Vaccine against animal colibacillosis

FIELD: medicine; veterinary science.

SUBSTANCE: vaccine includes inactivated adhesive antigens E.coli K88, E.coli K99, E.coli F-41, E.coli 987P and an adjuvant - aluminum hydrate. As the inactivated adhesive antigens E.coli use inactivated filamentous adhesive antigens E.coli K88 ab, E.coli K88 ad, E.coli K88 ad, E.coli K99, E.coli F-41, E.coli Att-25, E.coli 987P with activity level in reaction of the passive hemagglutination peer as 1:2048-4096; 1:1024-2048; 1:256-512; 1:1024-2056; 1:1024-2056; 1:32-64; 1:256-512, accordingly, at a following parity of components, wt. %: inactivated filamentous adhesive antigens E.coli K88 ab, E.coli K88 ad, E.coli K88 ad, E.coli K99, E.coli F-41, E.coli Att-25, E.coli 987P, taken in mass parities 1:(0.8-1.2:(0.8-1.2:(0.8-1.2:(0.8-1.2:(0.8-1.2:(0.8-1.2, accordingly - 55-65, 5.8-6.2%-s' aluminium hydrate the rest.

EFFECT: rising of safety of livestock of agricultural animals.

2 cl, 1 tbl, 8 ex

 

The invention relates to veterinary Microbiology and biotechnology, can be used in the development of specific prevention, and, in particular, funds for the fight against colibacillosis animals.

Known vaccine escherichiosis animals, including adhesive antigens of E. coli K88, E. coli K99, E. coli F-41, E. coli 987P and adjuvant is aluminum hydroxide (RF Patent No. 2043771, "Vaccine escherichiosis animals", bull. No. 26, 20.09.1995, MKI AC 39/108). In addition, the vaccine contains thermostable and thermolabile toxoids, somatic antigen and capsular polysaccharide antigens in the form of a cell suspension of Escherichia broth of Hottinger and phosphate-urea buffer. The vaccine used twice intramuscular intervals 14-20 days 5 cm3for injection.

The disadvantages of the known vaccine is the low quality of the target product, identified with low antigenic and immunogenic activity of strains of Escherichia, and the absence of immunity from incomplete antigenic composition of E. coli strains with adhesive antigen included in the vaccine. In addition, the vaccine has increased reactogenicity due to the content of highly toxic strains of serogroups O78, O141, designed to create immunity to the O-antigens and the introduction of this vaccine in a large dose of 10 cm3in one animal (two is dy 5 cm 3), which leads to sensitization of the animal organism.

Objective of the claimed technical solution is to improve the quality of the target product by expanding its antigenic spectrum and the enrichment of its main protective adhesive antigens, increasing their antigenic and immunogenic, and reduce non-specific load on the immune system of the animals and consequently reduce reactogenicity.

The problem is solved in the vaccine escherichiosis animals, including inactivated adhesive antigens of E. coli K88, E. coli K99, E. coli F-41, E. coli 987P and adjuvant is aluminum hydroxide, the fact that as inactivated adhesive antigens of E. coli using inactivated fimbrial adhesive antigens of E. coli K88 ab, E. coli K88 AC, E. coli K88 ad, E. coli K99, E. coli F-41, E. coli Att-25, E. coli 987P with the level of activity in the reaction of the passive haemagglutination equal to 1:2048-4096; 1:1024-2048; 1:256-512; 1:1024-2056; 1:1024-2056; 1:32-64; 1:256-512 respectively, in the following ratio, wt.%:

inactivated fibreline
adhesive antigens of E. coli K88 ab
E. coli K88 AC, E. coli K88 ad
E. coli K99, E.
coli F-41, E. coli Att-25,
E. coli 987P, taken in mass
ratios
1:(0,8-1,2):(0,8-1,2):(0,8-1,2):(0,8-
1,2):(0,8-1,2):(0,8-1,2) respectively55-65
6%aluminum hydroxiderest

The problem is solved in the vaccine escherichiosis animals that additionally contains formalin at a final concentration of 0.12 to 0.15%.

It is known to obtain and use fimbrial of adhesins to obtain antisera (U.S. patent No. 4769240, CL AC 39/02, 1988, RF patent №2077337, AK 39/02, bull. 11, 20.04.1997).

In the patent and scientific literature is not well-known technical solutions containing elements similar to the claimed, i.e. the proposal meets the criterion of "novelty".

The proposal was feasible in laboratory and industrial conditions, aimed at solving real technical problem, i.e. the sentence "industrially applicable".

Existing vaccines escherichiosis animals made from bacterial mass Escherichia grown without regard to the protective characteristics of the components of the cells of the pathogen that are functionally related to pathogenicity factors. Reactor cultivation of Escherichia until the stationary phase of bacterial growth is carried out without taking into account the definition of the Oia maximum level of specific activity, save this activity and distribution in the growth medium protective fimbrial of adhesins. The cultivation process is not controlled for factors of pathogenicity mode. We first established that in obtaining maximum yield of biomass, as it is currently adopted in industrial production of vaccines against escherichiosis animals, decreases as antigenic material, as upon reaching a certain phase of the development of culture under the action of its own enzymes cells in the aging process it happen destructive changes of the components parts fimbrial of adhesins that leads to loss of immunologically active material and unacceptable in the production of an effective vaccine against escherichiosis animals. In addition, we found that cultivation of Escherichia containing adhesive K88 antigens ab, K88 AC, K88 ad, K99, F-41, Att-25, R allow you to get the vaccine, it is coincident with the epizootic strains of the pathogen colibacillosis animals.

First proposed a method of obtaining a vaccine escherichiosis animals, in which, instead of defining criterion of the number of cells of Escherichia physico-chemical and biological parameters, as well as for evaluating the content of adhesive antigens, the maximum level of production of antigens is determined by the level of activity of the Phi is briannah of adhesins Escherichia reaction passive haemagglutination in the culture fluid, released from the cell Bakassi that allows you to get obvious positive effect of improving the quality of the target product and the acceleration of the method, i.e. the proposal meets the criteria of "novelty" and "inventive step".

The method is illustrated by the following examples.

Example 1.

To obtain vaccine escherichiosis animals selected seven production of E. coli strains containing adhesive K88 antigens ab, K88 AC, K88 ad, K99, F-41, Att-25, R (strains No. 729, 661/16, 798, 957, 688/19, 4 and 745), which is grown in a broth of Hottinger containing amine nitrogen to 150 mg%, tryptophan 50 mg%, at pH of 7.8. Each strain is grown separately. In every culture in the process of its growth determine the maximum activity fimbrial of adhesins Escherichia, which in the sample of culture by centrifugation at 3000 rpm for 20 min to separate the culture fluid from the bacterial mass. Bacterial mass resuspension 2 M phosphate-urea buffer with pH 7.4, the cell concentration was adjusted to 100 billion microbial cells in 1 ml and heated at a temperature of 60°C for 20 minutes. Next, a suspension of microbial bodies cooled to 5°C and centrifuged at 13,000 rpm for 20 minutes the resulting supernatant used for determination of activity fimbrial of adhesins in the reaction of the passive haemagglutination. After 5 hours under the Oia at the maximum value of credits adhesins Escherichia, containing fimbrial adhesive K88 antigens ab, K88 AC, K88 ad, K99, F-41, Att-25, 987P, as equal 1:2048; 1:1024; 1:256; 1:1024; 1:1024; 1:32; 1:256 accordingly, the cultivation of the above strains ceased. Separate the bacterial mass by centrifugation at 3000 rpm for 20 min, which resuspension 2 M phosphate-urea buffer with pH 7.4, the cell concentration was adjusted to 100 billion microbial cells in 1 ml and heated at a temperature of 60°C for 20 minutes. Next, a suspension of microbial bodies cooled to 5°C and centrifuged at 10,000 rpm for 20 minutes resulting supernatant each strain can formalin at a final concentration of 0.3%, then concentrate in 3 times and mixed in a mass ratio 1:0,8:0,8:0,8:0,8:0,8:0,8 respectively. Further to 55 grams of a mixture of inactivated fimbrial adhesive antigens of E. coli K88 ab, E. coli K88 AC, E. coli K88 ad, E. coli K99, E. coli F-41, E. coli Att-25, E. coli 987P add 45 g of 5.8%aluminum hydroxide, receiving structure 1, in the following ratio, wt.%:

inactivated fibreline
adhesive antigens of E. coli K88 ab, E. coli K88 AC
E. coli K88 ad, E. coli K99, E. coli F-41,
E. coli Att-25, E. coli 987P,
taken in the mass ratios
1:0,8:0,8:0,8:0,8:0,8:0,8 respectively55
5,8%aluminum hydroxiderest

Example 2.

To obtain vaccine escherichiosis animals selected seven production strains of E. coli containing adhesive K88 antigens ab, K88 AC, K88 ad, K99, F-41, Att-25, 987P (strains No. 729, 661/16, 798, 957, 688/19, 4 and 745), which is grown in a broth of Hottinger containing amine nitrogen to 150 mg%, tryptophan 50 mg%, at pH of 7.8. Each strain is grown separately. In every culture in the process of its growth determine the activity of fimbrial of adhesins Escherichia, which in the sample of culture by centrifugation at 3000 rpm for 20 min to separate the culture fluid from the bacterial mass. Bacterial mass resuspension 2.1 M phosphate-urea buffer with a pH of 7.3, the cell concentration was adjusted to 120 billion microbial cells in 1 ml and heated at a temperature of 65°C for 25 minutes Then suspended microbial cells is cooled to 4°and centrifuged at 15,000 rpm for 25 min. and the resulting supernatant used for determination of activity fimbrial of adhesins in the reaction of the passive haemagglutination. After 6 h of cultivation at the maximum value of credits fimbrial of adhesins Escherichia containing adhesive K88 antigens ab, K88 AC, K88 ad, K99, F-41, Att-25, 987P, as equal 1:4096; 1:2048; 1:512 1:2056; 1:2056; 1:64; 1:512 accordingly, the cultivation of the above strains ceased. Separate the bacterial mass by centrifugation at 3000 rpm for 20 min, which resuspension 2.1 M phosphate-urea buffer with a pH of 7.3, the cell concentration was adjusted to 120 billion microbial cells in 1 ml and heated at a temperature of 65°C for 25 minutes. Next, a suspension of microbial bodies cooled to 4°and centrifuged at 15,000 rpm for 25 minutes Received supernatant each strain can formalin at a final concentration of 0.5%, then concentrate in 4 times and mixed in a mass ratio 1:1,0:1,0:1,0:1,0:1,0:1,0 respectively. Next, 60 g of a mixture of inactivated fimbrial adhesive antigens of E. coli To 88 ab, E. coli 88 AC, E. coli To 88 ad, E.coli K, E. coli F-41, E.coli Att-25, E. coli 987P add 40 g of 6%aluminum hydroxide receiving part 2, in the following ratio, wt.%:

inactivated fimbrial adhesive
antigens of E. coli To 88 ab, E. coli To 88 speakers
E. coli To 88 ad, E.coli K, E. coli F-41,
E. coli Att-25, E. coli 987P, taken in mass
ratios 1:1,0:1,0:1,0:1,0:1,0:1,0 respectively60
6%aluminum hydroxidethe steel

Example 3.

To obtain vaccine escherichiosis animals selected seven production strains of E. coli containing adhesive antigens To ab 88, 88 AC, 88 ad, C, F-41, Att-25, 987P (strains No. 729, 661/16, 798, 957, 688/19, 4 and 745), which is grown in a broth of Hottinger containing amine nitrogen to 150 mg%, tryptophan 50 mg%, at pH of 7.8. Each strain is grown separately. In every culture in the process of its growth determine the activity of fimbrial of adhesins Escherichia, which in the sample of culture by centrifugation at 3000 rpm for 20 min to separate the culture fluid from the bacterial mass. Bacterial mass resuspension in 2,05 M phosphate-urea buffer with a pH of 7.35, the cell concentration was adjusted to 110 billion microbial cells in 1 ml and heated at a temperature of 62.5°With over 22,5 minutes Later suspended microbial cells is cooled to 3°C and centrifuged at 10,000 rpm for 22 minutes the resulting supernatant used for determination of activity fimbrial of adhesins in the reaction of the passive haemagglutination. After 6 hours of cultivation at the maximum value of credits fimbrial of adhesins Escherichia containing adhesive K88 antigens ab, K88 AC, K88 ad, K99, F-41, Att-25, 987P, as equal 1:4096; 1:2048; 1:512; 1:2056; 1:2056; 1:64; 1:512 accordingly, the cultivation of the above strains ceased. Separate the bacterial mass zentrifugenbau is receiving at 3000 rpm for 20 min, which resuspension 1.9 M phosphate-urea buffer with a pH of 7.35, the cell concentration was adjusted to 110 billion microbial cells in 1 ml and heated at a temperature of 62°C for 22 minutes. Next, a suspension of microbial bodies cooled to 3°C and centrifuged at 13,000 rpm for 20 minutes resulting supernatant each strain can formalin at a final concentration of 0.3%, then concentrated 5-fold and mixed in a mass ratio 1:1,2:1,2:1,2:1,2:1,2:1,2 respectively. Further to 65 g of a mixture of inactivated fimbrial adhesive antigens of E. coli K88 ab, E.coli K88 AC, E. coli K88 ad, E.coli K99, E. coli F-41, E. coli Att-25, E. coli 987P add 35 g of 6.2%aluminum hydroxide receiving part 3, in the following ratio, wt.%:

inactivated fimbrial adhesive
antigens of E. coli K88 ab, E. coli K88 AC
E. coli K88 ad, E.coli K99, E. coli F-41,
E. coli Att-25, E. coli 987P, taken in mass
ratios 1:1,2:1,2:1,2:1,2:1,2:1,2 respectively65
6,2%aluminum hydroxiderest

Example 4.

To obtain vaccine escherichiosis animals selected seven production strains of E. coli containing adhesive antigen is K88 ab, K88 AC, K88 ad, K99, F-41, Att-25, 987P (strains No. 729, 661/16, 798, 957, 688/19, 4 and 745), which is grown in a broth of Hottinger containing amine nitrogen to 150 mg%, tryptophan 50 mg%, at pH of 7.8. Each strain is grown separately. In every culture in the process of its growth determine the maximum activity fimbrial of adhesins Escherichia, which in the sample of culture by centrifugation at 3000 rpm for 20 min to separate the culture fluid from the bacterial mass. Bacterial mass resuspension 2 M phosphate-urea buffer with pH 7.4, the cell concentration was adjusted to 100 billion microbial cells in 1 ml and heated at a temperature of 60°C for 20 minutes. Next, a suspension of microbial bodies cooled to 5°C and centrifuged at 13,000 rpm for 20 minutes the resulting supernatant used for determination of activity fimbrial of adhesins in the reaction of the passive haemagglutination. After 5 h of cultivation at the maximum value of credits adhesins Escherichia containing fimbrial adhesive K88 antigens ab, K88 AC, K88 ad, K99, F-41, Att-25, 987P, as equal 1:2048; 1:1024; 1:256; 1:1024; 1:1024; 1:32; 1:256 accordingly, the cultivation of the above strains ceased. Separate the bacterial mass by centrifugation at 3000 rpm for 20 min, which resuspension 2 M phosphate-urea buffer with pH 7.4, the cell concentration was adjusted to 100 billion microbial cells in 1 is l and is heated at a temperature of 60° C for 20 minutes. Next, a suspension of microbial bodies cooled to 5°C and centrifuged at 10,000 rpm for 20 minutes resulting supernatant each strain can formalin at a final concentration of 0.3%, then concentrate in 3 times and mixed in a mass ratio 1:0,8:0,8:0,8:0,8:0,8:0,8 respectively. Further to 55 grams of a mixture of inactivated fimbrial adhesive antigens of E. coli K88 ab, E. coli K88 AC, E. coli K88 ad, E. coli K99, E. coli F-41, E. coli Att-25, E. coli R add 45 g of 5.8%aluminum hydroxide, getting composition, in the following ratio, wt.%:

inactivated fimbrial adhesive
antigens of E. coli K88 ab, E. coli K88 AC
E. coli K88 ad, E.coli K99, E. coli F-41, E. coli Att-25,
E.coli R taken in mass
ratios 1:0,8:0,8:0,8:0,8:0,8:0,8 respectively55
5,8%aluminum hydroxiderest

Next add formalin at a final concentration of 0.12%, receiving part 4.

Example 5.

To obtain vaccine escherichiosis animals selected seven production strains of E. coli containing adhesive K88 antigens ab, K88 AC, K88 ad, K99, F-41, Att-25, R (strains No. 729, 661/16, 798, 957, 688/19, 4 and 745), who grow the broth of Hottinger, containing amine nitrogen to 150 mg%, tryptophan 50 mg%, at pH of 7.8. Each strain is grown separately. In every culture in the process of its growth determine the activity of fimbrial of adhesins Escherichia, which in the sample of culture by centrifugation at 3000 rpm for 20 min to separate the culture fluid from the bacterial mass. Bacterial mass resuspension 2.1 M phosphate-urea buffer with a pH of 7.3, the cell concentration was adjusted to 120 billion microbial cells in 1 ml and heated at a temperature of 65°C for 25 minutes Then suspended microbial cells is cooled to 4°and centrifuged at 15,000 rpm for 25 min. and the resulting supernatant used for determination of activity fimbrial of adhesins in the reaction of the passive haemagglutination. After 6 h of cultivation at the maximum value of credits fimbrial of adhesins Escherichia containing adhesive K88 antigens ab, K88 AC, K88 ad, K99, F-41, Att-25, 987P, as equal 1:4096; 1:2048; 1:512; 1:2056; 1:2056; 1:64; 1:512 accordingly, the cultivation of the above strains ceased. Separate the bacterial mass by centrifugation at 3000 rpm for 20 min, which resuspension 2.1 M phosphate-urea buffer with a pH of 7.3, the cell concentration was adjusted to 120 billion microbial cells in 1 ml and heated at a temperature of 65°C for 25 minutes. Next, a suspension of microbial bodies cooled to 4°and the center of pageroot at 15,000 rpm for 25 minutes The resulting supernatant each strain can formalin at a final concentration of 0.5%, then concentrate in 4 times and mixed in a mass ratio 1:1,0:1,0:1,0:1,0:1,0:1,0 respectively. Next, 60 g of a mixture of inactivated fimbrial adhesive antigens of E. coli K88 ab, E.coli K88 AC, E. coli K88 ad, E.coli K99, E. coli F-41, E. coli Att-25, E. coli 987P add 40 g of 6%aluminum hydroxide, getting composition, in the following ratio, wt.%:

inactivated fimbrial adhesive
antigens of E. coli K88 ab, E. coli K88 AC
E. coli K88 ad, E.coli K99, E. coli F-41,
E. coli Att-25, E. coli 987P,
taken in the mass ratios
1:1,0:1,0:1,0:1,0:1,0:1,0 respectively60
6%aluminum hydroxiderest

Next add formalin at a final concentration is 0.135%, receiving part 5.

Example 6.

To obtain vaccine escherichiosis animals selected seven production strains of E. coli containing adhesive K88 antigens ab, K88 AC, K88 ad, K99, F-41, Att-25, 987P (strains No. 729, 661/16, 798, 957, 688/19, 4 and 745), which is grown in a broth of Hottinger containing amine nitrogen to 150 mg%, tryptophan 50 mg%, at pH of 7.8. It is jdy strain grown separately. In every culture in the process of its growth determine the activity of fimbrial of adhesins Escherichia, which in the sample of culture by centrifugation at 3000 rpm for 20 min to separate the culture fluid from the bacterial mass. Bacterial mass resuspension in 2,05 M phosphate-urea buffer with a pH of 7.35, the cell concentration was adjusted to 110 billion microbial cells in 1 ml and heated at a temperature of 62.5°With over 22,5 minutes Later suspended microbial cells is cooled to 3°C and centrifuged at 10,000 rpm for 22 minutes the resulting supernatant used for determination of activity fimbrial of adhesins in the reaction of the passive haemagglutination. After 6 hours of cultivation at the maximum value of credits fimbrial of adhesins Escherichia containing adhesive K88 antigens ab, K88 AC, K88 ad, K99, F-41, Att-25, 987P, as equal 1:4096; 1:2048; 1:512; 1:2056; 1:2056; 1:64; 1:512 accordingly, the cultivation of the above strains ceased. Separate the bacterial mass by centrifugation at 3000 rpm for 20 min, which resuspension 1.9 M phosphate-urea buffer with a pH of 7.35, the cell concentration was adjusted to 110 billion microbial cells in 1 ml and heated at a temperature of 62°C for 22 minutes. Next, a suspension of microbial bodies cooled to 3°C and centrifuged at 13,000 rpm for 20 minutes resulting supernatant each strain conse wirhout formalin at a final concentration of 0.3%, next, concentrated 5-fold and mixed in a mass ratio 1:0,8:0,8:0,8:0,8:0,8:0,8 respectively. Further to 65 g of a mixture of inactivated fimbrial adhesive antigens of E. coli K88 ab, Esoi K88 AC, E. coli K88 ad, E.coli K99, E. coli F-41, E. coli Att-25, E. coli 987P add 35 g of 6.2%aluminum hydroxide, getting composition, in the following ratio, wt.%:

inactivated fibreline
adhesive antigens of E. coli K88 ab
E. coli K88 AC, E. coli K88 ad
E. coli K99, E. coli F-41,E. coli Att-25, E. coli 987P,
taken in the mass ratios
1:0,8:0,8:0,8:0,8:0,8:0,8 respectively65
6,2%aluminum hydroxiderest

Next add formalin at a final concentration of 0.15%, receiving part 6.

Example 7.

Used compositions obtained in the following way. Form 4 groups of 100 pregnant sows were selected according to the principle of analogues. Table 1 shows the technical data of vaccines, obtained according to known (prototype) and the proposed (compounds 1-6, obtained according to examples 1-6) technical solutions.

As shown by the results of experimental studies proposed the vaccine may escherichiosis animals can improve the safety of the animals of 29.5-29.8 per cent.

Example 8.

Used compositions obtained in the following way. Form 4 groups of 50 pregnant cows that were selected according to the principle of analogues. Table 2 shows the technical data of vaccines, obtained according to known (prototype) and the proposed (compounds 1-6, obtained according to examples 1-6) technical solutions.

As shown by the experimental results, the proposed vaccine escherichiosis animals can improve the safety of the animals of 6.8 to 11.3%.

As shown by the experimental results, the proposed vaccine escherichiosis animals can improve the safety of the animals, as well as reducing the time required for vaccine escherichiosis animals for 18-42 hours, because the duration of cultivation according to a known method is 24-48 hours (in the proposed method is 5-6 hours).

Table 1.
IndicatorsComposition 1Part 2Part 3Part 4Part 5Part 6The placeholder
Vaccinated pregnant sows100100100100100 100100
The number of piglets obtained from pregnant sows1250128012861264127513091230
The number of pigs cases of colibacillosis156135164172152142386
The number of piglets that died from escherichiosis30283234283285
Livestock safety97,6of 97.897,597,3of 97.897,668
Table 2.
IndicatorsComposition 1Part 2Part 3Part 4Part 5Part 6The placeholder
Vaccinated pregnant cows50505050505050
The number of calves obtained from pregnant cows47464846454845
The number of calves, diseased ash is richison 1091089821
The number of calves that died from escherichiosis3322216
Livestock safety93,6for 93.495,895,695,597,986,6

1. Vaccine escherichiosis animals, including inactivated adhesive antigens of E. coli K88, E. coli K99, E. coli F-41, E. coli 987P and adjuvant - aluminum hydroxide, characterized in that as inactivated adhesive antigens of E. coli used inactivated fimbrial adhesive antigens of E. coli K88 ab, E.coli K88 AC, E.coli K88 ad, E.coli K99, E. coli F-41, E.coli Att-25, E. coli 987P with the level of activity in the reaction of the passive haemagglutination equal to 1:2048-4096; 1:1024-2048; 1:256-512; 1:1024-2056; 1:1024-2056; 1:32-64; 1:256-512, respectively, in the following ratio, wt.%:

inactivated fimbrial adhesive antigens
E.coli K88 ab, E.coli K88 AC, E.coli K88 ad, E.coli K99,
E. coli F-41, E.coli Att-25, E. coli 987P
taken in the mass ratios
1:0,8-1,2):(0,8-1,2):(0,8-1,2):(0,8-1,2):
(0,8-1,2):(0,8-1,2), respectively55-65
5.8 and 6.2%aluminum hydroxiderest

2. Vaccine escherichiosis animals according to claim 1, characterized in that it further comprises formalin at a final concentration of 0.12 to 0.15 wt.%.



 

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1 tbl

FIELD: medicine; pharmacology.

SUBSTANCE: agent possessing wound-healthing, antiinflammatory, antibacterial, immunomodulating, anaesthetising and antitumoral activity on a basis of terpenoids, contains a capsule extract of plants of Pinaceae bloodline exposed to short-term stressful influence, enriched with monoterpenes, obtained from a capsule extract. The pharmaceutical composition possessing wound-healthing, antiinflammatory, antibacterial, immunomodulating, anaesthetising and antitumoral activity, contains the above described agent in effective quantity and the target additive. Application of the above described agent for preparation of a medicinal preparation for treatment of pyoinflammatory diseases.

EFFECT: increased wound-healthing; antiinflammatory; antibacterial; immunomodulating; anaesthetising and antitumoral activities.

8 cl, 1 dwg, 6 tbl, 8 ex

FIELD: medicine.

SUBSTANCE: agent contains Rifabutin sorbated in polymeric nanoparticles matrix, potassium cholesterylphosphate, or sodium glycocholate, or hexadecyl dihydrogen phosphate, or a-tocopheryl succinate, water-soluble polymeric stabiliser and bulking agents. Polymeric nanoparticles sized 100-800 nm include lactic acid polymer/polymers and/or lactic and glycolic acid copolymer/copolymers at glycolic acid content in specified copolymers up to 50 mole %. Molecular weight of specified polymers and copolymers is 5 to 300 kDa. Molecular weight of water-soluble polymeric stabiliser is no more than 70 kDa and is selected from the group including polyvinyl alcohol, polyvinylpyrrolidone, polysorbate and seralbumin.

EFFECT: new agent provides durable action of Rifabutin; higher bioavailability of Rifabutin and efficiency of bacterial infection treatment.

3 dwg, 1 tbl, 7 ex

FIELD: medicine; veterinary science.

SUBSTANCE: preparation consists of viral-bacterial polyvalent blood serum for diarrhea with antibody titre 1:300-1:600 and sodium hypochlorite solution concentrated 300-350 mg/l at ratio of serum: sodium hypochlorite 7:3. Additionally contains serum maral blood preparation in ratio 2:1, where 2 portions are serum maral blood preparation. Method implies that preparation is injected in dosage 4 ml preventively. Therapeutically preparation is injected intramuscularly in dose 4-5 ml per 10 kg 2 or 3 times every 48 or 48, 72 hours respectively.

EFFECT: medical product and application method allow for lowered consumption of hyperimmune serum while preserving high medical preventive efficiency.

4 cl, 4 tbl, 1 ex

FIELD: medicine, microbiology.

SUBSTANCE: invention concerns nucleotide sequence coding TolC, and also to certain amino-acid sequence which is built in in permissive, located from lateral aspect of a membrane area of TolC. In the invention is described a plasmid, containing a nucleotide sequence, for an expression of the merged protein or merged peptide. The invention concerns also the transformed bacterium, in particular enterobacteria, to its use as a part of a pharmaceutical composition for stimulation of the immune response and in a diagnostic set for detection of interesting substance in an organism. The framed transport system provides high efficiency of presentation of a product of an expression in an external cellular membrane of a bacterium.

EFFECT: provision of high efficiency of presentation of product of expression in external cellular membrane of a bacterium.

13 cl, 1 dwg, 5 ex

FIELD: veterinary.

SUBSTANCE: escherichia coli cells containing adhesive gene K 88 ab, K 88 ac, K 88 ad, K 99, F-41, Att-25, 987P are cultivated in liquid nutrient medium to produce vaccine. Physical, chemical and biological indicators are investigated in the process of bacterial bulk growth. In addition, activity of fimbrial adhesin in passive hemagglutination of cellular bacterial bulk is determined. Further cultivation is terminated when fimbrial adhesins titers values are K 88 ab, K 88 ac, K 88 ad, K99, F-41, Att-25, 987P equal to: 1:2048-4096; 1:1024-2048; 1:256-512; 1:1024-2056; 1:1024-2056; 1:32-64; 1:256-512, accordingly. Biological bulk is then concentrated and spinned at 3000-5000 rev/min for 20-25 minutes to separate culture broth from cellular bacterial bulk. Then vaccine is inactivate.

EFFECT: improvement of target product quality.

2 tbl, 5 ex

FIELD: veterinary medicine.

SUBSTANCE: it is necessary to select the affected organs from dead nutrias out of local epizootic focus, prepare the suspension out of pathological material to inoculate onto differential-diagnostic media, isolate pure cultures of Escherichia coli, Salmonella typhimurium, Streptococcus pneumoniae and Streptococcus fecalis, grow separately the isolated cultured in beef-extract nutritive medium at addition of 0.2%-glucose to achieve the concentration of microbial cells of about 4-5 bln/cu/ cm, inactivate with formalin up to 0.4-0.6%-final concentration, keep at 37°C for about 72-96 h, mix the cultures at equal ratios, add 3%-aluminum hydroxide solution at the quantity of 20% against the volume of the culture to be thoroughly mixed. Then the vaccine obtained should be packed and sealed. The innovation is simple in usage, moreover, it enables to increase specificity and immunogenicity of the obtained vaccine.

EFFECT: higher efficiency.

1 tbl

FIELD: veterinary medicine.

SUBSTANCE: the vaccine suggested contains as antigens: the cell suspension of pure cultures of Escherichia coli, Salmonella typhimurium and Streptococcus fecalis obtained due to selecting the affected organs from dead nutrias out of local epizootic focus, preparing the suspension, inoculation onto differential-diagnostic media, isolating pure cultures of the agents of Escherichia coli, Salmonella typhimurium and Streptococcus fecalis. The obtained pure cultures of microorganisms should be separately grown in beef-extract broth with glucose up to the concentration of microbial cells being 4-5 bln/cu. cm to be then mixed at equal ratios. The vaccine, also, contains formalin, glucose and aluminum hydroxide at the following ratio of the components, weight%: cell suspension of Escherichia coli pure culture isolated from the affected organs out in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0; cell suspension of Salmonella typhimurium pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0, cell suspension of Streptococcus pneumoniae pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu/ cm 18.0-21.0 cell suspension of Streptococcus fecalis pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0, glucose 2.0-1.0; formalin 2.0-1.5; aluminum hydroxide - the rest. The vaccine in question is of high specificity, safety and immunogenicity.

EFFECT: higher efficiency.

5 ex, 1 tbl

FIELD: veterinary science.

SUBSTANCE: down-calving cows should be immunized simultaneously with vaccine "Coli-Vac C99" and formulated alum vaccine against salmonellosis in calves at 10-d-long interval 1.5 mo before calving. The first dosage corresponds to 10 cu. cm, the second one - 15 cu. cm. Simultaneously with vaccination it is necessary to inject Ligavirin for cows at the dosage of 5 ml/cow. As for calves born in these cows they should be once injected intramuscularly with Ligavirin for 0.5-1 h after their birth at the dosage of 1 ml/calf. The innovation enables to create more tense specific immunity in cows before calving and colostral immunity in calves born in these cows.

EFFECT: higher efficiency of prophylaxis.

3 ex, 5 tbl

FIELD: medicine, pediatrics, otorhinolaryngology.

SUBSTANCE: the present innovation deals with conservative treatment and rehabilitation in children in case of chronic adenoiditis (CA) and in ARVI-suffering ones. It is necessary to isolate the following groups of patients: with CA and purulent exudates, at CA at the background of allergic rhinitis and at CA at the background of intracellular infections. One should carry out conservative therapy along with washing a patient's nasopharynx and differentiated therapy for each group of patients. Therapy includes the intake of mucoregulating preparations, and differentiated therapy for patients with CA and purulent exudates additionally includes local antibacterial therapy with applying Bioparox for 7 d. For patients at CA at the background of allergic rhinitis - application of local antihistamine preparations or mast cells stabilizers. For patients at CA at the background of intracellular infections - two courses of systemic antibacterial therapy with macrolides and Bioparox locally followed by introducing interferonogens or interferon inductors. In about 7-8 d after the onset of therapy course all the patients should undergo 10 seances of quantum therapy with "Rikta" apparatus along with the intake of antioxidants, 4 times annually it is important to conduct rehabilitation with IRS-19, with biopreparation "Narine" endonasally for 10 d, inside - bifidumbacterin per 3 wk by alternating with "Narine". The innovation provides individual curative program for patients with CA of different groups at stable clinic effect of decreased hypertrophy of pharyngeal tonsil up to degree I and restoring microbiocenosis and values of local immunity in nasopharynx.

EFFECT: higher efficiency of therapy.

2 tbl

FIELD: medicine.

SUBSTANCE: method involves intranasally administering immunomodulator preparation of bacterial origin with one dose placed in each nasal passage twice a day during 14 days repeated in obligatory way every 3 months during one year.

EFFECT: stable repair of injured immunity chains; hindered fixation and reproduction of pathogenic organisms.

FIELD: medicine.

SUBSTANCE: method involves making instillations and applications with Cholisal gel 2-3 times a day. Imudon is introduced at a dose of 6 pills a day during 20 days. Polysorb MP is introduced at a dose of 1 tea spoon diluted with 100 ml of water once a day during 20 days.

EFFECT: enhanced immunotropic, detoxifying effectiveness together with bactericide and anti-inflammatory action; increased organism resistance.

3 tbl

FIELD: veterinary microbiology, biotechnology.

SUBSTANCE: the innovation deals with developing live avirulent vaccine against colibacteriosis in calves for intramuscular injection, being simple in manufacturing and application. The vaccine is being the suspension and contains an antigen "B-5" out of E. coli strain as an antigen at the content of 1.0x1010-1.2x1010 microbial cells/cu. cm physiological solution. The vaccine is avirulent, nonreactogenic and safe.

EFFECT: higher efficiency of application.

5 ex, 4 tbl

FIELD: veterinary medicine.

SUBSTANCE: it is necessary to select affected organs in dead nutrias from local epizootic focus to prepare the suspension followed by inoculation onto differentially-diagnostic media; then one should isolate pure culture of Escherichia coli agent to be grown in beef-infusion-peptone broth at addition of glucose up to 0.2%-concentration at the titer being about 5-6 bln. microbial cells/cu. cm followed by inactivation due to introducing formalin up to 0.5-0.6%-final concentration to be kept at 37° C for about 72-96 h. Then on should add 3%-aluminum hydroxide solution at the quantity of 20% against culture volume to be thoroughly mixed, packed and sealed. The method is simple and enables to obtain a highly efficient vaccine.

EFFECT: higher efficiency.

1 ex, 1 tbl

FIELD: veterinary microbiology.

SUBSTANCE: invention proposes the strain Escherichia coli B-5 that produces thermolabile toxin eliciting immunogenic properties being without pathogenic property.

EFFECT: valuable properties of strain.

1 ex

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