The first aim of this study was to detect antiviral activities of substances isolated from natural products against tick-borne encephalitis virus in in vitro model. Resveratrol isolated from plant material and adamantane derivatives were studied in this regard. The maximum tolerated concentrations of the investigated substances were determined for the glioblastoma cell line used in the experiments using flow cytometry and subsequently. Next, the number of viral particles produced by infected cells after incubation with the studied substances was determined using plaque titration. Possible antibacterial effects of the studied materials against standard strains of bacteria Staphyloccocus aureus, Staphyloccocus epidermidis, Escherichia coli and selected strains of Borrelia burgdorferi spirochetes were examined.

We examined the influence of the host genetic background on the pathogenesis of tick-borne encephalitis. We determined virus titers in organs and serum in different time intervals post-infection for different ways of inoculation. We also stated mean survival times and antibody production in different strains of mice infected with tick-borne encephalitis virus. Moreover, differences in expression of immunologicaly important genes in brains of infected mice were compared.

This thesis deals with infectious diseases transmitted by arthropods in the Czech Republic in period of 2002 - 2011. Meeting the objectives and answer the research questions, I managed using secondary analysis of reported cases infectious diseases transmitted by arthropods in the information system for reporting and recording of infectious diseases - EPIDAT and literature review and subsequent analysis of scientific articles. Important additional informations were provided by State Veterinary Institute in Prague and the National Reference Laboratory for arboviruses in Ostrava. Thesis does not deal with number of single values, but it is focused on epidemiological severity of analyzed cases, severity of clinical course and risk of introducing infection in the Czech Republic. Theoretical part of the thesis describes selected arboviruses infections. Tick-borne encephalitis is one of the most important diseases transmitted by arthropods in our surroundings. Number of reported cases of tick-borne encephalitis was quite constant in monitored period 2002 - 2011, according to my research and statistical verification. Highest morbidity was in 2006. The same year the highest incidence of disease Tick-borne encephalitic was reported in region of Vysočina. This number is far closer to number of reported cases in region of South Bohemia which was the most affected region thought the entire period. Prevention exists against tick-borne encephalitis. It is vaccination which is recommended for people staying in natural foci of infection in the Czech Republic and abroad. In the Czech republic becomes more important West Nile fever. Increased incidence of this disease is recorded in the USA, but also in many European countries (eg Greece, Hungary, Italy) in recent years. One of the goals of this thesis is to characterize the surveillance of West Nile fever in the Czech republic. Monitoring antibody titer against West Nile virus in the serum of horses is one of the basic elements of surveillance of this disease in our country. 5 samples of the blood serum was increased titers of antibodies to West Nile virus. Various geographic distribution of horses and high specific antibody titers indicate increasing activity of West Nile virus. Clinical disease of horses not being recorded in the Czech republic. In the field of human medicine have been reported three imported cases of West Nile fever. Other arboviruses infections, that appeared in our country, is dengue disease and viral disease Chikungunya. In the monitored period there were a total of 98 reported cases of imported disease. Most cases were imported from India. Three cases of Chikungunya disease were reported in 2006 in connection with a stay on the island of Mauritius, where was the epidemic at this time. People can travel freely and there is the risk of importing diseases from the tropics and subtropics. Some infections transmitted by arthropods are in areas where it was previously absent recently. Prevention is essential. Specific and non-specific prevention. Another important element of prevention is strict adherence to methods for surveillance of disease.Viral infections transmitted by arthropods are among the most important emergent infectious diseases. In addition to malaria, malnutrition and helminth infection are the largest public health problem, not only in third world countries.

The thesis is devoted to the issue of vaccination problematics and awareness of tick-borne encephalitis in a selected population group in South Bohemia. For the needs of research, the selected group was composed of students of secondary schools and grammar schools in the South Bohemian Region in the age group from 13 to 19 years old. The objectives of this work was to analyze the incidence of tick-borne encephalitis in the Czech Republic and the South Bohemian Region between 2000 and 2015, to compare the vaccination in the Czech Republic with the other European countries, to find the vaccination of a selected age group in the South Bohemian Region and to analyze the level of awareness of the risks of tick-borne encephalitis by the polling method and the attitude of the age group to vaccination against this disease. To achieve the objectives was used a secondary analysis of the data from the EpiDat system, where the incidence of the disease was detected in the period in the South Bohemian Region and the Czech Republic. The questionnaire survey was used to obtain awareness on observed disease and attitude of the study group towards vaccination against tick-borne encephalitis. The conclusions of the questionnaire survey were obtained on the basis of statistical processing in the MS Excel spreadsheet. For my significant findings from my research, I consider 80% of the vaccination in the monitored group. Information on tick-borne encephalitis in most cases did not get the respondents from a doctor, but from family members, from the internet or from television. However, information on vaccination is given to respondents to a greater extent. Contributions by insurance companies do not motivate more than half of respondents to inoculation. Research has also shown, that respondents have insufficient information about the amount of the contribution and whether their insurance company provide the contribution at all. In view of these circumstances, I consider the increased promotion of this vaccination by insurance companies to be very desirable.

West Nile virus (WNV) infections were first detected in Germany in 2018, but information about WNV seroprevalence in horses is limited. The study’s overall goal was to gather information that would help veterinarians, horse owners, and veterinary-, and public health- authorities understand the spread of WNV in Germany and direct protective measures. For this purpose, WNV seroprevalence was determined in counties with and without previously registered WNV infections in horses, and risk factors for seropositivity were estimated. The cohort consisted of privately owned horses from nine counties in Eastern Germany. A total of 940 serum samples was tested by competitive panflavivirus ELISA (cELISA), and reactive samples were further tested by WNV IgM capture ELISA and confirmed by virus neutralization test (VNT). Information about potential risk factors was recorded by questionnaire and analyzed by logistic regression. A total of 106 serum samples showed antibodies against flaviviruses by cELISA, of which six tested positive for WNV IgM. The VNT verified a WNV infection for 54 samples (50.9%), while 35 sera neutralized tick-borne encephalitis virus (33.0%), and eight sera neutralized Usutu virus (7.5%). Hence, seroprevalence for WNV infection was 5.8% on average and was significantly higher in counties with previously registered infections (p = 0.005). The risk factor analysis showed breed type (pony), housing in counties with previously registered infections, housing type (24 h turn-out), and presence of outdoor shelter as the main significant risk factors for seropositivity. In conclusion, we estimated the extent of WNV infection in the resident horse population in Eastern Germany and showed that seroprevalence was higher in counties with previously registered equine WNV infections.

No description available.

Tick-borne encephalitis is a serious disease of the central nervous system. It is caused by tick-borne encephalitis virus, which is transmitted by ticks. The Czech Republic is one of the countries with the highest prevalence of this disease. Tick-borne encephalitis virus is able to replicate in several cell types. In this work we focused on macrophage line IC-21 and dendritic cells, because these cells are the first, which encounter the virus and support its spreading in the host at early stage of infection. So far there is not known any specific receptor for virus entry into cells or which signaling pathways activates. Therefore, we decided to investigate the activation of selected signaling pathways after infection with tick-borne encephalitis virus and influence of tick saliva on this activation. We employed methods of dual luciferase reporter assay, immunosandwich assay and western blot. The obtained results showed that in virus infected IC-21 cells are activated phosphatidyl-inositol pathway, NF-κB pathway, signaling molecule Erk1/2 and others. Testing of tick saliva effect revealed significantly decreased activity of NF-κB, AP-1 and CREB.

No description available.

This thesis is concerned with the level of parents´ awareness and attitudes to above-standard vaccination of children from 0-3 years in Příbram. Among the optional vaccination of children from 0-3 years belong vaccination against pneumococcal disease, rotavirus infections, meningococcal invasive disease (caused by meningococcal of group C, A+C, or A, C, W 135 and Y, now also of group B), tick-borne encephalitis, varicella smallpox, influenza and hepatitis A (or a combination of type A + B). The first part deals with the issue in a theoretical perspective. It foreshadows the basic characteristics of the diseases against which the premium vaccination of children from 0-3 years is offered. Further it is focused on vaccines against these diseases, which can occur in the Czech Republic. The survey is summarized in the research. There were parents of children attending one of the 12 kindergartens in Příbram in the sample of the research. tely 40% of the parents (or mothers) of these children in each kindergarten.The results were summarized in schedules of absolute and relative frequencies, or graphs. It was set three basic goals that were met with five hypotheses. The first of these was to monitor the attitudes of parents to above-standard vaccination of children from 0-3 years in Příbram. To this target relate hypothesis H1: Parents are interested in above-standard vaccination of children from 0-3 years in Příbram, H2: Parents with higher educational level have significantly higher interest in above-standard vaccination of children from 0-3 years and H3: Parents are statistically significantly more interested in vaccination against tick-borne encephalitis than other extra vaccination of children from 0-3 years, due to an endemic area of tick-borne encephalitis in Příbram. These hypotheses were not confirmed statistically. The second aim investigated the main reason for the possible lack of interest in extra vaccination of children from 0-3 years in Příbram. The target was filled with hypothesis H4: The main reason for the possible lack of parents´ interest in extra vaccination of children from 0-3 years is high price of vaccines, which was statistically refuted. The third goal was to explore parents' knowledge about diseases against which the extra vaccination of children from 0-3 years is offered. To this goal was set the hypothesis H5: Parents have sufficient information about the diseases against which the extra vaccination of children from 0-3 years is offered. This hypothesis was confirmed. The hypotheses were verified by ?chi-square? test at a significance level of 5%, which is an instrument of verification or falsification of hypotheses. This work may be used in practice as a preview to the parents´ awareness and interest in extra vaccination of children from 0-3 years. As well as cumulative information materials for professionals and the public about the above-standard vaccination of children from 0-3 years.

Expanding urbanization, climate change, and population growth contribute to increased transmission and spread of arthropod-borne viruses (arboviruses), many of which cause severe disease in humans. Pathogenic arboviruses include dengue, Zika, tick-borne encephalitis, and sindbis viruses, which together threaten more than half the global population. Thus, there is a constant need for safe, specific, and sensitive molecular tests to identify early-stage infections for accurate diagnosis and molecular epidemiological data for disease prevention and control. The study tested the biosafety of using FTA™ cards when working with pathogenic arboviruses by conducting an infectivity assay using sindbis virus. Conditions for RNA extraction and storage of arboviruses on FTA were analyzed by measuring viral RNA (vRNA) stability using a SYBR-Green, Pan-Flavi RT-qPCR method composed of degenerate primers able to detect a variety of flaviviruses. Data from a Pan-Flavi RT-qPCR study comprising of 222 clinical blood and serum samples collected from a 2018 dengue virus outbreak in Hanoi (Vietnam) was analyzed to establish applicability of FTA for molecular epidemiology and diagnosis. Results showed that sindbis virus infectivity was inhibited by FTA-adsorption. FTA-adsorbed arboviruses were extracted with the highest yield using Trizol extraction and were preserved at storage at 4-20ºC for up to 30 days. The results showed that clinical blood samples acquired higher yields of vRNA for molecular testing than serum samples and that it may be possible to perform sequencing for genomic analysis. The study suggests that FTA cards may facilitate the storage and transportation of adsorbed arboviruses for downstream molecular epidemiological and diagnostic tests.