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Genotipska opredelitev ESBL in AmpC pozitivnih sevov bakterij vrste Escherichia coli, izoliranih iz perutnine v izbranih rejah
ID Šabić, Edita (Author), ID Ambrožič Avguštin, Jerneja (Mentor) More about this mentor... This link opens in a new window, ID Zorman Rojs, Olga (Co-mentor)

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Abstract
Escherichia coli je komenzalna bakterija v prebavnem traktu toplokrvnih živali, ki lahko s pridobitvijo genskih zapisov za različne dejavnike virulence kolonizira tudi druga tkiva in organske sisteme ter povzoči raznolika bolezenska stanja. Vse več patogenih sevov E. coli ima poleg dejavnikov virulence tudi gene, ki jim omogočajo odpornost proti protimikrobnim učinkovinam. Še posebno problematične so ß-laktamaze iz skupin ESBL, AmpC in karbapenemaze. Cilj magistrske naloge je bil ugotoviti prisotnost, porazdelitev in dinamiko širjenja genskih zapisov za encime iz skupine ESBL in AmpC v izbranih perutninskih rejah. Izolate iz zbirke fenotipsko opredeljenih ESBL- in AmpC-pozitivnih sevov E. coli, vzorčenih v perutninskih rejah v Sloveniji, smo uvrstili v klonalne in filogenetske skupine in pri vsakemu preverili prisotnost 20 genov za dejavnike virulence. Za 64 izolatov, ki smo jih uvrstili v 39 klonalnih skupin, smo genotipsko potrdili prisotnost zapisov za encime iz skupine ESBL in AmpC. Največ (40,3 %) izolatov je imelo alel blaSHV-12, za 29,2 % izolatov pa smo potrdili prisotnost blaCMY-2. Poleg tega smo v 28 izolatih potrdili prisotnost plazmidno zapisanega gena qnrS, ki je povezan z nizko ravnjo odpornosti proti fluorokinolonom. Večina qnrS-pozitivnih izolatov je hkrati imelo tudi alel blaSHV-12, zato smo predvidevali, da je plazmidno zapisan in se širi predvsem s horizontalnim genskim prenosom. S konjugacijo smo ugotovili relativno nizko (10-4-10-8) frekvenco prenosa konjugativnih plazmidov. Iz transkonjugant smo izolirali pDNA, jo razrezali in med seboj primerjali plazmidne restrikcijske profile. Ugotovili smo, da se plazmid s qnrS in blaSHV-12 širi v rejah predvsem horizontalno, s konjugacijo. Geni blaCTX-M se znotraj rej širijo predvsem vertikalno.

Language:Slovenian
Keywords:Escherichia coli, ESBL, AmpC, genotipizacija, konjugacija, perutnina
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Publisher:[E. Šabić ]
Year:2019
PID:20.500.12556/RUL-111078 This link opens in a new window
UDC:579.25:579.842.1/.2:577.2
COBISS.SI-ID:5100920 This link opens in a new window
Publication date in RUL:22.09.2019
Views:1894
Downloads:407
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Secondary language

Language:English
Title:Genotyping of ESBL and AmpC positive Escherichia coli isolates from selected poultry flocks
Abstract:
Escherichia coli is a commensal bacteria found in the gastrointestinal tract of warm-blooded animals. By aquisition of various virulence factor genes it can colonise other tissues and organs, thus becoming pathogenic. Additionally, the number of antimicrobial resistant strains is steadily increasing in the last two decades. The most concerning β-lactamases are ESBLs, AmpC and carbapenemases. The aim of our study was to determine the presence, distribution and the spread of ESBL and/or AmpC-producing E. coli in and among animals from selected poultry farms. Strains phenotypically positive for ESBL and AmpC enzymes were isolated from different poultry farms in Slovenia, have been grouped into 39 different clonal groups based on ERIC-PCR. Sixty-four isolates that were confirmed as ESBL- and AmpC-positive by molecular approaches, were further assigned to phylogenetic groups and screened for presence of different resistance and 20 virulence genes. The majority of the isolates (40,3 %) carried the blaSHV-12 allele and only 29,2 % the blaCMY-2 allele. Twenty-eight were positive for presence of qnrS gene. Twenty-six of them also carried the blaSHV-12 allele. Several conjugation experiments revealed a different low conjugation frequencies (10-4-10-8), depending on the donor strain and the selected resistotype. Restriction profile analysis of plasmid DNA from transconjugants confirmed the presence and more frequently vertical spread of different plasmids encoding AmpC- and CTX-M- β-lactamase genes and the predominantly horizontal spread of 2, almost identical, plasmids encoding blaSHV-12 and qnrS genes in and between poultry farm included in the survey.

Keywords:Escherichia coli, ESBL, AmpC, genotyping, conjugation, poultry

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