Detection of antibiotic resistance genes in wastewater treatment plant : molecular and classical approach

• Autorzy: Ziembińska-Buczyńska A. , Felis E. , Folkert J. , Meresta A. , Stawicka D. , Gnida A. , Surmacz-Górska J.

Identyfikatory

DOI

10.1515/aep-2015-0035

Warianty tytułu

PL

Wykrywanie genów oporności na antybiotyki w oczyszczalni ścieków : podejście klasyczne i biologii molekularnej

• Języki publikacji: EN

Abstrakty

EN

Antibiotics are a group of substances potentially harmful to the environment. They can play a role in bacterial resistance transfer among pathogenic and non-pathogenic bacteria. In this experiment three representatives of medically important chemotherapeutics, confirmed to be present in high concentrations in wastewater treatment plants with HPLC analysis were used: erythromycin, sulfamethoxazole and trimethoprim. Erythromycin concentration in activated sludge was not higher than 20 ng L⁻¹. N-acetylo-sulfamethoxazole concentration was 3349 ± 719 in winter and 2933 ± 429 ng L⁻¹ in summer. Trimethoprim was present in wastewater at concentrations 400 ± 22 and 364 ± 60 ng L⁻¹, respectively in winter and summer. Due to a wide variety of PCR-detectable resistance mechanisms towards these substances, the most common found in literature was chosen. For erythromycin: erm and mef genes, for sulfamethoxazole: sul1, sul2, sul3 genes, in the case of trimethoprim resistance dhfrA1 and dhfr14 were used in this study. The presence of resistance genes were analyzed in pure strains isolated from activated sludge and in the activated sludge sample itself. The research revealed that the value of minimal inhibitory concentration (MIC) did not correspond with the expected presence of more than one resistance mechanisms. Most of the isolates possessed only one of the genes responsible for a particular chemotherapeutic resistance. It was confirmed that it is possible to monitor the presence of resistance genes directly in activated sludge using PCR. Due to the limited isolates number used in the experiment these results should be regarded as preliminary.

PL

Antybiotyki to grupa związków potencjalnie szkodliwych dla środowiska. Odgrywają one rolę w procesach transferu antybiotykooporności pomiędzy patogenami i bakteriami niechorobotwórczymi. Wykorzystując metodę wysokosprawnej chromatografii cieczowej (HPLC) wykazano obecność erytromycyny, sulfametoksazolu i trimetoprimu w miejskiej oczyszczalni ścieków w następujących stężeniach: dla erytromycyny < 20 ng L⁻¹, N-acetylo-sulfametoksazolu 3349 ± 719 i 2933 ± 429 ng L−1, a trimetoprimu 400 ± 22 i 364 ± 60 ng L⁻¹, odpowiednio: zimą i latem. Ponieważ antybiotykooporność bakteryjna może być stymulowana obecnością antybiotyków w środowisku, istnieje możliwość pojawienia się wielu szlaków opornościowych u bakterii narażonych na działanie tych związków. Dlatego też podjęto próbę detekcji wybranych genów oporności na badane chemioterapeutyki metodą łańcuchowej reakcji polimerazy (PCR). Obecność elementów genetycznych badano zarówno w szczepach bakteryjnych, u których udowodniono oporność na badany związek bakteriobójczy, jak i w próbce osadu czynnego, z którego te bakterie izolowano. Do badań wybrano najczęściej występujące geny oporności: dla erytromycyny erm i mef, dla sulfametoksazolu: sul1, sul2, sul3, a dla trimetoprimu dhfrA1 i dhfr14. Wykazano, że wartość minimalnego stężenia inhibitującego (MIC), nie koresponduje z obecnością większej liczby mechanizmów oporności. Większość szczepów opornych wykazywała tylko jeden z badanych mechanizmów oporności na antybiotyk niezależnie od wartości MIC. Potwierdzono również możliwość monitorowania obecności genów oporności na antybiotyki metodą PCR bezpośrednio w osadzie czynnym. Ze względu na ograniczona liczbę izolatów użytych w tym eksperymencie wyniki uzyskane w pracy powinny być traktowane jako wstęp do dalszych badań.

Słowa kluczowe

EN

DGGE; denaturing gradient gel electrophoresi; PCR; polymerase chain reaction; sulfamethoxazole; trimethoprim resistance; erythromycin resistance; WWTP; wastewater treatment plant

PL

antybiotyki; PCR; reakcja łańcuchowa polimerazy; sulfametoksazol; geny oporności; odporność na trimetoprim; odporność na erytromycynę; oczyszczalnia ścieków

• Wydawca: Institute of Environmental Engineering, Polish Academy of Sciences
• Czasopismo: Archives of Environmental Protection
• Rocznik: 2015
• Tom: Vol. 41, no. 4
• Strony: 23--32
• Opis fizyczny: Bibliogr. 32 poz., rys., tab., wykr.

Twórcy

autor

Ziembińska-Buczyńska A.

• The Silesian University of Technology, Poland The Faculty of Environmental Engineering and Energy Environmental Biotechnology Department

autor

Felis E.

• The Silesian University of Technology, Poland The Faculty of Environmental Engineering and Energy Environmental Biotechnology Department

autor

Folkert J.

• The Silesian University of Technology, Poland The Faculty of Environmental Engineering and Energy Environmental Biotechnology Department

autor

Meresta A.

• The Silesian University of Technology, Poland The Faculty of Environmental Engineering and Energy Environmental Biotechnology Department

autor

Stawicka D.

• The Silesian University of Technology, Poland The Faculty of Environmental Engineering and Energy Environmental Biotechnology Department

autor

Gnida A.

• The Silesian University of Technology, Poland The Faculty of Environmental Engineering and Energy Environmental Biotechnology Department

autor

Surmacz-Górska J.

• The Silesian University of Technology, Poland The Faculty of Environmental Engineering and Energy Environmental Biotechnology Department

Bibliografia

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