Comparison of Cefotaxime-Resistant <i>Escherichia coli</i> and <i>sul</i>1 and <i>int</i>I1 by qPCR for Monitoring of Antibiotic Resistance of Wastewater, Surface Water, and Recycled Water
Awareness of the need for surveillance of antimicrobial resistance (AMR) in water environments is growing, but there is uncertainty regarding appropriate monitoring targets. Adapting culture-based fecal indicator monitoring to include antibiotics in the media provides a potentially low-tech and acce...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-07-01
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| Series: | Antibiotics |
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| Online Access: | https://www.mdpi.com/2079-6382/12/8/1252 |
| _version_ | 1797585775075786752 |
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| author | Krista Liguori Jeanette Calarco Gabriel Maldonado Rivera Anna Kurowski Ishi Keenum Benjamin C. Davis Valerie J. Harwood Amy Pruden |
| author_facet | Krista Liguori Jeanette Calarco Gabriel Maldonado Rivera Anna Kurowski Ishi Keenum Benjamin C. Davis Valerie J. Harwood Amy Pruden |
| author_sort | Krista Liguori |
| collection | DOAJ |
| description | Awareness of the need for surveillance of antimicrobial resistance (AMR) in water environments is growing, but there is uncertainty regarding appropriate monitoring targets. Adapting culture-based fecal indicator monitoring to include antibiotics in the media provides a potentially low-tech and accessible option, while quantitative polymerase chain reaction (qPCR) targeting key genes of interest provides a broad, quantitative measure across the microbial community. The purpose of this study was to compare findings obtained from the culture of cefotaxime-resistant (cefR) <i>Escherichia coli</i> with two qPCR methods for quantification of antibiotic resistance genes across wastewater, recycled water, and surface waters. The culture method was a modification of US EPA Method 1603 for <i>E. coli</i>, in which cefotaxime is included in the medium to capture cefR strains, while qPCR methods quantified <i>sul</i>1 and <i>int</i>I1. A common standard operating procedure for each target was applied to samples collected by six water utilities across the United States and processed by two laboratories. The methods performed consistently, and all three measures reflected the same overarching trends across water types. The qPCR detection of <i>sul</i>1 yielded the widest dynamic range of measurement as an AMR indicator (7-log versus 3.5-log for cefR <i>E. coli</i>), while <i>int</i>I1 was the most frequently detected target (99% versus 96.5% and 50.8% for <i>sul</i>1 and cefR <i>E. coli</i>, respectively). All methods produced comparable measurements between labs (<i>p</i> < 0.05, Kruskal–Wallis). Further study is needed to consider how relevant each measure is to capturing hot spots for the evolution and dissemination of AMR in the environment and as indicators of AMR-associated human health risk. |
| first_indexed | 2024-03-11T00:11:50Z |
| format | Article |
| id | doaj.art-b9332906d6334f6fae202cf31e7e3d8a |
| institution | Directory Open Access Journal |
| issn | 2079-6382 |
| language | English |
| last_indexed | 2024-03-11T00:11:50Z |
| publishDate | 2023-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibiotics |
| spelling | doaj.art-b9332906d6334f6fae202cf31e7e3d8a2023-11-18T23:59:01ZengMDPI AGAntibiotics2079-63822023-07-01128125210.3390/antibiotics12081252Comparison of Cefotaxime-Resistant <i>Escherichia coli</i> and <i>sul</i>1 and <i>int</i>I1 by qPCR for Monitoring of Antibiotic Resistance of Wastewater, Surface Water, and Recycled WaterKrista Liguori0Jeanette Calarco1Gabriel Maldonado Rivera2Anna Kurowski3Ishi Keenum4Benjamin C. Davis5Valerie J. Harwood6Amy Pruden7Via Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA 24060, USADepartment of Integrative Biology, University of South Florida, Tampa, FL 33620, USAVia Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA 24060, USAVia Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA 24060, USAVia Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA 24060, USAVia Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA 24060, USADepartment of Integrative Biology, University of South Florida, Tampa, FL 33620, USAVia Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, VA 24060, USAAwareness of the need for surveillance of antimicrobial resistance (AMR) in water environments is growing, but there is uncertainty regarding appropriate monitoring targets. Adapting culture-based fecal indicator monitoring to include antibiotics in the media provides a potentially low-tech and accessible option, while quantitative polymerase chain reaction (qPCR) targeting key genes of interest provides a broad, quantitative measure across the microbial community. The purpose of this study was to compare findings obtained from the culture of cefotaxime-resistant (cefR) <i>Escherichia coli</i> with two qPCR methods for quantification of antibiotic resistance genes across wastewater, recycled water, and surface waters. The culture method was a modification of US EPA Method 1603 for <i>E. coli</i>, in which cefotaxime is included in the medium to capture cefR strains, while qPCR methods quantified <i>sul</i>1 and <i>int</i>I1. A common standard operating procedure for each target was applied to samples collected by six water utilities across the United States and processed by two laboratories. The methods performed consistently, and all three measures reflected the same overarching trends across water types. The qPCR detection of <i>sul</i>1 yielded the widest dynamic range of measurement as an AMR indicator (7-log versus 3.5-log for cefR <i>E. coli</i>), while <i>int</i>I1 was the most frequently detected target (99% versus 96.5% and 50.8% for <i>sul</i>1 and cefR <i>E. coli</i>, respectively). All methods produced comparable measurements between labs (<i>p</i> < 0.05, Kruskal–Wallis). Further study is needed to consider how relevant each measure is to capturing hot spots for the evolution and dissemination of AMR in the environment and as indicators of AMR-associated human health risk.https://www.mdpi.com/2079-6382/12/8/1252antimicrobial resistancestandard methodswastewater monitoring |
| spellingShingle | Krista Liguori Jeanette Calarco Gabriel Maldonado Rivera Anna Kurowski Ishi Keenum Benjamin C. Davis Valerie J. Harwood Amy Pruden Comparison of Cefotaxime-Resistant <i>Escherichia coli</i> and <i>sul</i>1 and <i>int</i>I1 by qPCR for Monitoring of Antibiotic Resistance of Wastewater, Surface Water, and Recycled Water Antibiotics antimicrobial resistance standard methods wastewater monitoring |
| title | Comparison of Cefotaxime-Resistant <i>Escherichia coli</i> and <i>sul</i>1 and <i>int</i>I1 by qPCR for Monitoring of Antibiotic Resistance of Wastewater, Surface Water, and Recycled Water |
| title_full | Comparison of Cefotaxime-Resistant <i>Escherichia coli</i> and <i>sul</i>1 and <i>int</i>I1 by qPCR for Monitoring of Antibiotic Resistance of Wastewater, Surface Water, and Recycled Water |
| title_fullStr | Comparison of Cefotaxime-Resistant <i>Escherichia coli</i> and <i>sul</i>1 and <i>int</i>I1 by qPCR for Monitoring of Antibiotic Resistance of Wastewater, Surface Water, and Recycled Water |
| title_full_unstemmed | Comparison of Cefotaxime-Resistant <i>Escherichia coli</i> and <i>sul</i>1 and <i>int</i>I1 by qPCR for Monitoring of Antibiotic Resistance of Wastewater, Surface Water, and Recycled Water |
| title_short | Comparison of Cefotaxime-Resistant <i>Escherichia coli</i> and <i>sul</i>1 and <i>int</i>I1 by qPCR for Monitoring of Antibiotic Resistance of Wastewater, Surface Water, and Recycled Water |
| title_sort | comparison of cefotaxime resistant i escherichia coli i and i sul i 1 and i int i i1 by qpcr for monitoring of antibiotic resistance of wastewater surface water and recycled water |
| topic | antimicrobial resistance standard methods wastewater monitoring |
| url | https://www.mdpi.com/2079-6382/12/8/1252 |
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