The vaccine pseudomonas fur-bearing animals
(57) Abstract:The invention is intended to obtain the means of specific prophylaxis. The vaccine contains a mixture of antigens from strains of Ps. aeruginosa VGNKI N 40 sixth sarovara, N 63 eighth of sarovara, N 77 fifth sarovara, No. 110 second sarovara, N 278 third sarovara. Strains take effective ratio at a concentration of 6-8 billion MT in 1 cm3broth of Hottinger in volume 33,33-75,00%. The vaccine also contains the necessary amount of aluminium hydroxide, formaldehyde and saline solution. The vaccine, Pseudopodia harmless for foxes and polar foxes. Safety among vaccinated advantaged and disadvantaged population in Pseudomonas fur-bearing animals is 100%. The average geometric titer agglutinins in serum foxes and foxes is 1:220-1:380. The vaccine is applied to Arctic foxes and foxes at the optimum time in effective doses. The use of the invention in the veterinary practice will improve the effectiveness of measures to combat Pseudomonas foxes and polar foxes. table 1. The invention relates to veterinary biotechnology, in particular, to means of specific prophylaxis of Pseudomonas fur-bearing animals. Pseudomonas pyas animals.For prevention of this disease have been developed and used biological products on the basis of the pathogen Pseudomonas fifth, sixth, eighth, and eleventh serovars according to the International classification.Famous:
the vaccine Pseudomonas mink, which contains the causative agent of Pseudomonas third sarovara and enzymes: protease and elastase ;
the vaccine Pseudomonas fur-bearing animals, containing the antigen of the pathogen Pseudomonas sixth sarovara, formalin, potassium alum and saline solution  and othersHowever, the biological products do not provide adequate preventive effect, as in the more common mixed infection course, they marginally protect against infection of foxes and polar foxes and are intended primarily to prevent Pseudomonas mink.Also known vaccine against Pseudomonas fur-bearing animals, mainly mink, containing a mixture of antigens from strain Ps. aeruginosa N 40 sixth sarovara, N 48 - eleventh of sarovara, N 63 - eighth of serovar and N 77 - fifth of sarovara, aluminum hydroxide, formaldehyde, broth of Hottinger and saline solution .This vaccine pathogens Pseudomonas other serological variants.The aim of the invention is to increase the immunological activity of the target product for foxes and foxes.This goal is achieved by the fact that it further comprises antigens from strains of Ps. aeruginosa VGNKI 110 N second serovar and N 278 third serovar taken in the mixture in an effective ratio, at a concentration of 6-8 billion microbial cells, 1 cm3broth of Hottinger, in the following ratio of components,%:
A mixture of antigens from strains of Ps. aeruginosa VGNKI N 40 sixth sarovara, N 63 eighth of sarovara, No. 110 second sarovara, N 77 fifth sarovara and N 278 third serovar taken in the effective ratio at a concentration of 6-8 billion, M. K. 1 cm3broth of Hottinger - 33,33-75,00
Aluminium hydroxide - 0,30-0,35
Formalin - 0,32-0,34
Saline - Rest
For the manufacture of vaccines using new strains of Ps. aeruginosu second and third serovars. Strains Of Ps. aeruginosu deposited in the collection of microorganisms of the all-Russian state research Institute for control, standardisation and certification of veterinary preparations and have a registration number 110-02 and 278-S respectively.Strain Ps. aeruginosa N 110 (lab. Ainaži) second serovar selected from patologicheskogo material fallen Fox in 1987Strains Of Ps. aeruginosa VGNKI N 110-02 and VGNKI N 278-03 are characterized by the following characteristics and properties.Morphological features
Microscopic examination of smears broth cultures celebrate Grams/negative motile Bacillus with slightly rounded edges and a length of 0.8-1.2 MMK and a width of 0.6-0.8 MMK, arranged singly or in pairs in stored crops or in the form of short chains in siepermann cultures. Spores and capsules do not form.Cultural properties
On solid nutrient media: MPA, the agar of Hottinger and Martin culture strains grow in the form of a round grey tinged mucous colonies with the formation of a pronounced blue-green zone (strain N 110-02) or yellow-green zone (N 278-03). Under cultivation on liquid nutrient media: BCH broth of Hottinger and Martin is characterized by the formation of a surface film of gray-white color with staining through 14-18 h environment in the blue-green color (culture of strain N 110-02) or yellow-green color (culture of strain N 278-03)
Culture strains are fermented without the formation of gas glucose, peptonize milk, have hemolytic, catalase, gelatinase and lecithinase activity.Sweat is to have the flagellar "H" somatic "O" antigens. When you identify with serotypeable sera according to the International classification strain N 110 refers to 02 serological variant and strain N 278 - 03 serological variant. On the introduction of formalin inactivated cultures of strains (Ps. aeruginosa in the body foxes, foxes and Minks are formed antibody-agglutinin in titers of 1:180-1:320.Virulent properties
LD50culture of strain Ps. aeruginosa VGNKI N 110-02 for nonlinear mice weighing 14-16 g is 38.9 million M. K., and culture of the strain Ps. aeruginosa VGNKI N 278-03 22.3 million m K. For Guinea pigs weighing 300-350 g - 135 and 269 million M. K., respectively.Immunogenic properties
Inactivated with 0.3% formalin daily culture of strain Ps. aeruginosa N 110-02 protects from infection control 50% of the Guinea pigs weighing 300-350 g at the dose of 33.8 million M. K., and culture of the strain Ps. aeruginosa N 278-03 dose 38.9 million m K. the Minimum immunizing dose of polyvalent antigen-based strains Ps. aeruginosa N 110, 278, 77, 63, 40 for Guinea pigs weighing 300-350 g at 19.9 million M. K.The introduction of a vaccine based on a mixture of antigens from strains of Ps. aeruginosa N 40, 63, 77, 110 and 278 foxes, Arctic foxes, Minks induces in 98% of the animals after 6 days the formation of persistent immunity duration n Example 1. Metrovia seed cultures of strains of Ps. aeruginosa VGNKI N 40 sixth sarovara, N 63 eighth of sarovara, N 77 fifth sarovara, No. 110 second serovar and N 278 third cepouapa grown in separate reactors in the broth of Hottinger when 36,5-37,5oC up to a maximum accumulation of bacterial mass 12-18 billion, M. K. 1 cm3. At the end of the cultivation using the saline concentration of the bacterial mass of each strain is reduced to 6-8 billion, M. K./cm3add the formalin from the calculation of 0.32-0.34 percent of the volume and inactivate for 12-14 days at a 6.5-37,5oC. After inactivation of the suspension of each strain in the effective ratio is mixed in a separate reactor, add 0,30-0,35% of the amount of aluminium hydroxide and bring the pH of the biological product to 7.2 to 7.4, tested for sterility and packaging.The finished vaccine is a homogeneous suspension of grayish-white color. When resting on the bottom of the vial, a precipitate of grayish-white color, easily broken during the shaking of the vial.The vaccine was incubated for 14 days at 16-20oC, and then tested for sterility, as well as on safety and immunogenicity.Example 2. Study the efficacy of a vaccine against p.
A mixture of antigens from strains of Ps. aeruginosa VGNKI N40 sixth sarovara, N 63 eighth of serovar N 77 fifth sarovara, No. 110 second serovar and N 278 third serovar taken in the effective ratio, at a concentration of 8 billion, M. K. 1 cm3broth of Hottinger - 75,00
Aluminium hydroxide - 0,30
Formalin - 0,32
Saline - Rest
Guinea pigs in the amount of 270 heads live weight of 300-350 g vaccine is injected subcutaneously in the region of the back at a dose of 3.0-3.5 cm3, Arctic foxes (120 heads) and foxes (75 goal.) intramuscularly at a dose of 5-6 cm3(2.5-3.0 cm3in the right and left hind limbs.The study showed that the vaccine is harmless to animals. As a result of observations during the 10 days the animals were observed deviations from the normal physiological state. All animals remained alive.Example 3. The activity obtained in example 1 vaccine Pseudomonas fur-bearing animals in comparison with known vaccine (prototype) test on Guinea pigs (235 goal.) live weight of 300-350 g With this purpose, Guinea pigs Guinea groups, once intramuscularly pre-titrated dose of the vaccine, containing the following components about. %:
The mixture is of sarovara, taken in the effective ratio, at a concentration of 7 billion microbial cells in 1 cm3broth of Hottinger - 40,00
Aluminium hydroxide - 0,33
Formalin - 0,33
Saline - Rest
The control group of Guinea pigs (30 goal.) introduce similar vaccine Pseudomonas fur-bearing animals, containing the antigens of the strains Ps. aeruginosa VGNKI N40 sixth sarovara, N 48 the eleventh of sarovara, N 63 eighth of serovar and N 77 fifth serovar (prototype).After 7-10 days in 50% of inoculated animals take blood and serum examined in RA with homologous strains of the pathogen Pseudomonas included in the vaccine.The results are presented in the table.The data show that the activity proposed vaccine significantly higher activity known vaccines, despite the same amount of antigen in each dose of the drug.Example 4. Checking immunogenic properties of the vaccine Pseudomonas fur-bearing animals, prepared according to example 1, containing the following components,%:
A mixture of antigens from strain Ps. aeruginosa N 40 sixth sarovara, N 63 eighth of sarovara, N 77 fifth sarovara, 110 second serovar and N 278 tryna of Hottinger - 33,33
Aluminium hydroxide - 0,35
Formalin - 0,34
Saline - Rest
Guinea pigs (675 goal. ) live weight of 300-350 g Vaccinium live vaccine of the specified structure once intramuscularly in doses 0,01; 0,05; 0,1; 0,5 cm3foxes and foxes are subjected to immunization by intramuscular injection at a dose of 1.0 and 1.5 cm3respectively. Control animals received the vaccine is not administered. 14 days after immunization vaccinated and control Guinea pigs infect podarowano dose of cultures of strains of Ps. aeruginosa VGNKI N40 sixth sarovara, N 63 eighth of sarovara, N 77 fifth sarovara, No. 110 second serovar and N 278 third sarovara.Installed 100% immunological protection of the vaccinated Guinea pigs at the death of the pathogen Pseudomonas 95% not vaccinated Guinea pigs. The deaths among the vaccinated population of foxes and polar foxes are also not installed. The foxes vaccinated at doses of 1.0 cm3and foxes vaccinated with doses of 1.5 cm3in serum determined that agglutinins in titer 1:220-1:380.Example 3. Vaccine Pseudomonas fur-bearing animals of the composition (example 4) use a preventive purpose in immunization of female foxes and polar foxes (total 1500 goal.) in fur, hoteis the dose of 1.0-1.5 cm3the foxes in the dose of 1.5-2.0 cm3. The control group (1) animals (500 goal. foxes and foxes) to impose a known vaccine (prototype) in the same doses. The animals are not vaccinated (control II - 503 females of Fox and Fox). For vaccinated and control animals are clinical and epidemiological surveillance during the year. The study found that in a group of foxes and polar foxes vaccinated offered by the vaccine, cases of complications and breakthrough immunity is not marked. In groups of animals vaccinated with vaccine (the prototype), abortion pseudomonotone etiology observed in 18 cases. Among unimmunized animals in the control group there were cases of abortion and resorption of fetuses in the first half of pregnancy in 38 cases.The proposed use of the vaccine in veterinary practice will improve the effectiveness of measures to combat Pseudomonas foxes and foxes.Sources of information
1. U.S. patent N 4120950, CL 424-87, 1978.2. USSR author's certificate N 575889, CL C 12 K 5/00, 1972.3. USSR author's certificate N 1066074, class A 61 K 39/104, 1982 (prototype). The vaccine Pseudomonas fur-bearing animals, mainly foxes and foxes, containing antigens from strains of Pseudomonas aerug the tion, formalin and saline solution, characterized in that it further comprises antigens from strains of Pseudomonas aeruginosa VGNKI 110 N second serovar and N 278 third serovar taken in the mixture in an effective ratio at a concentration of 6 to 8 billion microbial cells in 1 cm3broth of Hottinger in the following ratio of components,%:
A mixture of antigens from strains of Pseudomonas aeruginosa VGNKI N 40 sixth sarovara, N 63 eighth of sarovara, N 77 fifth sarovara, No. 110 second serovar and N 278 third serovar taken in the effective ratio at a concentration of 6-8 billion microbial cells in 1 cm3broth of Hottinger - 33,33 - 75,00
Aluminium hydroxide - 0,30 - 0,35
Formalin - 0,32 - 0,34
Saline - Rest
FIELD: medicine, infectious diseases.
SUBSTANCE: invention relates to a method for preparing capsule antigen of meliodosis pathogen eliciting anti-phagocytic activity. Method involves culturing the strain B. pseudomallei, inactivation, isolation of glycoprotein by gel-filtration and ion-exchange chromatography methods resulting to preparing glycoprotein with molecular mass 200 kDA consisting of 90% of carbohydrates, 10% of proteins and comprising 2.17% of nitrogen, 46.4% of carbon and 8.09% of hydrogen. Advantage of method involves preparing the purified glycoprotein eliciting anti-phagocytic activity.
EFFECT: improved method for preparing, valuable properties of antigen.
2 dwg, 5 ex
FIELD: medicine, veterinary science.
SUBSTANCE: the suggested preparation consists of formalin-inactivated suspension of B.mallei deep culture sorbed upon aluminum hydroxide. The content of components, 1 cu. cm of the preparation: inactivated cell B.mallei 0.1-10 billion, aluminum ions up to 2.5 mg, formaldehyde - not more than 0.005%. Vaccination should be performed once annually, stable immunity should be formed to 7-14 d.
EFFECT: increased immunogenicity.
FIELD: veterinary medicine.
SUBSTANCE: the vaccine suggested contains cell suspension of pure cultures of Pseudomonas aeruginosa agent as antigens that belong to ten sero-groups N18(01); N47(02); N37(03); N19(04); N2(06); N13(010), N70(011); N66(013); N29(018); N28(019) obtained due to selecting parenchymal affected organs from dead animals out of local epizootic focus, inoculating onto differential-diagnostic media and isolating pure cultures of agents. Raising of pure cultures of agents should be carried out separately in meat-peptone broth and agar up to concentration of every sero-group of microbial cells to be about 9.5-10.5 bln/ml, then one should mix them in equal ratios. As a conservant one should introduce formalin, as an adjuvant - aluminum hydroxide at the following ratio of components, weight%: cell suspension of pure culture of Pseudomonas aeruginosa agent of ten sero-groups N18(01); N47(02); N37(03); N19(04); N2(06); N13(010), N70(011); N66(013); N29(018); N28(019) isolated out of local epizootic focus in nutritive medium at the titer being about 9.5-10.5 bln microbial cells/ml 84.6-89.5; formalin 0.4-0.5; aluminum hydroxide - the rest. The suggested vaccine is of high immunogenic activity.
EFFECT: higher efficiency.
5 ex, 1 tbl
FIELD: veterinary medicine.
SUBSTANCE: the innovation deals with cultivation of antigen, inactivation with formalin and introduction of aluminum hydroxide. As antigen one should apply epizootic strains of Pseudomonas aeruginosa of serogroups O1, O3, O4, O6, O11, O19 that provide "S"-forms of colonies, then one should prepare suspension, fulfill inoculation onto differentially-diagnostic media, isolate pure cultures of Pseudomonas aeruginosa agents of O1, O3, O4, O6, O19 serogroups that provide "S"-shaped forms of colonies. Then they should be grown separately for about 24-36 h upon Hottinger's broth at pH being 7.2-7.4 and dissolved at 36-37°C with physiological solution up to concentration of 10 bln microbial cells/ml. Then comes inactivation with formalin that contains 36% formaldehyde, not less to achieve final concentration ranged 0.3-0.4% to be kept for 14 d at about 37-37.5°C. Moreover, the culture should be mixed at equal volumes and at uninterrupted mixing one should introduce 3%-aluminum hydroxide solution at not more than 25% against the volume of cultures mixture to be thoroughly mixed, packed and sealed. The innovation enables to obtain highly immunogenic vaccine that forms tense immunity of 9 mo duration, not less and causes no post-vaccinal complications.
EFFECT: higher efficiency of manufacturing.
FIELD: medicine; pharmacology.
SUBSTANCE: present method of bacteria outer membrane vesicle preparation of Neisseria bacteria implies destruction of bacteria shell and gathering of outer membrane vesicles at deoxycholate solution < 0.05%. Bacteria express surplus NspA. Invention allows to keep significant bacterial immunogenic components, specifically (i) protective surface protein NspA, (ii) protein '287' and (iii) protein '741'.
EFFECT: significant immunogenic components keeping.
17 cl, 2 tbl, 2 dwg
FIELD: medicine, veterinary science.
SUBSTANCE: invention concerns veterinary medicine. Target affected organs of fallen nutrias are taken from a local epizootic nidus, and suspension is prepared from them. Inoculation in differentially diagnostic media is carried out. Pure culture of the germ is evolved. Pseudomonas aeruginosa is grown in meat infusion broth with titre of 5-6 milliard microbe cells per 1 cm3 and inactivated by adding formalin to the end concentration of 0.4-0.5%. After that the culture is exposed to 37°C for 48-72 hours. Aluminium hydroxide (20% by volume) is added to the culture. The components are thoroughly mixed, packed and sealed.
EFFECT: method is simple and allows of obtaining highly immunogenic safe vaccine.
1 tbl, 1 ex
FIELD: medicine, veterinary science.
SUBSTANCE: invention concerns veterinary medicine. vaccine contains inactivated antigen and adjuvant. The antigen contained in the vaccine is cell suspension of germ Pseudomonas aeruginosa pure culture. The culture is obtained by selecting target affected organs of fallen nutrias are taken from a local epizootic nidus, preparing suspension, inoculating in differentially diagnostic media, evolving pure culture of the germ and growing the culture in meat infusion broth until microbe cell concentration of 5-6 milliard per 1 cm3 is achieved. The vaccine also contains formalin and aluminium hydroxide in the following wt % ratio: cell suspension of germ Pseudomonas aeruginosa pure culture evolved from target affected organs of fallen nutria from a local epizootic nidus in meat infusion broth with titre of 5-6 milliard of microbal cells per 1 cm3- 83.0-85.5, formalin - 1.0-2.0, the remainder being aluminium hydroxide.
EFFECT: vaccine is safe, highly immunogenic and stable during storage.
1 tbl, 5 ex
SUBSTANCE: method involves sterilising a culture of the avirulent strain Burkholderia pseudomallei 107, suspending the dried bacterial mass in NaCl 0.15ml, exposing the suspension to ultrasound centrifuging the damaged cell suspension followed by separating the supernatant and salting out a glycoprotein fraction, which is used to prepare a diagnosticum.
EFFECT: invention enables the early identification of the specific actinobacillus mallei and melioidosis antibodies by indirect haemagglutination test.
2 cl, 1 dwg, 2 tbl, 3 ex
SUBSTANCE: Pseudomonas aeruginosa O11 strain is deposited in the collection of theFederal State Budgetary Institution VGNKI under the name "Pseudomonas aeruginosa No. 10" and the registration number "No. 10-DEP".
EFFECT: strain has high persistent immunogene activity therefore it can be used for production of the vaccine against pseudo-monose of pigs.
3 tbl, 4 ex