The aim of this study was to design and optimize new PCR primers for detection of potential cyanobacterial producers of cytotoxic lipopeptides puwainaphycins and minutissamides in environmental samples. Samples from two distinct localities were tested, as suggested based on preliminary data. The first set of samples consisted of cyanobacterial soil biofilms from sheep pastures affected by Alveld illness in Norway. The other one contained samples of planktic cyanobacaterial blooms from Protected Landscape Area Třeboň and its vicinity. Three different approaches were used for evaluation of the presence of cyanobacterial lipopeptide producers: microscopy, PCR with the designed primeres, and liquid chromatography-mass spectrometry analysis. Results of this study confirmed the specificity of the newly designed PCR primers. The presence of producers of puwainaphycins/minutissamides was proven at both tested localities.

Good quality of bathing water is an increasingly important factor taken into account when we choose summer vacations, a place to stay for a weekend or a weekend trip destination. In the Czech Republic, bathing waters may be divided according to the legal status into outdoor swimming pools, surface waters used for bathing (swimming areas), other water surfaces or artificial pools. In my thesis I dealt only with outdoor swimming pools and swimming areas. The difference between them lies in the fact that outdoor swimming pools have their own operators who monitor water quality and provide other services in the waterside. Usually there is a fee charged. Swimming areas have no operators and water quality monitoring is placed under the responsibility of regional public health authorities. In the Czech Republic, at present, 188 outdoor swimming pools and swimming areas have been monitored on a regular basis each bathing season. For my research I chose four sites located in the South Bohemian region and four sites in the South Moravian region. For each of these eight sites, I collected results of laboratory analyses, including categories of water quality in bathing seasons of the past five years, it means from the year 2005. With these obtained data I assessed the water quality development both during the bathing season of 2009 and its evolution over the past five years. On the basis of particular categories of water quality I also tried to compare water quality in the South Bohemian and the South Moravian region and also to compare water quality between individual sampling points in one locality. Another aim was to evaluate some measures and interventions made in order to improve water quality. I focused particularly on the pond Olšovec in the South Moravian region and the VN (water reservoir) Orlík in the South Bohemian region. The results do not show any prominent changes in water quality both during the bathing season 2009 and over the past five years. Pronounced fluctuations in water quality were recorded only at certain locations during the 2007 bathing season. Not even between individual regions significant differences in water quality were found. The proportion of individual water quality categories in the two regions did not differ by more than 4%. We can therefore say that water quality in selected localities in the South Bohemian and South Moravian regions is comparable. As for certain measures and interventions implemented to improve water quality, it is important to focus also on the wider neighborhood of the locality, because it could also be a source of substances worsening water quality.

Karotenoidy jsou přírodní pigmenty vyskytující se v mikroorganismech jako jsou řasy, kvasinky a sinice. Představují nejrozšířenější skupinu antioxidantů s významným biologickým účinkem. V současnosti vzrůstá zájem o karotenoidy vzhledem k jejich příznivým vlivům na lidské zdraví. Chlorofyly jsou zelená fotosyntetická barviva, která nacházejí uplatnění v potravinářství jako intenzivní zelená barviva. Koenzym Q je znám svým pozitivním vlivem pro správnou funkci řady orgánů v lidském těle. Ergosterol je nedílnou součástí membrán kvasinek a hub. Je to provitamin D2, který je důležitou součástí imunitního systému. Mikrobiální lipidy, nebo také ‚‚Single cell oils‘‘ jsou charakteristické vysokým obsahem zdraví prospěšných nenasycených mastných kyselin, které lze využít ve farmacii či kosmetice. Mikrobiální lipidy jsou dále studovány jako alternativa pro výrobu biopaliv. Dizertační práce byla zaměřena na studium a možnosti optimalizace produkce lipidů a lipidických látek vybranými kmeny karotenogenních kvasinek, mikrořas a sinic. V rámci práce byly testované kvasinky rodu Rhodotorula, Rhodosporidium, Cystofilobasidium a Sporidiobolus podrobené kultivacím na sérií médií s různými C/N poměry v rozsahu 13 až 100, obsahujících upravené odpadní substráty z potravinářského průmyslu. Vybrané kmeny byly poté kultivovány v bioreaktorech v médiu obsahujícím kombinaci odpadních substrátů. Kultivace mikrořas rodu Desmodesmus, Scenedesmus, Chlorella, Coccomyxa, Chlamydomonas, Botryococcus se zabývaly optimalizací jednotlivých komponent média a aplikací různých stresů s cílem navýšení produkce studovaných metabolitů. V rámci experimentů s extrémofilní mikrořasou Coccomyxa byly provedeny pilotní velkoobjemové kultivace v otevřených nádržích. V závěrečné části byl provedeny pilotní screeningové a velkoobjemové bioreaktorové experimenty zaměřené na možnosti kokultivace karotenogenních kvasinek a mikrořas. Testované kmeny kvasinek byly s rozdílnou úspěšností schopny utilizovat média obsahující hydrolyzované odpadní substráty. Nejlepším kmenem byl Sporidiobolus pararoseus, který na médiích dosahoval nejvyšších produkcí biomasy i sledovaných metabolitů. Z testovaných odpadních substrátů byla nejlepší kombinace odpadního fritovacího oleje a hydrolyzátu kávové sedliny. Úspěšná optimalizace složení hlavních komponent minerálního média vedla k zvýšené produkci studovaných metabolitů. Největší vliv měl optimální poměr P/N a aplikace oxidačního stresu. Nejlepších výsledků dosáhly mikrořasy rodu Desmodesmus a Scenedesmus. Velkoobjemové kultivace Coccomyxy onubensis potvrdily rezistenci kultury proti kontaminaci vnějšími vlivy a schopnost růstu za vysoké teploty a intenzity světelného záření. Kokultivační experimenty potvrdily schopnost symbiotického růstu kvasinek a mikrořas. Nejlepších výsledků dosahovaly všechny testované kvasinky s mikrořasami rodu Demsodesmus a Scenedesmus a v menší míře i rodu Coccomyxa.

The theoretical part of this master’s thesis is focused on the characteristics and forms of phosphorus, occurrence in natural waters and the problems associated with higher phosphorus concentrations, especially in the development of cyanobacteria and algae. There are also described the possibilities of determining the degree of pollution of natural waters by nutrients and the effects of eutrophication. There are also described methods for the determination of phosphates, especially spectrophotometric determination in ternary systems with dyes. Like other methods are mentioned AAS, ion chromatography, electrophoresis and izotachophoresis, electrochemical methods or enzymatic methods. In the experimental part of this work deals with the selection and optimalization of suitable ternary system with organic dyes for spectrophotometric determination of orthophosphates. The system with phosphovanadomolybdic acid is studied too. For analytical procedure with malachite green oxalate, which was found to be most useful, were studied interfering ions, commonly occurring in natural waters . The chosen procedure was finally tested on real water samples.

The House of Memory combines the functions of a cyanobacteria purifier, museum and a winery, in reaction to the conditions of the place.

Morphological diversity of 61 Anabaena populations of 13 morphospecies was described under the field conditions of Czech fishponds and reservoirs. Polyphasic approach was then applied in classification of 45 clonal strains isolated from those populations. Detailed morphological analyses were performed and partial 16S rRNA gene sequences were obtained for 33 of the strains, and secondary metabolite production was evaluated in 20 strains. Plasticity of morphological characteristics under varied conditions of light, temperature, nitrogen and phosphorus was studied in selected strains, as well as their temperature and light growth requirements. The results were then discussed with respect to the delimitation of single Anabaena morphospecies. A new genus Sphaerospermum was defined for the morphospecies Anabaena kisseleviana, A. reniformis and Aphanizomenon aphanizomenoides, whose phenotypic and genotypic features differed considerably from all other Anabaena morphospecies. Unique information was provided on the occurrence and distribution of A. reniformis and Aph. aphanizomenoides in the Czech Republic.