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Application of separation methods for the determination of selected pharmaceuticals in waters
Burešová, Jitka ; Opatřilová, Radka (referee) ; Vávrová, Milada (advisor)
Antibiotics are widely used pharmaceuticals in human and veterinary medicine. These compounds are biologically active. They decrease efficiency of biological processes in wastewater treatment plants. Antibiotics are not eliminated from sewage water completely and they are discharged as contaminants into the receiving waters. Several methods exist for the determination of antibiotics in sewage water. In the first place liquid chromatography, gas chromatography and capillary electrophoresis are used. The aim of this thesis was to developed a suitable izolation technique and an optimal analytical method for identification and determination of penicillins in wastewater. For determination was selected amoxicillin, ampicillin and benzylpenicillin (penicillin G). These are very often used penicillins. An optimized method was used for determination of these penicilins in real samples from a wastewater treatment plant situated in Veterinary and pharmaceutical university in Brno and from the large-scale wastewater treatment plant in Brno-Modřice. Real samples were concentrated using SPE (solid phase extraction). For penicillins determination were used high performance liquid chromatography with diode array detection (HPLC/DAD).
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The Use of Liquid Chromatography for Determination of Drug Residues
Dvořáková, Petra ; Hajšlová, Jana (referee) ; Chýlková, Jaromíra (referee) ; Sokol, Jozef (referee) ; Vávrová, Milada (advisor)
This work is based on the occurrence of drug residues in the environment. This study is focused on the development and optimization methods for determination of selected drugs in the surface water, aquatic sediment and sewage sludge from waste water treatment plant. From the group of drugs were chosen antibiotics. Antibiotics presented in the environment can cause adverse effects including toxic effects, immunity disorders and indirect bioalteration effects. Sulfonamide antibiotics, which are used in the treatment of urinary and respiratory tract infections as well as in the treatment of other infectious diseases, were chosen as a target compounds. Three optimized analytical methods for determination of sulfonamide antibiotics were developed. For the optimization of extraction were tested: solid phase extraction, pressurized solvent extraction, microwave extraction and ultrasonic extraction. For the final analysis was used liquid chromatography with two detectors - diode array detector and mass spectrometer. These optimized methods were applied for the analysis of real samples. The surface water and sediment samples were collected from two Moravian rivers (the Svratka river and the Svitava river). Samples of sewage sludge were collected from waste water treatment plant Brno-Modřice. Fish samples from the Svratka river were also collected. It was observed that all the selected sulfonamide antibiotics are present in real sediment samples (ug.kg-1). Simultaneously presence of some target analytes in real surface water (ug.l-1) and in sewage sludge (ug.kg-1) samples has been confirmed. In samples of surface water from the Svitava river and in fish samples sulfonamide antibiotics were not detected or their concentrations were below the limit of detection.
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Determination of antibiotics in the soil ecosystem
Brož, Tomáš ; Zlámalová Gargošová, Helena (referee) ; Mravcová, Ludmila (advisor)
This work deals with a current issue of increasing drug concentrations and thus the growing number of resistant bacteria in the soil. One of the problem helping this issue is the fertilization of agricultural fields with animal faeces, in which antibiotics occur in an unchanged and still active form, as well as the insufficient effectiveness of conventional methods used in wastewater treatment plants. In an effort to improve the monitoring of this problem, a method for the determination of these antibacterial agents in the soil matrix, using solid phase extraction, and also a UPLC/MS method for their evaluation have been developed and optimized. From the group of sulfonamide antibiotics, sulfacetamide, sulfadiazole, sulfamerazine, sulfamethazine, sulfamethoxazole, sulfapyridine, sulfasalazine, sulfathiaosal and trimethoprim were selected. The lowest limit of detection for the method is 0.3 µg/g for sulfathiazole and the highest of 8.1 µg/g for sulfasalazine. The highest average yields were achieved with sulfamethoxazole, namely 63 ±8 %.
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Study of the medium influence on the pollutants sorption on microplastics
Hamplová, Marie ; Kučerík, Jiří (referee) ; Mravcová, Ludmila (advisor)
With the increasing production of plastic waste, the pollution of the environment by persistent microplastics is increasing simultaneously. Other environmental pollutants, such as various pharmaceuticals, can be sorbed on microplastics. This increases the toxicity of microplastics and their risk to living organisms. Nowadays, the use of so-called biodegradable polymers, which are supposed to degrade spontaneously, is expanding. However, their degradation can also produce microplastics. Therefore, this bachelor thesis deals with the sorption of drugs onto microplastics from biodegradable polyhydroxybutyrate (PHB) and with the influence of the surrounding aqueous environment on the rate and mechanism of sorption. Sorption of drugs onto microplastics made of synthetic polyethylene terephthalate (PET) was also carried out for comparison. The quantity of drugs was analysed by UPLC-ESI-TQ. Sorption was observed from the drug mixture for antibiotics tetracyclines. The pH value of the surrounding environment influenced the sorption rate of tetracyclines, the highest sorption occurred at acidic pH 4. According to the results of evaluation of sorption by Langmuir and Freundlich adsorption isotherms, PHB microplastics have a higher sorption capacity than PET microplastics. Therefore, PHB microplastics could be a risk for living organisms.
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Evaluation of antibiotic-induced mitochondrial superoxide production in adherent cells
Ingrová, Kateřina ; Chmelíková, Larisa (referee) ; Zumberg, Inna (advisor)
The theoretical part of this bachelor´s thesis contains a description of the effect of reactive oxygen species on oxidative stress by mitochondria and the consequences of antibiotics use in cell line culturing. The cell line studied in this bachelor´s thesis is the mesenchymal stem cells (MSCs). The practical part describes the procedure of the experiment including cell culturing, passaging and cell labeling. The proposed experiment was repeated with sufficient number of repetitions. Finally, confocal microscopy images were processed in the MATLAB programming environment.
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Determination of selected drugs in water
Tran Xuan, Tiep ; Tulková, Tereza (referee) ; Vávrová, Milada (advisor)
Submitted thesis tackles the problem of numerous of antibiotics in waste water. Specifically, trimethroprim azithromycin and clarithomycin. In all of those cases we are dealing with a massive ecological threat at hand, causing harm to the enviroment. To isolation the specific kind of antibiotics the method of SPE was chosen by prior consultation with the supervisor. The method of high performance liquid chromatography with mass spectrometry detection was deemed superrior for the pinpointing of the exact pharmaceuticals in waste watters. The waste waters in and out flow was observed for twelve days with the maintainance of all necessary procedures listed above at water treatment plants Brno-Modřice.
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Antimikrobiální rezistence a její šíření v potravinovém řetězci
Maršová, Nikol
The bachelor thesis deals with the resistance of microorganisms to antimicrobial agents and its subsequent spread in the food chain. It focuses mainly on the resistance of microorganisms to antibiotics, in relation to the types of antibiotics according to their chemical structure and their mechanism of action. It describes the basic principles of agents used against viruses, yeasts, fungi and parasites. Characterises the nature of the emergence and mechanisms of resistance. It also discusses the methods used to determine the resistance of microorganisms to antimicrobial agents. Describes when resistance of microorganisms began to emerge historically and its occurrence in different sections of the food chain. In relation to the spread of antimicrobial resistance in the food chain, it describes the most serious zoonotic agents today.
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Výskyt antimikrobiální rezistence u streptokoků (převážně beta-hemolytických) z ran pacientů
Kuropata, Daniel
Recently, a frequent occurrence of antibiotic resistance has been observed, and is still increasing. Therefore, it is important to constantly monitor this resistance, which was the aim of this work for Streptococcus genera. The literature review is focused on the selected species of streptococci (S. agalactiae, S. dysga-lactiae subsp. equisimilis, S. pyogenes, S. mitis and S. oralis), antibiotics, including their mechanism of action, and antibiotic resistance with an emphasis on its emergence, spread, mechanisms, determining genes and the risks it presents. In the experimental part, the occurrence of resistance in streptococci (especially beta-haemolytic) isolated from patients’ wounds at the Trauma Hospital in Brno was monitored. For this purpose, determination of the minimum inhibitory concentration (MIC), analysis of the resistance phenotype to macrolides, lincosamides and streptogramin B (MLS) using the D-test and resistance gene screening by polymerase chain reaction (PCR) were used. Among the 31 clinical isolates the prevalence of resistance to cotrimoxazole (96.8%), ciprofloxacin (71%), tetracycline (29%), erythromycin (25.5%), clidamycin (16 .1%), ampicillin (3.2%) and oxacillin (3.2%) was found by using MIC. Resistance to chloramphenicol, gentamicin, vancomycin, teicoplanin, linezolid, tigecycline and nitrofurantoin has not been detected. The D-test was performed with erythromycin-resistant isolates. The constitutive (cMLS) phenotype was detected in 62.5% of erythromycin-resistant isolates, inducible (iMLS) in 25% and M phenotype in 12.5%. The PCR method confirmed the presence of selected resistance genes, namely ermA (16.1%), ermB (6.5%), mefA (3.2%), tetO (3.2%), tetM (25.8%) and intTn (32.3%). This study demonstrated that all streptococcal isolates from patients’ wounds are resistant to some of the tested antibiotics. Moreover, multiresistance was detected in some isolates.
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Vláknité mikromycety (plísně) a jejich biotechnologické využití
Beniačová, Michaela
The bachelor thesis focuses on filamentous micromycetes and their biotechnological utilization. This paper describes the nutrition, morphology, sexual and asexual reproduction of filamentous micromycetes and the optimal conditions for their growth and habitat. Following sections also contain the taxonomic distribution and characteristics of important species along with their short morphological description. The second part of the thesis categorizes and describes biotechnologies, following usage of filamentous micromycetes in the food, agricultural and pharmaceutical industries. Furthermore, this part touches utilization for environmental protection and use of industrially made substances produced by the metabolism of fungi.
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Antibiotická rezistence patogenních bakterií u vybraných druhů zájmových zvířat
Kaliská, Tereza
Some strains of pathogenic bacteria are capable of resistance to antibacterial agents. This resistance makes it difficult to treat diseases in both human and veterinary medicine. The most well-known pathogens of domestic animals, mainly dogs and cats, are mentioned in the context of the bachelor thesis. The thesis highlights the importance of understanding the mechanisms of resistance. It describes mobile elements, the principle of efflux pumps and spontaneous chromosomal mutations. Prevention should be the basis for preventing resistance from spreading further in the population. The observance of good hygiene habits, the appropriate use of antibiotics in the correct dosage and legislative adjustments on agricultural farms should be implemented on a global scale. The development of antimicrobial substances, including chemotherapeutics, should continue. At the same time, more empha-sis should be placed on the search for new alternatives. Mainly plant phytobiotics look promising.
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