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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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Pathogenic trichomonads in cats and dogs
Vobořilová, Pavlína ; Tachezy, Jan (advisor) ; Hampl, Vladimír (referee)
Trichomonads are anaerobic flagellated protists that are either parasites or commensals. They frequently inhabit digestive, respiratory, and urogenital tracts of vertebrates, including domestic cats and dogs. In these hosts, four trichomonad species has been described: Tetratrichomonas canistomae and Tetratrichomonas felistomae that are commensals of the host oral cavity; Pentatrichomonas hominis, a commensal of intestinal tract that could be found in dogs and cats but also in other mammals including humans; and pathogenic Tritrichomonas foetus that causes, in addition to cattle infection, feline intestinal trichomonosis. Although, trichomonads in dogs and cats are probably of cosmopolitan distribution we have no information about their presence in Czech Republic. The first aim of this study was to distinguish types of trichomonads present in the oral cavity of dogs and cats and to get preliminary epidemiological data. The second aim was to demonstrate the presence of Tritrichomonas foetus in cats and dogs in the Czech Republic and to identify potential risk factors for the disease. Cultivation and nested PCR were used to determine the presence of trichomonads in dogs and cats. Sequencing and phylogenetic analysis based on ITS1-5.8rRNA-ITS2 gene sequence was used to identify species of isolated...
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Practical Exercises in Biology of Unicellular Organisms (Protista) for Elementary Schools with Emphasis on the Use of Staining Techniques
Šalounová, Jana ; Mourek, Jan (advisor) ; Říhová, Dagmar (referee)
4 Abstract The main goal of my thesis was to create our own proposals for laboratory work in biology of unicellular organisms (Protista). I designed the laboratory tasks, so that they can be mastered by the pupils of sixth year of primary school. Creating the proposals for laboratory exercises was preceded by the work in the laboratory at Charles University in Prague, Faculty of Education, the Department of Biology and Environmental Studies. In the laboratory, I verified the methods of cultivation of selected representatives of ciliates (Paramecium caudatum), amoebae (Amoeba proteus) and euglenids (Euglena gracilis) and selected methods of dyeing their different organels. Key words: protists, Paramecium caudatum, Amoeba proteus, Euglena gracilis, cultivation, dying
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Laboratory diagnosis of dermatophytosis
Drnková, Nela ; Vejsová, Marcela (advisor) ; Křivčíková, Lucie (referee)
Author: Nela Drnková Title of the bachelor thesis: Laboratory diagnosis of dermatophytosis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Degree course: Medical laboratory technician My bachelor thesis is focused on laboratory diagnosis of dermatophytes. The purpose of this work is created summary of laboratory methods, which are used for diagnosis of dermatophytosis. Beginning of thesis is given to common characteristics of dermatophytes, taxonomony, nomenclature and morphology of individual genera. The big part of thesis creates summary of laboratory methods. There are described individual types of microscopy, cultural techniques, molecular methods and immunological methods. The end of the thesis is dealted with infections, which are caused by dermatophytes and their treatment. Light microscopy and culture are used most frequently. Key words: dermatophytes, Trichophyton, Microsporum, Epidermophyton, laboratory diagnosis, tinea
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Analysis of adherent cells confluency in 2D culture
Bracková, Michaela ; Čmiel, Vratislav (referee) ; Chmelíková, Larisa (advisor)
First part of this semester work dealing with the theoretical description of adherens cells, particularly structure and functions of this cells. Subsequently work is devoted to description of cell culturing, with mention of conditions which are necessary for cell culturing, following by substances which promote cell growth. Last part of theoretical research is concern with microscopy technique that is suitable for studying of adherent cells. Subsequently it is about fluorescence and confocal microscopes. Practical part dealing with cell culturing of adherent cells and the evaluation of realized experiments.
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