Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
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ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
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Advances in Clinical and Experimental Medicine

2012, vol. 21, nr 2, March-April, p. 187–192

Publication type: original article

Language: English

ESBL-Producing Escherichia coli Isolated from Children with Acute Diarrhea – Antimicrobial Susceptibility, Adherence Patterns and Phylogenetic Background

Szczepy Escherichia coli wytwarzające β-laktamazy ESBL izolowane od dzieci z ciężką biegunką – wrażliwość na leki, typy adhezji i pochodzenie filogenetyczne

Roman Franiczek1,, Beata Sobieszczańska1,, Michał Turniak1,, Urszula Kasprzykowska1,, Barbara Krzyżanowska1,, Katarzyna Jermakow1,, Grażyna Mokracka-Latajka1,

1 Department of Microbiology, Wroclaw Medical University, Wrocław, Poland

Abstract

Background. Escherichia coli remains the principal bacterial pathogen in childhood diarrhea and constitutes an important public health problem, especially in developing countries. Diarrheagenic E. coli strains often display resistance to β-lactams due to the production of extended-spectrum β-lactamases (ESBLs).
Objectives. A total of thirty ESBL-producing E. coli strains colonizing the gastrointestinal tracts of children with acute diarrhea were studied in order to determine their antimicrobial susceptibility, adherence patterns to the HEp-2 cell line and phylogenetic background.
Material and Methods. ESBL production was detected by the double disk synergy test (DDST). The minimal inhibitory concentrations (MICs) of antibacterial drugs were determined by an agar dilution technique on MuellerHinton agar. The presence of blaTEM, blaSHV and blaCTX-M determinants in the strains studied was ascertained by polymerase chain reaction (PCR).
Results. The strains displayed the resistance pattern typical of ESBL producers. The majority of them (23 out of 30) were found to produce CTX-M-type ESBLs conferring a high level of resistance to oxyimino-β-lactams, especially to cefotaxime and ceftriaxone. In many cases, the strains exhibited resistance to non-β-lactam antimicrobials, such as gentamicin, amikacin, co-trimoxazole and tetracycline. On the other hand, these strains were uniformly susceptible to carbapenems, to oxyimino-β-lactams combined with clavulanic acid and to tigecycline. The E. coli strains were distributed among the four main phylogenetic groups: A, B1, B2 and D. The in vitro adhesion assay revealed that all but two of the strains adhered to the HEp-2 epithelial cell line. Aggregative and diffuse adherence patterns were found to be the most prevalent.
Conclusion. CTX-M-type enzymes were the most prevalent ESBLs among the strains studied. As many as 40% of the diarrheagenic E. coli isolates were found to belong to phylogenetic group D, which usually comprises E. coli strains associated with extraintestinal infections. The effectiveness of tigecycline against ESBL-producing E. coli strains was similar to that of imipenem and meropenem.

Streszczenie

Wprowadzenie. Escherichia coli pozostaje nadal głównym bakteryjnym patogenem biegunek u dzieci, stanowiąc istotny problem zdrowotny, zwłaszcza w krajach rozwijających się. Szczepy E. coli wywołujące biegunki często wykazują oporność na większość β-laktamów, wynikającą z wytwarzania β-laktamaz o rozszerzonym spektrum substratowym (ESBL).
Cel pracy. Przedmiotem badań było 30 szczepów E. coli wytwarzających ESBL, kolonizujących przewód pokarmowy dzieci z ostrą biegunką. Celem badań było oznaczenie wrażliwości na leki przeciwbakteryjne, typu adhezji do linii komórkowej HEp-2 oraz pochodzenia filogenetycznego.
Materiał i metody. Wytwarzanie ESBL wykrywano testem synergizmu dwóch krążków (DDST). Minimalne stężenia hamujące (MIC) leków przeciwbakteryjnych oznaczono metodą seryjnych rozcieńczeń w podłożu agarowym Mueller-Hintona. Występowanie genów blaTEM, blaSHV i blaCTX-M w badanych szczepach oznaczono metodą PCR.
Wyniki. Badane szczepy odznaczały się typowymi dla producentów ESBL wzorcami oporności. Większość z nich (20 spośród 30) wytwarzała enzymy ESBL typu CTX-M, warunkujące wysoki poziom oporności na oksyimino-β- -laktamy, a zwłaszcza cefotaksym i ceftriakson. W wielu przypadkach badane szczepy wykazywały oporność na inne niż β-laktamy leki przeciwbakteryjne, takie jak: gentamycyna, amikacyna, kotrimoksazol i tetracyklina. Ponadto wszystkie szczepy cechowały się wrażliwością na karbapenemy, oksyimino-β-laktamy skojarzone z kwasem klawulanowym oraz tigecyklinę. Badane szczepy E. coli należały do 4 grup filogenetycznych: A, B1, B2 i D. Przeprowadzone in vitro badania nad adhezją wykazały, że wszystkie szczepy E. coli, z wyjątkiem dwóch, adherowały do nabłonkowej linii komórkowej Hep-2. Dominującymi wzorami adhezji były adhezja typu agregacyjnego i rozproszonego.
Wnioski. Wśród badanych szczepów enzymy CTX-M stanowiły najczęstszy typ ESBL. Aż 40% biegunkowych izolatów E. coli należało do filogenetycznej grupy D zawierającej zazwyczaj szczepy E. coli związane z zakażeniami pozajelitowymi. Skuteczność tigecykliny wobec szczepów E. coli wytwarzających ESBL była zbliżona do aktywności imipenemu i meropenemu.

Key words

E. coli, ESBL, antimicrobial resistance, adherence, phylogenetic groups

Słowa kluczowe

E. coli, ESBL, oporność na leki przeciwbakteryjne, adhezja, grupy filogenetyczne

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