The Transferable Resistome of Produce

ABSTRACT Produce is increasingly recognized as a reservoir of human pathogens and transferable antibiotic resistance genes. This study aimed to explore methods to characterize the transferable resistome of bacteria associated with produce. Mixed salad, arugula, and cilantro purchased from supermarke...

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Autores principales:	Khald Blau, Antje Bettermann, Sven Jechalke, Eva Fornefeld, Yann Vanrobaeys, Thibault Stalder, Eva M. Top, Kornelia Smalla
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	Escherichia coli IncF IncI antibiotic resistance horizontal gene transfer real-time PCR Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/307f9dd6446b4cfbaaa5dc095712805d
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spelling	oai:doaj.org-article:307f9dd6446b4cfbaaa5dc095712805d2021-11-15T15:52:18ZThe Transferable Resistome of Produce10.1128/mBio.01300-182150-7511https://doaj.org/article/307f9dd6446b4cfbaaa5dc095712805d2018-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01300-18https://doaj.org/toc/2150-7511ABSTRACT Produce is increasingly recognized as a reservoir of human pathogens and transferable antibiotic resistance genes. This study aimed to explore methods to characterize the transferable resistome of bacteria associated with produce. Mixed salad, arugula, and cilantro purchased from supermarkets in Germany were analyzed by means of cultivation- and DNA-based methods. Before and after a nonselective enrichment step, tetracycline (TET)-resistant Escherichia coli were isolated and plasmids conferring TET resistance were captured by exogenous plasmid isolation. TET-resistant E. coli isolates, transconjugants, and total community DNA (TC-DNA) from the microbial fraction detached from leaves or after enrichment were analyzed for the presence of resistance genes, class 1 integrons, and various plasmids by real-time PCR and PCR-Southern blot hybridization. Real-time PCR primers were developed for IncI and IncF plasmids. TET-resistant E. coli isolated from arugula and cilantro carried IncF, IncI1, IncN, IncHI1, IncU, and IncX1 plasmids. Three isolates from cilantro were positive for IncN plasmids and blaCTX-M-1. From mixed salad and cilantro, IncF, IncI1, and IncP-1β plasmids were captured exogenously. Importantly, whereas direct detection of IncI and IncF plasmids in TC-DNA failed, these plasmids became detectable in DNA extracted from enrichment cultures. This confirms that cultivation-independent DNA-based methods are not always sufficiently sensitive to detect the transferable resistome in the rare microbiome. In summary, this study showed that an impressive diversity of self-transmissible multiple resistance plasmids was detected in bacteria associated with produce that is consumed raw, and exogenous capturing into E. coli suggests that they could transfer to gut bacteria as well. IMPORTANCE Produce is one of the most popular food commodities. Unfortunately, leafy greens can be a reservoir of transferable antibiotic resistance genes. We found that IncF and IncI plasmids were the most prevalent plasmid types in E. coli isolates from produce. This study highlights the importance of the rare microbiome associated with produce as a source of antibiotic resistance genes that might escape cultivation-independent detection, yet may be transferred to human pathogens or commensals.Khald BlauAntje BettermannSven JechalkeEva FornefeldYann VanrobaeysThibault StalderEva M. TopKornelia SmallaAmerican Society for MicrobiologyarticleEscherichia coliIncFIncIantibiotic resistancehorizontal gene transferreal-time PCRMicrobiologyQR1-502ENmBio, Vol 9, Iss 6 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Escherichia coli IncF IncI antibiotic resistance horizontal gene transfer real-time PCR Microbiology QR1-502
spellingShingle	Escherichia coli IncF IncI antibiotic resistance horizontal gene transfer real-time PCR Microbiology QR1-502 Khald Blau Antje Bettermann Sven Jechalke Eva Fornefeld Yann Vanrobaeys Thibault Stalder Eva M. Top Kornelia Smalla The Transferable Resistome of Produce
description	ABSTRACT Produce is increasingly recognized as a reservoir of human pathogens and transferable antibiotic resistance genes. This study aimed to explore methods to characterize the transferable resistome of bacteria associated with produce. Mixed salad, arugula, and cilantro purchased from supermarkets in Germany were analyzed by means of cultivation- and DNA-based methods. Before and after a nonselective enrichment step, tetracycline (TET)-resistant Escherichia coli were isolated and plasmids conferring TET resistance were captured by exogenous plasmid isolation. TET-resistant E. coli isolates, transconjugants, and total community DNA (TC-DNA) from the microbial fraction detached from leaves or after enrichment were analyzed for the presence of resistance genes, class 1 integrons, and various plasmids by real-time PCR and PCR-Southern blot hybridization. Real-time PCR primers were developed for IncI and IncF plasmids. TET-resistant E. coli isolated from arugula and cilantro carried IncF, IncI1, IncN, IncHI1, IncU, and IncX1 plasmids. Three isolates from cilantro were positive for IncN plasmids and blaCTX-M-1. From mixed salad and cilantro, IncF, IncI1, and IncP-1β plasmids were captured exogenously. Importantly, whereas direct detection of IncI and IncF plasmids in TC-DNA failed, these plasmids became detectable in DNA extracted from enrichment cultures. This confirms that cultivation-independent DNA-based methods are not always sufficiently sensitive to detect the transferable resistome in the rare microbiome. In summary, this study showed that an impressive diversity of self-transmissible multiple resistance plasmids was detected in bacteria associated with produce that is consumed raw, and exogenous capturing into E. coli suggests that they could transfer to gut bacteria as well. IMPORTANCE Produce is one of the most popular food commodities. Unfortunately, leafy greens can be a reservoir of transferable antibiotic resistance genes. We found that IncF and IncI plasmids were the most prevalent plasmid types in E. coli isolates from produce. This study highlights the importance of the rare microbiome associated with produce as a source of antibiotic resistance genes that might escape cultivation-independent detection, yet may be transferred to human pathogens or commensals.
format	article
author	Khald Blau Antje Bettermann Sven Jechalke Eva Fornefeld Yann Vanrobaeys Thibault Stalder Eva M. Top Kornelia Smalla
author_facet	Khald Blau Antje Bettermann Sven Jechalke Eva Fornefeld Yann Vanrobaeys Thibault Stalder Eva M. Top Kornelia Smalla
author_sort	Khald Blau
title	The Transferable Resistome of Produce
title_short	The Transferable Resistome of Produce
title_full	The Transferable Resistome of Produce
title_fullStr	The Transferable Resistome of Produce
title_full_unstemmed	The Transferable Resistome of Produce
title_sort	transferable resistome of produce
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/307f9dd6446b4cfbaaa5dc095712805d
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The Transferable Resistome of Produce

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