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Population genetics of grey wolf in Western Carpathians
Veselovská, Lenka ; Hulva, Pavel (advisor) ; Suchomel, Josef (referee)
The grey wolf is a top predator that plays a keystone role in maintaining the ecological balance of forested habitats. This master thesis focuses on the population structure and demography of the grey wolves in Slovakia, using data collected over two winter seasons between 2017 and 2019. Special attention is directed to the Western Carpathians. In Slovakia, the range of the wolf may cover up to 60% of the whole area. The population size and structure of grey wolves are influenced by both natural factors, such as mountain ranges isolation and dispersal, and anthropogenic influences, such as population fragmentation caused by linear infrastructure and increased mortality from traffic collisions. Additionally, the annual culling quotas for grey wolves have had an impact on their populations until recently. Mainly a non-invasive genetics approach utilizing selected 10 nuclear microsatellite markers, a marker for sex detection and mitochondrial sequences was used. Firstly, the species determination was verified and data quality filtering was carried out under statistical control. The sex ratio of the examined sample was 1:1,3. Mitochondrial data were assigned to haplotypes to help infer population origin. Bayesian clustering analysis of microsatellite data using the STRUCTURE program was used to...
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The use of cell-free nucleic acids in maternal plasma for non-invasive prenatal diagnosis of monogenic diseases, placental insufficiency-related complications and Down syndrome
Veselovská, Lenka ; Hromadníková, Ilona (advisor) ; Schierová, Michaela (referee)
Since the discovery of cell-free fetal DNA in peripheral blood of pregnant women, cell-free nucleic acids in maternal plasma are explored in relation to non-invasive prenatal diagnosis of various fetal conditions and pregnancy complications. Non-invasive prenatal diagnosis of monogenic diseases represented by TSC1-linked tuberous sclerosis could be achieved by detection of paternally-inherited mutant allele in the pool of maternal alleles in plasma. Reliability of detection of mutant allele could be improved by simultaneous mutation haplotype analysis or detection of universal fetal marker. None of the 3 methods (allele- specific real-time PCR, SNaPshot minisequencing and quantitative fluorescent PCR) evaluated using artificial mixtures and maternal plasma samples reliably and accurately detected low-frequency allele distinguished by point mutation, SNP or microsatellite in TSC1 gene or in its close proximity. We developed a strategy for prediction of proportion of informative couples for panel of SNPs of interest that can be applied to any monogenic disease. Exploiting differential methylation of promoters of genes RASSF1A, HLCS and OLIG2 in maternal and fetal genome, we failed to establish functional fetal marker. MicroRNAs of placental origin released into plasma could serve as biomarkers of...
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Phylogeography and adaptive evolution of the grey wolf
Veselovská, Lenka ; Hulva, Pavel (advisor) ; Fornůsková, Alena (referee)
Grey wolf is a highly mobile top predator, a keystone and umbrella species within ecosystems throughout the Holarctic area. The occurrence of wolves' populations is influenced by glacial history, environmental conditions and human activity. Nowadays, wolves are returning to a man-altered country where they were exterminated, and they are adapting to human dominated landscape. People have largely contributed to its extinction in many areas around the world, resulting in a decline in genetic diversity. Due to different demographic and environmental conditions, many different lineages have evolved, which can be distinguished based on morphological and genetic analyses. Climatic factors can result in the formation of ecotypes, which become heritable and genetically distinguishable. The aim of this thesis is to provide a comprehensive overview of the phylogeography and adaptive evolution of the grey wolf in the context of genetic, geographical and morphological combination data. Keywords: grey wolf, Canis lupus, phylogeography, ecotypes, adaptive evolution
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The use of cell-free nucleic acids in maternal plasma for non-invasive prenatal diagnosis of monogenic diseases, placental insufficiency-related complications and Down syndrome
Veselovská, Lenka ; Hromadníková, Ilona (advisor) ; Schierová, Michaela (referee)
Since the discovery of cell-free fetal DNA in peripheral blood of pregnant women, cell-free nucleic acids in maternal plasma are explored in relation to non-invasive prenatal diagnosis of various fetal conditions and pregnancy complications. Non-invasive prenatal diagnosis of monogenic diseases represented by TSC1-linked tuberous sclerosis could be achieved by detection of paternally-inherited mutant allele in the pool of maternal alleles in plasma. Reliability of detection of mutant allele could be improved by simultaneous mutation haplotype analysis or detection of universal fetal marker. None of the 3 methods (allele- specific real-time PCR, SNaPshot minisequencing and quantitative fluorescent PCR) evaluated using artificial mixtures and maternal plasma samples reliably and accurately detected low-frequency allele distinguished by point mutation, SNP or microsatellite in TSC1 gene or in its close proximity. We developed a strategy for prediction of proportion of informative couples for panel of SNPs of interest that can be applied to any monogenic disease. Exploiting differential methylation of promoters of genes RASSF1A, HLCS and OLIG2 in maternal and fetal genome, we failed to establish functional fetal marker. MicroRNAs of placental origin released into plasma could serve as biomarkers of...
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