SU1706631A1 - Method for preparation anatoxin vaccine against bird staphylococcus - Google Patents

Abstract

The invention relates to microbiology, in particular to the preparation of a toxoid vaccine for the specific prevention of avian staphylococcus. The purpose of the invention is to simplify the technology and increase the activity of the target product. The method consists in growing staphylococci on a nutrient medium and inactivating them with detoxifying toxin to toxoid with formalin. The essence of the invention lies in the fact that the suspension of staphylococci isolated from patients with staphylococcal birds of birds, after growing on a liquid nutrient medium with 5-7% sodium chloride and glycerol for 7-10 days to a concentration of 5-7 ml of microbial cells C 1 ml are sterilized at 85-1 СО ° С turning of 1.0-1.5 h with detoxification of exo- and endotoxins with 0.6-0.8% formalin solution during exposure for 7-10 days. Use aerosol. I, l | HL C

Description

This invention relates to microbiology. D in particular, to the method of obtaining anatok-syn-psktsyna for the specific prevention of staphyloconage: oz-birds.

Methods of obtaining toxoids for the prevention of tetanus, staphylococcal infection, diphtheria are tested. formolsaccine against diplococcoda septicamin t, piglets, salmonellosis. based on detoxification of the toxin and inactivation of bacteria, viruses, formalin 1Ji-f 2.

A method of obtaining staphylococcal toxoid is known, which involves the identification of staphylococci on the kzozoin hydrolizate medium with the addition of blood serum and installations for the supply of sterile carbon dioxide, oxygen, followed by detoxification of the toxin and inactivation of staphylococci with 0.4-0.6% formalin 3 solution.

The disadvantages of the method are the lack of sterilization by the progress of staphylococci, the need to supply sterile gases, the duration of detoxification, the weak activity of the drug due to the use of only the product of autolysis and the biosynthesis of staphylococci, i.e. exotoxin, and its internal use.

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SHIROCCH; Г Г: СЛрОСТр:}: Г 1П 0 G. TOTF / LOCOK. .O ia birds ng g ;:; tsofoOri:; g with; requires (50.:,; l. tpsziogo, T jyhr..JUST PR J .; ..; 1Ј TICH; CH.) SL / LM PZ 113

Shgammoo .lg /; plako / .k.C ::;, syuy l oidi ;; zg: - Solsiani at pg; c,

Tspl and: 5p t - :; NPP - at l DPRK5 technology and noiH-iLL tjnr.Q activity of the target product.

According to method aa: lyuchuyuchnusnu in stavilokokk yg, nutritious with re to, and;: pntoyrsaan l them with detox, toxoid with formalin, use staphylococcus, separated from birds suffering from stigfilokko-zsm, and burning vehicles: sigg: nymi and ilizmikoo.migiruyuschimy SEYOSTOZMI who grew up ..;, 07 on;: and about 7--10 days at. 23-100C to 5-7 d. microbial cells i; . with the following.) sterilization of microorganisms at 85-100 ° С s for 1.0-1.5 h and toxicity of 0.6-0.8% with its formalin at 45-53 ° С for 7- 10 days, and the finished product is applied twice with an interval of 7-10 days by aerosol spraying for 40 minutes for chickens of 10–20 days at a rate of 1.0–1.2 l of zmatoxmn-vaccine per poultry house with a volume of 5000.

In connection with the resistance of staphylococcal toxin to cypcenic dotoxication of the toxin with formlean prssodlt at 45-55 ° C after the preliminary sterilization of microorganisms, the pH is 85-100 ° C for 1.0-1.5 h for g. completeness of inactivation of staphylococci and sydsleg: is.ch zndoxnna at ood o-tsrmicheskogo gidropmze cultures.

Re: - ppm of the staphylococcus staphylocation contributes to the elimination of the toxin-protein complex and iionv.CcXDpii cs with a cultured liquid up to 0.0-0.7 mg / ml instead of 0.20-0.25 to aptoklespropim above m - n. the biological activity of the drug.

A distinctive feature of the preparation and absorption of oxygen is the use of connotative toxicity in place of the transport route of the subcutaneous and internal hypodermic tissue to prevent 95-97% of the population of 10-30-years of age from somalizing hundred: l:; s:; eog: .. 1.

Example 1. One rep. Eiko rpviroTor: iv-whether mmso ;; gz: - :::;,.; This is about synthetic cpe, -j from 5% corner. - .. ssgo lptrm , and mountain s..uchg- 1% g., 1. About about old with MO mln gl1, cs: ;;;: - and. .tsi about cf: d ustanaoli-ammi -: .-; l. -Or PCOX cases up to 7.7-7.8, h; chole pyro; :; l. For the PSS-GCU: from the PTS. , ::. 1. shu .-) t m hooo f f. -iitc tnuyu kulm1 ;, pf rd.alpnye strains

CTC J -H / icKOKKOoSt.aur & n St.epldprriilflls. FLP improvement: sc.

The growth of staphylococci in all environments takes place (, urn for 5 - H days: or medium sludge

the process basically ceases, s with glycerol continues to a concentration of 10-IF5 billion: i ml. The reaction of culture: liquid with a suspension of staphylococci after seven days of incubation is

Sredoh with SGHARA 6.2, o with glycerin - 6.8. EXAMPLE 2 Staphylococcus, expressed for 7–10 days on a liquid nutrient medium, was progressed to 85–100 ° C for one hour, followed by the addition of formalin to a 0.7% formaldehyde concentration to detoxify the toxin to toxoid. at 45-55 ° C for 7 days. Before aerosol spraying, glycerin was added to the total volume of staphylococcal anatox-packin. Aerosol spraying of toxoid vaccines was carried out twice with an interval of 7 days in 1 liter of the preparation per treatment for 40 minutes. Chicken t disease with staphylococcosis was not observed.

The safety of the birds vaccinated with the staphylococcal toxoid vaccine is 97% with an increase of 15%. However, during the commission check and the breeding of raised birds 15 heads were found

with patkartina characteristic of staphylo-coccosis.

Note 3. After a seven-day incubation of staphylococci on a casein-hydrolyzate medium with glycerol

the suspension of microorganisms is sterilized at 100 ° C for one hour and detoxified with 0.7% formalin at 55 ° C for 7 days.

After testing for sterility, the absence of allergenic and allergenic properties in guinea-pigs, a toxoid-vaccine suspension is used to double for 7 days with an aerosol spray B for 40 minutes based on

1.0 1.2 liters per poultry house with 40 thousand chickens of 15-month old. The incidence of staphylococcal birds and the padezh from him to srednegl is 50-75 per each bird house.

Example 4 Cultivation of staphylococcosis is carried out on liquid nutrient soda with glycerin for 7 days to a concentration of 5 billion microbial cells in 1 ml. To obtain anatoxin-vaccine, suspension of staphylococci is inactivated at 35 ° C for 1.5 hours, and then deactivation is performed with a 0.5% formalin solution at 45 ° C for 10 days.

AFTER NJ IO: pKH H3 .10. EAGLES 1 L L L, TOXICITY iij, kgg before.-I.-poiio / ibiiiiM by spraying with a 5% glyceri - divider v-rastorom with: ohm in soy. Error 1: 5. Aerosol sprayed for 40 minutes and. rgg.chote 1.6 /: N;: TIER CI of the original connotation-pzhtsy HI o / tick avian volume COO MJ with 40 tms chomi chickens 15 days of age.

A re-run was performed after 10 days. After the use of staphylococcal anztohsin-wag.roshi, zgbollegnne and chickens waste from staphylococcosis is 1.7%. When using natox.im-packs of a negative g-ml or the drug, it is noted that the chickens of mushroom sesene are 12-16% more compared to the control ones.

Example 5. A staphylococcal culture grown on liquid nutrient medium with glycerin for 10 days to 7 billion microbial cells in 1 ml is sterilized at 100 ° C for one hour, formalin is added to detoxify to 0, 7% final concentration. Detoxification was carried out at 55 ° C for 7 days. After testing for sterility, the absence of toxin transfer again to the toxin, the preparation was used for a two-time spray with an interval of 10 days at the rate of 1.5 l per one 5,000-meter house with 40 thousand chickens of 20 days of age. Before spraying, the preparation is diluted with a physician with 5% glycerol in a ratio of 1: 5. The protective effect of staphylococcus toxoid-vaccine is 97.5%.

Example 6. For the growth of staphylococci, a combination of x-containing hydrolytic and synthetic media with glycerol was used without carbon dioxide and oxygen being supplied. Staphylococci are cultivated at 38CC in flow1; 10 days, the suspension is sterilized at 10 ° C for one hour. Formaldehyde is added to the inactivating suspension of staphylococci to 0.8% concentration and kept in a thermostat at 55CC for 7 days with occasional stirring. Zztem propertyut sterility for completeness ipakt ioztsi-l.

Anato Sin-Zaccina is sprayed with an aerosol about twice at an interval of eight days at expo. /u/.vi zi min, Zaoevanil birds stafiloko. OOSM p for 2.5 mine. but noted. Control infection of 60 golos (podse from each gpichnikl) suspension of staphylococci (aerosol discharging / e and kokokhno: 1 sssdnii;}) but samy za zlkhanii and pado: .-: and birds.

PRI me R 7. Dh prashisani p prigotoslon1 stat- and l o ls; Oiioi about apatoxins

on synthetic with. glyceg type was used strain Si.riurcns. A patient with staphylococcal poultry. On the spot of the stump day, -ch, ;; -.- tnig bacterial mass accumulation is 4 billion micro-5 Ony cells and 1 ml, on the seventh day - 5 k; lrd. microbial cells in 1 ml, and on the tenth day - billion microbial cells from 1 ml. In the following days, imultiirog concentration and mi ;; horopismo does not increase.

0 Aatlocvirus at 80 ° C of a staphyrophage / yucoccop culture does not ensure complete inactivation of microorganisms. and when the regime of the ptohlsvirooani 85 ° C for 1.5 h and sterilization 100 ° C for

5 one hours full microbial injection and the maximum release of toxins into the culture fluid.

The preparation of the toxoid vaccine was carried out using a 0.8% formalin solution at 50 ° C for 10 days. The lack of toxic properties of the culture liquid was checked by subcutaneous administration to white mice by

5 0.2-0.5 ml and guinea pigs - 0.80 ml. Aerosol treatment is carried out in accordance with examples 2-6.

Example The seeding of the mother suspension of the St.aurens strain isolated from farm animals is carried out in ml of casein hydrolyzate medium with glycerol. On the fifth day, the accumulation of bacterial mass is 4.5 billion, microbial cells in 1 ml, on the seventh day - 5, 2

5 billion microbial cells in 1 ml, on the tenth day - 7 billion microbial cells in 1 ml. The preparation and use of toxoid is carried out according to the mode of Example 7.

Thus, the advantages of the method of obtaining anatoxin-vaccine against avian staphylococcus are the use of them in casein hydrolysis or synthetic medium with glycerol without sulfur and the supply of sterile carbon dioxide, heat and heat inactivation of the suspension of staphylococci, toxins, to them, or in groups, they have been subject to them, they are inactivated by staphylococcal suspension, staphylococci, doxytoxins, and those who have received sterile carbon dioxide. . Nutrient media without sugars during the sterilization process

0 provide rapid growth of staphylococci. and subsequent sterilization with the use of pyroplasmone at 85-1CO ° С 1.0-1.5 h contributes to the additional release of the toxin.

Claims (1)

5 Formula of Invention The method of obtaining anatsksin-n.ktsiny against birds staphylococcus, including the Bird 4 culture of Art. F: - about the environment, the historical level 11.1 Hc omog.gi; 7, 170SC31 8 POST-1: CV), еТ (JS: 1КЈ1.: П ТИКСМНП, КОГ .. MLR.D.MIKRP THEM raspberries. about t l n h p-o y; and with g; so that, with O-C - t ml, sterilization is carried out at the purpose of ulitschznzhnich tekhnogogg. and FLASH NU Jo - U j ° C for i- t, 5 h, and de.ts-hiktstsiyu I will come: -71p, g; yrsd vyr.-ssho tg. Fri 0,8-0, In% solution shivakigm p sgsg.u ds gut 4 o5.% glg.io-5 forg- “npvi exposure 45-l5 ° C and tscherina. Airschis culture: from a turning 7-10 down 7-10 days-z.