National Repository of Grey Literature 7 records found  Search took 0.01 seconds. 
Molecular detection of cercarial dermatitis causative agents
Procházka, Jan ; Leontovyč, Roman (advisor) ; Baláž, Vojtech (referee)
Human cercarial dermatitis (CD), also called Swimmer's itch appears as a skin rash caused by skin penetration by larval stages of avian schistosomes. In the Czech Republic, the most common causative agens is genus Trichobilharzia. Since 2021, monitoring of CD is a part of a mandatory testing of bathing water quality. The official method used for CD detection is collecting and examination of aquatic snails under a light source. However, this method has many disadvantages, such as time demands, low sensitivity and difficult parasite determination. These disadvantages can be overcome by usage of alternative approaches, such as molecular methods based on environmental DNA (eDNA) filtration and isothermal amplification. Hence, the main goal of this thesis was development of molecular method based on eDNA filtration and cLAMP isothermal amplification for detection of CD causative agent in water. Based on molecular analysis a conserved region of target DNA was selected to create a cLAMP assay specific for genus Trichobilharzia. The cLAMP assay was confirmed to specifically amplify the DNA of at least four species of the genus Trichobilharzia. The laboratory experiments showed a sufficient sensitivity of cLAMP assay for eDNA detection, based on DNA dilution series and cercariae filtration from water. The...
Methods of molecular diagnostics of parasitic helminths
Dejmková, Tereza ; Leontovyč, Roman (advisor) ; Škorpíková, Lucie (referee)
Helminthiases represent a significant health risk, therefore the use of effective and rapid diagnostic methods is essential, including molecular methods, which are summarized in presented thesis. The biology of important human helminth pathogens is summarized in the introductory part. The DNA isolation, visualisation and molecular methods are described in the following parts. The section about molecular methods is divided into two parts. The first part is focused on PCR and its modifications, the second part describes methods of isothermal DNA amplification, which represent more practical and effective alternative to PCR. The advantages, disadvantages of all methods and examples of helminthiases to which they have been applied are also discussed. The final part is about the impact of the COVID-19 pandemic on the development of molecular diagnostic methods, during which were previously established protocols modified or new molecular methods were applied to increase the efficiency. The presented thesis brings an overview of molecular methods that were used to this day for diagnosis of helminthiases. At the same time the thesis discusses the impact of the COVID-19 pandemic on the development of molecular diagnostics and shows the possible future development of molecular diagnostic methods in the field of...
Transcriptomic analysis of Mesocestoides corti
Korená, Lucie ; Leontovyč, Roman (advisor) ; Převorovský, Martin (referee)
Some species of parasites, including helminths, can inhibit carcinogenesis in their hosts. The antitumoral effect has been discovered in the tapeworms Taenia crassiceps and Echinococcus granulosus, which genes associated with cancer regression have been identified. The effect of melanoma suppression has also been observed in tapeworm Mesocestoides corti by the Laboratory of Helminthology, Department of Parasitology, Faculty of Science, Charles University, however the mechanism-of-action, remains unknown. For the upcoming research it was essential to have the complex molecular data such as transcriptome of the developmental stage s of M. corti. This work is focused on the transcriptomic profiling of the tapeworm M. corti and the differential gene expression in two different strains of murine hosts (inbred and outbred) using the RNA-Seq. The main goal was to identify upregulated transcripts in the tapeworms from the murine hosts that could have a potential effect on cancer regression. Differential gene expression analysis was performed, and the results showed that tapeworms in murine hosts (regardless of strain) had more upregulated transcripts than tapeworms cultured in vitro. Analysis of highly upregulated transcripts in the tapeworms that were grown in the murine hosts identified several...
Construction and quality assessment of the genome assemblies
Korená, Lucie ; Leontovyč, Roman (advisor) ; Vorel, Jiří (referee)
Detailed information of the genome of the studied organism is crucial for many fields of modern research. Actual sequencing technologies are not able to read the whole DNA molecule at once therefore only fragments of the genetic information are obtained, which are not sufficiently informative on their own. The goal of the genomic-bioinformatic approach is to assemble these fragments into complete original information - genome assembly. The process of the genome assembly is demanding in terms of computational power, software equipment and expert staff. Many assemblers - programs for genome assembly are available differing in performance, size of the analyzed genome or target organism. The quality of final assembly is fully dependent on assembler and setting of inner parameters. In practice, multiple assemblies are constructed and their quality evaluated according to the technical and biological parameters. The presented thesis describes current high throughput sequencing technologies, different approaches and algorithms for genome assembly and methodology for their quality assessment. The practical part is focused on assembly and its quality assessment using Illumina data of the bird fluke Trichobilharzia szidati.
Molecular adaptations of neurotropic and visceral bird schistosomes during the infection of the avian definitive host
Leontovyč, Roman ; Kašný, Martin (advisor) ; Grevelding, Christoph G. (referee) ; Franta, Zdeněk (referee)
Genus Schistosoma is one the most studied group of helminths due to the importance of several representatives in terms of veterinary and human health. The advent of the modern sequencing technologies, as well as the increasing computational capacities, enabled large-scale screening of nucleic acids and thus deep exploration of complex transcriptome and genome information. To date the main attention of leading molecular parasitological "Schistosoma" research teams was focused on serious human pathogens Schistosoma mansoni, Schistosoma japonicum and Schistosoma haematobium. In the term of molecular/biochemical research, the other schistosomatids were mostly neglected and general knowledge was limited to characterization of particular genes/proteins without further link to biological functional complex. Presented thesis summarises the first large-scale insights into the molecular basis of biological principles of two bird schistosomes Trichobilharzia regenti and Trichobilharzia szidati during their invasion of the definitive avian host. While T. szidati uses the "classical" visceral way of migration - bloodstream and lungs (same as human schistosomes), T. regenti, migrates trough the peripheral nerves and spinal cord. Neurotropic migration is unique among schistosomes and it is also extremely rare...
Fasciolid flukes: from genes to diagnostic tools
Ježková, Monika ; Leontovyč, Roman (advisor) ; Sojka, Daniel (referee)
Liver flukes of the family Fasciolidae are parasites of mammals including human. Fascioloides magna and Fasciola hepatica are considered as a veterinary and medically important species occurring also in the Czech Republic. Fascioloides magna and F. hepatica infect wide spectrum of wild and domestic ruminants and in case of F. hepatica human can be also infected. Both flukes are responsible for damage of liver tissue and/or bile-ducts of their definitive hosts causing weight lose, anemia, reduced productivity and in specific cases the death of the host. Effective diagnosis plays the key role in control of F. hepatica and F. magna infections. Current diagnostics is predominantly based on serodiagnostic methods using specific antigens e.g. from excretory-secretory products (ESPs). Due to heterogenity of ESPs, such diagnostic markers can lack the specificity and also the reproducibility of the method is poor. Particular proteins of ESPs are often used in diagnostics of fasciolid flukes. Such approach requires biological material and laboratory procedures associated with identification, purification and antigenicity testing of selected proteins. Recent development of parallel sequencing technologies results in huge amount of genomic, transcriptomic and proteomic data, which are publicly available. Such...
Species of the genus Radix in transmission of trematodes in the Czech Republic
Leontovyč, Roman ; Horák, Petr (advisor) ; Hlaváč, Jaroslav (referee)
Snails of the genus Radix play an important role in the transmission of many species of trematodes which can represent dangerous parasites of animals and humans. Taxonomical situation within the genus Radix is unclear. It is caused by morphological plasticity of the shells of particular species. In the past the shell shape was the most frequently used determination criterion for new species description. Due to new data brought by molecular taxonomy, validity of some species became doubtful. In our contribution, modern methods (phylogenetic analysis based on gene sequences) and classical approaches (determination by conchological features and anatomy of reproductive organs) were used for determination of snails belonging to the genus Radix. For phylogenetic analysis two genes were characterized (mitochondrial 16S rDNA and nuclear ITS2 rDNA); in the samples 5 species of the genus Radix (R. auricularia, R. labiata, R. lagotis, R. peregra a R. ampla) were confirmed. A thorough morphological determination was based on 11 conchological characters which allowed to identify all selected snails; conchological and molecular identifications were in agreement. For determination by means of reproductive organ morphology, shape and position of bursa copulatrix and its duct were used. According to these criteria,...

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