magistrsko delo
Polona Lindič (Author), Mateja Pirš (Reviewer), Jerneja Ambrožič (Mentor), Andrej Golle (Co-mentor)

Abstract

Proti antibiotikom odporne bakterije predstavljajo enega izmed glavnih zdravstvenih problemov 21. stoletja, še posebej bakterije z geni za encime ESBL in karbapenemaze. K razvoju in širjenju odpornosti proti antibiotikom vplivajo tako človeški, živalski kot okoljski rezervoarji. V Sloveniji je malo znanega o prisotnosti bakterij z zmanjšano občutljivostjo za antibiotike v izvenbolnišničnem okolju, še predvsem na odlagališčih. Namen naloge je bila osamitev bakterij, ki so fenotipsko odporne proti betalaktamom z razširjenim spektrom delovanja in karbapenemom, iz površinskih in podzemnih vod, povezanih z izbranim odlagališčem odpadkov ter genotipska opredelitev fenotipske odpornosti, ki temelji na izločanju encimov za betalaktamaze z razširjenim spektrom delovanja (ESBL) in karbapenemaz. Bakterije iz vzorcev podzemnih in površinskih vod smo osamili z uporabo 3 izbranih selektivno diferencialnih gojišč in jih identificirali z metodo MALDI TOF. Encime ESBL in karbapenemaze smo najprej potrdili s fenotipskimi testi (metoda kombiniranih diskov za potrditev ESBL in CIM testi), nato s tržno dostopnimi kiti določili njihove gene. Ugotovili smo, da so izolati iz površinskih vod odporni proti več antibiotikom, kot izolati iz podzemnih vod. S pomočjo molekularnih testov smo določili gene za odpornost proti betalaktamom pri dveh izolatih, ki smo ju izolirali iz površinskih vod. Pri prvem izolatu Klebsiella oxytoca (ZZV20-11045), izoliranem iz površinskih padavinskih vod, smo potrdili gen za encim CTXM-1 in gen za encim OXA-48. Pri drugem izolatu Pseudomonas aeruginosa (ZZV20-11064), ki smo ga izolirali iz površinskih izcednih vod pa smo potrdili prisotnost gena za encim iz skupine VIM. Iz površinskih voda smo osamili več proti betalaktamom in karbapenemom odpornih bakterij kot v podzemnih vodah. Pri obeh odpornih mikroorganizmih smo potrdili prisotnost genov za encime iz skupine ESBL in karbapenemaze (dokazali smo gene blaVIM, blaCTXM-1 and blaOXA-48).

Keywords

mikrobiologija vode;površinske vode;podzemne vode;odlagališča odpadkov;odpornost proti antibiotikom;encimi;betalaktamaze;ESBL;karbapenemaze;enterobakterije;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [P. Lindič]
UDC: 579.68:628.472.3:615.015.8
COBISS: 112827139 Link will open in a new window
Views: 73
Downloads: 24
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Other data

Secondary language: English
Secondary title: Bacteria harbouring ESBL and carbapenemase genes in waters associated with a single landfill
Secondary abstract: Antibiotic resistance is one of the major health problems of the 21st century. The development and spread of antibiotic resistance is influenced by human, animal and environmental reservoirs. Little is known in Slovenia about the presence of bacteria with reduced sensitivity to antibiotics in the non-hospital environment. Among the major resistant bacteria that pose a threat to human health are bacteria with genes for the enzymes ESBL and carbapenemase. The aim of the study was to isolate bacteria that are phenotypically resistant to betalactams with extended spectrum and carbapenem from surface and groundwater associated with the selected landfill and genotypic definition of phenotypic resistance based on the secretion of enzymes for betalactamase (ESBL) and carbapenemases. Bacteria from groundwater and surface water samples were isolated by three specific selective-differential media and identified using MALDI-TOF. ESBL enzymes and carbapenemases were first confirmed by phenotypic tests (combined disk method to confirm ESBL and CIM tests), then their genes were determined by commercially available kits. Surface water isolates were found to be resistant to more antibiotics than groundwater isolates. Molecular tests confirmed the presence of resistance genes in two isolates isolated from surface waters. In the case of Klebsiella oxytoca isolate (ZZV20-11045) isolated from surface rainwater, the gene for the enzyme CTXM-1 and the gene for the enzyme OXA-48 were confirmed. In the isolate of Pseudomonas aeruginosa (ZZV20-11064), which was isolated from surface leachate, the presence of the gene for the enzyme from the VIM group was confirmed. More betalactam and carbapenem resistant bacteria were isolated from surface waters than in groundwater. In both resistant microorganisms, the presence of genes for ESBL and carbapenemase enzymes were confirmed (genes blaVIM, blaCTXM-1 and blaOXA-48 were detected).
Secondary keywords: water microbiology;surface water;groundwater;landfills;antibiotic resistance;enzymes;betalactamases;ESBL;carbapenemases;enterobacteria;
Type (COBISS): Master's thesis/paper
Study programme: 0
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Oddelek za mikrobiologijo
Pages: XII, 64 f., [4] f. pril.
ID: 15720974