Specific Appurtenance, Numbers, and Dynamics of Interaction of Acanthamoeba from Soils of Gorno-Altai High-Mountain Plague Focus with Yersinia pestis Strains

Objective of the study is to analyze the appurtenance, numbers, and dynamics of interaction of acanthamoeba from soils of GornoAltai plague focus with Yersinia pestis 367 strain, isolated in 2016 in enzootic territory of this focus. Materials and methods. Utilized were soil amoeba from Gorno-Altai high-mountain focus and the strain Y. pestis 367 of the main subspecies of antique biovar, isolated there in 2016. Determination of systematic relation of the isolated amoeba was carried out using PCR with genus specific primers and sequencing of the obtained PCR fragments followed by identification of nucleotide sequences against GenBank database. Localization of Y. pestis cells in acanthamoeba was performed using fluorescent antibody technique by means of Axio Imager Z2 (Carl Zeiss, Germany). Results and conclusions. For the first time ever established has been the presence of Acanthamoeba castellanii in soils of burrows of Marmota altaica in the numbers of up to 300000 cells/gr in Gorno-Altai high-mountain focus. Investigated has been the dynamics of interaction of these microorganisms. Preservation of the agent in vacuoles of endoplasmatic reticulum within 14 days has been revealed. It is an indicative of the possibility of Y. pestis persistence in amoeba of Acanthamoeba subspecies in soil biocoenosis of Gorno-Altai high-mountain plague focus.

plague agent, acanthamoeba, natural plague foci, mechanisms of persistence, Acanthamoeba, Yersinia pestis

1. Domadarsky I.V. [Is not plague a protozoan disease? (The role of Protozoa in epizootiology of plague)]. Med. Parazitol. 1999; 2:10–3.

2. Koshel’ E.I., Anisimova L.V., Novichkova L.A., Vidyaeva N.A., Guseva N.P., Eroshenko G.A., Kutyrev V.V. [Determination of specific appurtenance of soil amoeba from plague foci of Pre-Caspian based on analysis of ribosomal operon regions]. Genetika. 2015; 51:30–45. DOI: 10.7868/S0016675815010051.

3. Koshel’ E.I., Eroshenko G.A., Anisimova L.V., Novichkova L.A., Shirokov A.A., Burov A.M., Kuznetsov O.S., Kutyrev V.V. [Exploratory study of the long-term persistence of Yersinia pestis in the cells of soil-inhabiting ameba – Acanthamoeba ssp.]. Probl. Osobo Opasn. Infek. 2016; 2: 69–74. DOI: 10.21055/0370-1069-2016-2-69-74.

4. Kutyrev V.V., Eroshenko G.A., Popov N.V., Vidyaeva N.A., Konnov N.P. [Molecular mechanisms of plague agent interaction with invertebrate animals]. Mol. Genet. Mikrobiol. Virusol. 2009; 4:7–12.

5. Kutyrev V.V., Popova A.Yu., Ezhlova E.B., Demina Yu.V., Pakskina N.D., Shchuchinov L.V., Mikhailov E.P., Mishchenko A.I., Rozhdestvensky E.N., Bazarova G.Kh., Denisov A.V., Sharova I.N., Popov N.V., Kuznetsov A.A. [Infection of an individual with plague in the Gorno-Altai high-mountain natural focus in 2014. Communication 1. Epidemiological and epizootiological peculiarities of plague manifestations in the Gorno-Altai high-mountain (Sailyugemsky) natural plague focus]. Probl. Osobo Opasn. Infek. 2014; 4:9–16.

6. Onishchenko G.G., Kutyrev V.V., editors. [Laboratory Diagnostics of Dangerous Infectious Diseases. Practice Guidelines]. M.: ‘Shiko”, CJSC; 2nd Edition, 2013. 560 p.

7. Popov N.V., Koshel E.I., Eroshenko G.A., Kutyrev V.V. [Formation of modern concepts on the mechanism of plague enzooty]. Probl. Osobo Opasn. Infek. 2011; 3(109):5–8.

8. Abd H., Johansson T., Golovliov I., Sandstrom G., Forsman M. Survival and growth of Francisella tularensis in Acanthamoeba castellanii. Appl. Environ. Microbiol. 2003; 69:600–6. PMID: 12514047; PMCID: PMC152416.

9. Greub G., Raoult D. Microorganisms resistant to free-living amoebae. Clin. Microbiol. Rev. 2004; 17:413–33. PMID: 15084508; PMCID: PMC387402.

10. Pelandakis M., Pernin P. Use of multiplex PCR and PCR restriction enzyme analysis for detection and exploration of the variability in the freeliving amoeba Naegleria in the environment. Appl. Environ. Microbiol. 2002; 68(4):2061–5. PMID: 11916734; PMCID: PMC123847.

11. Schroeder J., Booton G., Hay J., Niszl I., Seal D., Markus M., Fuerst P., Byers T. Use of subgenic 18S ribosomal DNA PCR and sequencing for genus and genotype identification of Acanthamoebae from humans with keratitis and from sewage sludge. Clin. Microbiol. 2001; 39(5):1903–11. DOI: 10.1128/JCM.39.5.1903-1911.2001.