The purpose of this project is to solve scientific and practical problems of biological safety of the population of Kyrgyzstan and neighbouring countries using molecular genetic methods to study the plague (Yersinia pestis) and combined with it bacterial pathogens: pseudotuberculosis (Yersinia pseudotuberculosis), bartonellosis (Bartonella), brucellosis (Brucella) and rickettsial diseases (Rickettsia spp.).
The data obtained from the proposed project will be very valuable and important to the health authorities and other state institutions in Kyrgyzstan; they will enable to detect, identify, and promptly report on disease outbreaks caused naturally or intentionally delivered pathogens, and provide a system to control them, which will be budget based and due to sustainable infrastructure in Kyrgyzstan. From the public health perspective it is necessary to have an accurate data on possible spread of pathogens within the territory of the country, and also, it is important to define what kinds of animals are the reservoirs of bacterial infections, and what kinds of ectoparasites transmit specific agents of infectious diseases.
State of affairs in the field of research. Kyrgyzstan is located in Central Asia. Zoonotic agents active over here have not been insufficiently characterized in comparison with the same species found in Europe, North America or East Asia (Biggins & Kosoy, 2001). As a result, their diversity can occur both on genotypic and phenotypic levels as compared with other parts of the world. For instance, PCR study that uses DNA sequences of chromosomes and plasmids that are typical to European or North American strains may fail to recognize strains from Central Asia because of the genetic difference. Therefore, strains isolated in Kyrgyzstan should be analyzed to improve diagnosis by recognizing differences found in the local strains. This project aims at developing and implementing a screening method for unknown pathogens that may be present in rodents and their ectoparasites.
The most dangerous of zoonoses in the country is plague. The disease is seen as a real threat to human populations, not only within the natural foci, but also globally (Gage & Kosoy 2005).
Three natural foci of plague are identified in this country: Tien Shan, Alai and Talas, which include 18 sites. The Tien-Shan focus is divided into three autonomous foci: Sarydzhaz, Verhnenarynskii and Aksai. They differ in their biological characteristics and epizootic activity (Ibragimov, 2005; 2010). To date, the boundaries and dimensions of natural foci have been defined more precisely. The total area of plague foci in Kyrgyzstan is 3 million 200 thousand hectares.
Implementation into the monitoring of plague including classical and novel microbiological and molecular genetic methods cans undoubtedly contribute to progress in plague diagnosis. The work in this direction can take a fresh look at one of the most dangerous bacterium as Y. pestis and significantly improve diagnostic capabilities of the Public Health system of the Republic. Of particular interest is a possibility of acquiring Y. pestis resistance against antibiotics associated with the recovery of natural plague foci. To date, in the plague focy in Kyrgyzstan observed a tendency of increasing the number of fleas (Altybaev, 2007). In Aksu district of Issyk- Kul region there was a registered case of infection of human by bubonic plague with lethal outcome in the summer of 2013.
Rickettsial diseases caused by small obligate intracellular bacteria of the Rickettsia genus are spread throughout the world. Our research will focus on R. typhi and R. feliswhich are carried by rodents and transmitted by fleas, that are the causative agents of murine (endemic) typhus and flea-borne. The clinical symptoms of human diseases caused by different Rickettsia spp. can be very similar. This underlines the importance of a rapid identification of specific Rickettsia pathogens by molecular typing
Bartonellosis is called by small facultative intracellular bacteria of the Bartonella genus (including B. elizabethae, B. grahamii, B. vinsonii, B. henselae). B. henselae is the causative agent of cat fever, and at least 10 species of Bartonella cause human disease and is treated as new pathogens to humans (Kosoy, 2010). In most regions of the world Bartonella spp. were investigated, but still remain unknown in Kyrgyzstan and neighbouring countries of the Central Asia.
We will perform the following tasks to achieve the objective:
1. Epizootological examination of the Kyrgyzstan’s territory to determine eco-epidemiological patterns of spreading of the bacterial pathogens Y.pestis, Y.pseudotuberculosis, Bartonella, Brucella and Rickettsia spp. in the rodents organisms and their ectoparasites;
2. Study of the genetic characteristics of strains Y.pestis, Y.pseudotuberculosis, Bartonella, Brucella and Rickettsia spp. in Kyrgyzstan, as well as the definition of epidemiological significance of various rodent species and related bacteria in the formation of these infections’ nosoareas;
3. Determination of risk factors for spreading pathogenic bacteria of Y.pestis, Y.pseudotuberculosis, Bartonella, Brucella and Rickettsia spp. to humans in Kyrgyzstan depending on the type of settlements of main rodent hosts and their ectoparasites.
4. Development of new approaches for monitoring pathogens of “new” bacterial infections and epidemiological surveillance of natural foci using new methods of genetic analysis.
As part of the project within three years there will be investigated enzootic territories of Kyrgyzstan. Collected field samples will be tested for the presence of Y.pestis, Y.pseudotuberculosis, Bartonella, Brucella and Rickettsia spp. The project leading and participating organizations - Osh Antiplague Department (OshAPD), the Center of Quarantine and Especially Dangerous Infections of The Department of Disease Prevention and State Sanitary and Epidemiological Surveillance (CQEDI DDP and SSES), will study bacterial pathogens within the southern territory of the country (Osh, Jalal-Abad, and Batken regions). Another institute-participant (Centre of quarantine and disastrous infections of DPSSEC), will explore the northern regions of the country - Chui, Issyk-Kul, Naryn and Talas. During the project, a PCR laboratory will be established on the territory of OshAPD and the field material from the northern part will be delivered to the existing research centre of the PCR laboratory in CQEDI DDP and SSES.
The impact of the proposed project on the progress in this field. The project will allow the Department of Disease Prevention and State Sanitary and Epidemiological Surveillance of the Republic, other medical and diagnostic organizations in Kyrgyzstan to obtain information about environmental and epidemiological patterns of spreading Y.pestis, Y.pseudotuberculosis, Bartonella, Brucella and Rickettsia spp. in rodents and their ectoparasites in Kyrgyzstan.
The competence of the project participants in this area. Project participants have extensive experience in conducting surveys on epizootic plague, which is confirmed by the available publications and participations at various international scientific conferences, extensive experience in production activity. The bacteriologists have collected a large number of strains of Y. pestis, brucellosis and others. Physicians, bacteriologists, medical biologists, parazitologyists, and laboratory workers repeatedly passed the trainings on improvement of the diagnosis of quarantine and especially dangerous infections in various antiplaque GIS institutions. The recent courses have been taken at Kazakh Scientific Centre for Quarantine and Zoonotic Infections named after M. Aykimbaev, conducted with the support of ISTC within the Project K- 1817. The participants were actively conduct the annual monitoring of natural focal plague areas.
The project conformity with the ISTC. The proposed project conforms with the objectives of the ISTC, as it aims at applied research and contributes to the solution of national and international issues related to counter biological threats. Also, the compliance to these goals are achieved by the planned involvement of scientists from the participating organizations in the global scientific community by providing information on the project at international conferences and seminars.
Expected Results and Their Application
The project will:
1. to define modern environmental and epidemiological patterns of dissemination of Y.pestis, Y.pseudotuberculosis, Bartonella, Brucella and Rickettsia spp. on the territory of Kyrgyzstan.
2. to define genetic characteristics of strains of Y.pestis, Y.pseudotuberculosis, Bartonella, Brucella and Rickettsia spp. found on the territory of Kyrgyzstan.
3. to determine epidemiological importance of various rodent species in the formation of parasitic system of Y.pestis, Y.pseudotuberculosis, Bartonella, Brucella and Rickettsia spp.
4. to find the typological characteristic of dissemination of human pathogenic bacteria Y.pestis, Y.pseudotuberculosis, Bartonella, Brucella and Rickettsia spp. in Kyrgyzstan, depending on the type of settlements of the main hosts – rodents and their ectoparasites.
5. to develop the strategy of new approaches for monitoring pathogens, which are new for Kyrgyzstan and epidemiological surveillance of natural foci of the infection by using new methods of genetic analysis, and improvement of the system of control of zoonoses in Kyrgyzstan.
Meeting ISTC Goals and Objectives
The proposed project is consistent with the objectives of the ISTC, as it aims at conducting the research and contributes to the solution of national and international issues related to counter biological threats. Also, compliance with these goals achieved by the planned involvement of scientists from the participating organizations with integration into the global scientific community by providing information on the project at international conferences and seminars.
Role of Foreign Collaborators
Michael Kosoy, Head of the Laboratory at the Canters for Disease Control and Prevention in Fort Collins, Colorado, USA, and Vladimir Motin, Professor, Department of Microbiology and Immunology, Faculty of Medicine of Texas showed their desire to act as part of the project the collaborators.
Technical Approach and Methodology
The studies will apply PCR techniques, at the same time, in order to isolate the plague pathogen cultures there will be used microbiological, biological and serological methods. Methods of field zoological parasitological studies, epidemiological analysis techniques of GIS technology, as well as the methodology and techniques of molecular epidemiology.
