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Monitoring of Blood-Sucking Flies


Molecular Genetic Monitoring of Blood-Sucking Flies (Diptera) as a Basis for Biological Control of Vectors of Dangerous Infectious Diseases and Precautions Against the Acts of Biological Terrorism

Tech Area / Field

  • BIO-CGM/Cytology, Genetics and Molecular Biology/Biotechnology
  • AGR-DIS/Disease Surveillance/Agriculture
  • BIO-MIB/Microbiology/Biotechnology
  • MED-DIS/Disease Surveillance/Medicine

8 Project completed

Registration date

Completion date

Senior Project Manager
Genisaretskaya S V

Leading Institute
Institute of Molecular Biology, Armenia, Yerevan

Supporting institutes

  • NGO Aquafauna, Armenia, Yerevan


  • Leibniz-Zentrum für Agrarlandschaftsforschung e.V. / Deutsches Entomologisches Institut, Germany, Müncheberg\nUniversity of Oxford / Oxford University Museum of Natural History, UK, Oxford\nBrock University, Canada, ON, St Catharines\nClemson University / Department of Entomology, Soils and Plant Sciences, USA, SC, Clemson

Project summary

Over the last 15 years in Armenia, like elsewhere in the world, transmissible diseases such as malaria, rabbit fever and anthrax have become increasingly widespread. In March of 2007 alone, 28 cases of rabbit fever have been reported from the north of Armenia. Previously in 2003 more than 300 cases of rabbit fever had been documented in the village of Sukhoy Fontan in Armenia. Overall, 1156 events of malaria were recorded in 1998 and 616 in 1999. Animal and human anthrax is widely spread in the north and south of Armenia. The epidemiological situation of these diseases in Armenia remains unfavorable today. This is largely caused by a rapid increase in the number of populations of these disease vectors and extension in their distribution range. Furthermore, it is noteworthy that vectors of infectious diseases are an effective and hazardous biological weapon during the acts of terrorism. The introduction into the country of these vectors is an important factor in the transmission of communicable diseases. Unfortunately, this phenomenon occurs everywhere (Adhami and Murati, 1987, Monath, 1988). That is exactly why the biological control of vectors of these diseases is one of primary factors responsible for the economic and political stability in countries. The proposed research is directed towards antiterrorism. The results of this study will allow to undertake the implementation of effective measures against the acts of biological terrorism.

In terms of morbidity and mortality caused by vector-borne diseases, mosquitoes and blackflies are the most dangerous animals confronting mankind. Mosquitoes and black flies are an enormous nuisance problem in many parts of the world. In some areas of the world, such as Canada, USA, Germany, Italy, they have a negative impact on economic development (Laird et al. 1982). Mosquitoes transmit about 100 viruses, in addition to filarial worms, bacteria, and protozoans such as malarial agents that infect humans throughout the world (Kettle, 1995; Beaty and Marquardt, 1995; Lehane, 1991). Malaria is one of the world’s leading health problems, with about half a billion people currently infected and 2 million deaths annually (Tishkoff et al. 2001). Black flies transmit blood-borne parasites, including protozoans that cause avian leucocytozoonosis; filarial worms that cause human onchocerciasis (river blindness), mansonellosis, and bovine onchocerciasis; and the virus that causes vesicular stomatitis in livestock (Adler 2005).

It can be stated that all around the world the risk for the re-introduction and transmission of mosquito-borne diseases has increased rapidly over recent decades, due to the large number of tourists arriving from the tropics and carrying parasites and pathogens, as well as aircrafts carrying vectors causing the so-called “airport malaria”. The introduction of Ae. albopictus and Ae. japonicus japonicus into the United States and Italy is well known (Adhami and Murati, 1987). The occurrence of the Asian tiger mosquito Ae. albopictus and other invasive species in North America and South Europe may be seen as evidence of the increased risk of the introduction of new and maybe more virulent mosquitoborne diseases in many countries of the world. The examples have shown that the introduction into the country of these vectors is an important factor in the transmission of communicable diseases which, in effect, may be used in the cause of bioterrorism acts.

The main aim of the project is the scientific innovations on biological control of vector populations and new knowledge on biology of the blood-sucking Diptera which will help to prevent the spread of virulent diseases and secure the health of population in Armenia and other countries.

Related activities currently under way. Our Institute had been carrying out research work in areas defined and commissioned by the Ministry of defense of the USSR. The project participants’ team is composed mainly of researchers who had been involved in the implementation of the “weapons” project.

Our laboratory and the collaborators (Dr. Adler, Dr. Werner, Dr. Pont) have worked together on the three previous CRDF projects and an ISTC project dealing with black flies in Armenia, specifically their taxonomy, utility as water-quality indicators, and centers of biopersity (Adler & Kachvoryan, 2001; Kachvoryan et al., 2004 a,b; Nelder, Adler & Kachvoryan 2005; Pont, Werner & Kachvoryan. 2005; Werner, Kachvoryan, & Richter, 2006).

Expected Results and Their Application. The proposed investigation falls into the two categories of basic and applied investigations.

In accordance with the stages of the project the following results are to be obtained:

  1. A database on the pest species persity of black flies and mosquitoes;
  2. New data will be obtained on selected pathogens;
  3. The results of the proposed research will provide a comprehensive genomic database for the black flies and mosquitoes of the Caucasus Region, an area of extraordinary endemicity;
  4. Experimental source data on the impact of microbial and chemical pesticides on biologically controlled organisms;
  5. Preventive measures and a monitoring program will be designed to control pest species; a public awareness program will be fulfilled and recommendations on pest monitoring and management programs will be submitted to educational institutes, radio and television.

The realization of this project is of prime importance both for Armenia and for neighbouring countries. Furthermore, the project will be focusing its research on groups of insects which includes vectors of diseases such as anthrax, onchocerciasis and leucocytosis. From this point of view, the project impinges upon the defences against bioterrorism which is a developing threat everywhere.

Results to be promoted. Framework for biological control of vectors of malaria, rabbit fever and anthrax will be developed on the regional level and their application will be tested in other parts of the world. Study of the effect of microbial pesticides on biologically controlled organisms and their adaptation mechanisms will help to reduce population numbers of vector species. Forecasts will be developed to predict future epidemiological situation of malaria, rabbit fever and anthrax. These results will be published in high-impact scientific journals.

Uniqueness of results. Unique results on the genome and adaptive response in the Simuliidae and Culicidae, and the pathogens in Armenia will be obtained for the first time. Pioneer results will also identify the true role of pest species in the region. Methods of control to be developed by us will be unique, since their application will be adapted to this region in the foremost.

Demand for results. The potential “end-users” of the project results will include the sanitary centers and veterinary stations within Armenia, as well as other medical institutions.

Expected income. The proposed research is a fundamental and applied biomedical experimental program designed to generate new knowledge that will lay the foundation for prevention of diseases such as rabbit fever, anthrax and malaria in humans and domestic animals.

IPR situation. Research ideas generated in the course of the project will be patented and copyright protected.

Additional developments. Results of the current research closely reflect the real market demands. Building upon the results of the research on pest species, we plan to make management recommendations to use new biological methods for vector species and new biological material.

Meeting ISTC Goals and Objectives. The project will be carried out by the scientists of the Institute of Molecular Biology, Armenian NAS, and NGO “Aquafauna”, formerly engaged in the area of defense. The current project offers these scientists an opportunity to channel their activity towards peaceful needs, specifically in the area of medical parasitology and epidemiology. Labor costs of the “weapons” personnel comprise about 60% of the total costs. The scientific teams in Canada, Italy, UK and Germany have agreed to join the research as foreign collaborators. Results generated by the project can be used in medicine and veterinary science, and be of practical and scientific importance to the international community.

Scope of Activities. The proposed investigation falls into the two categories of basic and applied investigations.

The main aim of the project will be accomplished through completion of the tasks outlined below:

Task 1. To determine the species-spectrum of the dipteran families Simuliidae and Culicidae;

Task 2. To establish the definitive identification of the abundant pest species. This is of particular importance in the Simuliidae and Culicidae, which are of great medical and economic significance;

Task 3. To provide a genomic analysis of the black flies and mosquitoes of Armenia, involving macrogenomic (i.e., cytogenetic) analyses in our laboratory, supplemented with microgenomic (i.e., molecular) analyses in the laboratories of cooperators and thus define species limits and reveal sibling species:

Task 4. To investigate the ecology of blood-sucking dipterans; a) study population structure of blood-sucking dipterans; b) study developmental cycles of blood-sucking species and their distribution range; c) correlate pest problems with the spatial and seasonal distributions of the black flies and mosquitoes; d) monitor the population numbers of blood-sucking mosquitoes and black flies; e) reveal selected pathogens; f) examine the impact of microbial pesticides on biologically controlled organisms;

Task 5. To develop prophylactic measures and propose a pest monitoring program: a) provide pest management recommendations; b) implement a public education program.

All tasks are correlated and mutually influence one another. Reports and publications will be generated and made available on each task. The fulfillment of the project is planned for three years.

Role of Foreign Collaborators. The proposed research will be carried out jointly with collaborators in Canada, USA, UK, Germany and Italy. Our laboratory will communicate and exchange information with collaborators. The collaborators will provide comments to the technical reports (quarterly, annual, final, etc.) due for submission to the ISTC. Preparations developed by us in the course of the project will be evaluated by collaborators. Joint symposium and workshops are also on the agenda. We also plan publications in peer-reviewed scientific journals with collaborators.

Technical Approach and Methodology. We will collect material throughout Armenia, emphasizing mosquito and black fly biopersity hotspots, particularly areas identified for outbreaks of rabbit fever and malaria.Larval and pupal mosquitoes and black flies (up to 100 – 150 larvae) will be collected into acetic ethanol, a cytological fixative and into 95% ethanol for molecular analysis. The material will be used to screen for pathogenic organisms. Preparation of polytene chromosomes will involve both Feulgen and aceto-orcein methods (Adler et al. 2004). We will complement our cytogenetic analyses with molecular barcoding, using the mitochondrial cytochrome c oxidase (COI) gene.

The inhibitory effects of microbial preparations on each single species will be tested both in the laboratory and in the field. We will draw experimental data protocols which will be used to develop a management mechanisms and prevention methods to control vector organisms.