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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...
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The skin immune response of mice infected with avian schistosomes
Revalová, Alena ; Macháček, Tomáš (advisor) ; Kolářová, Libuše (referee)
Invasion of the mammalian skin by cercariae of avian schistosomes of the genus Trichobilharzia is associated with the skin pathology called cercarial dermatitis (CD). Although the manifestation of CD, characterized by the presence of maculo-papular rash and pruritus, is believed to be linked with the previous sensitization, the role of the infection dose and the specific causative agent is rarely considered. Therefore the skin immune response in mouse pinnae infected by 100, 1000 or 4x100 (repeated infections) cercariae of two Trichobilharzia species - T. regenti and T. szidati - was examined in this work. To compare the effect of these factors (the infection dose and species), a complex approach was ensured by employing both in vivo and in vitro methods. Firstly, histopathological changes in the pinnae were examined by classical histology and infiltrated leukocytes were characterized by flow cytometry. Sera of mice were used to detect the systemic production of cytokines and parasite-specific antibodies. In vitro cultivation of mouse pinnae was used to detect the local production of cytokines. The most remarkable observation was that the infection dose largely impacted the course of the immune response, both locally and systematically. In primary infected mouse pinnae, 100 cercariae of both...
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Larval Stages of Flukes (Platyhelmintes: Trematoda) in Secondary School Education
Šulcová, Hana ; Říhová, Dagmar (advisor) ; Podroužková, Štěpánka (referee)
The parasites are one of the most important factors that shape relationships in the nature. This thesis deals mainly with developmental stages of trematodes (Platyhelmintes: Trematoda), especially the cercariae of so-called Schistosomes and sporocysts of Leucochloridium paradoxum, as well as with and their intermediate host - freshwater snails. Introductory chapters are focused on general introduction into the topic, such as basic terminology or occurrence issue of trematodes (mainly schistosomes) in the world and in the Czech Republic. In order to determine larval stages of flukes and their morphological types, the research was conducted in four Prague localities with known presence of aquatic snails. Only in one site, Kunratická tůň Pond, the constant presence of echinostomous cercariae (and in lesser extent also furcocercariae) in Radix labiata was recorded during summer season of 2015. In small pond in the Botanical Garden of the Natural Sciences Faculty of Charles University and in Modřanské tůně Ponds, xiphidocercarie in Lymnaea stagnalis were found. No cercarie were detected in the pond in the Genetic garden of Charles University. The presence of Leucochloridium paradoxum in the European Amber Snail Succinea putris was confirmed in the vicinity of Modřanské tůně Ponds. The verified...
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Avian schistosomes and cercarial dermatitis in Czech Republic: distribution, diverzity and factors influencing their occurrence
Pokrupová, Zuzana ; Bulantová, Jana (advisor) ; Soldánová, Miroslava (referee)
Cercarial dermatitis (CD) is manifested as a strong itchy rash that usually occurs after bathing in the natural water bodies where it makes recreation uncomfortable. As a consequence, the natural swimming areas and be closed because of inconvenient water quality, what subsequently leads to the financial losses. The infection can also affect people working in the natural water bodies as lifeguards (at the natural swimming areas), people monitoring water organisms or water quality etc.). Nowadays, CD in Czech Republic occurs more frequently than in the past. For this reason, the topic of avian schistosomes and CD is very attractive not only for scientists, but newly also for employees of hygienic stations and health institutes. Based on the Act No. 258/2000 Coll. on protection of public health and related executive Decree No. 238/2011 Coll. approved this year their duty will be regular monitoring of official natural swimming areas for causative agents of CD. For the comprehensive overview about the occurrence of the avian schistosomes and CD, up to now, at the localities in the Czech Republic the specialized overview map with the marked catches of avian schistosomes and CD was created in the program ArcGIS Online. This map was made with use the records obtained from the scientific articles, final...
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Larval Stages of Flukes (Platyhelmintes: Trematoda) in Secondary School Education
Šulcová, Hana ; Říhová, Dagmar (advisor) ; Podroužková, Štěpánka (referee)
The parasites are one of the most important factors that shape relationships in the nature. This thesis deals mainly with developmental stages of trematodes (Platyhelmintes: Trematoda), especially the cercariae of so-called Schistosomes and sporocysts of Leucochloridium paradoxum, as well as with and their intermediate host - freshwater snails. Introductory chapters are focused on general introduction into the topic, such as basic terminology or occurrence issue of trematodes (mainly schistosomes) in the world and in the Czech Republic. In order to determine larval stages of flukes and their morphological types, the research was conducted in four Prague localities with known presence of aquatic snails. Only in one site, Kunratická tůň Pond, the constant presence of echinostomous cercariae (and in lesser extent also furcocercariae) in Radix labiata was recorded during summer season of 2015. In small pond in the Botanical Garden of the Natural Sciences Faculty of Charles University and in Modřanské tůně Ponds, xiphidocercarie in Lymnaea stagnalis were found. No cercarie were detected in the pond in the Genetic garden of Charles University. The presence of Leucochloridium paradoxum in the European Amber Snail Succinea putris was confirmed in the vicinity of Modřanské tůně Ponds. The verified...
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