritis developed at the Institute of Poultry UAAS (2). However, this vaccine contains a pathogenic strain of parvovirus XM-99 isolated in Ukraine. In addition, when developing a vaccination scheme for parent geese herds using this inactivated biological product, it was tested both in combination with a live vaccine (14 days after vaccination with a live vaccine) and independently, using it once or twice with an interval of 14 days. However, to confirm the immunizing effect of the use of a single or double vaccination with an inactivated vaccine, further studies are necessary, according to the authors.

A vaccine against the viral disease of geese caused by parvovirus is known, which is a prototype of the claimed invention, which contains a pathogenic virus inactivated with β-propiolactone, which does not exclude the possibility of its reactivation (3).

The problem to be solved within the framework of the claimed group of inventions is the creation of an inactivated vaccine against parvovirus infection of geese, used in industrial goose breeding, farms, creating long-term intense immunity in geese of the parent herd for the entire reproductive period, as well as an economical and technological way of vaccination against parvovirus infection geese, reducing the number of vaccinations in order to reduce stress and load on the bird's immune system while maintaining harmlessness, antigenic activity and storage duration of the vaccine preparation.

The problem is solved by creating a vaccine containing the dimeretyleniminin inactivated goose parvovirus antigen of the Strain Goosa Parvovirus VGNKI No. 71, clone 6 with an activity of 6.5-7.5 lg TCD _{50 /} cm ³ in a mixture with oil adjuvant Montanide ISA-70, taken in the ratio 1: 1-1: 4, and the concentration of dimethylenimine is at least 0.1%.

The vaccination method against parvovirus infection of geese includes intramuscular administration of the claimed vaccine preparation to the parent flock bird once 30 days before the expected egg laying in a dose of 2.5 · 10 ⁴ -2.5 · 10 ⁵ TCD ₅₀ / cm ³ . As a rule, the indication for the use of the vaccine is a disadvantage in farms with parvovirus infection of geese.

The strain Goosa parvovirus VGNKI No. 71, clone 6 is used to prepare vaccines against geese viral enteritis (4).

The strain is deposited in the collection VGNKI of veterinary drugs at number 71. Vaccine strain No. 71 obtained by selection. The strain has the following morphological, physiological and cultural characteristics.

Virions are small spherical with a size of 20 ± 2 nm, the nucleocapsid contains DNA. In cell culture, agar plating induces the formation of plaques with even edges 1-2 mm in size. The virus reproduces in the culture of embryos goose cells, causing acute form of infection with a pronounced cytopathic effect after 3-4 days in titers 6.5-7.0 logarithm TCD _50/05 ml. The strain is apatogenous for developing goose embryos, goslings, laboratory mammals when administered parenterally in doses 100 times higher than the vaccine, does not have hemagglutinating and hemadsorbing activity, is resistant to ether and chloroform, can withstand temperatures of 60 ° C for 60 minutes, has complete antigenic affinity with the virulent virus causing the disease in goose farms.

The content and quantitative ratio of the ingredients in the vaccine is dictated by its functional features.

Changing the ratio of antigen and adjuvant in the direction of increasing the amount of antigen entails a reduction in the shelf life of the vaccine and an excessive consumption of antigen, which is not economically viable. With a higher content of adjuvant with respect to the antigen, the activity of the vaccine decreases and insufficient immunity is generated to protect against the disease.

To obtain a vaccine, the components included in its composition are mixed in a homogenizer for 5-10 minutes at a speed of 3000 rpm.

The vaccine is a homogeneous emulsion of a pinkish-whitish color with high stability during storage at t = + (2-8) ° C.

The claimed method of vaccination of birds is the introduction of the aforementioned inactivated vaccine into the body of a bird intramuscularly once in a dose of 2.5 · 10 ⁴ -2.5 · 10 ⁵ TCD ₅₀ / cm ³ .

As a rule, the indication for the use of the vaccine is a disadvantage for parvovirus infection of geese. All clinically healthy birds of the parent herd are vaccinated 30 days before the start of the reproductive period. The drug is administered intramuscularly in a volume of 0.5 cm ³ / head.

The essence and practical applicability of the claimed group of inventions is illustrated by the following examples.

Example 1: Preparation of inactivated vaccine in various ratios of 1: 4; 1: 2.33; 1: 1.5; 1: 1

The vaccine against parvovirus infection of geese is prepared according to the standard method for producing inactivated emulsion preparations. 400, 233, 150 and 100 volumes of Montanid ISA-70 oil adjuvant, respectively, are added to 100 volumes of inactivated antigen, respectively. The resulting mixture is thoroughly mixed and poured into bottles, stored at a temperature of + (2-8) ° C.

The vaccine was used to prevent parvovirus infection of geese in the conditions of the experimental base of the GNU VNIVIP.

Immunogenic activity was monitored by detecting specific antibodies to goose parvovirus in the neutralization reaction and by enzyme immunoassay according to the Guidelines for the laboratory diagnosis of viral enteritis (parvovirus infection) of geese by enzyme immunoassay (5, 6).

The obtained test results are presented in table 1.

Table 1 The effect of the ratio of Ag: Hell on the immunological parameters of the vaccine (antibody titer in ELISA) No. p / p Number of birds Ag: Hell ratio Antibody titer before vaccination Antibody titer after vaccination on (days) Immune bird,% 28 42 one 10 1: 4 414 ± 31 3521 ± 67 3476 ± 136 80 2 10 1: 2.33 414 ± 31 11299 ± 92 10220 ± 61 one hundred 3 10 1: 1,5 414 ± 31 12050 ± 132 11825 ± 125 one hundred four 10 1: 1 414 ± 31 1200 ± 142 10382 ± 252 one hundred 5 5 the control 414 ± 31 414 ± 31 414 ± 31 0 Note: Ag is an antigen; Hell - adjuvant

As follows from the above results, the optimum ratio of antigen and adjuvant is 1: 2.33. The drug is harmless to poultry, has a pronounced antigenic and immunogenic activity.

Example 2

The resulting vaccine was used in the conditions of the experimental base of the GNU VNIVIP. To prevent parvovirus infection of geese, the vaccine preparation was administered in a volume of 0.5 cm ³ / head once at a dose of 2.5 · 10 ⁴ -2.5 · 10 ⁵ TCD ₅₀ / cm ³ intramuscularly in the body of the bird of the parent herd 30 days before the start of reproductive period. The obtained test results are presented in table 2.

table 2 The effect of various doses of the vaccine on immunological parameters No. p / p Number of birds The dose of the vaccine, TCD ₅₀ / 0.5 cm ³ ELISA antibody titer * Immune bird,% before vaccination 28 days after vaccination 42 days after vaccination one 10 2.5 · 10 ⁴ 414 ± 31 7011 ± 67 8671 ± 41 one hundred 2 10 2.5 · 10 ^4.5 414 ± 31 9030 ± 72 8390 ± 82 one hundred 3 10 2.510 ⁵ 414 ± 31 11299 ± 92 10220 ± 61 one hundred four 10 the control 414 ± 31 414 ± 31 414 ± 31 0 Note: * - the value of the titles in inverse values

As follows from the above data, the vaccine in tested doses causes intense immunity in geese.

The drug in a 5-fold dose (2.5 cm ³ / goal.) Is harmless to poultry. A single use of an inactivated vaccine compared to a live virus vaccine (1) is more economical and technological and causes long-term immunity for the entire period of laying.

Information sources

1. Patent of the Russian Federation No. 21118539, 09/10/1998.

2. Development of a vaccination scheme for geese against viral enteritis using an inactivated vaccine / A.V. Beletskaya et al. // Achievements in modern poultry farming: research and innovation. Materials of the XVI Conference of VNAP. Sergiev Posad, 2009 .-- S.336-337.

3. French patent No. 7406981, 1974.

4. SU No. 1499917, 10/15/1994.

5. Guidelines for the laboratory diagnosis of viral enteritis (parvovirus infection) of geese by enzyme-linked immunosorbent assay / B. B. Trefilov, N. V. Nikitina, D. Maslov. - SPb., 2005. - P.12.

6. Patent of the Russian Federation No. 2323743, 10.12.2007.

Claims (2)

1. A vaccine against parvovirus infection of geese containing an inactivated parvovirus antigen and adjuvant, characterized in that it contains a goose parvovirus antigen inactivated by dimethylenimine, of the strain Goosa Parvovirus VGNKI No. 71 clone 6 with an activity of 6.5-7.5 lg TCD ₅₀ / cm ³ in a mixture with oil adjuvant Montanide ISA-70, taken in a ratio of 1: 1-1: 4, and the concentration of dimethylenimine is at least 0.1%. 2. The method of vaccination against parvovirus infection of geese by introducing a vaccine preparation into the body of a bird, characterized in that the vaccine used is the vaccine obtained according to claim 1, which is administered to the bird of the parent herd once 30 days before the intended egg laying intramuscularly at a dose of 2, 5 · 10 ⁴ -2.5 · 10 ⁵ TCD ₅₀ / cm ³ .