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Study of the occurrence of colistin in soil
Veldamonová, Aneta ; Zlámalová Gargošová, Helena (referee) ; Mravcová, Ludmila (advisor)
Colistin is a polypeptide antibiotic used as a "last resort" effective against Gram-negative bacteria. However, increase in its consumption in veterinary medicine in the last 30 years has led to the development of bacterial resistance even to colistin. The application of slurry to soil containing unmetabolized colistin and resistant bacteria poses a risk, because the resistance of bacteria to colistin can be further developed and spread to other components of the environment. Therefore, this work was focused on the creation and optimization of extraction and analytical methods for soil samples containing colistin. Colistin was detected by liquid chromatography in connection with mass spectrometry (HPLC/MS and UPLC/MS/MS). Many extraction solutions have been tested in connection with ultrasonic extraction and purification on various SPE columns, yet colistin has not been successfully extracted from the soil. Colistin was successfully detected only in the slurry extract. The reason why colistin was not detected in soil extracts could be the sorption of colistin on soil organic matter or the formation of complexes of colistin with humic substances.
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Significance, occurrence and determinants of horizontally transmissible colistin resistance in Gram negative bacteria
Kislíková, Karolína ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
Colistin, also known as polymyxin E, is antibiotics active against most of Gram-negative bacteria. In the pas decade, emergency of multidrug-resistant bacteria led to increase of colistin administration as a last resort antibiotic for human infections. The first plasmid-mediated colistin resistance gene mcr-1 was identified in 2015 in animals in China and after first detection, additional mcr genes: mcr-2, mcr-3, mcr-4, mcr-5, mcr-6, mcr-7 a mcr-8 were described throughout the world. The aim of this thesis was to clarify whether there is horizontal transmission colistin resistance encoded by the mcr genes in gram-negative bacteria isolated from the environment, animals and their breeding and food. The mcr-1 gene was detected in 2 strains Escherichia coli isolated from waste water. The mcr-4 gene was detected in 1 strain Shewanella putrefaciens isolate obtained from the lake. The environment is the most important source and way of spreading this type of resistance in the Czech Republic.
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Study of the occurrence of colistin in soil
Veldamonová, Aneta ; Zlámalová Gargošová, Helena (referee) ; Mravcová, Ludmila (advisor)
Colistin is a polypeptide antibiotic used as a "last resort" effective against Gram-negative bacteria. However, increase in its consumption in veterinary medicine in the last 30 years has led to the development of bacterial resistance even to colistin. The application of slurry to soil containing unmetabolized colistin and resistant bacteria poses a risk, because the resistance of bacteria to colistin can be further developed and spread to other components of the environment. Therefore, this work was focused on the creation and optimization of extraction and analytical methods for soil samples containing colistin. Colistin was detected by liquid chromatography in connection with mass spectrometry (HPLC/MS and UPLC/MS/MS). Many extraction solutions have been tested in connection with ultrasonic extraction and purification on various SPE columns, yet colistin has not been successfully extracted from the soil. Colistin was successfully detected only in the slurry extract. The reason why colistin was not detected in soil extracts could be the sorption of colistin on soil organic matter or the formation of complexes of colistin with humic substances.
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Colistin resistance in clinically important Enterobacteriaceae
Smělíková, Eva ; Tkadlec, Jan (advisor) ; Ježek, Petr (referee)
Colistin is a last-resort antibiotic used to treat serious infections caused by Enterobacteriaceae and other multidrug resistant gram-negative bacteria. Recently discovered plasmid-borne colistin resistance, mediated by the mcr genes, poses a serious risk to colistin therapy. The aim of this diploma thesis was to map the occurrence of Enterobacteriaceae carrying the mcr-1 to 8 genes in hospitalized patients, travellers, prospective colistin-resistant clinical isolates and in a retrospective collection of Enterobacteriaceae using a combination of selective cultivation and qPCR. Isolates with a detected mcr gene were characterized by Whole-Genome Sequencing. The localization of mcr genes was determined and other resistance genes and plasmids were identified. Furthermore, the physiological profile of selected colistin- resistant Escherichia coli isolates was characterized. In the presence of a subinhibitory amount of colistin, a strain carrying the mcr-1 gene may be favored. Later, the mcr-9 gene was described and its occurence was subsequently tested retrospectively. Enterobacter spp. isolates carrying the mcr-9 gene were mostly colistin-sensitive but, in some cases, resistance was induced after exposure to sublethal doses of colistin. The results of the study show that the incidence of plasmid-mediated...
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Significance, occurrence and determinants of horizontally transmissible colistin resistance in Gram negative bacteria
Kislíková, Karolína ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
Colistin, also known as polymyxin E, is antibiotics active against most of Gram-negative bacteria. In the pas decade, emergency of multidrug-resistant bacteria led to increase of colistin administration as a last resort antibiotic for human infections. The first plasmid-mediated colistin resistance gene mcr-1 was identified in 2015 in animals in China and after first detection, additional mcr genes: mcr-2, mcr-3, mcr-4, mcr-5, mcr-6, mcr-7 a mcr-8 were described throughout the world. The aim of this thesis was to clarify whether there is horizontal transmission colistin resistance encoded by the mcr genes in gram-negative bacteria isolated from the environment, animals and their breeding and food. The mcr-1 gene was detected in 2 strains Escherichia coli isolated from waste water. The mcr-4 gene was detected in 1 strain Shewanella putrefaciens isolate obtained from the lake. The environment is the most important source and way of spreading this type of resistance in the Czech Republic.
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