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Evolution of mixed cultures of the pathogenic yeasts Candida albicans and Candida guilliermondii
Hirko, Dominik ; Heidingsfeld, Olga (advisor) ; Schierová, Michaela (referee)
The objective of this thesis was to investigate how cocultures of Candida albicans and Candida guilliermondii change overtime under control conditions and under the influence of fluconazole. These species are opportunistic fungal pathogens but widely differ in their susceptibility to antimycotic of interest - fluconazole. After a brief introduction to special commonalities, the mechanism of pathogenesis, and the treatment of infection, this work explores each organism's growth curves under selected conditions and the process of artificial evolution using the model of passaging of cocultures. Afterwards, these populations of C. albicans and C. guillieromondii were investigated using qPCR and chromogenic media. qPCR analysis revealed that under control conditions, C. albicans (CA) prevails; the possible reason behind this is a 20% shorter generation time, as revealed by the growth curve. In the presence of fluconazole, two trends occurred. One is related to the innate resistance of C. guilliermondii (CG), where CG was dominant by the end of passaging. The second trend led to CA being the dominant one, despite its susceptibility. This is a result of a heightened resistance, where minimal inhibitory concentration 50 (MIC50) increased almost 10-fold, possibly due to mutations. The change in populations...
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Complete genome sequence analysis of TBE virus, strain Hypr, in different passages in host \kur{in vivo}.
ŠIMONOVÁ, Zuzana
I have compared genomic sequences of three variants of tick-borne encephalitis virus, strain Hypr. The variants used were Hypr5 {--} virus passaged five times in suckling mice brains, Hypr50/V {--} a variant highly passaged in vivo in Vienna, and Hypr50/CB {--} a variant highly passaged in vivo in České Budějovice. Both high-passage variants were passaged in suckling mouse brains as well. I was interested in changes in nucleotide sequence of genomes of high-passage variants Hypr50/V and Hypr50/CB compared with Hypr5 and also consequential changes in amino-acid sequences of viral proteins. I have found several identical changes in amino-acid sequences of E protein, NS3 and NS5 proteins in both high-passage variants and also a number of substitutions unique to one of the variants in proteins E, NS1, NS2A, NS2B, and NS3. A nucleotide substitution was found also in 5{\crq}UTR of the viral genome.
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