RU2021818C1 - Associated vaccine against hay fever and campylobacteriosis of cattle - Google Patents

Abstract

FIELD: biotechnology. SUBSTANCE: vaccine contains antigens of leptospiras of serological groups Pomona, Tarassovi, Grippotyphosa sejroe and antigens of campylobacteria of strains VGNKI No.43/3 and VGNKI No.169/17 selected in optimal relations, formalin as inactivator and aluminium hydroxide, sheep' blood serum protein and albumin as adjuvant. Vaccine produces highly-efficient immunity both against leptospirosis and campylobacteriosis of cattle. EFFECT: minimizing material losses incurred by the above diseases. 3 tbl

Description

 The invention relates to biotechnology and is an associated vaccine against leptospirosis and campylobacteriosis in cattle.

 After brucellosis, leptospirosis and campylobacteriosis of cattle are the most significant diseases causing reproductive dysfunction due to abortion in the second half of pregnancy, metritis, vaginitis, and sexual cycle disorders. Leptospirosis and campylobacteriosis often occur as a mixed infection.

 Chemotherapy of leptospirosis and campylobacteriosis is not effective enough, time-consuming and expensive. Therefore, the main and almost the only effective method of combating leptospirosis and campylobacteriosis is vaccination, which prevents both abortion and other clinical manifestations of the disease, as well as the carriage of the pathogen.

 Vaccination of cattle against leptospirosis is carried out with the multivalent vaccine VGNKI against leptospirosis of animals or the vaccine against leptospirosis of animals containing antigens from the leptospira strains of serogroups Pomona (VGNKI-6), Tarassovi (VGNKI-4), Grippotyphosa (Irhcti) VGNKI-2), Canicola (VGNKI-3), Hebdomadis (VGNKI-5), Sejroe (Hardjo), adjuvant, preservative and water [1].

 The use of these vaccines prevents leptospirosis in cattle and naturally does not create immunity against campylobacteriosis. The additional use of a vaccine against bovine campylobacteriosis for immunization of animals, containing a mixture of antigens from Campylobacter fetus subspecies intes - tinalis strains VGNKI NP-10/16 and subspecies fetus VGNKI N 43/3 and adjuvant [2], helps to increase the yield of calves, but not provides prevention of campylobacteriosis caused by Campylobacter jejuni in newborn calves. In addition, carrying out two vaccinations doubles the cost of labor and stretches the timing of vaccination, which significantly reduces its effectiveness in mixed infections.

 Also associated vaccine against leptospirosis and campylobacteriosis of cattle, including antigens of leptospira serogroups Pomona, Sejroe (Hardjo), Canicola, Grippotyphosa, Icterohaemorrhagiae and Campylobacter Vibrio fetus antigen, inactivator and aluminum hydroxide [3].

 The disadvantage of this vaccine is the inability to cause immunity to leptospira serogroup Tarassovi and Campylobacter jejuni due to their absence in the vaccine.

 The aim of the invention is to increase the immunogenic activity of the target product.

This is achieved by the fact that the vaccine contains the VGNKI-6 strain as leptospira serogroup antigens, the VGNKI-1 strain leptospira serogroup antigens, and the Tarassovi serogroup leptospira antigens-VGNKI-4 strain taken in the mixture in equal proportions at a concentration of 750 million microbial cells in 1 cm ^{3 of} phosphate buffer, and a mixture of antigens from Campylobacter fetus subsp. strains as campylobacter antigens. venerialis VGNKI N 43/3 and Campylobacter jejuni VGNKI N 169/17, taken in equal proportions at a concentration of 150 billion microbial cells in 1 cm ^{3 of} physiological saline, as formalin inactivator and additionally as an adjuvant, sheep serum protein and albumin in the following ratio of components. about.%:
A mixture of antigens from
leptospira strains
serogroups: Pomona
VGNKI-6, Tarassovi
VGNKI-4, Grippotyphosa
VGNKI-1, Sejroe "Hardjo",
taken in equal proportion
at a concentration of 750 million

microbial cells
in 1 cm ³ phosphate buffer 49.12-50.23
A mixture of antigens from
Gampylobacter strains
fetus subsp. venerialis
VGNKI N 43/3 and
Campylobacter jejuni
VGNKI N 169/17 taken
in equal proportion with
concentration of 150 billion

microbial cells in
1 cm ^{3 of} physiological solution 33.43-34.23 Aluminum hydroxide 14.00-16.00 Formalin 0.29-0.31 Serum protein 1.11-1.13 Albumin 0.05-0.10
The Campylobacter ferus subsp. Strain used to make the associated vaccine. jejuni VGNKI N 169/17 was previously used as a production one for the manufacture of diagnosticum and specific anti-Campylobacteriosis serum [4].

According to Bergey's "Sistematic Bacteriology" USA, 1984, v. 1, p. 114-115 [5], the names of the campylobacter strains used to make the associated vaccine are as follows:
instead of Campylobacter fetus subsp. fetus VGNKI N 43/3 - Campylobacter fetus subsp. venerialis VGNKI N 43/3;
instead of Campilobacter fetus subsp. jejuni VGNKI N 169/17 - Campylobacter jejuni VGNKI N 169/17.

PRI me R 1. Production strains of leptospira serogroups Pomona VGNKI-6, Tarassovi VGNKI-4, Grippotyphosa VGNKI-1, Sejroe "Hardjo" are grown separately in cylinders or reactors in a medium based on phosphate buffer enriched with sheep serum protein and albumin for 5-10 days before the accumulation of at least 75 million / cm ³ leptospira, the culture of leptospira is mixed in equal proportions, formalin is added to a concentration of 0.1-0.15%, incubated for 1-2 hours, concentrated by ultrafiltration on the UPV installation -6 to the content of 750 million microbial cells in 1 cm ³ phosphate buffer, add formalin to a final concentration in the medium of 0.23-0.27%.

Campylobacter fetus subsp. Campylobacter bacterial production strains venerialis VGNKI N 43/3 and Campylobacter jejuni VGNKI N 169/17 are grown separately on meat and peptone agar for 1-3 days. Formalinized culture is rinsed with saline at a concentration of 0.5% formalin and inactivate them for 3 days at ³⁷ ° C, adjusted concentration 150 Bln. / Cm ³ of the microbial cells, followed by mixing them in equal proportions.

 Antigens from leptospira strains are mixed with antigens from campylobacter strains in a ratio of 1.5: 1, 6% aluminum hydroxide gel is added.

 The resulting associated vaccine against leptospirosis and campylobacteriosis of cattle is packaged in bottles and controlled for appearance, the presence of impurities, the concentration of hydrogen ions, sterility, safety and activity.

 Get 3 series of the associated vaccine against leptospirosis and campylobacteriosis of cattle in the following ratio of components, vol.% (Table 1).

 The finished vaccine is a yellowish liquid in appearance with a loose precipitate that easily breaks when shaken into a homogeneous suspension.

 PRI me R 2. The activity of the associated vaccine against leptospirosis and campylobacteriosis series 1-3 is tested on rabbits weighing 3-3.5 kg in comparison with the vaccine against leptospirosis in cattle and with a mixture of formalized culture of campylobacter included in the associated vaccine . The rabbits are given once intravenously titrated minimal doses of the drugs, causing them a complete immune response. The serum of rabbits was examined after 23-25 days in PMA and RA to determine the titer of antibodies to leptospira and campylobacter. The results are presented in table.2.

 From the materials of the table. 2 it can be seen that the associated vaccine induces a more active immune response in animals, while different antigens do not inhibit each other, and leptospira have a synergistic effect with respect to campylobacter bacteria.

PRI me R 3. Conduct studies to determine the immunizing dose of the associated vaccine series 2 against leptospirosis and campylobacteriosis for cattle and immunity in vaccinated animals. In a farm that is safe for leptospirosis and campylobacteriosis, 30 pregnant cows are vaccinated with this vaccine. The vaccine is administered to animals once intramuscularly at doses of 4, 6 and 8 cm ³ , using 10 cows per dose, 10 cows (control) are vaccinated with the animal leptospirosis vaccine intramuscularly once at a dose of 2 cm ³ and 10 cows (control) - a mixture of formalinized campylobacter culture that are part of the proposed associated vaccine intramuscularly once at a dose of 6 cm ³ .

 The results are presented in table.3.

 From the data in Table 3 it can be seen that the associated vaccine against leptospirosis and campylobacteriosis in cattle causes the formation of more intense immunity in animals both to leptospirosis and campylobacter than individual antigenic preparations.

The associated vaccine against leptospirosis and campylobacteriosis of cattle is used on farms once intramuscularly in the following doses:
calves from two months of age and older - 4 cm ³ ;
cows and heifers - 6 cm ³ ;
manufacturing bulls - 10 cm ³ .

 Cows are vaccinated one month before coverage or in the first third of pregnancy. Animals are not subject to vaccination in the last month of pregnancy and within a month after calving.

 The use of the proposed vaccine in veterinary practice allows to minimize the material damage caused by leptospirosis and campylobacteriosis in cattle.

Claims (1)

ASSOCIATED VACCINE AGAINST LEPTOSPIROSIS AND CAMPYLOBACTERIOSIS OF CATTLE, Including Leptospira antigens of serogroups Pomona, Grippotyphosa and Sejros (Hardjo), Campylobacter antigens, serum inactivator and antioxidant are different from , as antigens leptospira serogroup Grippotyphosa strain VGNKI-1 and additional antigens leptospira serogroup TaraSSovi strain VGNKI-4, taken in a mixture in equal proportions at a concentration of 750 million microbial cells in 1 m ³ phosphate buffer as antigens Campylobacter - a mixture of antigens from strains of Campylobacter fetus Subsp venerialis VGNKI N 43/3 and Campylobacter jejuni VGNKI N 169/17, taken in equal proportions at a concentration of 150 billion microbial cells per 1 cm ³ of physiologic solution, in as an inactivator, formalin and, optionally, as an adjuvant, sheep serum protein and albumin in the following ratio of components, in vol.%:
A mixture of antigens from the serum group Letospira strains of Pomona serogroups - VGNKI-6, TaruSSovi VGNKI-4, Grippotyphosa VGNKI-1, Sejros (Hardjo) taken in equal proportions at a concentration of 750 million microbial cells in 1 cm ^{3 of} phosphate buffer 49.12 - 50.23
A mixture of antigens from strains of campylobacter fetus Subsp. venerialis VGNKI N 43/3 and Campylobacter jejuni VGNKI N 169/17 taken in equal proportions at a concentration of 150 billion microbial cells in 1 m ³ physiological saline 33.43 - 34.23
Aluminum hydroxide 14.00 - 16.00
Formalin 0.29 - 0.31
Sheep Serum Protein 1.11 - 1.13
Albumin 0.05 - 0.10

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