TY - JOUR
T1 - Comparison of culture-and quantitative PCR-based indicators of antibiotic resistance in wastewater, recycled water, and tap water
AU - Rocha, Jaqueline
AU - Fernandes, Telma
AU - Riquelme, Maria V.
AU - Zhu, Ni
AU - Pruden, Amy
AU - Manaia, Célia M.
N1 - Funding Information:
The Portugal co-authors acknowledge the Fundação Luso Americana para o Desenvolvimento-FLAD/NSF-“Programa de Redes de Investigação Portugal/EUA 2015”-Ciência Ambiental-Proj 2015/298 and the European Regional Development Fund (FEDER), through the North Regional Operational Program (North PO), under the project DEPCAT.: Demonstration of new equipment involving integrated catalytic processes for the treatment of organic pollutants and disinfection of waters (NORTE-01-0247-FEDER-033330). JR was supported by the International PhD Programme in Biotechnology – BIOTECH. DOC (NORTE-08-5369-FSE-000007). Funding was also provided by the U.S. National Science Foundation Partnership in International Research and Education award OISE 1545756. The authors thank Suraj Gupta and Mathew Chan of Virginia Tech, and the UWTP operators, who assisted with the US and PT sample collection.
Funding Information:
Funding: The Portugal co-authors acknowledge the Fundação Luso Americana para o Desenvolvimento - FLAD/NSF - “Programa de Redes de Investigação Portugal/EUA 2015” - Ciência Ambiental - Proj 2015/298 and the European Regional Development Fund (FEDER), through the North Regional Operational Program (North PO), under the project DEPCAT.: Demonstration of new equipment involving integrated catalytic processes for the treatment of organic pollutants and disinfection of waters (NORTE-01-0247-FEDER-033330). JR was supported by the International PhD Programme in Biotechnology – BIOTECH. DOC (NORTE-08-5369-FSE-000007). Funding was also provided by the U.S. National Science Foundation Partnership in International Research and Education award OISE 1545756.
Publisher Copyright:
© 2019 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2019/11/1
Y1 - 2019/11/1
N2 - Standardized methods are needed to support monitoring of antibiotic resistance in environmental samples. Culture-based methods target species of human-health relevance, while the direct quantification of antibiotic resistance genes (ARGs) measures the antibiotic resistance potential in the microbial community. This study compared measurements of tetracycline-, sulphonamide-, and cefotaxime-resistant presumptive total and fecal coliforms and presumptive enterococci versus a suite of ARGs quantified by quantitative polymerase chain reaction (qPCR) across waste-, recycled-, tap-, and freshwater. Cross-laboratory comparison of results involved measurements on samples collected and analysed in the US and Portugal. The same DNA extracts analysed in the US and Portugal produced comparable qPCR results (variation <28%), except for blaOXA-1 gene (0%–57%). Presumptive total and fecal coliforms and cefotaxime-resistant total coliforms strongly correlated with blaCTX-M and intI1 (0.725 ≤ R2 ≤ 0.762; p < 0.0001). Further, presumptive total and fecal coliforms correlated with the Escherichia coli-specific biomarkers, gadAB, and uidA, suggesting that both methods captured fecal-sourced bacteria. The genes encoding resistance to sulphonamides (sul1 and sul2) were the most abundant, followed by genes encoding resistance to tetracyclines (tet(A) and tet(O)) and β-lactams (blaOXA-1 and, blaCTX-M), which was in agreement with the culture-based enumerations. The findings can help inform future application of methods being considered for international antibiotic resistance surveillance in the environment.
AB - Standardized methods are needed to support monitoring of antibiotic resistance in environmental samples. Culture-based methods target species of human-health relevance, while the direct quantification of antibiotic resistance genes (ARGs) measures the antibiotic resistance potential in the microbial community. This study compared measurements of tetracycline-, sulphonamide-, and cefotaxime-resistant presumptive total and fecal coliforms and presumptive enterococci versus a suite of ARGs quantified by quantitative polymerase chain reaction (qPCR) across waste-, recycled-, tap-, and freshwater. Cross-laboratory comparison of results involved measurements on samples collected and analysed in the US and Portugal. The same DNA extracts analysed in the US and Portugal produced comparable qPCR results (variation <28%), except for blaOXA-1 gene (0%–57%). Presumptive total and fecal coliforms and cefotaxime-resistant total coliforms strongly correlated with blaCTX-M and intI1 (0.725 ≤ R2 ≤ 0.762; p < 0.0001). Further, presumptive total and fecal coliforms correlated with the Escherichia coli-specific biomarkers, gadAB, and uidA, suggesting that both methods captured fecal-sourced bacteria. The genes encoding resistance to sulphonamides (sul1 and sul2) were the most abundant, followed by genes encoding resistance to tetracyclines (tet(A) and tet(O)) and β-lactams (blaOXA-1 and, blaCTX-M), which was in agreement with the culture-based enumerations. The findings can help inform future application of methods being considered for international antibiotic resistance surveillance in the environment.
KW - Antibiotic resistance monitoring
KW - Antibiotic resistant coliforms
KW - Water quality
UR - http://www.scopus.com/inward/record.url?scp=85074343266&partnerID=8YFLogxK
U2 - 10.3390/ijerph16214217
DO - 10.3390/ijerph16214217
M3 - Article
C2 - 31671709
AN - SCOPUS:85074343266
SN - 1661-7827
VL - 16
SP - 1
EP - 13
JO - International Journal of Environmental Research and Public Health
JF - International Journal of Environmental Research and Public Health
IS - 21
M1 - 4217
ER -
