Mixed vaccine based on antigens of microorganism moraxella bovis and herpes virus type i for specific prophylaxis of infectious keratoconjunctivitis in cattle

FIELD: veterinary microbiology, virology, biotechnology.

SUBSTANCE: invention proposes vaccine that comprises as antigens the following components, vol. %: inactivated suspension of cells of hemolytic strains of microorganism M. bovis with the total concentration 100-120 billion cells in 1 cm3: strain M. bovis "G97-VNIVI", 4.0-5.0; strain M. bovis "SHZ-01", 4.0-5.0; inactivated cultural suspension of the strain Herpesvirus bovis type I "TKA-VIEV-V2" with infectious titer 107.0 - 107.5 TCD50/ml, 78.0-83.0; 6% solution of aluminum hydroxide gel, the balance. Vaccine enhances accumulation of specific protective antibodies in serum blood of vaccinated animals and provides the development of immunity against infectious keratoconjunctivitis that is retained for one year.

EFFECT: enhanced effectiveness and valuable properties of vaccine.

4 tbl, 6 ex

 

The invention relates to the field of veterinary Microbiology, Virology and biotechnology, in particular the production of vaccines for specific prophylaxis of infectious keratoconjunctivitis in cattle. It is established that the main etiological agents of the disease are hemolytic forms of bacteria Moraxlla bovis in the face of intense solar UV radiation. Infectious keratoconjunctivitis widespread in all countries of the world. The frequency of its appearance in the Russian Federation is highest among the young cattle in feedlots with a high stocking density and intensity of development of epizootic process depends on the degree of exposure of animals during the summer months, extreme solar UV rays and disease comprises from 25 to 90% of the population.

Along with this, it is proved that the virus of infectious bovine rhinotracheitis bovine herpesvirus type 1) can also cause acute conjunctivitis in this species. However, the inflammatory process is not progressing to corneal opacity and ulceration, as is the case with infectious keratoconjunctivitis (Jensen R., McKay D. Diseases of cattle in industrial fattening. M.: Kolos, 1977, - s-129; Abinanti FR, Plumer G.J.: The isolation of Infectious Bovine Rhinotracheitis Virus from Cattle Affected with Conjunctivitis-Oservations on the Experimental Infection. // Am. J.Vet Res., 1961, 22, - p.13-17; Hughes JP et al. Keratoconjunctivitis Associated with Infectious Bovine Rhinotracheitis. // Am. J. Vet. Med. Ass., 1964, 45, - p.32-42; George W. et al. Bovine Infectious Keratoconjunctivitis: Interaction of Moraxella bovis and Infectious Rhinotrachitis Virus. // Am. J. Vet. Res., 1970, 31, No. 4 - p.653-662).

The results of studies conducted in the 80-ies on the problem of infectious keratoconjunctivitis in cattle have shown that the virus of infectious bovine rhinotracheitis is one of the agents, which together with the bacteria Moraxella bovis can cause this disease (Straub OF Infection of cattle caused by the herpes virus. - M.: Kolos, 1981, - pp.33-35).

Other literary sources also indicate that the virus of infectious bovine rhinotracheitis may be the primary causative agent of conjunctivitis, but not keratoconjunctivitis (Rebnun W. et al. Anoutbreak of the form of conjunctivitis infectious bovine rhinotraheitis. // Cornell. Veter., 1978, 68, 3, - p.297-307). In several papers it has been suggested that the virus of infectious bovine rhinotracheitis in cattle creates a very favorable environment for pathogenic action of the main exciter keratoconjunctivitis - bacteria Moraxella bovis.

It was later proven that the less pronounced the etiological role in infectious keratoconjunctivitis in cattle can also play chlamydia (Wehr I. et al. Untersuchungen zur Bedeutung der Chlamidien bei der infektiosen Keratokonjunctivitis des Rindes. // Wissensch. Humbold. Univ. Berlin, 1980, 29, 1, - p.61-65), Mycoplasma bovoculi (Langford E.V. Characterization of ycoplasma isolated from infectionus bovine keratoconjunctivitis: M. bovoculi sp. nov. // Can. J. Environ., 1973, 19, - p.1435-1444), rickettsiae and conditionally pathogenic bacteria, participating mainly in the formation of mixed infections.

Prossimamente etiology of infectious keratoconjunctivitis in our country, the absence of any normative documents on TB diagnosis and control measures, the import of breeding cattle from Western European countries - hidden carriers of the pathogen, has led to the emergence permanently affected with infectious keratoconjunctivitis large pockets and saving trends in the further spread of the disease, causing significant economic losses due to reduced milk yield and liveweight gain of animals due to partial or complete loss of vision (here VK Infectious keratoconjunctivitis in cattle in farms Gomelskoj area. // Disease C. agricultural animals and birds, their prevention and treatment./Sbornik works LVI, issue 39. - L., 1974, - s-179; Andresen V.B. have been Infectious keratoconjunctivitis in cattle in the North-Western zone of the RSFSR // proceedings of the LVI. - L., 1988, - p.6-11; Cocolin I.E. Infectious keratoconjunctivitis calves in the Irkutsk region. // And the book: Prevention of diseases of young animals C. farm animals Siberia and the Far East. Novosibirsk, 1982, - s-75; Gaffarov HS et al. Epidemiological aspects of infectious keratoconus Evita cattle. // Materials of the International scientific conference dedicated to the 125th anniversary of KHAWM. Kazan, 1998, - p.29-31; Gaffarov HZ, Spiridonov G.N., Valerna L.V. Strain of bacteria Moraxella bovis "G-unive used for the manufacture of diagnostic kits and vaccines against infectious keratoconjunctivitis in cattle. // Patent RF №2145353, priority dated 15.09.1998,; Gaffarov HZ, Velebny L.V., G. Spiridonov. Infectious keratoconjunctivitis in cattle in the region of the Middle Volga and Ural region. // Actual problems of diseases of young animals in modern conditions. Materials of international scientific-practical conference 23-25 September 2002, Voronezh, - s-191).

Made epizootological monitoring in relation to the KIC 48 farms in 12 districts of the Republic of Tatarstan and 29 farms in 8 districts of the Republic of Bashkortostan and Udmurtia showed that the incidence of calves 1-5 months of age is 50-85%in calves older age - 20-25% among dairy cattle it ranges from 12-15% (Gaffarov HZ, Velebny L.V., G. Spiridonov. Infectious keratoconjunctivitis in cattle in the region of the Middle Volga and Ural region. // Actual problems of diseases of young animals in modern conditions. Materials of international scientific-practical conference 23-25 September 2002, Voronezh, - s-191).

Uch is given, infectious keratoconjunctivitis is widespread and affects all age groups of animals, especially young animals, vaccination is a very effective method of dealing with it. It should be noted that the problem of specific prophylaxis of infectious keratoconjunctivitis in many countries of the world have been solved in the 70-90-ies of the last century (Hughes D.et. al. Experimenally Induced Infectious Bovine Keratoconjunctivitis: Effectivenes of Intramuscular Vaccination with Viable Moraxella bovis Culture. // Am. J. Vet. Res., 1971, 32, №6, - p.879-886; Pugh G.W. et al. Infectious bovine keratoconjunctivitis in cattle vaccinated and medicated against Moraxella bovis befare parturition. // Am. J. Vet. Res., 1982, 43 No. 2, - p.320-325; R. Gwin Medicament and method for prodicing the immunity of infectious bovine keratoconjunctivitis. // U.S. patent No. 4539201 September 3, 1985). Therefore, the problem of creating a vaccine against infectious keratoconjunctivitis in cattle in Russia is very important. Analogues of vaccines against infectious keratoconjunctivitis in cattle, made on the basis of hemolytic forms of bacteria Moraxella bovis to our research does not exist. The closest analogue in relation to the proposed "Associated vaccines for specific prophylaxis of infectious keratoconjunctivitis in cattle based on the antigens of the bacteria Moraxella bovis and herpesvirus type 1" may be found in U.S. patent "Moraxella bovis infectious bovine keratoconjunctivitis steam-killed bacterin" 3401219 ot, which describes the fact that the use immunities antigen Moraxella bovis, in particular bacteria, which is used as a preventive measure against infectious keratoconjunctivitis in cattle. Bacteria are inactivated fluid vapor within one hour, the suspension of bacteria Moraxella bovis. Bacteria are able to develop immunity against infectious keratoconjunctivitis in 6 months. The disadvantage of this vaccine is that it does not protect animals from keratoconjunctivitis herpetic etiology.

The objective of the invention is the creation of associated vaccine based on hemolytic forms of bacteria Moraxella bovis and herpesvirus type 1, intended for creation of specific prophylaxis of infectious keratoconjunctivitis in cattle in the territory of the Russian Federation, and the increase in the duration of immunity up to 12 months.

The technical result of the invention consists in obtaining the associated vaccine with high immunogenic activity, and the expansion of its immunogenic spectrum.

The invention consists in the following: the associated vaccine against infectious keratoconjunctivitis in cattle contains antigens hemolytic forms of bacteria Moraxella bovis and herpesvirus cattle, of deposited is consistent on aluminium hydroxide, in the following ratio of components,%:

Inactivated cell suspension
the M. bovis bacteria concentration
100-120 billion, M. K.:
1 cm3: PCs "G-unive"- 4,0-5,0
pieces"s-01"4,0-5,0
Inactivated culture
suspension herpesvirus
large
cattle PCs "TKA-VIEW-B2"
infectious titer of 107,0-107,5
TCD50/ml78,0-83,0
Gel aluminium hydroxide, 6% solutionrest

For the manufacture of associated vaccine for the specific prophylaxis of infectious keratoconjunctivitis in cattle based on the antigens of the bacteria Moraxella bovis and herpesvirus type 1 uses a patented strain of bacteria Moraxella bovis "G-unive" (RF Patent for the invention №2145353 priority from, 15.09.1998) and a new strain of bacteria Moraxella bovis "s-01"obtained from the patient to the infectious keratoconjunctivitis calf. The strain of bacteria Moraxella bovis "s-01" the character of ersuade the following properties: it is a short thick sticks with a length of 1.5-2.0 and a diameter of 0.5-1.0 μm with rounded ends, located in pairs or in short chains do not form spores, fixed, stained gram-negative. On blood MPA at pH 7,2-7,6 after 24 hour incubation at 37°form colonies with a diameter of from 1 to 3 mm with an area of β-hemolysis width of 0.5 to 2.0 mm Colonies are smooth, round, shiny, grayish-white. On BCH 24-48 hour incubation at 37°observed turbidity of the broth with the formation of shallow draft. The strain of the facultative aerobe, not fermented sugars, does not restore the nitrate to nitrite, does not form indole, thins gelatine, i.e. possesses proteolytic activity, peptonized litmus milk, gives a positive reaction for oxidase, which are characteristic for bacteria of the family Neisseriaceae, of the genus Moraxella. The strain is virulent for white mice, causes their death within 24 hours after intraperitoneal administration of them daily agar culture in a dose of 500 million M.K.

Example 1. Getting motroway the seed of each strain of bacteria Moraxella bovis.

For the manufacture of vaccines take capsules with liofilizirovannami crops production strains .bovis; "G-unive" and "s-01 ", which are dissolved in 0.5-2.0 cm3BCH and transferred to Petri dishes with 10%blood MPA. The cultivation is carried out in a period of 16-18 hours, then the culture is checked for compliance with morphological, cultures is selected, enzymatic and virulence properties. If they match hemolytic forms of bacteria .bovis, then use them to get bacterial mass needed for the manufacture of associated vaccine.

Strains were stored in lyophilized form. A suspension of the daily culture of the bacteria Moraxella bovis with a concentration of 50 billion, M. K. 1 cm3mix a protective environment (skim milk, sucrose 5%, gelatinous of 1.5%) in the ratio 1:1, filled into ampoules of 1 cm3, dried by sublimation, sealed ampoules under vacuum and stored at a temperature of 2-8°C.

Example 2. Cultivation of bacterial mass of each strain of bacteria Moraxella bovis separately and its inactivation.

To obtain biomass crops M. bovis use MPA, containing 10% defibrinating the blood of the lamb. With this purpose, the specified environment in a sterile environment is poured into 1.5 liter mattresses of 250-300 cm, after solidification of the medium doing the sowing of these strains by making each mattress 2 cm3a bacterial suspension containing 1 billion, M. K. 1 cm3. Crops incubated at 37 ° °C. After 36-48 h grown colony cultures .bovis wash 0,02%solution of sodium chloride, to prepare a suspension with a concentration of 100-120 billion M.K. 1 cm3for bacterial or optical turbidity standard of gisk named after. That is asevich. The culture is checked for purity, morphological and serological typicality.

Inactivation of bacterial suspension exercise formalin. To do this in the microbial suspension add formaldehyde content 37-38% active formaldehyde 0.5 cm3100 cm3suspension. Mixed and placed in a thermostat at 72 hours Then take a sample for completeness of inactivation, which is carried out by inoculation of inactivated microbial suspensions on blood MPA.

Example 3. Preparation of native suspension herpesvirus cattle and its inactivation.

Herpesvirus bovine pieces "TKA-VIEW-B2") propagated in human cell cultures of tracheal embryo cow (line TR)grown in three-liter roller bottles at a combined growth medium consisting of medium 199 - 10%, CLAY - 40%, eagle medium MEM - 40% and bovine serum at 10%. The culture is grown for 3-4 days at a seeding concentration of 140-150 thousand cells in 1 cm3environment. Growth medium is drained, make a support medium (medium 199 - 20%, CHAP 40% and the Needle MEM - 40%) in the number of 350-370 cm3that pre-make herpesvirus rate of 10 cm3500 cm3Wednesday, 3%solution of glutamine - 5 cm3and antibiotics (streptomycin 100 µg/cm3penicillin - 100 U./cm3). Bottles incubated in roller installation pre-38° C for 3-4 days. Collecting the culture fluid produced by three freezing and thawing during the period of pronounced cytopathic effect of maximum accumulation of the virus. Received the virus suspension combine with the purpose of virus inactivation add formalin to a concentration of active formaldehyde 37-38% rate of 0.5 cm3100 cm3viral suspension, mix and leave for 24-36 h at room temperature, and then tested for bacterial and fungal contamination. For preparation of the vaccine take viral suspension without any contamination. Control of completeness of inactivation of herpesvirus cattle is carried out by 3 successive passages samples inactivated virus suspension used for producing the associated vaccine keratoconjunctivitis in cattle, transplantable cell cultures of tracheal embryo cow (line TR) with an interval of 5-6 days. The viral suspension is considered inactivated, if the third passage is not detected by cytopathic effect of the virus.

Example 4. Preparation of associated vaccine for the specific prophylaxis of infectious keratoconjunctivitis in cattle based on the antigens of the bacteria Moraxella bovis and herpesvirus type 1.

The vaccine is prepared from antigen is of herpesvirus cattle with infectious titer of 10 7,0-7,5TCD50/ml and industrial strains of bacteria Moraxella bovis: "G-unive", "s-01".

Take herpes virus suspension, there was added 100-120 billion-ing the suspension of the bacteria Moraxella bovis on the basis of 4-5 cm3each strain 100 cm3vaccine preparation, as well as 6%gel of aluminium hydroxide at the rate of 10 cm3100 cm3the vaccines. Bring the pH of the vaccine to 7.2-7.4 and Packed with careful periodic mixing vials, which are then closed with rubber stoppers, running-metal caps and etecetera. Table 1 shows the composition of the vaccine.

Table 1

The composition of associated vaccine for the specific prophylaxis of infectious keratoconjunctivitis in cattle based on the antigens of the bacteria Moraxella bovis and herpesvirus type 1
The vaccine componentsTheir content in 1 liter of vaccine (ml)
Antigens .bovis containing 1 cm3100-120 billion M.K.:
PCs. "G-unive"40-50
PCs. "S-01"40-50
Antigen Bovine Herpesvirus pieces"TKA-VIEW-B2"infectious titer of 107,0-7,5TCD50/ml780-830
6%solution gel aluminium hydroxide Rest

Example 5. Control vaccines for sterility, safety and antigenic activity.

To test for sterility of the 6 vials of vaccine each series make crops 0.2 cm3at BCH, IPA, black IPA, MPB under vaseline oil and agar Saburo 2 tubes on each bottle. Crops with all the media incubated at 37°and agar Saburo - at 20-22°C for 15 days. The nutrient medium should be sterile.

To test for harmlessness select the 10 white mice live weight of 17-18 g and introduce them the vaccine subcutaneously at a dose of 0.5 cm3. The vaccine is considered harmless if the mouse within 10 days after the vaccine remain alive and healthy.

Antigenic activity of vaccine control in 3 rabbits live weight of 2.0 to 2.5 kg, which the drug is administered subcutaneously at a dose of 2 cm3twice with an interval of 14 days. 14 days after the last vaccination in rabbits from the ear veins take blood and test serum with a view to establishing the level of specific antibodies to the herpes virus type 1 and antigens of the bacteria Moraxella bovis. The titers of antibodies to herpesvirus cattle must be in the range of 1:320-1:640 in ELISA, to strains of the bacteria Moraxella bovis - 1:400-1:1600 in response ELISA.

The inspection results of the 24 experimental series vaccine showed that all series biopro the Arata were sterile, harmless and antigenically-active. Indicators antigenic activity associated vaccines are presented in table 2.

Production testing of experimental batches of the vaccine were conducted in 8 farms of the Republic of Tatarstan and one of the Chuvash Republic, affected with infectious keratoconjunctivitis in cattle, where the incidence of calves under 6 months of age was 24-56%, youngsters up to 12 months of age 19-35% of youngsters over the years and the adult population 8-22%. All the cattle population in these farms were vaccinated with the associated vaccine before pasture to pasture twice with an interval of 21 to 30 days in doses: calves under 6 months of age - 3 cm3, youngsters from 6 to 12 months. - 5 cm3, young animals over one year of age and the adult population is 10 cm3. The results of vaccine trials in these farms are presented in table 4. From the table it follows that the associated vaccine has a high immunogenic activity. Its use prophylactically in 9 stationary disadvantaged farms allowed to completely eradicate the disease in 6 households, and the remaining three - to reduce the disease to individual cases.

Example 6. The purpose of General information and the application procedure associated vaccines for specific prevention infecciones the keratoconjunctivitis in cattle based on the antigens of the bacteria Moraxella bovis and herpesvirus type 1.

The vaccine is intended for the specific prophylaxis of infectious keratoconjunctivitis in cattle caused by a mixed infection caused by the bacteria Moraxella bovis and virus infectious bovine rhinotracheitis (herpesvirus type 1) on the background of increased solar ultraviolet radiation in the summer months.

The vaccine is a mixture of concentrated antigens hemolytic forms of bacteria .bovis and culture suspension herpesvirus type 1 in the optimal proportions.

In appearance the vaccine is deposited hydroxide of aluminum, an opaque liquid with a white precipitate, which upon shaking the container can be easily broken, forming a homogeneous suspension pale pink color.

The vaccine is produced in glass bottles of 100, 200 and 500 cm3with tightly closed with rubber corks, rolled metal caps, and etecetera in accordance with the specifications.

The vaccine is suitable for use within 12 months from the date of manufacture, provided that storage and transportation at a temperature of 2-10°C.

The vaccine is used for prophylactic purpose in threatened and permanently affected farms in infectious keratoconjunctivitis in cattle. Vaccinium all emerging young at the age of 30-35 days regardless of the time of year, as well as all the livestock to the total cattle before pasture to pasture. The vaccine is injected subcutaneously in the middle third of the neck twice with an interval of 21 to 30 days at doses of:

- 3 cm3- calves aged 1 to 6 months;

- 5 cm3the youngsters aged 6 months. to one year;

10 cm3the young animals over one year of age, cows and bulls producers.

The vaccine provides creation in young and adult cattle immunity that persists for years.

For the period 1999-2003 made 814,5 liters associated vaccine, which is used in the farms of the Republic of Tatarstan, Mari El, Bashkortostan, Udmurtia and Chuvashia, affected with infectious keratoconjunctivitis in cattle (table 3).

Table 2

Indicators antigenic activity associated vaccines for specific prophylaxis of infectious keratoconjunctivitis in cattle based on the antigens of the bacteria Moraxella bovis and herpesvirus type 1 in experimental conditions.
AnimalAntibody titers (M±t) in ELISA
km. bovis units T-unive"to M.hovis pieces "s-01"to herpesvirus pieces "TC-VIEW-B2"
Prior to vaccinationAfter 1 vaccinate the After 2 vaccinationsPrior to vaccinationAfter 1 vaccinationAfter 2 vaccinationsPrior to vaccinationAfter 1 vaccinationAfter 2 vaccinations
Rabbits-280,00±82,152720,00±1117,32-1360,00±558,565440,00±1073,0-160,00±0,00576,00±208,61
Cattle5280,00±1252,1910240,00±1752,006400±0,008320,00±2146,00160,00±80,00896,00±175,27960,00±226,27

Table 3

Unfavorable items on infectious keratoconjunctivitis in cattle, which was conducted extensive production testing of the associated vaccine.
№ p/pThe name of the farmsThe number of used vaccine in litres
123
The Republic of Tatarstan
1KP " Menger" atna district4,0
2KP"avant-garde" Aktanysh region to 12.0
3KP them. Lenin Buinsk district10,0
4LLC "Jalil" Bugulma district25,0
5Ochos KHAWM Vysokogorsky district25,0
6ACP "Zilina" Drozhzhanovsky district18,0
7Drozhzhanovsky Hospitalhennie26,0
8HUM "Metriopharm" Kazan320,0
9KP Chinanski" Zelenodolsk district30,0
10KP "Vegetable" Zelenodolsk district18,0
11SHPK "Board" Zelenodolsk district10,0
12APC imparitive Kukmor district20,5
13PE "Sadykov oil on canvas," Laishevsky district20,0
14SHP "Devyatovsky" Laishevsky district15,0
15LOC "Capsle" Laishevsky district5,0
16TNV "Vafin and" Laishevsky district15,0
17JSC Nurmanka" Laishevsky district20,0
18 LOC "Range" Laishevsky district5,0
19LLC "Idel" Laishevsky districtto 12.0
20LLC "birch" Laishevsky district10,0
21SCK "imlitala "Nurlat district10,0
22KP AK-YAR" Novosheshminskoe district6,0
23FSC " Utata" Yutazinsky district4,0
24SEC "tugai" Yutazinsky district10,0
Diurtiulinsk district Republic of Bashkortostan
25SPK "Koszul"20,0
26SEC "Plemzavod them. Lenin"20,0
27SEC "them. Enikeeva"30,0
28SEC " Plemzavod Victory"30,0
The Republic of Udmurtia
29KP "Bolshevik" Analsaga district20,0
30KP "Young guard" Analsaga district10,0
31The KDP, "May 1" Novo-Tarasovskogo district of the Republic of Mari El14,0
32SHPK " Star" of the Mariinsky Posad area is and the Republic of Chuvashia 20,0
Total814,5

Table 4

Indicators of efficiency of application of associated vaccine for the specific prophylaxis of infectious keratoconjunctivitis in cattle based on the antigens of the bacteria Moraxella bovis and herpesvirus type 1
№p/pThe name of the farmLivestock cattle headsIncidence, %
before vaccinesafter applying vaccines
calves under 6 months of agecalves up to 12 months. ageyoung animals over 12 months of age and cowscalves under 6 months of agecalves up to 12 months. ageyoung animals over 12 months of age and cows
1SHPK "Devyatovsky"131935-4528163-4--
2LOC "Capsle"17338358---
3 TNV "Vafin K"1148322713---
4JSC Nurmanka"2560563422---
5LOC "Range"213241914---
6LLC "Idel"884383219---
7SHPK "them. Vakhitova"297035-5025-2710-152-32-
8SHPK "Star"1237353020---
9"Ochos KHAWM"119840-4520-2510-122-3--

The associated vaccine for the specific prophylaxis of infectious keratoconjunctivitis in cattle based on the antigens of the bacteria Moraxella bovis and herpesvirus type I, characterized in that the tumor is on contains inactivated suspension of cells hemolytic strains of bacteria Moraxella bovis "G-unive" and Moraxella bovis "s-01", with a total concentration of 100-120 billion M.K. 1 cm3, inactivated culture suspension herpesvirus type I PCs "TKA-VIEW-B2" infectious titer of 107,0-7,5TCD50/ml adjuvant - 6%solution gel of aluminium hydroxide in the following ratio of components,%:

Inactivated suspension of cells hemolytic

bacterial strains M. bovis, with a total concentration of

100-120 billion, M. K. 1 cm3:
pcs. M. bovis "G-unive"4,0-5,0
pcs. M. bovis "s-01"4,0-5,0

Inactivated culture suspension

Herpesvirus bovis type I "TKA-VIEW-B2"

infectious titer of 107,0-7,5TCD50/ml78,0-83,0
Gel aluminium hydroxide, 6% solutionRest



 

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3 ex

FIELD: biotechnology, medicine.

SUBSTANCE: invention proposes the vaccine strain A/17/Panama/99/242 (H3N2) that represents reassortant prepared by breeding the epidemic strain A/Panama/200799 (H3N2) with cold-adopted temperature-sensitive virus A/Leningrad/134/17/57 (H2N2) as the attenuation donor that is harmless for adults. The strain A/17/Panama/99/242 (H3N2) multiplies actively in developing chicken embryos at optimal temperature 33-34oC and characterized by sensitivity to temperature and adaptation to cold. From the epidemic virus the reassortant inherited two genes encoding surface proteins (hemagglutinin and neuraminidase) and six genes encoding non-glycosylated proteins from the attenuation donor. The strain A/17/Panama/99/242 is areactogenic for adults in intranasal administration and satisfies to immunogenicity indices. The vaccine strain A/17/panama/99/242 (H3N2) satisfies to requirements by reactogenic indices to Pharmacopoeia article 42-3569-98 for vaccine strains of live anti-influenza vaccine used for intranasal applying in adults.

EFFECT: valuable properties of strain and vaccine.

1 dwg, 2 tbl, 1 ex

FIELD: microbiological industry, microbiology, insecticides.

SUBSTANCE: invention relates to methods for producing agents for plants protection and against harmful insects. Invention proposes a method for preparing insecticide preparation and preparation based on microorganisms Bacillus thuringiensis. Method involves culturing microorganisms Bacillus thuringiensis, isolation of delta-endotoxin from biomass by addition of a solvent, removal of precipitate and mixing solution with a filling agent and sticker followed drying the end product. Insecticide preparation obtained by proposed method comprises the following components, wt.-%: active component - dissolved delta-endotoxin from Bacillus thuringiensis, 10-20%; alkali (NaOH or KOH), 3-8%; reducing agent - sodium sulfite or sodium sulfide, 1-8%; sticker - a stabilizing agent of working suspension molasses or dry buttermilk, or pectin, or whey, or hydrolyzed chitin, or ground fish glue, or pectin from beet or apple pulp, 3-20%; filling agent, for example, NaCl - up to 100%. Method provides preparing highly effective and rapidly acting insecticide preparation.

EFFECT: improved preparing method, valuable properties of preparation.

2 cl, 5 tbl, 6 ex

FIELD: microbiological industry, microbiology, insecticides.

SUBSTANCE: invention relates to methods for producing agents for plants protection and against harmful insects. Invention proposes a method for preparing insecticide preparation and preparation based on microorganisms Bacillus thuringiensis. Method involves culturing microorganisms Bacillus thuringiensis, isolation of delta-endotoxin from biomass by addition of a solvent, removal of precipitate and mixing solution with a filling agent and sticker followed by drying the end product. Insecticide preparation obtained by proposed method comprises the following components, wt.-%: active component - dissolved delta-endotoxin from Bacillus thuringiensis, 10-20%; alkali (NaOH or KOH), 3-8%; reducing agent - thiourea, 1-8%; sticker - a stabilizing agent of working suspension molasses or dry buttermilk, or pectin, or whey, or hydrolyzed chitin, or ground fish glue, or pectin from beet or apple pulp, 3-20%; filling agent, for example, NaCl - up to 100%. Method provides preparing highly effective and rapidly acting insecticide preparation.

EFFECT: improved preparing method, valuable properties of preparation.

2 cl, 5 tbl, 6 ex

FIELD: microbiological industry, microbiology, insecticides.

SUBSTANCE: invention relates to methods for producing agents for plants protection and against harmful insects. Invention proposes a method for preparing insecticide preparation and preparation based on microorganisms Bacillus thuringiensis. Method involves culturing microorganisms Bacillus thuringiensis, isolation of delta-endotoxin from biomass by addition a solvent, removal of precipitate and mixing solution with a filling agent and sticker followed by drying the end product. Insecticide preparation obtained by proposed method comprises the following components, wt.-%: active component - dissolved delta-endotoxin from Bacillus thuringiensis, 10-20%; alkali (NaOH or KOH), 3-8%; reducing agent - β-mercaptoethanol, 1-8%; sticker - a stabilizing agent of working suspension molasses or dry buttermilk, or pectin, or whey, or hydrolyzed chitin, or ground fish glue, or pectin from beet or apple pulp, 3-20%; filling agent, for example, NaCl - up to 100%. Method provides preparing highly effective and rapidly acting insecticide preparation.

EFFECT: improved preparing method, valuable properties of preparation.

2 cl, 5 tbl, 6 ex

FIELD: microbiological industry, microbiology, insecticides.

SUBSTANCE: invention relates to a method for producing agents for plants protection and against harmful insects. Invention proposes a method for preparing preparation and preparation based on microorganisms Bacillus thuringiensis. Method involves culturing microorganisms Bacillus thuringiensis, isolation of delta-endotoxin from biomass by addition of solvent, removal of precipitate and mixing solution with a filling agent and sticker followed by drying the end product. Insecticide preparation obtained by proposed method involves the following components, wt.-%: active component - dissolved delta-endotoxin from Bacillus thuringiensis, 10-20%; alkali (NaOH or KOH), 3-8%; reducing agent - sodium hydrosulfite, 1-8%; sticker - a stabilizing agent for working suspension molasses or dry buttermilk, or pectin, or whey, or hydrolyzed chitin, or ground fish glue, or pectin from beet or apple pulp, 3-20%; filling agent, for example, NaCl, up to 100%. Method provides preparing highly effective and rapidly acting insecticide preparation.

EFFECT: improved preparing method, enhanced effectiveness of preparation.

2 cl, 5 tbl, 6 ex

FIELD: biotechnology, veterinary science, microbiology, vaccines.

SUBSTANCE: invention relates to a strain that is deposited in collection of pathogenic strains causing diseases in fur animals in the museum of bacterial and viral cultures of Science-Research institute of fur farming and rabbit breeding (NIIPZK) named for V. A. Afanasjev and has registered number 289 M-positive. The strain relates to streptococci of group A. The strain is culture in beef-extract broth, in Todd-Hevitt's medium, Pike's medium at 37°C for 18-20 h. After control of grown culture for virulence the separated biomass is subjected for inactivation and further extraction by addition of hydrochloric acid followed by addition of aluminum hydroxide as an adjuvant and packaging. Invention provides prevention losses in reproduction of fur animals in farms with streptococcus infection.

EFFECT: valuable properties of strain.

3 ex

FIELD: microbiology, biochemistry.

SUBSTANCE: invention relates to methods for treatment of sewage waters, in particular, to microbiological methods. Sewage waters are neutralized with a mixture of calcium oxide and calcium carbonate followed by treatment with microorganisms. The consortium of microorganisms Bacillus brevis and Arthrobacter species IB DT-5 is used as microorganisms. Method is used in treatment of organic substances from sewage waters formed in manufacturing isopropyl alcohol.

EFFECT: improved method for treatment.

1 tbl, 1 ex

FIELD: biotechnology, food processing industry, medicine.

SUBSTANCE: claimed consortium is obtained from bifidus-bacterium strains with excellent superoxide dismutase, antioxidant and antibiotic activity, namely: Bifidobacterium bifidum, Bifidobacterium longum, Bifidobacterium adolescentis, Bifidobacterium infantis, Bifidobacterium breve. Consortium has superoxide dismutaseactivity not less than 24.9 unit/mg of microbial protein mass. Consortium is able to utilize wide amino acid spectrum and has high antibiotic resistance and increased resistance to aggressive media of gastrointestinal tract.

EFFECT: bifidus-bacterium consortium with increased colony forming activity for manufacturing of products with excellent probiotic and healthy effect.

2 tbl, 5 ex

FIELD: biotechnology, food processing industry, medicine.

SUBSTANCE: strain Bifidobacterium bifidum is obtained by sequential cycles of lyophilic drying and reducing on thioglycol cultural medium (50 cycles). Clones with excellent antioxidant activity have been selected according to maximum accumulation of bifidus-bacteria biomass in sterile defatted milk not less than lg 9 after cultivation for 16 h. Strain has superoxide dismutase activity not less than 19.7 unit/mg of microbial protein mass. Strain Bifidobacterium bifidum can effectively growth in cultivation media and accumulate necessary titer of viable bacteria, has high acid-forming activity, antagonistic action against pathogenic and conditional pathogenic microflora, and excellent antioxidantaction. Culture has marked probiotic action cased by its superoxide dismutase activity. Strain is useful in preparation of functional foodstuff, bioactive supplement, cosmetic and toiletry agents.

EFFECT: bifidus-bacterium strain for manufacturing of products with excellent probiotic and healthy effect.

2 tbl, 5 ex

FIELD: biotechnology, food processing industry, medicine.

SUBSTANCE: strain Bifidobacterium longum is obtained by sequential cycles of lyophilic drying and reducing on thioglycol cultural medium (50 cycles). Clones with excellent antioxidant activity have been selected according to maximum accumulation of bifidus-bacteria biomass in sterile defatted milk not less than lg 9 after cultivation for 16 h. Strain has superoxide dismutase activity not less than 25.9 unit/mg of microbial protein mass. Strain B. lognum can effectively growth in cultivation media and accumulate necessary titer of viable bacteria, has high acid-forming activity, antagonistic action against pathogenic and conditional pathogenic microflora, and excellent antioxidantaction. Culture has marked probiotic action cased by its superoxide dismutase activity. Strain is useful in preparation of functional foodstuff, bioactive supplement, cosmetic and toiletry agents.

EFFECT: bifidus-bacterium strain for manufacturing of products with excellent probiotic and healthy effect.

2 tbl, 5 ex

FIELD: biotechnology, food processing industry, medicine.

SUBSTANCE: strain Bifidobacterium infantis is obtained by sequential cycles of lyophilic drying and reducing on thioglycol cultural medium (50 cycles). Clones with excellent antioxidant activity have been selected according to maximum accumulation of bifidus-bacteria biomass in sterile defatted milk not less than lg 9 after cultivation for 16 h. Strain has superoxide dismutase activity not less than 25.9 unit/mg of microbial protein mass. Strain B. onfantis can effectively growth in cultivation media and accumulate necessary titer of viable bacteria, has high acid-forming activity, antagonistic action against pathogenic and conditional pathogenic microflora, and excellent antioxidantaction. Culture has marked probiotic action cased by its superoxide dismutase activity. Strain is useful in preparation of functional foodstuff, bioactive supplement, cosmetic and toiletry agents.

EFFECT: bifidus-bacterium strain for manufacturing of products with excellent probiotic and healthy effect.

2 tbl, 5 ex

FIELD: pharmaceutics, in particular, curative agent and device for its preparation.

SUBSTANCE: the invention is in the fact that the curative agent on the basis of a sodium chloride solution includes ions of copper within 6.0 to 24.0 mg/l; citric acid - 100 mg/l; and ions of silver, the amount of silver makes up at least 1/1,000 part of the amount of copper. The ionator for preparation of a curative agent of antimicrobic effect has two short-circuited of antimicrobic effect has two short-circuited electrodes, one electrode is made of copper, and the second electrode - of silver. The area of the surface of the copper electrode relative to the area of the surface of the silver electrode makes up within 1:0.8 to 1:1.

EFFECT: enhanced antimicrobic effect, improved regenerative processes and accelerated recovery.

5 cl, 1 tbl, 1 dwg

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