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Morphological and molecular diversity of the free-living representatives of the family Metopidae and the discovery of a new lineage of anaerobic ciliates
Rotterová, Johana ; Čepička, Ivan (advisor) ; Fiala, Ivan (referee)
Anoxic sediments host a wide variety of ciliates. Although it is known that anaerobiosis has independently arisen in several lineages of the main 11 lineages of Ciliophora and anaerobes have been found in at least eight lineages of ciliates, the diversity of anaerobic ciliates of the class Armophorea is severely understudied. Similarly, not much is known about their hydrogenosomes. To deepen our knowledge about the diversity of Armophorea, we have cultivated more than 100 armophorid strains from fresh water, brackish, and marine anoxic sediments worldwide. We determined their SSU rDNA sequences, performed protargol staining techniques, and studied light-microscopic morphology. In addition, we used transmission electron microscopy to assess the ultrastructure of some of the strains. Several novel clades of metopids, the free-living anaerobic ciliates of the class Armophorea, were identified. Importantly, a new deep lineage of marine anaerobic ciliates, muranes, was discovered. According to the SSU rDNA analysis, it is related to SAL group (Spirotrichea, Armophorea, and Litostomatea) with Cariacotrichea, but forms a separate lineage, possibly a novel class. We conclude that anoxic sediments harbour a high diversity of undescribed anaerobic ciliates. Key words: Anaerobiosis, Armophorea, ciliates,...
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Transmission and detection of the crayfish plague pathogen under experimental conditions
Svoboda, Jiří ; Petrusek, Adam (advisor) ; Fiala, Ivan (referee)
The crayfish plague pathogen, Aphanomyces astaci, is one of the most serious threats to European indigenous crayfish species, e.g., the noble crayfish (Astacus astacus). The only way to protect susceptible crayfish species from the disease is to prevent the dispersion of the pathogen to their populations. One of the most important sources of the crayfish plague pathogen in Central Europe is the spiny-cheek crayfish (Orconectes limosus), a species of North American origin, which can carry the parasite in its cuticle for years. Some literature sources claimed that the pathogen dispersion from the American vectors is restricted to periods of moulting or to the time before and after the crayfish death. However, experimental evidence for such hypotheses was lacking. The main aim of my thesis was to test these predictions, and the alternative scenario that the crayfish plague pathogen can be transmitted from the infected spiny-cheek crayfish also in other periods. For this purpose, experiments were set up to investigate A. astaci transmission from infected spiny-cheek crayfish to non-infected spiny-cheek or noble crayfish. As expected, the pathogen was transmitted to noble crayfish much more easily than to the uninfected American host. Nevertheless, we succeeded in the pathogen transmission also among spiny-cheek...
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The method of DNA barcoding and its use in protists
Rotterová, Johana ; Čepička, Ivan (advisor) ; Fiala, Ivan (referee)
DNA barcoding is a molecular method of species identification based on short regions of DNA sequences. These DNA barcodes should be unique for each species. The sequences within species should be identical or very much alike, while between species they should display a significant amount of differences. The sequences of an undetermined specimen can be compared with sequences vouchered to a particular species in a global DNA barcode database. The database was developed with the aim to simplify traditional taxonomy, eliminate its problems and accelerate its approaches. However, DNA barcoding has its own limitations that may hinder achieving its original goals and a compliance with the concept. The aim of this bachelor thesis consists of explaning the evolution of DNA barcoding through time and assessing the importance of this method at present, particularly in protists. Species identification may be very difficult in many protist groups, which can be explained by several reasons. To appraise this method completely is a hard task, because creation of the database has not yet been finished and official DNA barcodes have not been chosed for many groups. We can only find out in the future, therefore i will only focus on factors that influence the success of DNA barcoding at present.
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Kinetoplastida of birds and bloodsucking Diptera: molecular characterization and life cycles
Zídková, Lenka ; Svobodová, Milena (advisor) ; Fiala, Ivan (referee) ; Modrý, David (referee)
We studied biodiversity of avian trypanosomes. We used newly obtained isolates of trypanosomatids from ornitophilic bloodsucking diptera (81) and from avian hosts (145). To investigate relationships among isolates we employed RAPD method and sequence analyses of SSU rRNA, eventually gGAPDH, 5S RNA or SL RNA. Additionally we used morphological characters; cell lengths and kinetoplast widths. All isolates obtained from biting midges belonged to monogenetic kinetoplastids. We described new species and a new genus Sergeia podlipaevi isolated from Culicoides (Oecacta) festivipennis a C. (O.) truncorum. We successfully infected with this species laboratory bred biting midge of different subgenus Culicoides (Monoculicoides) nubeculosus. We described isolate from Culicoides truncorum as new species Herpetomonas trimorpha. Its sister species was H. ztiplika described previously also from biting midges. We performed the most extensive study of biodiversity of avian trypanosomes based on comparison of wide spectrum of isolates obtained from different hosts. We confirmed that avian trypanosomes are polyphyletic and form three independent lineages within the genus Trypanosoma. We identified at least 11 species of avian trypanosomes. We found that ornitophilic mosquitoes belong among important vectors of avian...
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