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Antivirotické a antibakteriální účinky biologicky aktivních látek z přírodních zdrojů a jejich potenciální využití proti klíšťaty přenášeným patogenům
LUDVÍKOVÁ, Nikola
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.
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Effect of infection with the tick-borne encephalitis virus on Ixodes ricinus tick activity
VÝLETOVÁ, Eva
The aim of this study was to examine the effect of tick infection with tick-borne encephalitis virus on its behaviour and development. The effect of infection on feeding performance, metamorphosis, locomotion or phototaxis was analysed. Despite the fact that we were not able to demonstrate any significant effect of infection on tick behaviour, the obtained results contribute to understanding transmission dynamics of the virus during tick life cycle including co-feeding and transovarial transmission.
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Measurement of signalling pathway activation in mouse macrophage line IC-21 and primery dendritic cells after infection with tick-borne encephalitis virus.
Kožantová, Jana ; Lieskovská, Jaroslava (advisor) ; Drbal, Karel (referee)
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.
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Diagnostics of Tick-borne Encephalitis: Present State and Future
Hustedová, Anna ; Španielová, Hana (advisor) ; Saláková, Martina (referee)
Tick-borne encephalitis is one of the most severe tick-borne neuroinfections in Europe and Asia. This thesis is focused on its source, Tick-borne encephalitis virus (TBEV). Diagnostics of TBEV is usually based on detection of specific anti-TBEV antibodies in patient's serum by using enzyme-linked immunosorbent assay. However antibodies that develop during the infection often cross-react with other flavivirus specific antibodies. As an antigen for ELISA tests the formalin inactivated virus is usually used, its production is restricted to laboratories with biosafety level 3. This thesis is seeking for possibilities of production of specific antigen for serological diagnostic of TBEV using heterologous expression systems. As the new antigens, parts of glycoprotein E and subviral particles were used. Production of antigen in the formo of subviral particles seems to be a good alternative to inactivated virus.
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Antiviral vaccination - tick borne encephalitis virus
Solarová, Pavlína ; Janštová, Vanda (referee) ; Drda Morávková, Alena (advisor)
Tick borne encephalitis is by one of the most severe disease of central nervous system. Agent of this disease is tick-borne encephalitis virus, which is transmitted mainly by tick. Clinical picture of TBE is considerably different and varied in individuals - from quite infection without symptoms up to severe cerebro-spinal meningitis, in such cases attacked persons by this virus leave hospital with whole-life handicap. We can prevent this disease by inoculation. We can see it in the neighboring Austria, where TBE almost became extinct because 90% of population is vaccinated. Vaccines are commercially available from 1979. Nevertheless only 17% population is vaccinated against TBEV. The reason for it could be fact that insurance companies do not provide the vaccines as well as not very effective health education. One of possibilities how we can this tendency reverse would be higher awareness of pupils at schools. In school educational programs there is contained prevention of viral disease, especially in general biology, biology of viruses and education to health.
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Comparison of various ways of tick infection with the tick-borne encephalitis virus
VÝLETOVÁ, Eva
The aim of the bachelor thesis was to compare various methods of Ixodes ricinus tick infection with the tick-borne encephalitis virus. For infection by immersion method, less virulent TBE virus strain Neudoerfl was used resulting in 5 % of infected ticks. Using more virulent strain Hypr provided 60 % of infected ticks. 75 % of ticks became infected with the Neudoerfl virus by capillary feeding. Infection of ticks by feeding on viraemic mice was unsuccessful. Applicability of tested methods for infection of tick life stages is discussed.
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Value of the Antibodies Testing in the Diagnostics of Tick Borne Encephalitis
CHADTOVÁ, Marie
The Diagnostic Value of the Antibody Level Testing in Tick Born Encephalitis. This bachelor thesis is focused on the antibody testing and its diagnostic value in Tick Born Encephalitis.Data for this thesis were obtained at the laboratory of Virology, Dpt. of Medical Microbiology of the Cenral Laboratories, Hospital České Budějovice. The data were collected between January 1st and December 31st 2014. The thesis is divided in two parts: the theoretical and the experimental part. The theoretical part covers the history of the disease, its geographic distribution and the ways of the transmission of the virus. The disease was first described by Austrian physician Hans Schneider in 1931 as a periodically occuring disease. He called it "Epidemische acute Meningitis serosa". The virus was first described as the cause of the disease in Russia in 1937. In Czechoslovakia, the virus was described in 1947 in Prague by virologists František Galia and Josef Rampas. The first succesful virus isolation in Czechoslovakia was from Beroun district, the next successful isolation was in Vyškov district made by Dr. Krejčí. The virus was later isolated in other European countries as well. The hypothesis, that the virus is transmitted by ticks, was proved by Dr. Rampas and Dr. Galia by the isolation of the virus directly from the ticks Ixodes ricinus, collected in the Beroun district. The transmission of the virus by alimentary route was first described in 1951 near east Slovakian town Rožňava. The virus was transmitted by drinking nonpasteurized sheep milk. The laboratory infection is possible, when laboratory staff works with the viable virus. The Tick Borne Encephalitis virus is part of the genus Flavivirus , family Flaviviridae. It is an enveloped virus with cubic symetry. Virus particles are round shaped, with diameter 5060 nm. The central part is nucleus, containing viral RNA, enveloped by protein capsid and lipoprotein envelope. Lipoprotein envelope is double lined and contains important antigens - glycoprotein E and menbrane protein M. Tick Borne Encephalitis is transmitted by ticks. The incidance is therefore closely connected to the activity of the ticks in nature. The activity of the ticks depends on the season, temperature, humidity and altitude. The presence of people in focal point is also essential. Tick Borne Encephalitis is an infection with the natural focal point. Its natural reservoir are mainly small rodents and other wild animals. Human activity in nature can lead to the transmission of the disese to humans. Human are a blind end of the transmission chain, no interhuman transmisson was described. The virus penetrates the organism via the tick bite. The virus penetrates to the skin from salivary glands of the tick through the small wound. The primary multiplication starts there. The virus is later spread by lymphatic system to the blood stream. This corresponds with the first phase of the infection. The second phase of the infection is accompanied by secondary viremia, the virus multiplies in various tissues and spreads into the central nervous system. The incubation period before the beginning of the first phase is 714 days. The first phase may proceed under the clinical picture of mild, flu- like virosis with the increased temperature, muscle pain and fatigue. This period lasts 1 to 2 days, maximum 6 days. After the first phase there is a period of approximately one week without any signs of the disease. After this period the second phase may occur, with sthe signs of meningitis, menigoencephalitis or myelitis. The treatment is symptomatic. To diminish the symptoms of the disease, analgetics, antipyretics , anticonvulsives and sometimes antiedematic treatment is indicated. The epidemiological mesures are based on education and vaccination. The vaccine is available since the eigties of the last century. Laboratory diagnostic may use direct or indirect detection of the virus. The methods of direct detect
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Vector-pathogen-host interaction on the example of spirochetes Lyme boreliosis disease (and tick-borne encephalistis virus)
VAVRUŠKOVÁ, Zuzana
This study was focused on vector-pathogen-host interaction. Questing ticks from field were tested for presence of Borrelia burgdorferi s.l. and host DNA. Small rodents were trapped, ticks were collected from them, infestation patterns were estimated regarding the species and stage of ticks and species, sex and body weight of the host. Ticks aquired from hosts were tested for presence of Borrellia burgdorferi s.l. and tick-borne encephalitis virus. Both results from identification of hosts and from detection of pathogens were compared to be able to investigate interactions between host, vector and pathogen.
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