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Molecular Characterisation of Parvalbumin Gene: Evolutionary Insights and Forensic Applications for Fish Species Identification and Authentication
Mukherjee, Subham ; Horká, Petra (advisor) ; Kalous, Lukáš (referee) ; Flajšhans, Martin (referee)
Abstract Parvalbumin (pvalb), a low molecular weight calcium-binding protein, plays a crucial role in regulating Ca2+ switching in fast-twitch muscle fibres and has been identified as a major cause of fish-induced food allergies. The molecular evolution of pvalb genes in teleost fish and its cause, duplication of the whole genome, was investigated, revealing high diversity and complex gene repertoires, making detection and identification challenging. This study provides robust genomic evidence of the complex evolution of parvalbumin genes in teleost fish. In addition to its role as a potent allergen, the pvalb gene, a nuclear gene, can serve as a valuable molecular marker. Keeping this in mind, a real-time PCR assay is developed to detect and quantify two European anglerfish species simultaneously, Lophius piscatorius and Lophius budegassa, which are susceptible to illegal species substitutions in the global seafood trade. The assay targets the intronic region of the pvalb gene, demonstrating high specificity, efficiency, and robustness, making it a potential forensic tool to prevent food fraud and ensure the accurate identification of fish species. Furthermore, a standardised quantitative PCR-based method is presented for the β-pvalb gene in Lophius piscatorius, utilising a plasmid DNA calibrator...
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Evolutionary history, genetic diversity and human impact on selected leuciscids from the southern Balkans
Viñuela Rodríguez, Nuria ; Vukićová, Jasna (advisor) ; Perea, Silvia (referee) ; Kalous, Lukáš (referee)
- ABSTRACT - Fishes represent the most diverse group of vertebrates on Earth, with freshwater fishes accounting for half of this diversity, and occupying a mere 0.01 % of the world's water surface. The Balkans are considered a European hotspot of biodiversity, and its southern section is recognized as centre of ichthyological diversity, characterized by its exceptional richness and endemicity. Within the southern Balkans, the Ionian ecoregion stands out as one of the most isolated ichthyological regions in Europe due to its very complex geological history and the variety of events that influenced its ichthyofaunal composition. All fish species inhabiting the Ionian region are endemic, and most of them display reduced distribution areas with a marked geographic pattern. The western Aegean region, on the contrary, is considered a species-poor ecoregion within the southern Balkans, despite also harbouring a significant number of endemic species. However interesting the region of the southern Balkans is in terms of its ichthyofauna, there is a lack of knowledge for many of the fish species inhabiting them. In this thesis, we address some of these knowledge gaps for two representative genera, Pelasgus and Telestes, of the most diverse family in the area (Leuciscidae), with the aim to better understand the...
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Phylogeography of the genus Squalius in Albania
Lerch, Zdeněk ; Švátora, Miroslav (advisor) ; Kalous, Lukáš (referee)
The thesis is focused on the issue of the Squalius genus phylogeography in Albania in the Balkan region. Phylogenetic analyses of sequence variation at mitochondrial genes (cytochrome b) were used to examine these issues for the freshwater fish genus Squalius from various river systems in the Adriatic Sea region. There were identified 3 genetic lineages of vague taxonomic position where the genetic variation amount to 1.6-2.1 %. The first lineage is distributed in north of Albania and in the whole Periadriatic region, the second lineage is especially spread to northern part of Albania and the third lineage covers especially the zone of the European ancient lake systems on the Balkan Peninsula (Lake Ohrid and Prespa), from where expands to the southern part of Albania. There sympatrically coincide the both lineage in the hydrological river-lake system of Drin and Skadar. The phylogenetic and taxonomic position of the Squalius genus in the region seems to be interesting topic for subsequent and more detailed study. Keywords: Squalius, Albania, cytochrome b, phylogeography
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Aquatic fauna of karstic fields in the Balkans
Doulíková, Tereza ; Vukićová, Jasna (advisor) ; Kalous, Lukáš (referee)
Dinaric Karst region of the Balkan Peninsula is one of the hot spots of biodiversity, and it is characterized by a high degree of endemism of freshwater fishes. Numerous geological and climatic events have influenced the formation of the area and speciation. Karst poljes are the specific features of Dinaric Karst. They represent a specific ecosystem full of endemic species, where hydrogeological processes play an important role. The periodic flooding appears on poljes. It leads to activation of various underground channels and their interconnection. Twenty two endemic fish species have adapted to this regime. They live in underground waters or in surface waters part of the year and they can migrate between poljes through karst channels. They show significant reduction in size and change in complexity of scaliness. Scales are small and they are barely visible in some species. All these species and abundant underground karst fauna are threatened primarily by pollution, habitat destruction, and drainage of poljes.
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The evolution of vision in ray-finned fishes
Truhlářová, Veronika ; Musilová, Zuzana (advisor) ; Kalous, Lukáš (referee)
Vision plays a key role in life of many vertebrates, and the performance of visual system is often adapted to specific environments inhabited by individual species. Fish colonized a wide range of habitats and adjusted their visual abilities to maximize their success rates in hunting, reproduction and predator avoidance. This thesis is focused on molecular mechanism of visual system, namely on genes for photoreceptor proteins, opsins, of two major groups of teleost fishes: African riverine cichlids (family Cichlidae, order Cichliformes, part of larger taxa Percomorpha), and European freshwater cyprinids (family Cyprinidae, order Cypriniformes, part of larger taxa Ostariophysi). Two types of photoreceptor cells are present on retina: the cones and the rods. Actinopterygian fishes in general have four cone opsin types (SWS1, SWS2, RH2 and LWS) used for colour (photopic) vision, and one rod opsin type (rhodopsins) for vision in deteriorated light conditions (scotopic vision). In the present thesis, I focus on 1) DNA sequence and amino acid substitutions of the opsin genes, and on 2) gene expression levels of opsins sensitive to various wavelengths of light spectrum. The results of my work show that both cichlids (family Cichlidae) and cyprinids (family Cyprinidae) have a complete set of opsin genes in...
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Evolutionary relationships within genus Barbus (Cyprinidae) with emphasis on Greece, and biogeography of Greek lineages
Kyralová, Eva ; Vukićová, Jasna (advisor) ; Kalous, Lukáš (referee)
Evolutionary relationships between Barbus species have been already studied in the past, but most of the studies were based only on mitochondrial DNA, there is no detailed study based on nuclear markers. This is mainly due to the fact that Barbus species are tetraploid. The western part of Greece has been studied insufficiently, and moreover a greater diversity of freshwater ichthyofauna is expected here. Greece is one of the hotspots of biodiversity. Greek river system is characterized by a significant proportion of endemic species of freshwater fishes. In this work evolutionary relationships between Barbus species were studied, with focus on diversity of Greek lineages. The geographical distribution of Barbus species has been also revised. In total, 192 individuals were analyzed, 156 individuals were from 40 Greek localities and another 36 Barbus species were from other areas of distribution of the genus. Dataset was supplemented with avaible published sequences. For the phylogenetic analysis mitochondrial gene cytochrom b, 1st intron of ribosomal protein gene S7 and 2nd intron of gene beta-actin were used. For both nuclear markers paralog-specific primers were used. The analysis of cytochrom b revealed several well-supported lines: 1. species of southwestern Mediterranean (B. meridionalis and B....
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Aquatic fauna of karstic fields in the Balkans
Doulíková, Tereza ; Vukićová, Jasna (advisor) ; Kalous, Lukáš (referee)
Dinaric Karst region of the Balkan Peninsula is one of the hot spots of biodiversity, and it is characterized by a high degree of endemism of freshwater fishes. Numerous geological and climatic events have influenced the formation of the area and speciation. Karst poljes are the specific features of Dinaric Karst. They represent a specific ecosystem full of endemic species, where hydrogeological processes play an important role. The periodic flooding appears on poljes. It leads to activation of various underground channels and their interconnection. Twenty two endemic fish species have adapted to this regime. They live in underground waters or in surface waters part of the year and they can migrate between poljes through karst channels. They show significant reduction in size and change in complexity of scaliness. Scales are small and they are barely visible in some species. All these species and abundant underground karst fauna are threatened primarily by pollution, habitat destruction, and drainage of poljes.
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Phylogeography of the genus Squalius in Albania
Lerch, Zdeněk ; Švátora, Miroslav (advisor) ; Kalous, Lukáš (referee)
The thesis is focused on the issue of the Squalius genus phylogeography in Albania in the Balkan region. Phylogenetic analyses of sequence variation at mitochondrial genes (cytochrome b) were used to examine these issues for the freshwater fish genus Squalius from various river systems in the Adriatic Sea region. There were identified 3 genetic lineages of vague taxonomic position where the genetic variation amount to 1.6-2.1 %. The first lineage is distributed in north of Albania and in the whole Periadriatic region, the second lineage is especially spread to northern part of Albania and the third lineage covers especially the zone of the European ancient lake systems on the Balkan Peninsula (Lake Ohrid and Prespa), from where expands to the southern part of Albania. There sympatrically coincide the both lineage in the hydrological river-lake system of Drin and Skadar. The phylogenetic and taxonomic position of the Squalius genus in the region seems to be interesting topic for subsequent and more detailed study. Keywords: Squalius, Albania, cytochrome b, phylogeography
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Microbiota of the digestive tract of bees and related insect and the influencing facftors
Hroncová, Zuzana ; Havlík, Jaroslav (advisor) ; Kalous, Lukáš (referee)
This thesis consists of six research chapters, out of which, five have already been published in research journals, 1 is in the process of submission and the last chapter presents original unpublished confidential data in the process of manuscript preparation.
The research presented in this thesis concerns the complex mechanisms of bees and wasps immune system focused on microbiota as component of immunity. As shown in the introduction, highly social and managed species of bees like honey bees and bumble bees, play key roles in natural and agricultural ecosystems worldwide. Recent losses of bees have been attributed to pesticide exposure, poor nutrition, increased parasite loads and habitat degradation. Over the past several years, governments, beekeepers, and the general public worldwide have become concerned by increased losses of honey bee colonies, calling for more research on how to keep colonies healthy. Our main aim was to explore the complex mechanisms of bees and related species gut bacterial populations, their links to insect immunity and investigate the opportunities for an intervention. Part of our aim was to characterise the bumble bee and wasp gut microbiota using 16S RNA sequencing in a search for potentially novel bacterial species.
We have tried to explain how microbiome interacts with the host and showed that major members of these communities appear to benefit the host. The simple gut communities of social bees present ideal model systems to investigate the underlying evolutionary and genetic processes of such interactions. Information based on our results may help in the design of proper probiotic supplementation strategies with respect to physiological conditions in the honey bee gut.
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Transboundary eel (Anguilla anguilla) management plans
Lehoučková, Sabina ; Kalous, Lukáš (advisor) ; Nechanská, Denisa (referee)
In the bachelor thesis are collected and compared data obtained from available materials that deal with the issue of declining state of European eel (Anguilla anguilla) in European waters. Population decline was recorded already in the 80s of last century and throughout the range, which includes most inland and coastal waters of Europe and North Africa. The first chapter describes the species of Anguilla anguilla, developmental stages and migration cycle, who during his life this species pass. In the following part of the work is information on the Council Regulation (EC) No. 1100/2007 of 18 September 2007 establishing measures for the recovery of the stock of European eel. This regulation requires the individual Member States of the European Union to draw up plans Management of European eel on their territory if the area falls between habitats eel. The next section discusses the management plans for the Czech Republic, England and Wales, the Netherlands, Denmark and Spain. The management plans are then selected individual chapters devoted to the definition of the basin, which measures for the recovery of the eel population concerning the procedure for determining the target escapement levels of 40 % of adults with eel in marine waters and further concrete measures implemented by means of the watercourses which should be the target level achieved. The final part is devoted to the discussion, which are summarized and compared the measures used by Member States in the individual management plans. Furthermore, this part contains data on the eel population after the introduction of primary measures for its renewal.
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