AP14870683 «Distribution and genetic characterization of causative agents of tick-borne relapsing fevers (Relapsing Fever Borrelia) in the southern region of Kazakhstan»


Tick-borne relapsing fever (TBRF) is a group of generalized infections caused by RF borrelia (Relapsing Fever Borrelia), accompanied by an infectious intoxication syndrome manifested by multiple bouts of fever interspersed with periods of apyrexia. The main problem of untreated borreliosis is a fairly frequent transition of the acute phase of the disease into a chronic form. Due to the presence of certain virulence factors in borrelia, immunity to them is not sterile, and the pathogen can persist in the body for a long time. Correct diagnosis and timely initiation of adequate antibiotic treatment are very important, because after the disease passes into a chronic state, its consequences can take a generalized character and lead to the disability of patients.

Species of tick vectors of TBRF agents are widespread in our country, and previously the presence of DNA of one of the representatives of RF-borreliae was shown in Eastern Kazakhstan. Both ixodes and argas ticks are common in the southern region of Kazakhstan. While Ixodes persulcatus, the main potential vector of B. miyamotoi, is predominantly distributed in the forested mountain ranges of the Zailiyskiy Alatau and Dzungarian Alatau (Altmati region), the argas ticks Ornithodoros spp. and Argas spp., which are the main potential vectors of TBRF, live mainly in desert and semi-desert zones of Turkestan and Zhambyl regions of Kazakhstan. It should be noted that the last large-scale studies concerning the establishment of tick distribution areas on the territory of the RK belonged to the Soviet period (70s of the XX century). At the same time, for the last three decades there have been significant climate changes in the territory of the country, especially in the southern region of Kazakhstan. And the climate changes that occurred during these years could affect the distribution areas of ticks-carriers of dangerous diseases. This is especially true for argas ticks, whose range may seriously expand northward due to the increase in average annual temperatures. In this regard, the data on the occurrence of certain species of argas and ixodid ticks in the southern region of Kazakhstan, obtained during this project, will be useful for understanding the current ranges of their distribution. Tick-borne infections such as tick-borne encephalitis, rickettsioses and tularemia are monitored quite well in Kazakhstan. Other tick-borne infections are not as well researched. At the same time, there is no epidemiologic surveillance of tick-borne infections in Kazakhstan. Currently, there are only mentions in the literature that B. miyamotoi DNA is detected in ticks caught in Kazakhstan. There is no information about other representatives of RF-borrelia in Kazakhstan at all. Studies of argas ticks for the presence of Borrelia DNA in them have not been conducted. No large-scale studies on the level of infection of ticks with these borreliae, as well as the level of prevalence of antibodies to RF-borreliae have been conducted so far.

Within the framework of the project, it is planned for the first time to carry out genetic characterization of Kazakhstani isolates of RF-borreliae by partial sequencing of their DNA at several genomic loci. This will allow phylogenetic analysis with respect to TBRF pathogens on the territory of the country. The project also aims to study the distribution of RF-borreliae in the southern region of Kazakhstan, genotyping of the pathogen and determination of seroprevalence of antibodies to this group of pathogens in the population of the southern region of the country. The results of the work will allow to justify the surveillance strategy, optimize methods of detection of RF-borrelia DNA, reduce the risk of spread of Ebola and increase the efficiency of diagnosis of generalized fevers of unclear genesis in Kazakhstan.

The aim

The aim of the project is to study the spread of RF-borreliae in the southern region of Kazakhstan, conduct their genotyping, determine the seroprevalence of antibodies to RF-borreliae in the population of the southern region of the country and develop a strategy for epidemiological surveillance of this group of infections in the Republic of Kazakhstan.

Expected Results

The results obtained will contribute to the introduction of epidemiologic surveillance of Ebola in Kazakhstan and should improve the quality of diagnostics of fevers in the south of the country. The results obtained in the course of the project realization can become the basis for improvement of epidemiological surveillance of natural focal infections on the territory of Kazakhstan, ultimately serving the welfare of citizens and reducing the costs of the state in the sphere of public health.

In the course of the project realization the following direct scientific results will be obtained:

– GIS maps of the collection points of the main vectors of TBRF will be created;

– phylogenetic trees will be constructed for TBRF isolates from the southern region of Kazakhstan;

– a standardized definition of an TBRF case (presumptive, probable and confirmed cases) will be developed;

– new primers for detection of RF-borreliae by nested PCR to the 16S rRNA gene region will be developed;

– recommendations for doctors will be developed to improve the quality of diagnostics of CFS in Kazakhstan.

Based on the results of scientific research conducted under the project, there will be published:

– at least 3 (three) articles and (or) reviews in peer-reviewed scientific publications indexed in Science Citation Index Expanded of Web of Science base and (or) having a percentile on CiteScore in Scopus base not less than 35 (thirty-five) presumably: in the journal “Virus Genes” (IF 2. 10; percentile 45 (Scopus); Q4); and at least 1 (one) article or review in a peer-reviewed foreign or domestic publication recommended by the CQAES (Presumably – in “Bulletin of KazNU. Series of biological and medical”);

– OR at least 2 (two) articles and (or) reviews in peer-reviewed scientific publications, included in the 1 (first) and (or) 2 (second) quartile on impact factor in the Web of Science database and (or) having percentile on CiteScore in the Scopus database not less than 65 (sixty-five) (presumably: in the journal “Indian Journal of Medical Research, Supplement “(IF 2.90; percentile 65 (Scopus); Q2 (WoS)).

– OR at least one (1) article or review in a peer-reviewed scientific publication with an impact factor in the first quartile of Web of Science and/or a CiteScore in Scopus of at least eighty (80) (presumably: PLOS Biology (IF 8.029; percentile 97 (Scopus); Q1 (WoS)).

Project manager

Skiba Yu.A. (1984) – project manager, candidate of biological sciences (molecular biology), deputy director of AF NCB, head of the Laboratory of Molecular Biology, biological security officer. Research interests: molecular-genetic typing and study of genetic determinants of drug resistance of pathogens of especially dangerous human and animal diseases, evolution and epidemiology of pathogenic microorganisms, development of diagnostic systems, recombinant technologies, structural and functional organization of eukaryotic genome, regulation of gene expression and genetic engineering. ORCID: 0000-0003-4895-1473. Scopus ID: 56677594900. Researcher ID: H-6528-2017.

Executive team members

Ostapchuk E.O. – Project Executive Officer, PhD (Biology), Associate Professor, Head of Laboratory. Research interests: molecular and cellular immunology, epidemiology, zoonotic infections. She has experience in the study of natural focal zoonotic infections. He is engaged in research of infectious agents of tick-borreliosis in Kazakhstan. ORCID: 0000-0002-3771-423X. Researcher ID: D-1254-2015. Scopus ID: 56823472400

Perfilieva Y.V. – PhD (Immunology), Associate Professor. Specialist in the field of molecular and cellular immunology. She has experience in the study of natural focal zoonotic infections. He is engaged in research in Kazakhstan on infectious agents of cu-fever and rickettsioses. ORCID: 0000-0001-6803-0773. Researcher ID: AAF-9666-2020. Scopus ID: 56823500600

Zhigailov A.V. – Candidate of Biological Sciences (molecular biology). Specializes in genetic engineering, research of molecular mechanisms of regulation of gene expression and protein biosynthesis, genetic characterization of viruses and bacterial tick infections. Trained to perform tick identification. ORCID: 0000-0002-9646-033X. Researcher ID: N-6073-2015. Scopus ID: 6508121286

Dmitrovsky A.M. – Doctor of medical sciences (epidemiology, zoonotic infections), professor, head of the laboratory, infectious diseases doctor. He is a specialist in the field of epidemiology, diagnostics and treatment of especially dangerous zoonotic human diseases. He was engaged in research of distribution in Kazakhstan of plague, anthrax, Crimean-Congo hemorrhagic fever, rickettsioses, Ku fever. ORCID: 0000-0003-4714-3079. Researcher ID: AAZ-2816-2020. Scopus ID: 57204864464

Mamadaliev S.M. (1948) – Doctor of veterinary sciences, professor, specialist in especially dangerous diseases of animals. In 1992-2010. S.M. Mamadaliev headed the RSE “Research Institute of Biological Safety Problems” KN MES RK, and from 2017 to the present time he is the Director of AF NCB. ORCID: 0000-0002-7767-0251. Researcher ID: N-8389-2017. Scopus ID: 37000092700

Maltseva E.R. (1983) – PhD-student, a specialist in molecular biology. At the AF NCB she is Head of the Biosafety and Biosecurity Division and Biosecurity Officer in the CRL. She has been involved in genotyping of human and animal pathogens such as brucellosis, tuberculosis, and borreliosis. E.R. Maltseva is part-time deputy chairman of the National Committee for the UNESCO program “Man and Biosphere”. ORCID: 0000-0001-9198-695X. Researcher ID: N-4309-2017. Scopus ID: 57202717826

Nizkorodova A.S. (1981) – Candidate of Biological Sciences (molecular biology). She specializes in genetic engineering, development of diagnostic test systems, detection of microbial nucleic acids. ORCID: 0000-0002-1597-7207. Researcher ID: AAY-1646-2020. Scopus ID: 57215971184

Abdollah N. (1985) – PhD (Immunology). Specializes in the study of molecular mechanisms of defense responses of human and animal organisms. ORCID: 0000-0002-4769-7824. Researcher ID: R-2193-2016. Scopus ID: 57194001982

Berdygulova J.A. (1985) – PhD-student (biotechnology), specialist in virology and molecular biology. She was engaged in the study of phages of pathogenic bacteria, as well as detection and molecular characterization of rickettsiae and borreliae. ORCID: 0000-0003-0379-2472. Researcher ID: I-2943-2018. Scopus ID: 23977664200

Mashzhan A.S. (1995) – PhD-student (biotechnology), a specialist in microbiology. He has experience in genomic and metagenomic research (next generation sequencing). ORCID: 0000-0001-6803-0773. Researcher ID: AAF-9666-2020. Scopus ID: 56823500600

Naizabaeva D.A. (1994) – PhD- student (biotechnology). In AF NCB she is responsible for registration, movement and storage of biological samples. She is responsible for compliance of works with the requirements of integrated management system and ISO. She has extensive experience in genotyping of dangerous and highly dangerous pathogens. ORCID: 0000-0002-0606-4289. Researcher ID: AAY-5696-2020. Scopus ID: 57218288692

Kuatbekova S.A. (1992) – Master’s degree, specialist in the field of veterinary medicine. She has experience in conducting monitoring studies on detection of human and animal pathogens. ORCID: 0000-0002-1091-0150. Researcher ID: ABA-3202-2020

Публикации и охранные документы научного руководителя проекта и членов исследовательской группы, касающиеся темы проекта

  1. Skiba, Y. (1-й автор, автор для корреспонденции), Mokrousov I., Nabirova D., Vyazovaya A., Maltseva E., Malakhova, N., Ismagulova G., Pole I., Ranka R., Sapiyeva Z., Ismailov S., Moffett D. Mycobacterium tuberculosis RD-Rio Strain in Kazakhstan. Emerging infectious diseases. 2019;25(3):604–606. https://doi.org/10.3201/eid2503.181179. Q1 (WoS); Scopus – 92%.
  2. Zhigailov A.V., Neupokoyeva A.S., Maltseva E.R., Perfilyeva Y.V., Bissenbay A.O., Turebekov N.A., Frey S., Essbauer S., Abdiyeva K.S., Ostapchuk Y.O., Berdygulova Z.A., Aimbetov R.S., Naizabayeva D.A., Dmitrovskiy A.M., Skiba Y.A. (автор для корреспонденции), Mamadaliyev S.M. The prevalence of Borrelia in Ixodes persulcatus in southeastern Kazakhstan. Ticks Tick Borne Dis. 2021;12(4):101716. https://doi.org/10.1016/j.ttbdis.2021.101716. Q2 (WoS); Scopus – 92%.
  3. Perfilyeva Y.V., Shapiyeva Z.Z., Ostapchuk Y.O., Berdygulova Z.A., Bissenbay A.O., Kulemin M.V., Ismagulova G.A., Skiba Y.A., Sayakova Z.Z., Mamadaliyev S.M., Maltseva E.R., Dmitrovskiy A.M. Tick-borne pathogens and their vectors in Kazakhstan – A review. Ticks Tick Borne Dis. 2020; 11(5):101498. https://doi.org/10.1016/j.ttbdis.2020.101498. Q2 (WoS); Scopus – 92%.
  4. Ostapchuk Y.O., Perfilyeva Y.V., Zhigailov A.V., Maltseva E.R., Neupokoyeva A.S., Bissenbay A.O., Berdygulova Z.A., Naizabayeva D.A., Nizkorodova A.S., Shapiyeva Z.Z., Yegemberdiyeva R.A., Kuznetsova T.V., Kuatbekova S., Akanova A., Ismagulova G.A., Mamadaliyev S.M., Dmitrovskiy A.M., Skiba Y.A. Monitoring of pathogenic Borrelia burgdorferi sensu lato in the Almaty oblast, Kazakhstan. Ticks Tick Borne Dis. 2021;12(4):101725. https://doi.org/10.1016/j.ttbdis.2021.101725. Q2 (WoS); Scopus – 92%.
  5. Mokrousov I., Vyazovaya A., Sinkov V. Skiba Y. et al. Practical approach to detection and surveillance of emerging highly resistant Mycobacterium tuberculosis Beijing 1071-32-cluster. Sci. Rep. 2021; 11:21392. https://doi.org/10.1038/s41598-021-00890-7. Q1 (WoS); Scopus – 93%.
  6. Yeh K.B., Tabynov K., Parekh F.K., Maltseva E., Skiba Y., Shapiyeva Z., Sansyzbai A., Frey S., Essbauer S., Hewson R., Richards A. L., Hay J. Building Scientific Capability and Reducing Biological Threats: The Effect of Three Cooperative Bio-Research Programs in Kazakhstan. Frontiers in public health. 2021;9:683192. https://doi.org/10.3389/fpubh.2021.683192. Q1-Q2 (WoS); Scopus – 56%.
  7. Mokrousov I., Shitikov E., Skiba Y., Kolchenko S., Chernyaeva E., Vyazovaya A. Emerging peak on the phylogeographic landscape of Mycobacterium tuberculosis in West Asia: Definitely smoke, likely fire. Molecular phylogenetics and evolution. 2017;116:202–212. https://doi.org/10.1016/j.ympev.2017.09.002. Q2 (WoS); Scopus – 93%.
  8.   Ostapchuk Y.O., Zhigailov A.V., Perfilyeva Y.V., Shumilina A.G., Yeraliyeva L.T., Nizkorodova A.S., Kuznetsova T.V., Iskakova F.A., Berdygulova Z.A., Neupokoyeva A.S., Mamadaliyev S.M., Dmitrovskiy A.M. Two case reports of neuroinvasive West Nile Virus infection in the Almaty region, Kazakhstan. IDCases. 2020;21:e00872. https://doi.org/10.1016/j.idcr.2020.e00872. Scopus25%.
  9.  Bissenbay, A.O., Zhigailov, A.V., Perfilyeva, Y.V., Naizabaeva D.A., Neupokoyeva A.S., Berdygulova Zh.A., Ostapchuk Ye.O., Maltseva E.R., Kuatbekova S.A., Nizkorodova A., Dmitrovsky A.M., Skiba Y.A., Mamadaliyev, S.M. Epidemiology and molecular genetic characteristics of Lyme borreliosis pathogens circulating in tick’s population in the Almaty oblast of the Republic of Kazakhstan. Zhurnal Mikrobiologii Epidemiologii i Immunobiologii. 2020;97(6):535–545. Scopus – 4%.
  10. Mukhatayev Z., Ostapchuk Y.O., Fang D., Le Poole I. C. Engineered antigen-specific regulatory T cells for autoimmune skin conditions. Autoimmunity reviews. 2021;20(3):102761. https://doi.org/10.1016/j.autrev.2021.102761. Q1. Scopus – 92%.

Achieved Results

2022 year

As part of the project implementation for 2022, a risk analysis and zoning of the territory of the southern region of Kazakhstan according to the risk of spread of RF-borreliae causing tick-borne relapsing fever (TBRF) was carried out. Since there is no epidemiological surveillance for TBRF in Kazakhstan, the zoning of the territory was carried out taking into account the analysis of such indicators as the prevalence of ixodes and argas ticks, potential vectors of RF-borrelia, population density in their habitats and data on attacks on humans in recent years, the presence of borders with other countries where TBRF is recorded, as well as climatic and geographical data. The most susceptible areas for TBRF caused by hard ticks are Almaty city and all districts of Almaty oblast, except Balkhash (lower reaches of Zailiyskiy Alatau), eastern districts of Zhetysu oblast (lower reaches of Zhetysu Alatau) and Sarkan district of Zhetysu oblast. Tension areas for TBRF caused by soft ticks are Iliyskiy (western spurs of Shu-Iliyskiy mountains) and Balkhashskiy districts of Almaty oblast (in lower reaches of Ile river) and all districts of Zhambyl, Turkestan and Kzylorda oblasts, except for foothill areas of southern districts. Based on the results of risk analysis, we developed a map of zoning of the territory of the southern region of the Republic of Kazakhstan, reflecting the potential risks in relation to the spread of RF-borreliae. For serologic monitoring we collected serum samples from 100 conditionally healthy people in Zhambyl oblast and 10 patients with clinical manifestations similar to those caused by Borrelia of the TBRF group from Almaty.

2023 year

Within the framework of the project implementation for 2023, the collection of ixodes and argas ticks, potential reservoirs of RF-borreliae, was carried out during the epidemic season 2023 in the territory of Zhambyl (193 ticks), Almaty (266 ticks), Turkestan (1507 ticks) and Zhetysu (64 ticks) oblasts. The collected ticks were accepted for storage and a database was prepared. A systematic identification of the collected ticks to species was carried out. From the collected ticks, 404 potential vectors of RF-borrelia pathogens were homogenized. Total DNA samples were isolated from the tick homogenates and analyzed for the presence of RF-borrelia DNA by PCR, 3.7% positive samples were detected. Genetic characterization of three RF-borrelia isolates was carried out. It was found that one of the isolates detected in a tick belonging to the species Argas persicus, collected in Ordabasinsk district, Turkestan region, belongs to B. anserina, two other isolates detected in ticks belonging to the species Ixodes persulcatus, collected in Almaty city, Almaty region, belong to B. miyamotoi. Also, blood serum samples from patients with fevers of unclear etiology in Almaty (26) and Shymkent (33), as well as conditionally healthy volunteer donors living in Almaty (82), Turkestan (105) and Zhambyl (142) oblasts were collected for serologic monitoring. Serologic analysis of blood serum samples from patients with fevers of unclear etiology for the presence of antibodies of IgM class against Borrelia revealed 10.2% of positive samples. Seroprevalence of IgG antibodies against Borrelia in the group of conditionally healthy donors amounted to 5.8%, including 9.8% in the studied sample in Almaty region, 1.9% in Turkestan region and 6.3% in Zhambyl region.

Main design and technical and economic parameters:

– Synthesized and purified primers for detection and genetic characterization of RF-borreliae.

– Standard operating procedures (SOPs) related to the project activities were developed (2 SOPs).

Project Publications

  1. Ostapchuk Y.O., Dmitrovskiy A.M., Pak E.A., Perfilyeva Y.V. A case of combined infection with tick-borne encephalitis and Lyme borreliosis with severe meningoencephalitis and complete recovery // Journal of Global Infectious Diseases. – 2023. – Vol. 15 (2). – P. 81-83. doi:10.4103/jgid.jgid_76_22. (Impact Factor-1.92, Q3 (35.08% Infectious Diseases) Web of Science, Journal Citation Reports, CiteScore – 4.6, Q2 (57% Infectious Diseases) Scopus).