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Optimization of cultivation of carotenogenic yeasts on mixed waste substrates
Holub, Jiří ; Mikulíková, Renata (referee) ; Márová, Ivana (advisor)
The master thesis addresses the issue of cultivation of selected strains of carotenogenic yeasts on waste materials of the food industry using a laboratory bioreactor. Carotenogenic yeasts are able to produce highly valuable metabolites during cultivation, which are located predominantly in the lipid part of the cells. Particularly, they are carotenoids, ergosterol, coenzyme Q and fatty acids. The thesis is divided into two main parts, the theoretical part and the practical part. The theoretical part describes individual yeast strains, types of waste materials, produced metabolites and methods of their analysis. The experimental part deals with the processing of waste materials of the food industry, specifically animal fat, whey and spent coffee grounds into the form of substrates usable as nutrition sources for yeast cultivation. Furthermore, cultivations focused on the recovery of the monitored metabolites and their analysis by using HPLC/PDA and GC/FID assemblies were studied as well. The yeast strains Rhodotorula mucilaginosa (CCY 19-4-6), Rhodotorula kratochvilae (CCY 20-2-26), Rhodosporidium toruloides (CCY 062-002-001), Sporidiobolus pararoseus (CCY 19-9-6) a Cystofilobasidium macerans (CCY 10-1-2) were used in this work. As one of the best producing strains Sporidiobolus pararoseus (CCY 19-9-6) was found, which achieved very high productions of carotenoids, coenzyme Q and ergosterol.
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Use of algae for production of biomass and bioproducts
Slonek, Jaroslav ; Máša, Vítězslav (referee) ; Brummer, Vladimír (advisor)
Submitted bachelor thesis is focused on description of use of algae for biomass and bioproducts production. The first part of the thesis is devoted to the taxonomic classification of algae and a basic summary of the characteristics of the different algal groups. The next part is aimed at bioproducts, which are obtained by algae cultivation and possibilities of further processing for energy purposes are also described. As for the cultivation, the emphasis is on modern methods in this area, i.e. on the possibility of using waste products in the culture process. In the next part, technological parameters of cultivation are discussed, for instance effects of technological parameters on growth and composition of biomass and production of desired products. The next chapter summarizes types of used culture bioreactors in which the growth of biomass occurs in a controlled environment. The information obtained and collected in the thesis is further discussed and summarized in the conclusion.
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Live food organisms for fish - an overview
MILLEROVÁ, Adéla
Feeding organisms are an important part of the fish's nutrition, they are used not only when foddering fish larvae, but also as a supplementary feed for adult fish. The addition of animal and vegetal ingredients in the feed of fish has a proven effect on their growth, reproduction, condition and the intensity of their coloration. To the group of feeding organisms used in fishkeeping belong protozoans, rotifers, crustaceans, nematodes, oligochaetes, molluscs and insects. A specific group of feeding organisms are algae and cyanobacteria, which are used not only as a supplementary feed for aquarium fish, but also as a food for some feeding organisms, for example rotifers, cladocera and copepods. The individual chapters of the thesis contain information about the usage of these specific organisms in fishkeeping, basic characteristics (size, occurrence, eventually also reproduction), breeding in home conditions (inclusive the list of needed things, advantages, disadvantages and the costs of breeding), also the method of obtaining and preservation of this specific type of nutriment.
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Diagnostics of the blood cultures from the departments of internal medicine in Nemocnice Tábor, a.s.
SMRŽOVÁ, Kamila
The thesis describes blood culture testing at internal medicine departments of Nemocnice Tábor, a.s. (Tábor Hospital). The theoretical part presents the most important terms related to infections of the bloodstream and sepsis. EARS-Net, an important European project is also mentioned. The practical part describes blood culture laboratory process in all phases (preanalytical, analytical and post-analytical) including the steps taken when a sample is positive. Furthermore, the study evaluates data on blood cultures done at internal medicine departments of Nemocnice Tábor, a.s. in 2018, determining the incidence of gram-negative bacteria in positive samples and seeking to describe the differences between community and hospitalised patient groups. Finally, the study also gives an overview of antibiotic resistance in the most commonly detected gram-negative rods including a comparison to EARS-Net data (mean value for internal medicine departments in the Czech Republic). In total, 527 blood culture vials were tested as positive in 2018. A total of 267 positive vials were analysed further after the removal of duplicities and contaminations; a total of 97 strains of gram-negative rods were cultured from the vials. The most commonly detected gram-negative bacterial strains found in positive samples included Escherichia coli (48.5%) and Klebsiella pneumoniae (22.7%). Differences between community and hospitalised patient groups were found only for Klebsiella pneumoniae; however, the pathogens spectra in both groups were not distinctively different. In terms of resistance assessment in the two most common bacterial strains, a higher frequency of incidence of Klebsiella pneumoniae strains resistant to third generation cephalosporins and ciprofloxacin is alarming, also in comparison to EARS-Net data (mean value for internal medicine departments in the Czech Republic).
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Využití kultivačních metod v diagnostice zánětů mléčné žlázy ve vybraném chovu dojnic
ULMOVÁ, Kateřina
The goal of my thesis was to evaluate origin of mastisis diseases and specify the best way to cure this disease with antibiotics. Secondary goal was to examin origin of things that cause these diseases, efficiency of the cure and how it is influenced by the stage of the illness. This research was made in cowshed, fron april to december 2018 with 136 specimen. During this period I took samples of milk, that had more than 800 000 somatic cells in 1ml of milk. Based of possitve NK test, samples of mammary gland were taken and cultivated on one used only Petri dish. Depending of the origin of the infection, the proper atibitics were chosen for the cure. After 7 to 14 days after antibiotics treatment, milk sample was taken from the cured part od mammary gland and It was tested to see, if the treatment was successful. 77% of the specimen were sick because of microorganisms and most of the specimen were afflicted the most during 1st lactation (36.8% to 47.1%). Following microorganisms caused mastisis - Staphylococcus spp. (KNS), Streptococcus uberis, Klebsiella oxytoca, Escherichia coli, Candida rugosa a Enterococcus faecalis. 36% of mastisis was caused by Staphylococcus spp. (KNS), followed by coliform bacteria (22%). 17% of mastisis was caused by Streptococcus uberis. 10.5% of specimen was infected by Enterococcus faecalis. Candida rugosa was found in 7.4% of the samples and in 9.5% of samples the diseasehad no specific microorganism responsible. From 105 infected cows, 68% of them fully recovered. At the end, based on the stage of the mastisis, efectivness of the treatmend was statistically evaluated. The evaluation showed, that stage of infection massively influence the result of the treatment.
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Kultivace klíšťaty přenášených babesií a charakterizace jejich proteasomu jako nového terapeutického cíle
REICHENSDÖRFEROVÁ, Dominika
Tick-transmitted parasites of the genus Babesia represent an important worldwide veterinary threat and an emerging risk to humans. In comparison to their malaria-causative relatives, these erythrocyte infecting Apicomplexa have been widely neglected and no specific treatment has been developed. Thus, this thesis is focused on the optimised cultivation of the two species: B. divergens in bovine erythrocyte cultures (in vitro) and B. microti in BALB/c mice (in vivo). Since the Babesia 26S proteasome has been recently validated by our laboratory for drug development we applied different strategies used to isolate the P. falciparum 26S proteasome to optimize its isolation from the two babesia species - ultrasonication, mechanical homogenization and ultracentrifugation. In addition, we used ion exchange chromatography to purify the 26S proteasome from B. microti lysates. Future steps will involve optimised protocols using either ion exchange, immunoprecipitation and/or ubiquitin affinity to purify volumes of B. microti proteasome suitable for substrate specifity profiling of the proteolytic subunits as well as for 3D protein studies involving Cryo-EM or conventional protein crystalography. The overal goal of the long term laboratory project is to produce and validate novel Babesia selective proteasome inhibitors that could be developed into yet missing specific treatment for babesiosis.
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Production of carotenoids grown on waste fat
Holub, Jiří ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
Carotenoids are lipid-soluble pigments which are contained in plants and microorganisms. Carotenoids are known for their antioxidant effects. Lipids are predominantly non-polar substances, which are essential and present in all types of organisms. The research was made by theoretical and practical form. It was dealing with production of carotenoids pigments and different lipophilic substances by yeasts of Rhodotorula glutinis, Rhodosporidium toruloides and Sporidiobolus metaroseus, cultivated in media containing waste fat or glycerol. Further, methods for obtaining lipid substances from microorganisms and with further analysis of samples by gas or liquid chromatography were optimized. The main task of the research was to find out which types of media and yeasts are the most suitable for production of lipid substances. For production of lipid substances in glycerol media, as the best producing strain Rhodosporidium toruloides was found, which simultaneously reported the highest concentrations of carotenoids on fatty and fatty hydrolyzed media.
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In vitro cultivation of the trematode species Trichobilharzia regenti
Vrbová, Kristýna ; Kašný, Martin (advisor) ; Skelly, Patrick (referee)
The class Trematoda includes many pathogenic representatives. Main subject of this thesis, avian schistosome Trichobilharzia regenti, is a close relative to the important human pathogen Schistosoma mansoni (family Schistosomatidae). In vitro cultivation of trematodes enables closer understanding of their biology and parasite- host interactions; however, no trematode species has been successfully kept in vitro from the egg stage to the adults producing eggs. Many studies are focused on the problematic of S. mansoni cultivation, but data concerning T. regenti cultivation remain scarce. Only the ability of T. regenti cercariae to transform into schistosomula in vitro was documented, with following survival in a culture medium for a few days. Comparison of eight transformation methods was performed with T. regenti cercariae. Based on the number of tailless cercarial bodies obtained, five transformation methods were selected for further evaluation of the early schistosomula characteristics (glycocalyx shedding, penetration glands emptying and survival in vitro). It was observed that the largest quantity of cercarial bodies can be obtained by using a syringe needle or the BeadBeater cell disrupter. The largest quantity of schistosomula meeting the criteria of early schistosomulum was recorded after...
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Microscopic Fungi in Practical Exercises at Lower Secondary School
Tejmlová, Kristýna ; Pavlasová, Lenka (advisor) ; Skýbová, Jana (referee)
Microscopic fungi (micromycetes) is a topic that is at lower secondary schools neglected and mentioned only marginally. Fungi are of great importance for humans in their everyday life, whether negative or positive. It is the reason to rethink how and when to inform pupils about them. The aim of my thesis is to create my own practical exercises that acquaint the pupils of lower secondary schools with the appearance, properties, occurrence, uses, cultivation methods, and last, but not least with the safety of micromycets. In the practical exercises are used basic methods of work with micromycets such as inoculation of the population from the natural environment to a sterile culture medium, the sedimentation method or the method of spreading a dilute suspension of the stick on the agar medium. The revaccination is used to eliminate secondary structures. Then we deal with the possibility of preparing culture media. Isolated cultures of microscopic fungi are identified by macroscopic characters. Due to the safety with micromycets, I recommend to teach microscopic characteristics in high school. The presented practical exercises were verified at the lower secondary school in Vinařice by pupils of the 6th class. The practical exercises proved to be suitable for pupils in lower secondary school. Securing...
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Cultivation of Trematodes
Vrbová, Kristýna ; Kašný, Martin (advisor) ; Chanová, Marta (referee)
Many of Trematode species are serious pathogens of human and animals; therefore they are the subjects of intensive investigation. The successful in vitro cultivation of these organisms could represent the first step for further sophisticated experiments focused on characterization of crucial biological processes related to the particular phases of fluke life cycle or for the studies focused on testing the anthelmintic effect of various compounds. This thesis reviews majority of information published in relation to in vitro cultivation of Trematodes. It shows that for many Trematode families is our knowledge of culturing methods very limited. In contrast to the first attempts to cultivate some species of Trematodes about 100 years ago, nowadays we dispose of technologies enabling to cultivate particular live cells isolated from different tissues of flukes. Some of the cultivation media originally used for cell cultures are now commercially available and they were successfully used for cultivation of whole worms. Nevertheless, we still haven't been able to continuously cultivate a Trematode species by using an egg at the beginning and finish with fully developed adult producing eggs. However, there are some partially successful experiments in which development of particular life stages of some flukes has been...
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