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J Water Health
. 2025 Jun;23(6):727-742.
doi: 10.2166/wh.2025.374. Epub 2025 May 9. Skilled nursing facility wastewater surveillance: a SARS-CoV-2 and antimicrobial resistance detection pilot study
Ariel Jose Santiago 1 , Maria Burgos Garay 2 , Mariya Campbell 3 , Yimu Cahela 4 , Rodney Donlan 2 , Paige Gable 2 , Christine Ganim Kyros 2 , Lauren Franco 3 , Leila Kartforosh 3 , Susanna Lenz 3 , Amanda K Lyons 2 , Jamari Moore 4 , Judith Noble-Wang 2 , Carrie Sanders 4 , Bethelhem Abera 5 , Colin H Adler 5 , Sophie Jones 2 6 , Magdalena Medrzycki 2 , Maroya S Walters 2 7 , Peter Cook 8 , Yan Li 8 , Ying Tao 8 , Jing Zhang 8 , Lakshmi Malapati 8 , Adam Retchless 8 , Suxiang Tong 8 , Angela D Coulliette-Salmond 2 7
Affiliations
The purpose of this study was to determine the feasibility of facility-level wastewater surveillance in the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in skilled nursing facility (SNF) wastewater using three concentration methods, as well as a proof-of-concept for antimicrobial resistance (AR) genes/organisms detection. Wastewater effluent samples were collected from an SNF over an 8-week period. Wastewater was concentrated using electronegative membrane filtration (enMF), polyethylene glycol precipitation, and Nanotrap® magnetic virus particles (NP). Quantification of the genome copy concentration from SARS-CoV-2 and bovine respiratory syncytial virus (BRSV), a SARS-CoV-2 surrogate spiked into all samples, was performed with droplet digital polymerase chain reaction (ddPCR). Wastewater sample aliquots were also enriched in microbiological culture media and screened for organisms with AR phenotypes on selective and differential agars. Multiplex real-time PCR was used to detect a broad array of carbapenem resistance genes. SARS-CoV-2 was detected and quantified from a single enMF-concentrated wastewater sample. The highest concentration of BRSV came from enMF-concentrated samples. Klebsiella, Enterobacter, Citrobacter, and Escherichia coli exhibiting AR phenotypes were successfully detected using culture-dependent approaches. Culture-independent, multiplex PCR indicated that blaKPC was the main carbapenemase gene detected in wastewater samples. Facility-level wastewater surveillance could be a useful strategy for SNFs.
Keywords: SARS-CoV-2; antimicrobial resistance; skilled nursing facility; wastewater surveillance.
. 2025 Jun;23(6):727-742.
doi: 10.2166/wh.2025.374. Epub 2025 May 9. Skilled nursing facility wastewater surveillance: a SARS-CoV-2 and antimicrobial resistance detection pilot study
Ariel Jose Santiago 1 , Maria Burgos Garay 2 , Mariya Campbell 3 , Yimu Cahela 4 , Rodney Donlan 2 , Paige Gable 2 , Christine Ganim Kyros 2 , Lauren Franco 3 , Leila Kartforosh 3 , Susanna Lenz 3 , Amanda K Lyons 2 , Jamari Moore 4 , Judith Noble-Wang 2 , Carrie Sanders 4 , Bethelhem Abera 5 , Colin H Adler 5 , Sophie Jones 2 6 , Magdalena Medrzycki 2 , Maroya S Walters 2 7 , Peter Cook 8 , Yan Li 8 , Ying Tao 8 , Jing Zhang 8 , Lakshmi Malapati 8 , Adam Retchless 8 , Suxiang Tong 8 , Angela D Coulliette-Salmond 2 7
Affiliations
- PMID: 40586337
- DOI: 10.2166/wh.2025.374
The purpose of this study was to determine the feasibility of facility-level wastewater surveillance in the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in skilled nursing facility (SNF) wastewater using three concentration methods, as well as a proof-of-concept for antimicrobial resistance (AR) genes/organisms detection. Wastewater effluent samples were collected from an SNF over an 8-week period. Wastewater was concentrated using electronegative membrane filtration (enMF), polyethylene glycol precipitation, and Nanotrap® magnetic virus particles (NP). Quantification of the genome copy concentration from SARS-CoV-2 and bovine respiratory syncytial virus (BRSV), a SARS-CoV-2 surrogate spiked into all samples, was performed with droplet digital polymerase chain reaction (ddPCR). Wastewater sample aliquots were also enriched in microbiological culture media and screened for organisms with AR phenotypes on selective and differential agars. Multiplex real-time PCR was used to detect a broad array of carbapenem resistance genes. SARS-CoV-2 was detected and quantified from a single enMF-concentrated wastewater sample. The highest concentration of BRSV came from enMF-concentrated samples. Klebsiella, Enterobacter, Citrobacter, and Escherichia coli exhibiting AR phenotypes were successfully detected using culture-dependent approaches. Culture-independent, multiplex PCR indicated that blaKPC was the main carbapenemase gene detected in wastewater samples. Facility-level wastewater surveillance could be a useful strategy for SNFs.
Keywords: SARS-CoV-2; antimicrobial resistance; skilled nursing facility; wastewater surveillance.