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Determination of optimum immunization dose of vaccines against anthrax from Bacillus anthracis ua-07 at guinea pigs
One of the main issues in the safety of animal and human health in our time is bioterrorism. One of the bacterial pathogens used as an agent is Bacillus anthracis. The causative agent causes anthrax disease and can be used for massive impressions, leading to significant economic losses for countries. Prevention of anthrax develops antisubmarine vaccines. Abroad use the licensed vaccine AVA (Anthrax Vaccine Adsorbed) from the strain V770-NP1-R (USA), the strain Langzhou A16R (China) and others.
In Ukraine, as a result of vaccination, anthrax occurs sporadically. According to the results of the research, it was found that the largest number of diseased animals in the year was observed in Volyn (1994), Luhansk (1994), Kherson (1999), Odessa (2000), and Kyiv (2001) regions.
The development of preventive study does not stop and scientists are looking for better indicators when using live vaccines. During the development of vaccine digs, various properties of strains are studied in laboratory animals, in particular in Guinea pigs. In determining the immune status of experimental animals with anthrax, it is necessary to take into account not only the humeral and cellular indices of the factors of protection.
The purpose of the research is to study the optimum immunization dose of vaccine against anthrax, produced from the strain of Bacillus anthracis UA–07 in laboratory animals.
The experimental series of the vaccine against anthrax of animals from the strain Bacillus anthracis UA–07 "Antravak" was used in this work. The control strain (strain punch) used strain Bacillus anthracis M-71 from the collection of the National Center for strains of microorganisms.
Determination of pathogenic activity of the test strain Bacillus anthracis M–71 was carried out on clams weighing 350-400 g. Concentration of the dispute was determined in the camera Guryev. The breeding of spore suspension strain was carried out in isotonic sodium chloride solution and subcutaneously from the inner surface of the right hip was infected with mollusks. Animal monitoring was carried out for 10 days. Fixed the death of experimental animals. To determine the optimum immunization dose of the live spore vaccine created, a series of vaccine No. 1 containing 13.03 million spores / cm3 was used.
It was found that all the mullets were killed in the groups that were administered 10000 and 12000 spores. Consequently, 1 MLD of the control strain of Bacillus anthracis M–71 was 1,106 spores, since the mortality of the Guinea pigs was 100% (2–3 days after the injection).
The results of the studies on the study of the optimum immunization dose of Bacillus anthracis UA–07 strain indicate that anthrax vaccine Anthravac manufactured from the strain Bacillus anthracis UA–07 did not cause an increase in body temperature above 1 oC from physiological norm, inhibition, anaphylactic shock, local reactions.
Introduction to experimental animals of Bacillus anthracis M–71 culture resulted in an increase (38,87 ± 0,54 oC) of body temperature and death in control animals on the second day. Vaccination of animals in different amounts of spores (7,82–11,73 million spores) resulted in a slight increase (within 37,70 ± 0,25 – 38,52 ± 0,21 oC) of body temperature.
The obtained data on the study of the optimal immunization dose on the Guinea pigs, indicate that during the period of observation after administration of the Bacillus anthracis M–71 culture, the control animals of the control animals died from the second to the fourth day (100%). Experimental groups of animals that received spores of Bacillus anthracis UA–07 strain in the amount of 7,82–23,45 million spores remained alive (100%).
The highest immune protection was in the Guinea pigs of 3–8 groups, which received 7,82–3,45 million spores of Bacillus anthracis UA–07.
It should be noted that the dose of 5,21 million spores caused protection in 60% of vaccinated animals, while the highest number of spores – 6,52 million was introduced in the body of Guinea pigs, caused 70% protection. In the third group of animals, the level of protection against the pathogenic strain of Bacillus anthracis M–71 was 80 %. An increase in the number of controversies entered to 23,45 million – has led to the protection of 100% of clams.
Thus, the strain Bacillus anthracis UA–07 in the dose from 7,82 to 23,45 million live spores provides immunity against the introduction of 1,0x106 live spores of the Bacillus anthracis M–71 strain. Anthravac vaccine studies have shown a high level of protection in vaccinated animals against the control strain Bacillus anthracis M–71 at a dose of 7,82–23,45 million spores.
Anthravac vaccine, made from the Bacillus anthracis UA–07 strain, provided protection for Guinea pigs at doses of 9,12, 10,42, 12,38, 13,03 and 23,45 million spores at a level of 100%. The protection of Guinea pigs at the level of 80% provided vaccine doses of 7,82 million spores.
1. Dumas E.K., Garman L., Cuthbertson H., Charlton S., Hallis B.,. Engler R.J, Choudhari S., Picking W.D., James J.A., Farris A.D. (2017). Lethal factor antibodies contribute to lethal toxin neutralization in recipients of anthrax vaccine precipitated. Journal Vaccine. Vol. 35 (26), pp. 3416–3422.
2. Jones R.M., Burke M., Dubose D., Chichester A.J., Manceva S., Horsey A., Streatfield S.J., Breit J,, Yusibov V. (2017). Stability and pre-formulation development of a plant-produced anthrax vaccine candidate Author links open overlay panel / R.M. Jones, // Journal Vaccine. Vol. 35 (41), pp. 5463–5470.
3. Kracalik I., Abdullayev R., Asadov K., Ismayilova R., Baghirova M., Ustun N., Shikhiyev M. (2014). Changing patterns of human anthrax in Azerbaijan during the post-soviet and preemptive livestock vaccination eras. Journal Plos neglected tropical disrases. Vol. 8(7), pp. 1–12.
4. Jula G.M., Sattari M., Banihashemi R., Razzaz H., Sanchouli A., Tadayon K. (2011). The phenotypic and genotypic characterization of Bacillus anthracis isolates from Iran. J. Trop. Anim. Health Prod. №43 (3), pp. 699–704. doi:10.1007/s11250-010-9756-2 18.
5. Li Y., Yin W., Hugh-Jones M. (2017). Epidemiology of Human Anthrax in China 1955-2014. Emerg Infect Dis. №23 m(1), pp. 14–21.
6. Zavіrjuha G.A., Slups'ka V.V., Javors'ka K.V. (2014). Protisibіrkovі vakcini ta perspektivi їh udoskonalennja. Veterinarna bіotehnologіja. № 24, pp. 56–63, http://nbuv.gov.ua/UJRN/vbtb_2014_24_12.
7. Friedlander A.M., Little S.F. (2009). Advances in the development of next-generation anthrax vaccines. Vaccine.
Vol. 27 (4), pp. 28–32, t-generation.
8. Merkel T.J., Perera Pin-Yu, Kelly V.K., Verma A., Llewellyn Z.N., Waldmann T.A., Mosca J.D., Perera L.P. (2010). Developmen to fahighly ef efficacious vaccinia-baseddual vaccine against small poxand anthrax, two important bioterrorentities. Proc. Natl. Acad. Sci, USA. Vol. 107 (42), pp. 18091–18096.
9. Rublenko І.O., Skripnik V.G. (2016). Analіz danih epіzootichnih spalahіv sibіrki na teritorії Ukraїni [Analysis of the data of epizootic outbreaks of anthrax on the territory of Ukraine]. (perіod 1994 – 2016 rr.). Nauk. vіsnik vet. med. Zbіrnik naukovih prac'. Vip.1 (127), Bіla Cerkva, №1 (127), pp. 87–95.
10. Derzhprodspozhivsluzhba vzhivaє zahodi shhodo nedopushhennja rozpovsjudzhennja spalahіv sibіrki [State Committee for Proprietary Services takes measures to prevent the spread of outbreaks of anthrax]. http://www.consumer.gov.ua/ News/2280/Derzhprodspozhivsluzhba_vzhivae_zakhodi_shchodo_nedopushchennya_rozpovsyudzhennya_spalakhiv_sibirki.
11. Chitlaru T. A., Israeli M., Rotem S., Elia U., Bar-aim E., Ehrlich S., Cohen O., Shafferman A. (2017). Novel live attenuated anthrax spore vaccine based on an acapsular Bacillus anthracis Sterne strain with mutations in the htrA, lef and cya genes. Journal Vaccine. Vol. 35 (44), pp. 6030–6040.
12. Bezymennyi M., Barro A., Skrypnyk A., Skrypnyk V., Blackburn J.K. (2014). Spatio-temporal patterns of livestock anthrax in Ukraine during the past centry (1913–2012). Journal Applied geography. Vol. 54, pp. 129–138.
13. Liang Y., Y.Liang, Coffin M.V., Manceva S.D., Chichester J.A., Mark R. (2016). Controlled release of an anthrax toxin-neutralizing antibody from hydrolytically degradable polyethylene glycol hydrogels. Inc. journal biomed. mater. Res. Part. №104A, pp. 113–123.
14. Remy K.E., Hicks C., Eichacker P.Q. (2016). Anthrax Infection. Human Emerging and Re-emerging Infections. Journal Viral and Parasitic Infections. Vol. I, pp. 773–794.
15. Missiakas D., Schneewind O. (2017). Assembly and Function of the Bacillus anthracis S-Layer. Journal Annu. Rev. Microbiol. Vol. 71, pp. 79– 98.
16. Kuznecov V.G. (2004). Sposoby vychislenija 50 % jeffektivnoj dozy biologicheski aktivnih agentov. Zhurnal mikrobiologii, jepidemiologii i immunologii. [Methods of calculating 50% of the effective dose of biologically active agents. Journal of Microbiology, Epidemiology and Immunology]. № 6, pp. 18–22.
17. Romanov G.I. (1977). Izuchenie virulentnyh svojstv sibirejazvennyh shtammov po LD50. Trudy gos. nauch.-kontr. in-ta veterinarnyh preparatov. [Study of virulent properties of anthrax strains according to LD50. Transactions of state. sci. -cont. in-t veterinary drugs]. M. Vol. 24–25, pp. 186–191.
18. (2004). Manual of diagnostic tests and vaccines for terrestrial animals (mammals, birds and bees) / World organization for animal health OIE fifth edition volume 1, Paris, pp. 133–144.
19. Anthrax Spore Vaccine (Live) for Veterinary Use: 01/2005:0441 // European Pharmacopeia 5.0. 715 p.
20. (2015). Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Quality control of vaccines // OIE Terrestrial manual. –. http://www.oie.int/fileadmin