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Production of magnetic particles by microorganisms
Chvalkovská, Eva ; Mgr. Martina Mikešová, Ph.D. (roz. Pravečková) (referee) ; Trachtová, Štěpánka (advisor)
This thesis focuses on the magnetic nanoparticles produced by bacteria. These are two types of bacteria. First of them we can find in an industrial environment that produce magnetic nanoparticles using magnetosomes. Magnetosomes are units containing nanocrystals and are surrounded by a lipid layer. They are made up of chains and work as a compass. In this thesis I deal with possibilities of cultivation od these bacteria, acquisition of magnetosomes and subsequent processing. The second type is bacteria living in the human body, they start to produce magnetic nanoparticles after the addition of trigger, such as silver nitrate. Bakteria which can produce silver or ferrous nanoparticles are Bacillus sp, Escherichia coli, Lactobacillus casei ssp. casei and Lactobacillus fermentum. Experimental part of bachelor thesis focused on the production of silver nitrate in concetration of 0,1; 1; 2; 4 mM. The bacteria that have been shown to produce silver nanoparticles are Lactobacillus casei ssp. casei CCM 7088T a Lactobacillus casei ssp. casei CCM 7089 a Lactobacillus rhamnosus CCM 1825T.
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Role of extracellular DNA in microbial biofilms
Gromov, Anton ; Chytilová, Aneta (referee) ; Obruča, Stanislav (advisor)
Abstract This bachelor’s thesis deals with defining the role of extracellular DNA in microbial biofilms. The thesis includes a part about the basic characteristics and functions of biofilms, as well as the characteristics and functions of extracellular DNA. The theoretic part additionally deals with the process of formation of biofilms, and the influence of different types of stresses on it. Also, the paper discusses the role of such stresses. The role of biofilm in bacteria and their reproduction methods are also discussed. An interesting section of the thesis outlines the impact of antibiotics on biofilms, or the ways to gain DNA or use DNase. The eDNA functions and their effects on bacterial defence will be discussed thoroughly. In the experimental part, the biofilm content was examined and compared with cells growing in a liquid and solid medium. Measurement of DNA content in media was carried out and the effects of salt and temperature stresses on biofilm were also measured. Stress conditions affected the production of PHB, which is used as a reserve form of energy and carbon. Total sugars were measured using an anthracite reagent to study the effects of stresses on the content of macromolecules. The presence of eDNA in biofilms and planktonic cells was firstly checked using gel electrophoresis and then accurate concentrations in planktonic and biofilm cells were determined using different spectrophotometric methods.
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Determination of Streptococcus pyogenes in throat swab by cultivation and imunoturbidimetry metode
SMOLOVÁ, Klára
Early identification of infections caused by bacteria Streptococcus pyogenes is very important in order to start antibiotic treatment and to prevent from so-called late consequences of streptococcal infections, including rheumatic fever and acute glomerulonefritis. Aim of my thesis is to compare two methods which are used for examination of throat swab specimen in Písek Hospital, INC. when presence of Streptococcus pyogenes is expected. The first method used by The Department of Clinical Microbiology is a cultivation, the second one is a method used by children's emergency. Main advantage of QuikRead go Strep A method is the speed of availability of the examination result and also the fact that it can be performed directly in the consulting room. However, compared to the cultivation method, QuikRead go Strep A method has its limits. It is unable to detect any other cause of the disease, it is not possible to use it subsequently to determine antibiotic sensibility and its reliability is reported as 93 %. That is why I concentrated on comparison of results of samples examined by both of the methods. In targeted period, i.e. year 2016, there were 218 samples examined by QuikRead go Strep A method, 143 of them were subsequently re-examined by the method of cultivation. A total of 62,9 % of the samples showed a consistent result with both methods. The QuikRead go Strep A method proved to be more reliable in case of a negative test result.The same result, e.i. negative presence of Streptococcus pyogenes in the sample, was measured at 94,7 % of samples, on the other hand, when a sample was detected as positive by QuikRead go Strep A method, match with cultivation method was only in 26,9 % of samples. A high error rate could have been caused by incorrect sampling for imunoturbidimetric examination. The method guide warns to the fact that if a swab touches the tongue, gums or cheeks, then a false positive result may occur. Therefore, the match between the two methods could be different if we obtained the samples of more cooperative adult patients.
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The diagnosis of pulmonary tuberculosis
CHALOUPKOVÁ, Lenka
Even in 21st century tuberculosis (TBC) pose a significant medical and social problem. Open borders intensified migrations, expansin of AIDS pandemic and related increase of patients with reduced immunity are all reasons why tubercuosis still remains one of the main causes of death all over the world. The disease originator is Mycobacterium tuberculosis, an acidoresistent bacteria that is able to survive in a human organism for a lot of years. Symptoms of the disease are not characteristic, that is why the correct diagnosis is essential to reveal the new cases of the disease. One of the standards to stay is bacteriological examination based on microscopic examination and cultivation of the material. Also rapid molecular genetic techniques and methods based on the immune response are availabe. This work uses data gaied from Plicní léčebna Nemocnice České Budějovice, a.s. (Pulmonary Department of Hospital in České Budějovice) for the period of 2008-2016, as well as data gained from Pracoviště imunologie (Immunolgy Department) in Nemocnice České Budějovice, a.s. The goal of the work was to create a statistical overview of patients with confirmed pulmonary tuberculosis according to age and gender for the period of 2008-2016 . Next it aims at comparison of tuberculin skin test and quantiferon test in patients with pulmonary tuberculosis for the period of 2008 - 2016. Implementation and evaluation of the quantiferon test in patients in 2016 are the end of my work. The methodology was mainly based on gathering the necessary data and their evaluation. Results show that pulmonary tuberculosis was confirmed in 81 patients in relation to data obtained from the Pulmonary Department. Of that 75% were male and only 25% female. For both sexes, the occurrence of tuberculosis was on record in all age categories. In men most often at the age of 31-50 and 51-70. Tuberculin skin test (PPD) and quantiferon (QF) test were also accomplished in 62% of patients. Only in 7% patients none of the tests was done. When the two tests were compared, both PPD and QF tests were positive in 64%. Also the results show that the quantiferon test is more sensitive than the tuberculin skin test. The quantiferon test was evaluated separately for the positive and negative results, positive result was compared with bacteriological examination. Next the QF test was evaluated from the point of view of gender, age, diagnosis and engaged department. A relatively signifcant finding was that even though the QF test is more sensitive, for more than a third of patients with the positive QF test result, mycobacterial identification was not required at all. Compared to bacteriology examination TB was cofirmed in 14% of patients. Despite the fact that the Czech Republic ranks among the countries with a low incidence of tuberculosis, this fact can not be underestimated. The public's feeling that there is no need to worry about this illness has also been supported by the approval of the cancellation of flat vaccinations, effective from 1. 11. 2010. Also, a number of physicians are no longer taking this disease into account while making diagnosis. However, education of lay public is essential above all in this issue.
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Vliv působení trávicího procesu zavíječe voskového (Galleria mellonella) na spory původce moru včelího plodu (Paenibacillus larvae).
MRÁZ, Petr
This diploma thesis deals with a serious honey bee (Apis mellifera) disease, the American foulbrood (AFB), and with possibilities of its control. The thesis is divided into two parts, the theoretical and the experimental. The theoretical part is written as a research and contains two big chapters. The first one describes American foulbrood disease and its causative agent, bacteria Paenibacillus larvae. The second part deals with the wax moth (Galleria mellonella). The experimental part then combines above mentioned themes in an effort to find the relationship between the wax moth caterpillars and bacteria P. larvae that could possibly help to control the AFB. The aim of this work was to determine whether the wax moth can disrupt resistant layers of P. larvae spores thanks to its well adapted digestive tract and whether it could change their germination.
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Interaction of the pathogenic yeasts of the genus Candida with the bacterium Pseudomonas aeruginosa
Mynářová, Lenka ; Hodek, Petr (advisor) ; Šťovíček, Vratislav (referee)
Pathogenic yeasts of the genus Candida and the bacterium Pseudomonas aeruginosa are opportunistic pathogens and they often occur in the human micoflora as harmless commensals. They do not mean a threat for healthy humans but they can represent a true hazard for a person with supressed immunity. Cystic fibrosis (CF) is an incurable genetic disease and among others, it causes supression of immunity. Most of the patients with CF suffer from the chronical lung infections caused by the bacterium Pseudomonas aeruginosa. The pathogenic yeasts can coexist with the bacteria both in lungs of the CF patients, and on various surfaces, e.g. catheters used in hospitals. Therefore it is important to explore the mode of their interaction in the host body, as well as in vitro. Both Candida and Pseudomonas secrete a range of proteins that act as virulence factors, and also small molecules, which mediate the interaction and communication between microorganisms. We therefore tested cocultivation of two Candida species (C. albicans and C. parapsilosis) with P. aeruginosa in three different chemically defined liquid media. We monitored viability and survival of the individual species and analyzed the proteins secreted to the media. We found that P. aeruginosa secreted several hydrolytic enzymes, for example aminopeptidase,...
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Interaction of Borrelia sp. with HL-60 cells and monocytes and cultivation of Anaplasma phagocytophilum in HL-60 cell culture
Marková, Lucie ; Hulínská, Dagmar (advisor) ; Beranová, Jana (referee)
Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum are causative agents of Lyme disease and human granulocytic anaplasmosis. Their common vector in Europe are the ticks from the genus Ixodes. In our work, we focused on interaction of innate immune cells with the causative agent of Lyme diseases, that are insubstitutable in their function in the early phase of the disease. Anaplasma phagocytophilum is hard to cultivate, the only possibility is to cultivate it in cell cultures. Successful cultivation of Anaplasma phagocytophilum acquired from patients in our geographic area is crucial for following experiments and for diagnostics too. In our experiments, we used validated cell cultures of HL-60 cells, canine monocytes DH82 and murine monocytes P388D1. During our studies of interaction of the causative agent of Lyme diseases with cells, we used two strains of different species Borrelia. Borrelia garinii M192 and Borrelia burgdorferi sensu stricto B31. These strains vary in virulence. The strain M192 is virulent, but the strain B31 lost its virulence by passages. We specialised in study of morphological changes using light microscopy (observation of dyed and fixed preparates and observation in dark field), eventually by transmision electron microscopy. During our experiments, we concluded that HL-60...
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