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Neutralization of B.anthracis Spores in Soil

#3177


Development of the Laboratory-Experimental Technology of Neutralization of B.anthracis Spores in Soil with Spore Germination Activators and Bacterial Phages

Tech Area / Field

  • BIO-MIB/Microbiology/Biotechnology
  • BIO-OTH/Other/Biotechnology
  • BIO-REM/Bioremediation/Biotechnology

Status
8 Project completed

Registration date
24.12.2004

Completion date
23.05.2011

Senior Project Manager
Crocker W E

Leading Institute
Institute of Immunological Engineering, Russia, Moscow reg., Lyubuchany

Supporting institutes

  • Center of Science of Biomedical Technologies, Russia, Moscow

Collaborators

  • Universitat Hohenheim / Institut fuer Umwelt- und Tierhygiene, Germany, Stuttgart\nImmune Network Ltd., Canada, BC, Vancouver\nMDM Group, Inc., USA, CA, Santa Clara

Project summary

The objective of the project is to develop a laboratory-experimental technology for neutralization B.anthracis spores in soil with spore germination activators and bacteriophages ; to determine a dependence of the process of neutralization on the type of soil and its chemical and physical compositions by modeling various natural conditions; to select the most effective anti-B.anthracis bacterial phages and spore activators; to set up an experimental site to produce B.anthracis bacterial phages and activators in quantities sufficient for conducting laboratory and subsequently field trials. (All works in the framework of the project will be performed by using vaccine strains of B.athracis).

The B.anthracis-associated anthropozoonosic infection is of nature-locus character. Therefore, for preventing the spread of the infection, measures aimed at decontaminating areas to eliminate the pathogen, namely B.anthracis spores, should be taken [1-7].

According to estimates (B.L. Tcherkassky, Academician, RAMS, Central Research Institute of Epidemiology, Ministry of Health, RF,), there are about 35,000 cattle burial grounds in Russia, which are considered potential reservoirs for this infection [20, 23, 24, 29-30]. The Institute is currently making up a land-survey of these sites. Most of them are located in regions of Northern Caucasus Mountains, the river Volga, Central Russia, as well as in some other Russian regions [29-32]. However, biological safety regimen with these grounds are not always followed in a proper way. So, some cattle burial grounds are located within agricultural areas that are actively explored. They naturally become a zone of high risk. The others are still unidentified, and corresponding records about them are lost [19, 20, 27, 30, 31]. Before the post-war period (2nd World War, 1941-1945), unregistered cattle burial grounds existed nearly in every village. Their locations remain unknown. It is often that construction of gas- and oil- pipelines, the digging of trenches, as well as construction of hydraulic engineering facilities is fulfilled without the involvement of the Ministry of Health [30].

Dr. E.N. Shlyakhov [16, 31, 32] offers his list of settlements, which is made up depending on the incidence of the disease, to elaborate differential measures for monitoring of B.anthracis:


- Favorable settlements, where cases of anthrax in animals and humans did not take place or were not registered for a long time.
- Unfavorable settlements, which are pided into 4 categories:
- “demonstrative” settlements, where cases of anthrax cases took place within recent 10 years, including: 1) “regular” settlements, where cases of anthrax is annually registered or nearly annually; 2) “relapsing” settlements, where the infection occurs from time to time (for example, every 3-5years);
- “conventional- demonstrative”, where cases of anthrax previously occurred every 2-3 years, but with interval of not more than 15 years;
- “non-demonstrative” settlements, where anthrax was previously registered, but the disease did not occur for a long time (3-5 years for newly formed settlements, 10 years for demonstrative, 15 years for conventional-demonstrative);
- unclassified or newly set up settlements, where anthrax is firstly registered, and 3-5-year monitoring is needed. Depending on outcomes of the monitoring, such settlements may be attributed either to demonstrative (relapsing) or to non-demonstrative sites.

All loci containing spores of anthrax are a potential source of this lethal infection. Worldwide there are thousands of unfavorable cattle burial grounds, which may release the pathogen because of rain falls, floods, and human activities. Spores remain viable for hundreds of years. As to application of anthrax spores by terrorists, then, according to data provided by the WHO, all countries of the world are highly vulnerable to this threat. It is not always possible and reasonable to apply disinfectants to liquidate natural reservoirs for anthrax.

In this connection, we propose to develop a method allowing neutralization of anthrax spores in soil with bacterial phages and spore germination activators; to develop a complex formulation to be used in practice.

Phages themselves are unable to degrade spores of B.anthracis. They affect either initial spores (germinated spores) or vegetative cells of B.anthracis. Therefore, the application of bacterial phages along with activators of trigger mechanisms of spore germination is more reasonable. Activators might be physicochemical and biological factors, such as temperature, humidity, рН of the environment, L-alanine, and surface active substances (SAS), etc.

Project participants have long experience in the field of especially dangerous infections, bacterial phages and specific activators for B.anthracis spores [9-26]. In Russia there are anti-B.anthracis phages, but so far they are out of practical usage as bacillus disactivators. Besides the development of protective and prophylactic measures, our work will contribute to solving the problem of biological terrorism.


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